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Sample records for cd25 expression controls

  1. Expression of regulatory CD4+CD25+ Treg,CD4+CD25higTreg cells and Foxp3 mRNA in wheezing infants and its clinical significance%CD4+CD25+、CD4+CD25hig调节性T细胞和Foxp3mRNA在婴幼儿喘息中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    彭力; 钟礼立; 黄寒; 厉娟; 梁沫; 李云

    2012-01-01

    Objective To evaluate the changes of CD4+ CD25+ Treg,CD4 +CD25hig Treg and Foxp3 mRNA in peripheral blood from wheezing infantsMethods Fifty-one wheezing infants and twenty healthy volunteers were included in this study. The proportion of CD4 + CD25 + Treg and CD4 + CD25hig Treg population in total T cells was evaluated by flow cytometric analysis. The expression of Foxp3 mRNA was tested by flow cytometry. Total serum IgE of wheezy infants was detected by enzyme immunoassay. Results Compared with those of healthy control, the frequency of CD4 + CD25 + Treg and CD4 + CD25hig Treg in the peripheral blood from wheezing infants showed a significant increase (6. 31 + 2. 96) % / (3.52 + 1.46) % ,P<0. 01 ,P<0. 01, respectively).The expression of CD4 + CD25+ Treg,CD4 + CD25hig Treg and Foxp3 mRNA in peripheral blood from wheezing infants with atopy burden was lower than those from non-wheezing infants(P<0. 05). The correlation analysis showed that CD4 + CD25hig Treg (r= -0. 75 , P<0.01) and Foxp3 mRNA(r= -0. 61,P<0. 01) had significantly positive relation with total serum IgE,while CD4+CD25 + Treg had significantly negative relation with total serum IgE(r=0. 36,P<0. 05). Conclusion CD4+CD25+ Treg, CD4+CD25hig Treg and Foxp3 mRNA play an important role in activation of wheezing infants.%目的 探讨婴幼儿喘息CD4+CD25+、CD4+CD25hig调节性T细胞(Treg)和叉头/翼状螺旋转录因子(Foxp3)mRNA的表达及意义.方法 采用流式细胞术检测51例首次喘息婴幼儿外周血CD4+CD25+Treg和CD4+CD25higTreg的比例,RT-PCR检测Foxp3 mRNA的表达量,酶联免疫吸附法(ELASA)检测总IgE水平,并与正常婴幼儿对照.结果 喘息婴幼儿外周血CD4+CD25+Treg、CD4+CD25higTreg占CD4+T细胞的百分比分别为(6.31+2.96)%和(3.52+1.46)%,均明显低于健康对照组(P<0.01);特应征喘息组CD4+CD25+Treg、CD4+CD25higTreg及Foxp3 mRNA表达均低于非特应征喘息组(P<0.05).喘息患儿CD4+CD25higTreg百分率及Foxp3 mRNA表达与

  2. Forkhead Transcription Factor FOXP3 Upregulates CD25 Expression through Cooperation with RelA/NF-κB

    OpenAIRE

    Camperio, Cristina; Caristi, Silvana; Fanelli, Giorgia; Soligo, Marzia; Porto, Paola Del; Piccolella, Enza

    2012-01-01

    Considerable evidence supports the prediction that CD25 is directly regulated by the forkhead transcription factor FOXP3. However, given that CD25 is normally upregulated in activated T cells, regardless of whether they express FOXP3, this issue has still to be definitively demonstrated. Here we describe that FOXP3, induced by CD28 signals in human CD4+CD25 − T lymphocytes, synergizes with RelA on a regulatory region of Cd25 promoter to mediate the transcriptional activation of Cd25 gene. We ...

  3. Bromelain treatment reduces CD25 expression on activated CD4+ T cells in vitro✩

    OpenAIRE

    Secor, Eric R.; Singh, Anurag; Guernsey, Linda A.; McNamara, Jeff T.; Zhan, Lijun; Maulik, Nilanjana; Thrall, Roger S.

    2009-01-01

    Bromelain (Br), an extract from pineapple stem with cysteine protease activity, exerts anti-inflammatory effects in a number of inflammatory models. We have previously shown that Br treatment decreased activated CD4+ T cells and has a therapeutic role in an ovalbumin-induced murine model of allergic airway disease. The current study was designed to determine the effect of Br on CD4+ T cell activation, specifically the expression of CD25 in vitro. CD25 is up regulated upon T cell activation, f...

  4. Interleukin-7 optimizes FOXP3+CD4+ regulatory T cells reactivity to interleukin-2 by modulating CD25 expression.

    Directory of Open Access Journals (Sweden)

    Federico Simonetta

    Full Text Available The vast majority of Foxp3 regulatory T cells (Treg exhibits constitutive expression of CD25 (IL-2Rα, which allows the constitution of the high affinity IL-2Rαβγ receptor, ensuring efficient IL-2 binding by Treg. Maintenance of CD25 expression at Treg surface depends on both cell intrinsic factors and environmental stimuli such as IL-2 itself. Whether other factors can participate to maintenance of CD25 expression in vivo is at present unknown. In the present work we demonstrated that IL-7, a gamma-chain cytokine exerting a crucial role in T cell development and homeostasis, is able and necessary to sustain the expression of high levels of CD25 at Treg surface. We demonstrated that, during in vitro cultures performed in the absence of IL-2, IL-7 is able to sustain CD25 expression at Treg surface through a transcriptional mechanism. By studying mice in which IL-7 signaling is either genetically impaired or increased and by employing adoptive transfer murine models, we demonstrated that IL-7 is necessary for sustained expression of CD25 at Treg surface in vivo. To ascertain the biological impact of IL-7 mediated modulation of CD25 expression, we demonstrated that IL-7 modulation of CD25 expression at Treg surface affected their ability to efficiently bind IL-2 and transduce IL-2 signaling. Finally, we demonstrated that IL-7 dependent modulation of CD25 associated with potentiated IL-2 induced expansion of Treg in vivo. Collectively, our results identify IL-7 as a necessary factor contributing to sustained CD25 expression at Treg surface in vivo thereby affecting their ability to efficiently react to IL-2.

  5. FOXP3, CBLB and ITCH gene expression and cytotoxic T lymphocyte antigen 4 expression on CD4(+) CD25(high) T cells in multiple sclerosis

    DEFF Research Database (Denmark)

    Sellebjerg, F; Krakauer, M; Khademi, M; Olsson, T; Sørensen, P S

    2012-01-01

    phenotype of CD4(+) CD25(high) T cells in MS by flow cytometry and its relationship with expression of the FOXP3, ITCH and CBLB genes. We found that untreated MS patients had lower cell surface expression of cytotoxic T lymphocyte antigen 4 (CTLA-4) on CD4(+) CD25(high) T cells and higher intracellular CTLA......-4 expression than healthy controls. Cell surface expression of CTLA-4 on CD4(+) CD25(high) T cells correlated with expression of FOXP3 mRNA in untreated patients and increased significantly with time from most recent injection in patients treated with IFN-β. FOXP3 mRNA expression correlated with...... CBLB and ITCH and T helper type 2 cytokine mRNA expression in MS patients. These data link expression of FOXP3, CBLB and ITCH mRNA and CTLA-4 expression on the surface of CD4(+) CD25(high) T cell in MS. We hypothesize that this may reflect alterations in the inhibitory effect of CTLA-4 or in regulatory...

  6. The control of CD4+CD25+Foxp3+ regulatory T cell survival

    Directory of Open Access Journals (Sweden)

    Lenardo Michael J

    2008-02-01

    Full Text Available Abstract CD4+CD25+Foxp3+ regulatory T (Treg cells are believed to play an important role in suppressing autoimmunity and maintaining peripheral tolerance. How their survival is regulated in the periphery is less clear. Here we show that Treg cells express receptors for gamma chain cytokines and are dependent on an exogenous supply of these cytokines to overcome cytokine withdrawal apoptosis in vitro. This result was validated in vivo by the accumulation of Treg cells in Bim-/- and Bcl-2 tg mice which have arrested cytokine deprivation apoptosis. We also found that CD25 and Foxp3 expression were down-regulated in the absence of these cytokines. CD25+ cells from Scurfy mice do not depend on cytokines for survival demonstrating that Foxp3 increases their dependence on cytokines by suppressing cytokine production in Treg cells. Our study reveals that the survival of Treg cells is strictly dependent on cytokines and cytokine producing cells because they do not produce cytokines. Our study thus, demonstrates that different gamma chain cytokines regulate Treg homeostasis in the periphery by differentially regulating survival and proliferation. These findings may shed light on ways to manipulate Treg cells that could be utilized for their therapeutic applications. Reviewers This article was reviewed by: Avinash Bhandoola, Fred Ramsdell (nominated by Juan Carlos Zuniga-Pflucker and Anne Cooke.

  7. T cells that cannot respond to TGF-β escape control by CD4+CD25+ regulatory T cells

    OpenAIRE

    Fahlén, Linda; Read, Simon; Gorelik, Leonid; Hurst, Stephen D.; Coffman, Robert L.; Flavell, Richard A.; Powrie, Fiona

    2005-01-01

    CD4+CD25+ regulatory T (T reg) cells play a pivotal role in control of the immune response. Transforming growth factor-β (TGF-β) has been shown to be required for T reg cell activity; however, precisely how it is involved in the mechanism of suppression is poorly understood. Using the T cell transfer model of colitis, we show here that CD4+CD45RBhigh T cells that express a dominant negative TGF-β receptor type II (dnTβRII) and therefore cannot respond to TGF-β, escape control by T reg cells i...

  8. Androgen receptor modulates Foxp3 expression in CD4+CD25+Foxp3+ regulatory T-cells

    OpenAIRE

    Walecki, Magdalena; Eisel, Florian; Klug, Jörg; Baal, Nelli; Paradowska-Dogan, Agnieszka; Wahle, Eva; Hackstein, Holger; Meinhardt, Andreas; Fijak, Monika

    2015-01-01

    CD4+CD25+Foxp3+ Treg cells are crucial for the maintenance of immunological homeostasis. Androgens significantly induce Foxp3 expression in humans and regulate the differentiation of Treg cells. A functional androgen receptor–binding site is identified within the Foxp3 locus leading to epigenetic changes of histone H4.

  9. CD4 + CD25 + Treg cell separate, phenotype identity and Foxp3 gene expression identity%CD4+CD25+Treg细胞的分选、表型鉴定及Foxp3表达鉴定

    Institute of Scientific and Technical Information of China (English)

    韩文杰; 史艳侠

    2010-01-01

    目的 为验证从C57BL/6小鼠脾脏中分离出高纯度CD4+CD25+Treg细胞及证实CD4+CD25+Treg细胞中Foxp3基因的表达.方法 使用免疫磁珠分选出CD4+CD25+Treg细胞,流式细胞仪检测纯度;使用TRIZOL抽提Foxp3基因mRNA,使用RT-PCR方法逆转录出Foxp3基因的cDNA.结果 从C57BL/6小鼠脾脏中分离出了纯度达到90%CD4+CD25+Treg.进一步应用RT-PCR技术克隆出Foxp3的cDNA,通过凝胶电泳证实了克隆出了Foxp3的cDNA.结论 使用免疫磁珠方法能够分离出C57BL/6小鼠CD4+CD25+Treg细胞,并进行了Foxp3基因表达的鉴定.%Objective To confirm high purity CD4 + CD25 + Treg cells can be separated from the spleen of C57BL/6 mice and Foxp3 gene can be express in CD4 + CD25 + Treg cells.Methods CD4 + CD25 + Treg cells are separated with immunomagnetic beads,and purity is detected by flow cytometry.Foxp3 gene mRNA is extracted using TRIZOL.Foxp3 gene cDNA is reverse transcription using RT-PCR technology.Results This study separated CD4 + CD25 + Treg cell of 90% purity from the spleen of C57BI/6 mice,to advance used technology of RT-PCR to make the clone of Foxp3 gene cDNA,and confirmed it is the cDNA of foxp3.Conclusion This study CD4 + CD25 + Treg cell can be separated from the spleen of C57BL/6 mice with immnnomagnetic beads,and identified foxp3 gene expression.

  10. CD25 expression on residual leukemic blasts at the time of allogeneic hematopoietic stem cell transplant predicts relapse in patients with acute myeloid leukemia without complete remission.

    Science.gov (United States)

    Ikegawa, Shuntaro; Doki, Noriko; Kurosawa, Shuhei; Yamaguchi, Tsukasa; Sakaguchi, Masahiro; Harada, Kaito; Yamamoto, Keita; Hino, Yutaro; Shingai, Naoki; Senoo, Yasushi; Hattori, Keiichiro; Igarashi, Aiko; Najima, Yuho; Kobayashi, Takeshi; Kakihana, Kazuhiko; Sakamaki, Hisashi; Haraguchi, Kyoko; Okuyama, Yoshiki; Ohashi, Kazuteru

    2016-06-01

    Recent studies have shown that CD25 expression at the time of diagnosis of acute myeloid leukemia (AML) may be associated with an unfavorable outcome. We focus on patients with AML without complete remission (CR) and examine the clinical correlation between surface CD25 expression at the time of transplant and subsequent transplant outcomes. We observed a significant difference in overall survival (OS), disease-free survival (DFS) and cumulative incidence of relapse (CIR) between CD25 positive (+) (n = 22) and negative (-) groups (n = 44) (2-year OS; CD25 (+) group: 5% vs. CD25 (-) group: 40%, p expression was an independent adverse factor for OS (p = 0.002) and relapse (p = 0.001). Patients with AML with residual CD25 positive blasts at the time of transplant may require additional therapy before or after transplant to improve survival. PMID:26422713

  11. Prognostic value of CD25 expression on lymphocytes and tumor cells in squamous-cell carcinoma of the head and neck

    NARCIS (Netherlands)

    Loose, David; Signore, Alberto; Bonanno, Elena; Vermeersch, Hubert; Dierckx, Rudi; Deron, Philippe; Van de Wiele, Christophe

    2008-01-01

    CD4(+)CD25(+) T-cells play a central role in initiating and maintaining anticancer immune response. On the other hand, CD25(+) is also expressed on tumor cells, the meaning of which is currently unclear. Therefore, this study was designed to determine the prognostic value of the presence of CD4(+)CD

  12. 吸毒人员外周血CD4+CD25+Foxp3调节性T细胞表达%Expression of CD4+CD25+Foxp3 regulatory T cells in the blood of drug abuse population

    Institute of Scientific and Technical Information of China (English)

    庞惠勇; 葛恒明; 陈晓芹; 刘小林; 李忠典; 张振宇; 张健

    2011-01-01

    Objective:This study was designed to investigate the effect of drug abuse on human immune function by examining the blood CD4+ CD25 + Foxp3 regulatory T cell expression. Methods: Blood samples were collected in 114 different drug taken route and period people. Flow cytometry was employed to examine the expression of CD4 + CD25 + Foxp3 regulatory T cells. Results: The expression of CD4 + CD25 + Foxp3 regulatory T cells was different among different taken route groups (Oral, 49.07% ± 14.88% > intravenous injection, 34.96% ± 13.41% > mixed routes, 26.72% ± 8.49% ). There were significant differences (P<0. 01) between any of the above two groups. We also examined the effect of drug taken period on the expression of CD4+ CD25 + Foxp3 regulatory T cells. For oral taken people, the expression was much lower in the people with taken period longer than 10 years (37. 14% ± 12.29%) compared with those people with shorter drug taken period ( ≤ 10 years) (51.79% ± 10.44%, P < 0. 01 ). For mixed taken route patients, however, the expression increased from 27.06% ± 8.99% in people with ≤ 10 - year drug taken period to 35.47% ± 11.02% in people with > 10 - year drug taken period( P < 0.01 ). There was no significant difference in the intravenous injection group( P >0.05 ). Conclusion: By examining the blood CD4 + CD25 + Foxp3 regulatory T cell expression in the drug abuse population, it was found that different drug taken routes and periods may induce different extents of injury to the body immune function. Our results provide not only an accurate, reliable monitoring index, but also a new approach to examine the immune function in drug abuse population.%目的:探讨外周血CD4+CD25+Foxp3调节性T细胞与吸毒人员机体免疫的关系.方法:采集114名吸毒人员外周血,根据不同吸毒方式和吸毒年限进行分组,应用流式细胞仪检测外周血中CD4+CD25+Foxp3调节性T细胞表达.结果:不同吸毒

  13. CD4+CD25+调节性T细胞在不同肝脏肿瘤中的表达及意义%Expression and Significance of CD4+ CD25+ Regulatory T Cells in Different Liver Tumors

    Institute of Scientific and Technical Information of China (English)

    王俊; 罗英; 颜秉菊

    2012-01-01

    目的:研究CD4+CD25+调节性T细胞(Treg)在不同肝脏肿瘤中的分布特点,评价其在肿瘤发生和发展中的作用.方法:根据病理诊断结果将80例肝脏肿瘤分为肝局灶性结节状增生(FNH)组10例、不典型腺瘤样增生(AAH)组10例以及原发性肝癌(HCC)组60例.另选取10例正常肝组织(肝血管瘤边缘肝组织)石蜡包埋标本为对照组.采用双重酶标免疫组化染色的方法测定不同肝脏肿瘤切片中Treg细胞的表达状况.对比分析Treg细胞在FNH、AAH和HCC各组中的表达特点,并进一步分析在HCC组中Treg细胞表达的影响因素.结果:对照组及FNH组中均未发现Treg细胞的表达.AAH组、HCC组中有Treg细胞的表达,且HCC组较AAH组增多(P<0.01).在癌旁组织中已有Treg 细胞浸润,但较肝癌组织中Treg细胞数量少(P<0.01).肝癌组织中不同患者性别、年龄、术前AFP水平的Treg细胞数量差异无统计学意义,而在不同肿瘤大小、肿瘤包膜是否完整及术前HBV-DNA水平是否升高中Treg细胞数量差异有统计学意义(P<0.05或P<0.01).结论:Treg细胞的表达与肿瘤的发生和发展有关,在肿瘤免疫中起负调节作用.%Objective:To investigate the distribution of CD4+CD25+ regulatory T cells(Treg) in different liver tumors, and the role in the process of tumor occurrence and development. Methods: According to the pathological result, 80 patients were divided into focal nodular hyperplasia group (FNH, n=10), atypical adenomatous hyperplasia group (AAH, n=10) and he-patocellular carcinoma group (HCC, n=60). Another 10 cases of normal liver tissue (liver hemangioina edge of the liver) were selected for the control group. The expression of Treg cells in different tumor slices was detected by double ELISA of immuno-histochemical staining. The expression of Treg cells was compared between FNH, AAH and HCC groups. The influencing factors of Treg cells were analyzed between groups in HCC groups. Results

  14. Expansion of CD4(+ CD25(+ and CD25(- T-Bet, GATA-3, Foxp3 and RORγt cells in allergic inflammation, local lung distribution and chemokine gene expression.

    Directory of Open Access Journals (Sweden)

    You Lu

    Full Text Available Allergic asthma is associated with airway eosinophilia, which is regulated by different T-effector cells. T cells express transcription factors T-bet, GATA-3, RORγt and Foxp3, representing Th1, Th2, Th17 and Treg cells respectively. No study has directly determined the relative presence of each of these T cell subsets concomitantly in a model of allergic airway inflammation. In this study we determined the degree of expansion of these T cell subsets, in the lungs of allergen challenged mice. Cell proliferation was determined by incorporation of 5-bromo-2'-deoxyuridine (BrdU together with 7-aminoactnomycin (7-AAD. The immunohistochemical localisation of T cells in the lung microenvironments was also quantified. Local expression of cytokines, chemokines and receptor genes was measured using real-time RT-PCR array analysis in tissue sections isolated by laser microdissection and pressure catapulting technology. Allergen exposure increased the numbers of T-bet(+, GATA-3(+, RORγt(+ and Foxp3(+ cells in CD4(+CD25(+ and CD4(+CD25(- T cells, with the greatest expansion of GATA-3(+ cells. The majority of CD4(+CD25(+ T-bet(+, GATA-3(+, RORγt(+ and Foxp3(+ cells had incorporated BrdU and underwent proliferation during allergen exposure. Allergen exposure led to the accumulation of T-bet(+, GATA-3(+ and Foxp3(+ cells in peribronchial and alveolar tissue, GATA-3(+ and Foxp3(+ cells in perivascular tissue, and RORγt(+ cells in alveolar tissue. A total of 28 cytokines, chemokines and receptor genes were altered more than 3 fold upon allergen exposure, with expression of half of the genes claimed in all three microenvironments. Our study shows that allergen exposure affects all T effector cells in lung, with a dominant of Th2 cells, but with different local cell distribution, probably due to a distinguished local inflammatory milieu.

  15. CTLA-4 is Required by CD4+CD25+ Treg to Control CD4+ T Cell Lymphopenia-Induced Proliferation

    OpenAIRE

    Sojka, Dorothy K.; Hughson, Angela; Fowell, Deborah J.

    2009-01-01

    CTLA-4 is constitutively expressed by CD4+CD25+Foxp3+ regulatory T cells (Treg) but its precise role in Treg function is not clear. Although blockade of CTLA-4 interferes with Treg function, studies using CTLA-4 deficient Treg have failed to reveal an essential requirement for CTLA-4 in Treg suppression in vivo. Conditional deletion of CTLA-4 in Foxp3+ T cells disrupts immune homeostasis in vivo but the immune processes disrupted by CTLA-4 deletion have not been determined. We demonstrate tha...

  16. Expression and significance of CD4 + CD25 + CD127 - regulatory T Cells in patients with chronic hepatitis C viral infection%慢性丙型肝炎患者中CD4+ CD25+ CD127-调节性 T 细胞的表达及意义

    Institute of Scientific and Technical Information of China (English)

    韩荔芬; 林国贤; 陈怡; 陈清; 邱荣仙; 郑玲

    2015-01-01

    Objective To explore the expression of CD4 + CD25 + CD127 - regulatory T Cells from patients with chronic hepatitis C viral infection,and to evaluate its clinical significance. Methods 67 patients with HCV and 32 healthy volunteers were included in this study. The proportion of CD4 + CD25 + CD127 - regulatory T cells in peripheral blood was detected by using flow cytometry. HCV RNA detection in the pe-ripheral blood of HCV patients were done by real - time fluorescent quantitative PCR. Results Comparing with healthy controls,drug abusing and non - drug abusing HCV - infected patients in peripheral blood had a lower percentage of CD4 + CD25 + CD127 - regulatory T cells. The proportion of CD4 + CD25 + CD127 - regulatory T cells to CD4 + T cells in peripheral blood from drug abusing and non - drug abusing HCV - infected patients (4. 44% ± 1. 67% )account for(3. 84% ± 1. 95% )and(4. 44% ± 1. 67% )respectively,which was significantly lower than those from healthy controls(6. 10% ± 1. 65% )( P 0. 05);The expression of CD4 + CD25 + CD127 - regulatory T cells in HCV group before antiviral treatment was evident-ly lower than that in treated HCV group and control group( P < 0. 01). Conclusion The expression of CD4 + CD25 + CD127 - regulatory T cells in the antiviral treatment HCV patients was evidently higher than those of untreated HCV patients,which explained that the regulatory T cell possi-bly hold a role in the activation of the immune response to HCV infection. In addition,the expression level of CD4 + CD25 + CD127 - regulatory T cells in peripheral blood may be monitoring the patientˊs body cell immune status and become the index of body anti - virus immune response.%目的:探讨 CD4+ CD25+ CD127-调节性 T 细胞在慢性丙型肝炎(HCV)患者中的表达及其临床意义。方法收集 HCV 患者67例,并根据是否吸毒分为吸毒丙肝组35例和非吸毒丙肝组32例;健康体检人员32例作为对照。采用流式细

  17. CD4+CD25+Foxp3+调节性T细胞与自身免疫性疾病%Expression and function of CD4 + CD25 + Foxp3 + in autoimmune diseases

    Institute of Scientific and Technical Information of China (English)

    王露; 张政

    2012-01-01

    自身免疫性疾病系由于机体免疫系统失衡,产生针对自身组织的免疫应答并导致自身组织、器官损害的一类疾病.调节性T淋巴细胞(regulatory T cell,Treg)具有免疫应答低下和免疫抑制特性,在维持机体免疫耐受和免疫应答稳态方面具有非常重要的作用,Treg的异常与多种自身免疫性疾病有关[1].Foxp3特异性表达于CD4+ CD25+ Treg细胞,与其发育、成熟以及抑制功能关系密切.但是目前关于该转录因子的表达调控机制却不清楚.本文拟就CD4+ CD25+ Foxp3 Treg细胞的研究进展及与多种自身免疫性疾病的关系作一综述.%Autoimmune Diseases are disorders caused by immune reactions against self - organs, as results of pathological imbalance of the immune system. New evidences indicates that regulatory T cells (Treg), characterized by immune annergy and immune suppression, have play critical roles in containing immune tolerance, immune response, and homeostasis. The expression of Foxp3, mainly in Treg cells, is thought to be the major factors determining the immune regulative function of Treg. However, F0XP3 regulation is poorly understood. In this article, we reviewed the recent progresses in researches of CD4 + CD25 + Foxp3 + Treg and its role in some autoimmune disease.

  18. The investigation of expression level of TSLP in thymus and the phenotype of CD4+CD25+Foxp3+ Treg cells in patients with myasthenia gravis%重症肌无力胸腺基质淋巴细胞生成素表达与CD4+CD25+Foxp3+Treg细胞表型的研究

    Institute of Scientific and Technical Information of China (English)

    孙延鹏; 卢祖能; 孙强; 杨超; 王云甫

    2012-01-01

    Objective To investigate the correlation between expression level of thymic stromal lymphopoietin (TSLP) in thymus and the expression of CD4+ CD25+ Foxp3+ Treg cells in patients with myasthenia gravis (MG). Methods The ratio of CD4+ CD25+ Foxp3+ Treg/CD4+ T cell were tested by flow cytometry from peripheral blood mononuclear cell which has been dealt with CD4+CD25+ antibody on the surface of the cell and Foxp3+ antibody into the cell in 16 patients with MG and 23 patients with congenital heart disease (control group). At the same time, thymuses cut from the corresponding patients were obtained to count the amount of TSLP positive Hassell corpuscles, the amount of TSLP positive Hassell corpuscles was compared between the two groups. The correlation between the amount of TSLP positive Hassall's corpuscles and CD4+ CD25+ Foxp3+ Treg cells expression was analyzed by logistic regression test. Results There was no statistical difference of the ratio of CD4+ CD25+ T/CD4+ T cells between the MG group [ (6. 24 + 0. 62) %] and the control group [ (6. 56 ±0. 65) %] (P>0. 05), but the ratio of CD4+ CD25+ Foxp3+ Treg/CD4+ T cells in MG group [ (6. 24 ± 0. 62)%] was significantly lower than that in the control group [ (5. 73 ±0. 56)%] (F<0. 01). The number of TSLP positive Hassell corpuscles in MG group was significantly fewer than that in the control group (6. 81 + 2. 17 versus 18. 87 + 3. 06, P<0. 01). The logistic regression analysis demonstrated that the expression of TSLP in the MG groups was linear related with the expression of Treg cells (R2 =0. 158, F= 13. 42, P< 0. 01). Conclusions The inadequate expression of TSLP is positively related to phenotype defiiency of the CD4+ CD25+Foxp3+ during Treg cell growth.%目的 探索重症肌无力患者胸腺基质淋巴细胞生成素(TSLP)表达水平与CD4+CD25+Foxp3+调节性T细胞(Treg)表型的相关性.方法 MG组(16例经胸腺切除的MG患者)及对照组(23例先天性心脏病心脏手术后患者)取

  19. Human T cells express CD25 and Foxp3 upon activation and exhibit effector/memory phenotypes without any regulatory/suppressor function

    Directory of Open Access Journals (Sweden)

    Godder Kamar

    2009-10-01

    Full Text Available Abstract Background Foxp3 has been suggested to be a standard marker for murine Tregs whereas its role as marker for human Tregs is controversial. While some reports have shown that human Foxp3+ T cells had no regulatory function others have shown their role in the inhibition of T cell proliferation. Methods T cell activation was performed by means of brayostatin-1/ionomycin (B/I, mixed lymphocyte reaction (MLR, and CD3/CD28 activation. T cell proliferation was performed using BrdU and CFSE staining. Flow cytometry was performed to determine Foxp3 expression, cell proliferation, viabilities and phenotype analyses of T cells. Results Both CD4+ and CD8+ T cells expressed Foxp3 upon activation in vitro. Expression of Foxp3 remained more stable in CD4+CD25+ T cells compared to that in CD8+CD25+ T cells. The CD4+CD25+Foxp3+ T cells expressed CD44 and CD62L, showing their effector and memory phenotypes. Both FoxP3- responder T cells and CD4+FoxP3+ T cells underwent proliferation upon CD3/CD28 activation. Conclusion Expression of Foxp3 does not necessarily convey regulatory function in human CD4+CD25+ T cells. Increased FoxP3 on CD44+ effector and CD44+CD62L+ memory T cells upon stimulation suggest the activation-induced regulation of FoxP3 expression.

  20. Expression and significance of CD4+ CD25+ Foxp3+ T cells in the acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation%骨髓或外周血中CD4+CD25+Foxp3+调节性T淋巴细胞与aGVHD的关系

    Institute of Scientific and Technical Information of China (English)

    王晓娟; 车传珍; 周昱男; 刘仲萍; 黄士昂

    2009-01-01

    reaction with aGVHD as the internal control gene, and the relative expression of Foxp3 protein by Western blot with α-tubulin as the internal control protein. Results aGVHD occurred in 13 out of 37 patients (aGVHI) group),and the remaining 24 had no aGVHD (no aGVHD group) within 100 days after transplantation. The percentage of CD4+ CD25+ Foxp3+ T cells was the lowest at 0 day of transplantation with the difference being statistically significant difference in comparison to that at other time points (-7, +30, +60,and +90 days) (P<0.01). The percentage of CD4+ CD25+ Foxp3+ T cells in no aGVHD group was significantly higher than in aGVHD group (P <0.01). The expression level of Foxp3 mRNA in aGVHD group was gradually increased after transplantation, but obviously lower than in no aGVHD group, especially at the post-transplantation day 60,and 90,there was significant difference between two group (P<0.01). Western blot revealed that the expression of Foxp3 protein in aGVHD group was significantly lower than in no aGVHD group. Conclusion CD4+ CD25+ Foxp3+ T cells may be of importance in preventing the development of aGVHD. Monitoring of the percentage of CD4+ CD25+ Foxp3+ T cells in bone marrow or peripheral blood can predict the occurrence of aGVHD after transplantation.

  1. CD4+CD25bright T cells in human intestinal lamina propria as regulatory cells.

    Science.gov (United States)

    Makita, Shin; Kanai, Takanori; Oshima, Shigeru; Uraushihara, Koji; Totsuka, Teruji; Sawada, Taisuke; Nakamura, Tetsuya; Koganei, Kazutaka; Fukushima, Tsuneo; Watanabe, Mamoru

    2004-09-01

    It is well known that immune responses in the intestine remain in a state of controlled inflammation, suggesting that not only active suppression by regulatory T cells plays an important role in the normal intestinal homeostasis, but also its dysregulation leads to the development of inflammatory bowel disease. In this study, we demonstrate that the CD4(+)CD25(bright) T cells reside in the human intestinal lamina propria (LP) and functionally retain regulatory activities. All human LP CD4(+) T cells regardless of CD25 expression constitutively expressed CTLA-4, glucocorticoid-induced TNFR family-related protein, and Foxp3 and proliferate poorly. Although LP CD4(+)CD25(-) T cells showed an activated and anergic/memory phenotype, they did not retain regulatory activity. In LP CD4(+)CD25(+) T cells, however, cells expressing CD25 at high levels (CD4(+)CD25(bright)) suppressed the proliferation and various cytokine productions of CD4(+)CD25(-) T cells. LP CD4(+)CD25(bright) T cells by themselves produced fewer amounts of IL-2, IFN-gamma, and IL-10. Interestingly, LP CD4(+)CD25(bright) T cells with regulatory T activity were significantly increased in patients with active inflammatory bowel disease. These results suggest that CD4(+)CD25(bright) T cells found in the normal and inflamed intestinal mucosa selectively inhibit the host immune response and therefore may contribute to the intestinal immune homeostasis. PMID:15322172

  2. CD4+CD25- T cells that express latency-associated peptide on the surface suppress CD4+CD45RBhigh-induced colitis by a TGF-beta-dependent mechanism.

    Science.gov (United States)

    Oida, Takatoku; Zhang, Xingmin; Goto, Masao; Hachimura, Satoshi; Totsuka, Mamoru; Kaminogawa, Shuichi; Weiner, Howard L

    2003-03-01

    Murine CD4(+)CD25(+) regulatory cells have been reported to express latency-associated peptide (LAP) and TGF-beta on the surface after activation, and exert regulatory function by the membrane-bound TGF-beta in vitro. We have now found that a small population of CD4(+) T cells, both CD25(+) and CD25(-), can be stained with a goat anti-LAP polyclonal Ab without being stimulated. Virtually all these LAP(+) cells are also positive for thrombospondin, which has the ability to convert latent TGF-beta to the active form. In the CD4(+)CD45RB(high)-induced colitis model of SCID mice, regulatory activity was exhibited not only by CD25(+)LAP(+) and CD25(+)LAP(-) cells, but also by CD25(-)LAP(+) cells. CD4(+)CD25(-)LAP(+) T cells were part of the CD45RB(low) cell fraction. CD4(+)CD25(-)LAP(-)CD45RB(low) cells had minimal, if any, regulatory activity in the colitis model. The regulatory function of CD25(-)LAP(+) cells was abrogated in vivo by anti-TGF-beta mAb. These results identify a new TGF-beta-dependent regulatory CD4(+) T cell phenotype that is CD25(-) and LAP(+). PMID:12594277

  3. Tim-3 mRNA Expression in Peripheral Blood Mononuclear Cells and Its Relationship with CD4+CD25+ Regulatory T Cells from Asthmatic Children%哮喘儿童外周血单个核细胞Tim-3mRNA的表达及其与CD4+CD25+调节性T细胞的关系

    Institute of Scientific and Technical Information of China (English)

    陆小霞

    2011-01-01

    (Treg) , explore clemcntarily the role of Tim-3 in the occurrence and dcvclopment of asthmatic inflammation. Methods Scventy-three asthmatic childrcn were enrolled in this study,including 35 children with acute asthma exacerbation(acute attack group)and 38 with asthma remission(remittent group). The Tim-3 mRNA expression was detected by using RTPCR. The ratio of CD4+ CD25+ Treg(CD4+CD25+ Treg/CD4+ T) was measured by using flow cytometry. The levels of interleukin-6(IL6)and trans forming growth factor-β ( TGF-β)were detcrmincd by using ELISA. The correlation among Tim-3 mRNA, CD4+ CD25+ Treg and IL-6 level was analyzed. Results The cxpression of Tim-3 mRNA in childrcn with acute asthma exacerbation was(0. 86± 0. 17) , significantly highcr than that in remission stage and normal control group(0. 39±0. 11 and 0. 06±0. 03 , both P<O. 05). The expression of Tim-3 mRNA in children with asthma remission was significantly higher than that in normal control group(P< 0. 05). The pcrccntage of CD4+ CD25+ Trcg in children with acute asthma exacerbation was( 8. 35± 1. 67)% , significantly lower than that in remission stage and normal control group [(10. 21±2. 04)% and ( 12. 43±2. 58)% , both P < 0. 05]. The level of IL- 6 in children with acute asthma exacerbation was( 78. 35± 14. 59) pg/mL, significantly higher than that in remission stage and normal control group(36. 48±9. 18 and 10. 24±3. 57 pg/mL, both P<0. 05). The level of IL-6 in children with asthma remis sion was significantly higher than that in normal control group(P<O. 05). There was no significant difference in level of TGF-β among three groups. The Tim-3 mRNA cxprcssion in asthma exaccrbation and rcmission stage was correlated negatively with the level of CD4+ CD25+ Treg( r= - 0. 81 , - 0. 79 , both P<O. 05) , and positively with the level of IL-6(r= 0. 87 , 0. 83 , both P<0. 01). Conclusion The increased expression of Tim3 mRNA in PBMC might take part in the occurrence and devclopment of asthmatic

  4. Curcumin blocks interleukin (IL)-2 signaling in T-lymphocytes by inhibiting IL-2 synthesis, CD25 expression, and IL-2 receptor signaling

    Energy Technology Data Exchange (ETDEWEB)

    Forward, Nicholas A.; Conrad, David M. [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia (Canada); Power Coombs, Melanie R.; Doucette, Carolyn D. [Department of Pathology, Dalhousie University, Halifax, Nova Scotia (Canada); Furlong, Suzanne J. [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia (Canada); Lin, Tong-Jun [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia (Canada); Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia (Canada); Hoskin, David W., E-mail: d.w.hoskin@dal.ca [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia (Canada); Department of Pathology, Dalhousie University, Halifax, Nova Scotia (Canada); Department of Surgery, Dalhousie University, Halifax, Nova Scotia (Canada)

    2011-04-22

    Highlights: {yields} Curcumin inhibits CD4{sup +} T-lymphocyte proliferation. {yields} Curcumin inhibits interleukin-2 (IL-2) synthesis and CD25 expression by CD4{sup +} T-lymphocytes. {yields} Curcumin interferes with IL-2 receptor signaling by inhibiting JAK3 and STAT5 phosphorylation. {yields} IL-2-dependent regulatory T-lymphocyte function and Foxp3 expression is downregulated by curcumin. -- Abstract: Curcumin (diferulomethane) is the principal curcuminoid in the spice tumeric and a potent inhibitor of activation-induced T-lymphocyte proliferation; however, the molecular basis of this immunosuppressive effect has not been well studied. Here we show that micromolar concentrations of curcumin inhibited DNA synthesis by mouse CD4{sup +} T-lymphocytes, as well as interleukin-2 (IL-2) and CD25 ({alpha} chain of the high affinity IL-2 receptor) expression in response to antibody-mediated cross-linking of CD3 and CD28. Curcumin acted downstream of protein kinase C activation and intracellular Ca{sup 2+} release to inhibit I{kappa}B phosphorylation, which is required for nuclear translocation of the transcription factor NF{kappa}B. In addition, IL-2-dependent DNA synthesis by mouse CTLL-2 cells, but not constitutive CD25 expression, was impaired in the presence of curcumin, which demonstrated an inhibitory effect on IL-2 receptor (IL-2R) signaling. IL-2-induced phosphorylation of STAT5A and JAK3, but not JAK1, was diminished in the presence of curcumin, indicating inhibition of critical proximal events in IL-2R signaling. In line with the inhibitory action of curcumin on IL-2R signaling, pretreatment of CD4{sup +}CD25{sup +} regulatory T-cells with curcumin downregulated suppressor function, as well as forkhead box p3 (Foxp3) expression. We conclude that curcumin inhibits IL-2 signaling by reducing available IL-2 and high affinity IL-2R, as well as interfering with IL-2R signaling.

  5. Curcumin blocks interleukin (IL)-2 signaling in T-lymphocytes by inhibiting IL-2 synthesis, CD25 expression, and IL-2 receptor signaling

    International Nuclear Information System (INIS)

    Highlights: → Curcumin inhibits CD4+ T-lymphocyte proliferation. → Curcumin inhibits interleukin-2 (IL-2) synthesis and CD25 expression by CD4+ T-lymphocytes. → Curcumin interferes with IL-2 receptor signaling by inhibiting JAK3 and STAT5 phosphorylation. → IL-2-dependent regulatory T-lymphocyte function and Foxp3 expression is downregulated by curcumin. -- Abstract: Curcumin (diferulomethane) is the principal curcuminoid in the spice tumeric and a potent inhibitor of activation-induced T-lymphocyte proliferation; however, the molecular basis of this immunosuppressive effect has not been well studied. Here we show that micromolar concentrations of curcumin inhibited DNA synthesis by mouse CD4+ T-lymphocytes, as well as interleukin-2 (IL-2) and CD25 (α chain of the high affinity IL-2 receptor) expression in response to antibody-mediated cross-linking of CD3 and CD28. Curcumin acted downstream of protein kinase C activation and intracellular Ca2+ release to inhibit IκB phosphorylation, which is required for nuclear translocation of the transcription factor NFκB. In addition, IL-2-dependent DNA synthesis by mouse CTLL-2 cells, but not constitutive CD25 expression, was impaired in the presence of curcumin, which demonstrated an inhibitory effect on IL-2 receptor (IL-2R) signaling. IL-2-induced phosphorylation of STAT5A and JAK3, but not JAK1, was diminished in the presence of curcumin, indicating inhibition of critical proximal events in IL-2R signaling. In line with the inhibitory action of curcumin on IL-2R signaling, pretreatment of CD4+CD25+ regulatory T-cells with curcumin downregulated suppressor function, as well as forkhead box p3 (Foxp3) expression. We conclude that curcumin inhibits IL-2 signaling by reducing available IL-2 and high affinity IL-2R, as well as interfering with IL-2R signaling.

  6. Effect of Lactobacillussalivariuson the Number of CD4+CD25+Foxp3+Treg Cells and the Expression of TGF-β1 in Asthma Balb/c Mice%唾液乳杆菌对哮喘小鼠CD4+CD25+Foxp3+Treg细胞数量及TGF-β1表达的影响

    Institute of Scientific and Technical Information of China (English)

    相云; 尚云晓; 李淼

    2015-01-01

    Objective To explore the effect of Lactobacillussalivariuson the number of CD4+CD25+Foxp3+Treg cells and expression of transform⁃ing growth factorβ1(TGF⁃β1)in asthma Balb/c mice. Methods Thirty⁃two female Balb/c mice were randomly divided into four groups:the nor⁃mal control group,the asthma group,the Lactobacillus salivarius group,and the asthma combined Lactobacillussalivariusgroup. Acute asthma mod⁃el was established by the ovalbumin challenge method. After extraction of primary spleen cells,flow cytometry was used to test CD4+CD25+Foxp3+Treg/CD4+T ratio in spleen lymphocytes. The levels of IL⁃4,IFN⁃γand TGF⁃β1 in the spleen cell culture supernatant were measured by ELISA method. Results The level of Th2 cytokine(IL⁃4)in the spleen cell culture supernatant of the asthma group was significantly higher than that of the control group(P<0.05),however,the level of Th1 cytokine(IFN⁃γ)was significantly lower than that of the control group(P<0.05). The ex⁃pression level of Th2 cytokine(IL⁃4)in the Lactobacillussalivariusintervention group was significantly decreased compared with the asthma group, and the Th1 cytokine(IFN⁃γ)expression level was elevated compared with the asthma group(P<0.05). The level of TGF⁃β1 in the Lactobacillus salivarius intervention group was higher than in the asthma group(P<0.05). The proportion of CD4+CD25+Foxp3+Treg/CD4+T in spleen lympho⁃cytes in the asthma group was lower than that in the control group(P<0.05),and was higher in the Lactobacillus salivarius intervention group than in the asthma group(P<0.05). Conclusion CD4+CD25+Foxp3+Treg was associated with the pathogenesis of asthma. Lactobacillus salivarius could adjust Th1/Th2 imbalance and reduce asthma inflammation through up⁃regulation of CD4+CD25+Foxp3+Treg and TGF⁃β1 expression.%目的:探讨唾液乳杆菌对哮喘Balb/c小鼠CD4+CD25+Foxp3+Treg细胞数量及转化生长因子β1(TGF⁃β1)表达的影响。方法将32只

  7. In ovo injection of anti-chicken CD25 monoclonal antibodies depletes CD4+CD25+ T cells in chickens.

    Science.gov (United States)

    Shanmugasundaram, Revathi; Selvaraj, Ramesh K

    2013-01-01

    The CD4(+)CD25(+) cells have T regulatory cell properties in chickens. This study investigated the effect of in ovo injection of anti-chicken CD25 monoclonal antibodies (0.5 mg/egg) on CD4(+)CD25(+) cell depletion and on amounts of interleukin-2 mRNA and interferon-γ mRNA in CD4(+)CD25(-) cells posthatch. Anti-chicken CD25 or PBS (control) was injected into 16-d-old embryos. Chicks hatched from eggs injected with anti-chicken CD25 antibodies had a lower CD4(+)CD25(+) cell percentage in the blood until 25 d posthatch. The anti-chicken CD25 antibody injection nearly depleted CD4(+)CD25(+) cells in the blood until 16 d posthatch. At 30 d posthatch, the CD4(+)CD25(+) cell percentage in the anti-CD25-antibody-injected group was comparable with the percentage in the control group. At 16 d posthatch, the anti-chicken CD25 antibody injection decreased CD4(+)CD25(+) cell percentages in the thymus, spleen, and cecal tonsils. Chickens hatched from anti-CD25-antibody-injected eggs had approximately 25% of CD4(+)CD25(+) cells in the cecal tonsils and thymus compared with those in the cecal tonsils and thymus of the control group. The CD4(+)CD25(-) cells from the spleen and cecal tonsils of chicks hatched from anti-chicken-CD25-injected eggs had higher amounts of interferon-γ and interleukin-2 mRNA than CD4(+)CD25(-) cells from the control group. It could be concluded that injecting anti-chicken CD25 antibodies in ovo at 16 d of incubation nearly depleted the CD4(+)CD25(+) cells until 25 d posthatch. PMID:23243240

  8. Orally-Induced Intestinal CD4+ CD25+ FoxP3+ Treg Controlled Undesired Responses towards Oral Antigens and Effectively Dampened Food Allergic Reactions.

    Directory of Open Access Journals (Sweden)

    Paola Lorena Smaldini

    Full Text Available The induction of peripheral tolerance may constitute a disease-modifying treatment for allergic patients. We studied how oral immunotherapy (OIT with milk proteins controlled allergy in sensitized mice (cholera toxin plus milk proteins upon exposure to the allergen. Symptoms were alleviated, skin test was negativized, serum specific IgE and IgG1 were abrogated, a substantial reduction in the secretion of IL-5 and IL-13 by antigen-stimulated spleen cells was observed, while IL-13 gene expression in jejunum was down-regulated, and IL-10 and TGF-β were increased. In addition, we observed an induction of CD4+CD25+FoxP3+ cells and IL-10- and TGF-β-producing regulatory T cells in the lamina propria. Finally, transfer experiments confirmed the central role of these cells in tolerance induction. We demonstrated that the oral administration of milk proteins pre- or post-sensitization controlled the Th2-immune response through the elicitation of mucosal IL-10- and TGF-β-producing Tregs that inhibited hypersensitivity symptoms and the allergic response.

  9. Expression of surface markers on peripheral CD4+CD25high T cells in patients with atopic asthma: role of inhaled corticosteroid

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Background CD4+CD25+ regulatory T cells (Tregs) mediate immune suppression through cell-cell contact with surface molecules,particularly cytotoxic T lymphocyte-associated antigen 4 (CTLA-4),glucocorticoid-induced tumor necrosis factor receptor family-related protein (GITR),and transforming growth factor β(TGF-β),but little is known about the exact role of Tregs in the pathogenesis of asthma.This study sought to characterize the expression of surface markers on peripheral blood mononuclear cells-derived Tregs in patients with atopic asthma and healthy subjects,and to investigate the effect of inhaled corticosteroid on them.Methods The expression of surface molecules on CD4+CD25hign Tregs was detected by flow cytometry.The effect of inhaled corticosteroid on expression of the surface molecules on Tregs was determined in vivo and in vitro.Total serum immunoglobulin E (IgE) and high-sensitivity C-reactive protein were measured by enzyme linked immunosorbent assay and latex enhanced immunoturbidimetric assay,respectively.Results Equivalent numbers of peripheral Tregs were found in patients with atopic asthma (stable and acute) and healthy subjects.Tregs preferentially expressed CTLA-4, GITR,toll-like receptor 4 (TLR4),latency-associated peptide (LAP/TGF-β1),and forkhead box P3 (FOXP3).Patients with acute asthma had decreased numbers of CD4+CD25high LAP+ T cells compared to healthy subjects and stable asthmatics.Inhaled corticosteroid enhanced the percentage of Tregs expressing LAP in vivo and in vitro dose-dependently.Furthermore,the percentages of Tregs expressing LAP were negatively correlated with total serum IgE levels and severity of asthma,but positively correlated with forced expiratory volume in one second percentage of the predicted value in patients with asthma.Concluslons The results suggest that membrane-bound TGF-β1 is a potential candidate for predicting the severity of asthma,and may contribute to the sustained remission of asthma,Strategies targeting

  10. Over-expression of Stat5b confers protection against diabetes in the non-obese diabetic (NOD) mice via up-regulation of CD4+CD25+ regulatory T cells

    International Nuclear Information System (INIS)

    Highlights: ► This is the first study to provide direct evidence of the role of Stat5b in NOD mice. ► Over-expression of wild type Stat5b transgene protects NOD mice against diabetes. ► This protection may be mediated by the up-regulation of CD4+CD25+ Tregs. -- Abstract: The signal transducers and activators of transcription (STAT) family of proteins play a critical role in cytokine signaling required for fine tuning of immune regulation. Previous reports showed that a mutation (L327M) in the Stat5b protein leads to aberrant cytokine signaling in the NOD mice. To further elaborate the role of Stat5b in diabetes, we established a NOD transgenic mouse that over-expresses the wild type Stat5b gene. The incidences of spontaneous diabetes as well as cyclophosphamide-induced diabetes were significantly reduced and delayed in the Stat5b transgenic NOD mice compared to their littermate controls. The total cell numbers of CD4+ T cells and especially CD8+ T cells in the spleen and pancreatic lymph node were increased in the Stat5b transgenic NOD mice. Consistent with these findings, CD4+ and CD8+ T cells from the Stat5b transgenic NOD mice showed a higher proliferation capacity and up-regulation of multiple cytokines including IL-2, IFN-γ, TNF-α and IL-10 as well as anti-apoptotic gene Bcl-xl. Furthermore, the number and proportion of CD4+CD25+ regulatory T cells were significantly increased in transgenic mice although in vitro suppression ability of the regulatory T-cells was not affected by the transgene. Our results suggest that Stat5b confers protection against diabetes in the NOD mice by regulating the numbers and function of multiple immune cell types, especially by up-regulating CD4+CD25+ regulatory T cells.

  11. Natural CD4~+CD25~+ regulatory T cells express α7-nicotinic acetylcholine receptor subunits%小鼠天然CD4~+CD25~+调节性T细胞表达α7烟碱型乙酰胆碱受体

    Institute of Scientific and Technical Information of China (English)

    王大伟; 周荣斌; 姚咏明

    2010-01-01

    Objective To investigate whether CD4~+ CD25~+ regulatory T cells (Treg) from C57BL/6J mice express alpha7 nicotinic acetylcholine receptor (α7nAChR). Methods CD4~+ CD25~+ regulatory T cells were isolated from mouse splenocytes with a CD4~+ CD25~+ regulatory T Cell isolation kit (Mihenyi Bio-tee). The purity of isolated Tregs was analyzed by flow eytometry. Expressions of α7nAChR in mouse CD4~+ CD25~+ Tregs were examined by immunofluorescence staining, Western blotting, and reverse transeription-PCR, respectively. Results It was revealed by flow cytometry that Tregs could bind alpha-bungarotoxin (α-BGT)-F/TC, a specific α7 nAChR antagonist. Moreover, a positive binding to α-Bgt was also observed on the cell surface of Treg, as viewed by fluorescent confoeal microscopy. In addition, a clear band of a7nAChR with a molecular mass of approximately 55 kD was found from Tregs by Western blotting analysis, and α7nAChR mRNA was expressed with the expected size of 199 bp from Tregs by reverse transcription-PCR. Conclusion Natural CD4~+ CD25~+ Tregs from mice express α7nAChR.%目的 探讨C57BL/6J小鼠的天然CD4~+CD25~+调节性T细胞(CD4~+CD25~+Treg)是否存在α7烟碱型乙酰胆碱受体(a7nAchR).方法 使用小鼠调节性T细胞试剂盒分离小鼠脾脏CD4~+CD25~+Treg,流式细胞术鉴定CD4~+CD25~+Treg的纯度.分别采用免疫荧光染色、共聚焦湿微镜、Western印迹和逆转录聚合酶链反应检测Treg表面α7nAchR蛋白/基因表达.结果 α-银环蛇毒素-FITC染色、流式检测显示Treg细胞表面结合α-银环蛇毒素-FITC;共聚焦显微镜成像观察到Treg细胞表面结合大量α-银环蛇毒素;Western印迹检测证实Treg细胞样本中检测到了清楚的α7nAchR条带,分子量大小约为55 kD;RT-PCR分析发现Treg细胞样本中检测到了199 bp大小的特异性α7nAchR目的 基因条带.结论小鼠天然CD4~+CD25~+Treg细胞表达α7nAChR.

  12. Prognostic Relevance of Cytokine Receptor Expression in Acute Myeloid Leukemia: Interleukin-2 Receptor α-Chain (CD25) Expression Predicts a Poor Prognosis

    Science.gov (United States)

    Nakase, Kazunori; Kita, Kenkichi; Kyo, Taiichi; Ueda, Takanori; Tanaka, Isao; Katayama, Naoyuki

    2015-01-01

    A variety of cytokine/cytokine receptor systems affect the biological behavior of acute leukemia cells. However, little is known about the clinical relevance of cytokine receptor expression in acute myeloid leukemia (AML). We quantitatively examined the expression of interleukin-2 receptor α-chain (IL-2Rα, also known as CD25), IL-2Rβ, IL-3Rα, IL-4Rα, IL-5Rα, IL-6Rα, IL-7Rα, the common β-chain (βc), γc, granulocyte-macrophage colony-stimulating factor (GM-CSF)Rα, G-CSFR, c-fms, c-mpl, c-kit, FLT3, and GP130 in leukemia cells from 767 adult patients with AML by flow cytometry and determined their prevalence and clinical significance. All cytokine receptors examined were expressed at varying levels, whereas the levels of IL-3Rα, GM-CSFRα, IL-2Rα, γc, c-kit, and G-CSFR exhibited a wide spectrum of ≥10,000 sites/cell. In terms of their French-American-British classification types, GM-CSFRα and c-fms were preferentially expressed in M4/M5 patients, G-CSF in M3 patients, and IL-2Rα in non-M3 patients. Elevated levels of IL-3Rα, GM-CSFRα, and IL-2Rα correlated with leukocytosis. In patients ≤60 years old, higher levels of these 3 receptors correlated with poor responses to conventional chemotherapy, but only IL-2Rα was associated with a shorter overall survival. By incorporating IL-2Rα status into cytogenetic risk stratification, we could sort out a significantly adverse-risk cohort from the cytogenetically intermediate-risk group. Analyses with various phenotypical risk markers revealed the expression of IL-2Rα as an independent prognostic indicator in patients with intermediate-risk cytogenetics. These findings were not observed in patients >60 years old. Our results indicate that several cytokine receptors were associated with certain cellular and clinical features, but IL-2Rα alone had prognostic value that provides an additional marker to improve current risk evaluation in AML patients ≤60 years old. PMID:26375984

  13. IL-2 regulates FOXP3 expression in human CD4+CD25+ regulatory T cells through a STAT-dependent mechanism and induces the expansion of these cells in vivo.

    Science.gov (United States)

    Zorn, Emmanuel; Nelson, Erik A; Mohseni, Mehrdad; Porcheray, Fabrice; Kim, Haesook; Litsa, Despina; Bellucci, Roberto; Raderschall, Elke; Canning, Christine; Soiffer, Robert J; Frank, David A; Ritz, Jerome

    2006-09-01

    IL-2 plays a critical role in the maintenance of CD4+CD25+ FOXP3(+) regulatory T cells (Tregs) in vivo. We examined the effects of IL-2 signaling in human Tregs. In vitro, IL-2 selectively up-regulated the expression of FOXP3 in purified CD4+CD25+ T cells but not in CD4+CD25- cells. This regulation involved the binding of STAT3 and STAT5 proteins to a highly conserved STAT-binding site located in the first intron of the FOXP3 gene. We also examined the effects of low-dose IL-2 treatment in 12 patients with metastatic cancer and 9 patients with chronic myelogenous leukemia after allogeneic hematopoietic stem cell transplantation. Overall, IL-2 treatment resulted in a 1.9 median fold increase in the frequency of CD4+CD25+ cells in peripheral blood as well as a 9.7 median fold increase in FOXP3 expression in CD3+ T cells. CD56+CD3- natural killer (NK) cells also expanded during IL-2 therapy but did not express FOXP3. In vitro treatment of NK cells with 5-aza-2'-deoxycytidine restored the IL-2 signaling pathway leading to FOXP3 expression, suggesting that this gene was constitutively repressed by DNA methylation in these cells. Our findings support the clinical evaluation of low-dose IL-2 to selectively modulate CD4+CD25+ Tregs and increase expression of FOXP3 in vivo. PMID:16645171

  14. 胃癌患者外周血及癌组织中CD4+CD25+调节性T细胞、转录因子Foxp3的表达及临床意义%Expression and clinical signification of CD4+CD25+ regulatory T cells,the transcription factor Foxp3 in peripheral blood and cancerous tissue of gastric cancer

    Institute of Scientific and Technical Information of China (English)

    陆威; 李永翔; 张尚鑫; 闫强

    2013-01-01

    目的 探讨胃癌患者外周血及胃癌组织中CD4+CD25+调节性T细胞(Tregs)和转录因子Foxp3表达及临床意义.方法 收集38例胃癌患者术前血液样本和20例健康体检者血液样本进行流式细胞仪检测,同时胃癌组中留取胃癌组织和癌旁组织(距肿瘤边缘>5 cm)进行免疫组化染色,检测CD4+CD25+Foxp3+Tregs表达,并与胃癌的TNM分期、肿瘤分化程度、淋巴转移、肿瘤所在部位、肿瘤直径等相关参数进行分析.结果 胃癌组外周血中CD4+CD25+Foxp3+Tregs的表达水平与对照组相比差异有统计学意义(P<0.01),且在胃癌TNM分期、淋巴转移组内比较差异有统计学意义(P<0.01).胃癌组织中Foxp3+Tregs的表达水平与癌旁组织相比,差异有统计学意义(P<0.01),且在胃癌TNM分期、分化程度、淋巴转移组内比较差异有统计学意义(P<0.01,P<0.05).外周血中CD4+CD25+ Foxp3+Tregs与胃癌组织中Foxp3+Tregs的表达水平呈显著正相关(r=0.786,P 5 cm ) from gastric cancer were subjected to immunohis-tochemistry to detect CD4 + CD25 + Foxp3 + Tregs expression, and the correlation analysis with some clinical related parameters was made. Results The percentages of CD4 + CD25 + Foxp3 + Tregs in peripheral blood of gastric cancer group were significantly higher than those from healthy control group( P <0. 01 ),and were highly correlated to TNM stage, lymph node metastases( P <0. 01 ). The percentages of Foxp3 + Tregs in gastric cancer tissue were significantly higher than those from para-tumorous tissue( P <0. 01 ),and were highly correlated to TNM stage,tumor differentiation,lymph node metastases( P < 0. 01 , P < 0. 05 ). The percentages of CD4 + CD25 + Foxp3 + Tregs in peripheral blood was positively correlated to Foxp3 + Tregs in gastric cancer tissue( r - 0. 786, P < 0. 01 ). Conclusion The proportion of CD4 + CD25 + Foxp3 + Tregs in Peripheral blood and Foxp3 + Tregs in gastric cancer tissue is significantly elevated in

  15. Proliferative index and expression of CD38, Zap-70, and CD25 in different lymphoid compartments of chronic lymphocytic leukemia patients

    Directory of Open Access Journals (Sweden)

    Olga Khoudoleeva

    2011-01-01

    Full Text Available Olga Khoudoleeva1 Eugeny Gretsov1 Natasha Barteneva2,3 Ivan Vorobjev11Hematology Scientific Center, Russian Academy of Medical Sciences, Moscow, Russia; 2Immune Disease Institute and Program in Cellular and Molecular Biology, Children Hospital of Boston, Boston, MA, USA; 3Department of Pathology, Harvard Medical School, Boston, MA, USAAbstract: Recent studies of chronic lymphocytic leukemia (CLL show that malignant B cells proliferate at a rate similar to normal B lymphocytes. This is in apparent contradiction to the very low proliferation rate found in blood specimens from CLL patients. To address this problem, we studied the expression of Ki-67, CD38, CD25, and Zap-70 in different compartments of CLL patients. Using triple-color flow cytometry, we examined the expression of CD38, CD25, Zap-70, and Ki-67 antigens in the peripheral blood, bone marrow, spleen, and lymph nodes biopsies of patients with CLL, splenic marginal zone lymphoma (SMZL, and nonmalignant diseases. In parallel probes of lymph node/spleen biopsies and blood taken from one and the same patient, Ki-67 expression was 17 times higher. Among the whole cohort, we also found significantly higher Ki-67 expression in biopsies from lymph nodes and spleen (4.95% ± 0.55%, compared with bone marrow (1.88% ± 0.32% and peripheral blood (0.45% ± 0.03%, P < 0.01. In CLL patients, there are statistically significant correlations between the expression of CD38 and Ki-67 in bone marrow (P ≤ 0.01, Zap-70 and Ki-67 in blood (P ≤ 0.01, and Zap-70 and CD38 in blood (P ≤ 0.01. Patients with SMZL also showed a significant correlation between Ki-67 and CD38 expression (P ≤ 0.01 and between Ki-67 and Zap-70 expression (P ≤ 0.01. We show for the first time that proliferation of B lymphocytes in CLL patients is associated primarily with lymph nodes/spleen. Malignant cells in the blood represent only a subpopulation of nonproliferating and less-activated B cells in this disease

  16. Involvement of CD4+CD25+ regulatory T cells in the pathogenesis of polycythaemia vera

    Institute of Scientific and Technical Information of China (English)

    ZHAO Wen-bo; LI Ying; LIU Xin; ZHANG Ling-yan; WANG Xin

    2008-01-01

    Background Regulatory T cells (Treg) have been shown to play an important role in the regulation of hematopoietic activity. However, there is no information about the effect of Treg cells in the pathogenesis of polycythaemia vera (PV).Methods In this study, we investigated the percentage and function of Treg cells in the peripheral blood of 21 PV patients and 25 healthy donors. Treg cells were identified and characterized as CD4+CD25+FOXP3+ by flow cytometry.The suppressive activity of CD4+CD25+ Treg cells was assessed by the proliferation and cytokine secretion of the co-cultured CD4+CD25- fractions.Results The results showed that the percentage of Treg cells in the peripheral blood of PV patients significantly increased compared to healthy controls ((10.93±4.02)% vs (5.86±1.99)%, P <0.05). Moreover, the mRNA and protein expression of FOXP3 was higher in CD4+CD25+ Treg cells. Coordinately, when co-cultured with the activated CD4+CD25-cells, the CD4+CD25+ Treg cells showed enhanced suppressive function in PV. Yet, the underlying mechanism for the increased frequency and function of CD4+CD25+ Treg cells is still to be clarified.Conclusion Treg cells expansion might account for the abnormal T cell immunity in PV patients and thus contribute to the pathogenesis of PV.

  17. Over-expression of Stat5b confers protection against diabetes in the non-obese diabetic (NOD) mice via up-regulation of CD4{sup +}CD25{sup +} regulatory T cells

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Yulan; Purohit, Sharad [Center for Biotechnology and Genomic Medicine, Georgia Health Sciences University, GA (United States); Department of Pathology, Medical College of Georgia, Georgia Health Sciences University, GA (United States); Chen, Xueqin; Yi, Bing [Center for Biotechnology and Genomic Medicine, Georgia Health Sciences University, GA (United States); She, Jin-Xiong, E-mail: jshe@georgiahealth.edu [Center for Biotechnology and Genomic Medicine, Georgia Health Sciences University, GA (United States); Department of Pathology, Medical College of Georgia, Georgia Health Sciences University, GA (United States)

    2012-08-10

    Highlights: Black-Right-Pointing-Pointer This is the first study to provide direct evidence of the role of Stat5b in NOD mice. Black-Right-Pointing-Pointer Over-expression of wild type Stat5b transgene protects NOD mice against diabetes. Black-Right-Pointing-Pointer This protection may be mediated by the up-regulation of CD4{sup +}CD25{sup +} Tregs. -- Abstract: The signal transducers and activators of transcription (STAT) family of proteins play a critical role in cytokine signaling required for fine tuning of immune regulation. Previous reports showed that a mutation (L327M) in the Stat5b protein leads to aberrant cytokine signaling in the NOD mice. To further elaborate the role of Stat5b in diabetes, we established a NOD transgenic mouse that over-expresses the wild type Stat5b gene. The incidences of spontaneous diabetes as well as cyclophosphamide-induced diabetes were significantly reduced and delayed in the Stat5b transgenic NOD mice compared to their littermate controls. The total cell numbers of CD4{sup +} T cells and especially CD8{sup +} T cells in the spleen and pancreatic lymph node were increased in the Stat5b transgenic NOD mice. Consistent with these findings, CD4{sup +} and CD8{sup +} T cells from the Stat5b transgenic NOD mice showed a higher proliferation capacity and up-regulation of multiple cytokines including IL-2, IFN-{gamma}, TNF-{alpha} and IL-10 as well as anti-apoptotic gene Bcl-xl. Furthermore, the number and proportion of CD4{sup +}CD25{sup +} regulatory T cells were significantly increased in transgenic mice although in vitro suppression ability of the regulatory T-cells was not affected by the transgene. Our results suggest that Stat5b confers protection against diabetes in the NOD mice by regulating the numbers and function of multiple immune cell types, especially by up-regulating CD4{sup +}CD25{sup +} regulatory T cells.

  18. 前列腺癌患者手术前后外周血调节性T细胞和FOXP3mRNA的表达及临床意义%Levels of CD4+CD25+Regulatory T Cells & Expression of FOXP3 mRNA after Operation in Peripheral Blood from Patients with Prostate Cancer and its Clinical Significance

    Institute of Scientific and Technical Information of China (English)

    章更生

    2011-01-01

    目的 探讨前列腺癌(Pca)患者手术前后外周血CD4+CD25+调节性T细胞(Tregs)及其特异标志物FOXP3mRNA的表达比例的变化及临床意义.方法 应用流式细胞术检测50例前列腺癌及50例正常对照组外周血单个核细胞(PBMC)中Tregs占CD4+T细胞的比例,应用RT-PCR技术检测人外周血PBMC FOXP3 mRNA的表达.结果 随着Gleason分级升高,CD4+CD25+Tregs/CD4+T比值和FOXP3 mRNA表达量均有升高趋势;Pca患者术前CD4+CD25+ Treg细胞占CD4+T细胞的比例和FOXP3 mRNA表达水平高于正常对照组(均P<0.05),术后水平与正常对照组差异无统计学意义;Pca组Tregs表达率与FOXP3 mRNA呈正相关(r=0.623,P<0.01).结论 Tregs及其特异标志物FOXP3具有维持自身免疫稳定和抑制肿瘤免疫作用,可能参与前列腺癌的发生.%Objective To explore the rate change of CD4+ CD25+ regulatory T cells and expression of F0XP3 mRNA after operation in peripheral blood from patients with prostate cancer and its clinical significance. Methods Flow cytometry was used to analyze the proportion of CD4+CD25+ regulatory T cells/CD4T cells in peripheral blood mononuclear cells (PBMC) among 50 patients with prostate cancer and 50 healthy controls. The expression of FOXP3 mRNA in PBMC was detected by RT-PCR. Results The value of CD4+CU25+Tregs/CD4+ T cells and expression of FOXP3 mRNA were increased with Gleason pathological parameters increased. And the value of CD4+CD25+ regulatory T cells/CD4+T cells and expression of FOXP3 mRNA in PBMC from patients with prostate cancer before operation were significantly higher than that of the healthy controls. But there were no statistical significance between patients with prostate cancer after operation and healthy controls. Correlative analysis showed that there was a positive correlation between the CD4+CD25+ regulatory T cells and the expression of F0XP3 mRNA in PBMC of the patients with prostate cancer (r=0.623,p<0.01). Conclusion CD4+CD25

  19. Separation and Amplification of CD4 + CD25 + Regulatory T Cells from Sensitized Mice%致敏小鼠CD4+CD25+调节性T细胞磁珠分选及体外扩增

    Institute of Scientific and Technical Information of China (English)

    潘莉; 翁文骏; 许吕宏; 魏菁; 方建培

    2012-01-01

    The aim of this study was to separate and amplify CD4 + CD25 + Treg cells from splenocytes of sensitized nrice. The percentage of CD4 + CD25 + Treg cells was detected by flow cytometty in sensitized and normal control mice. CD4 + T, CD4 + CD25 + Treg and CD4' CD25" T cells were isolated from mouse splenocytes by MACS. CD4 + CD25+ Treg cells were expanded in vitro cultures in addition of CD3/CD28 MACSiBead and IL-2. The activity of cells was detected with 0.4% trypan blue staining. The purity of cells after sorting, the main surface marker and the level of Foxp3 were detected by flow cytometry. The results showed that CD4 + CD25 + Treg cell proportion was higher in sensitized mice than normal control mice ( P 0.05). It is concluded that the sorting of CD4 + CD25 + Treg cells is isolated successfully by MACS without affecting the vitality of target cells. The amplification of CD4 + CD25 + Treg cells is successral in vitro. Expression of surface markers and Faxp3 gene does not obviously change after amplification, so that to establish a practical method to recover and enlarge the amount of CD4 + CD25 + Treg cells in good condition.%本研究探讨致敏小鼠CD4+ CD25+调节性T细胞的分选及体外扩增.流式细胞术检测致敏小鼠及正常小鼠体内CD4+ CD25+ Treg细胞水平,免疫磁珠分选方法从小鼠脾细胞中分选出CD4+T细胞、CD4+ CD25+ Treg细胞和CD4+ CD25-T细胞,负载抗CD3/CD28单克隆抗体MACSiBead联合IL-2共同刺激CD4+ CD25+ Treg细胞进行体外扩增培养,用0.4%台盼蓝染色并计数检测细胞的活性,流式细胞术检测分选后细胞纯度、主要表面标记及Foxp3基因的表达.结果表明:致敏小鼠体内CD4+ CD25+ Treg水平较正常小鼠升高(P<0.05).分选出CD4+ CD25+Treg细胞纯度平均达到87%,细胞活性大于97%,高表达Foxp3基因.体外扩增2周后细胞数扩增倍数能够达到42倍,CD4+ CD25+ Treg细胞所占比例为85.32%,Foxp3表达由(76.92±1.72)%稍下降至(75

  20. Role of Circulating CD4+ CD25high Foxp3+ Regulatory T-Cells in Paediatric Asthma

    Directory of Open Access Journals (Sweden)

    Ensaf Khalil Mohammed*, Zeinab Farag Asheiba

    2011-04-01

    Full Text Available Background: The role of T-Helper 2 (Th2 cells in the pathogenesis of allergy and asthma has been well described. However, the immunologic mechanisms that down modulate and protect against the development of these disorders are poorly characterized. A spectrum of CD4+ T cells, including, FOXP3-positive CD4+CD25+ T regulatory cells (Tregs might play a critical role in regulating these diseases. Objective: To investigate the role of CD4+CD25high FoxP3 Tregs in the pathogenesis of pediatric asthma. Methods: The study included 24 asthmatic children, 12 had mild intermittent asthma and 12 were of severe persistent asthma . In addition, 12 healthy subjects were used as controls. All patients were subjected to clinical examination and laboratory investigations including complete blood count with differential leucocytic and absolute eosinophilic count, serum total IgE level by ELIZA and flow cytometry was used to study the frequency of Tregs in peripheral blood lymphocytes of all studied groups using specific markers: cell-surface CD25 and CD4 expression and cytoplasmic FoxP3 expression. Results: It was noticed a significant decrease in CD4+CD25+ % and CD4+CD25 high % in both mild intermittent cases and severe persistent asthmatic patients when compared to healthy controls. FoxP3 expression in Tregs was significantly lower in CD4+CD25high T-cells of mild asthmatic patients when compared to control group. While the FoxP3 expression in Tregs was non- significantly lower in CD4+CD25high T-cells of severe asthmatic patients .Tregs cells % was correlated significantly with mild asthma .While it did not show correlation with severe asthma . An inverse correlation between FoxP3 protein expression was revealed within CD4+CD25high T-cells and total serum IgE when analyzed for all subjects. However, when correlation analysis was performed in each studied group separately, no significant correlation was found between FoxP3 expression and total serum IgE levels and

  1. Expression and signifiacnce of CD4 +CD25 +regulatory T cells(Treg) in the pathogenesis for the pneumonia patients with leucopenia%白细胞降低性肺炎患者 CD4+CD25+调节性T 细胞的表达与临床意义

    Institute of Scientific and Technical Information of China (English)

    林燕梅

    2015-01-01

    目的:探索CD4+CD25+调节性T细胞( Treg)在白细胞( WBC)降低性肺炎( LP)中的表达及临床意义。方法对40例LP患者进行血常规及Treg检测和痰培养,将Treg与WBC及病原体类型进行相关性分析。结果 LP组患者 Treg、WBC 及单核细胞( MON )均明显降低( P<0.05)。痰培养显示肺炎支原体( MP)占26例并均予阿奇霉素治疗,发现第3天Treg频率显著高于第1天( P=0.028)。 Treg与WBC及MON比例均呈正相关性( P<0.05),但与中性粒细胞及淋巴细胞均呈负相关性( P<0.05);Treg与MP感染存在显著负相关性( P<0.05)。结论 Treg频率降低是LP的重要免疫表现,有助于支原体肺炎的早期诊断及疗效判断。%Objective To explore the expression and significance of CD 4 +CD25 +regulatory T cells ( Treg) for the patients with leucopenia pneumonia ( LP) .Methods Forty patients with LP were enrolled and their blood rou -tine examination ,Treg testing and sputum cultivation were done .The correlations between Treg ,white blood cell ( WBC) and pathogens were analyzed .Results Compared with the control group , Treg, WBC and monocytes (MON) decreased significantly(P<0.05).Sputum cultivation showed that there were 26 patients suffering from my-coplasma(MP) infection and they were all treated with azithromycin .The frequencies of Treg on the 3rd day of the MP patients were significantly higher than those on the 1st day during azithromycin treatment (P=0.028).Correlation a-nalysis showed that Treg was positively related to WBC and MON ( P<0.05 ) , whereas Treg was negatively related to neutrophil granulocyte and lymphocyte , as well as the MP ( P<0.05 ) .Conclusion Decreasing of Treg is one of the important immunological features for patients with LP , which can help the early diagnosis and curative effect evalua-tion for the MP.

  2. INFLUENCE OF FK 506 (TACROLIMUS) ON CIRCULATING CD4+ T CELLS EXPRESSING CD25 AND CD45RA ANTIGENS IN 19 PATIENTS WITH CHRONIC PROGRESSIVE MULTIPLE SCLEROSIS PARTICIPATING IN AN OPEN LABEL DRUG SAFETY TRIAL

    OpenAIRE

    Lemster, B.; Huang, L.L.; Irish, W.; Woo, J.; Carroll, P B; Abu-Elmagd, K.; Rilo, H.R.; Johnson, N; Russell-Hall, R.; Fung, J. J.; Starzl, T.E.; Eidelman, B.; Thomson, A. W.

    1994-01-01

    We have taken the opportunity of a clinical trial of the potential efficacy and safety of FK 506 (tacrolimus) in chronic progressive multiple sclerosis (MS) to examine the influence of this potent new immunosuppressant on circulating T-lymphocytes in an otherwise healthy non-transplant population. Peripheral blood levels of subsets of CD4+ T lymphocytes expressing the activation molecule interleukin-2 receptor (p55 &α chain: CD25) or the CD45RA isoform were determined sequentially in 19 patie...

  3. BAY 50-4798, a novel, high-affinity receptor-specific recombinant interleukin-2 analog, induces dose-dependent increases in CD25 expression and proliferation among unstimulated, human peripheral blood mononuclear cells in vitro.

    Science.gov (United States)

    Matthews, Lynn; Chapman, Sherita; Ramchandani, Meena S; Lane, H Clifford; Davey, Richard T; Sereti, Irini

    2004-12-01

    Interleukin-2 administration induces CD4 T cell expansion in HIV-infected patients, however, toxicity can limit dosing. BAY 50-4798 is a recombinant IL-2 analog with >1000-fold specificity for the high-affinity IL-2 receptor. The effects of this compound on unstimulated human PBMC were evaluated. PBMC from HIV(-) and HIV(+) donors were cultured in vitro with incremental doses of BAY 50-4798 or aldesleukin. CD25 expression and proliferation were evaluated with flow cytometry. Cytokine levels were measured by ELISA in culture supernatants. BAY 50-4798 induced dose-dependent increases in CD25 expression and proliferation of T cells, NK, and B cells and showed selectivity for CD4 T cells expressing CD25. Induction of pro-inflammatory cytokines was also dose-dependent and was observed at the concentrations of BAY 50-4798 with the highest biologic activity. These data suggest that BAY 50-4798 can induce proliferation of unstimulated T cells but loss of T cell selectivity and induction of pro-inflammatory cytokines occur at concentrations exerting the highest biologic activity. PMID:15507389

  4. 自身免疫性血小板减少性紫癜患者外周血中CD4+CD25+调节性T细胞、sFas和sFasL的表达及临床意义%Expression and Clinical Significance of CD4 + CD25 + Treg Cells, sFas and sFasL in Peripheral Blood of Patients with Autoimmune Thrombocytopenic Purpura

    Institute of Scientific and Technical Information of China (English)

    贾瑞萍; 赵雪芸

    2011-01-01

    本研究通过检测成人自身免疫性血小板减少性紫癜(AITP)外周血中CI4+CD25+调节性T细胞(Treg)、sFas和sFasL的表达,探讨它们在AITP发病机制中的作用及临床意义,为寻求AITP治疗的有效方法提供理论依据.采用流式细胞术分别检测30例AITP患者和18例正常对照者外周血CD4+T、Treg、CD4+ CD25-T细胞表达率及Treg/CD4+T比值;用酶联免疫吸附法(ELISA)检测AITP患者治疗前后及对照组外周血sFas、sFasL表达水平.结果表明,AITP组外周血中CD4+T细胞表达率低于对照组(p<0.05),Treg细胞表达率及Treg/CD4+T比值明显低于正常对照组(p<0.0l).AITP组患者治疗前外周血中sFas和sFasL水平明显高于治疗后和正常对照组(p<0.01),AITP组治疗后与正常对照组外周血sFas、sFasL水平差异无统计学意义(p>0.05).AITP患者治疗前Treg细胞表达率、Treg/CD4+T细胞比值与血小板计数呈正相关;AITP患者外周血中sFas和sFasL水平呈正相关;CD4+T细胞、CD4+CD25 -T细胞表达率,sFas、sFasL浓度与血小板计数无明显相关;Treg细胞表达率和sFas、sFasL浓度间没有明显相关性.结论:Treg在AITP的发病机制中发挥一定作用;Treg细胞水平与AITP病情的严重程度有关;sFas和sFasL水平异常参与了AITP的免疫病理过程.%This study was aimed to detect the expression of CD4 * CD25 * regulatory T cells (Treg) , sFas and sFasL in patients with autoimmune thrombocytopenic purpura ( AITP) , and to explore their roles in the pathogenesis of AITP and clinical significance, so as to provie a theoretical basis for effective treatment for AITP. The expressions of CD4 * T, Treg, CD4 * CD25" T, Treg /CD4 * T in peripheral blood of 30 the patients with AITP and 18 controls were detected by flow cytometry, and enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of sFas and sFasL in peripheral blood of controls and the patients with AITP before and after treatment. The

  5. Hemeoxygenase-1 expression effect on the up-regulation of peripheral blood CD4+CD25+CD127low/- regulatory T cells mediated by mesenchymal stem cells in asthma patients%血红素加氧酶1在间充质干细胞上调哮喘患者外周血调节性T细胞中的作用

    Institute of Scientific and Technical Information of China (English)

    温冰; 颛孙永勋; 颜富德; 陈瑞; 张蔚; 冯素玲; 李建国

    2011-01-01

    BACKGROUND: Up-regulating CD4+CD25+CD127low/- regulatory T cells is a new target in the treatment of asthma. Human bone marrow mesenchymal stem cells (MSCs) can up-regulate CD4+CD25+CD127low/- regulatory T cells in vitro, while the mechanism is not clear.OBJECTIVE: To study the effect of hemeoxygenase -1 (HO-1) expression in the up-regulation of peripheral blood CD4+CD25+CD127low/- regulatory T cells mediated by MSCs in asthma patients.METHODS: Real-time PCR was used to examine the expression of HO-1 mRNA in MSCs pretreated with 0, 15, 30, 45,60 μmol/L Hemin (the revulsive of HO-1) and 0, 5, 10, 15, 20 μmol/L ZnPP (the inhibitor of HO-1) respectively. Peripheral blood mononuclear cells (PBMCs), which were isolated from 10 cases of asthma patients with acute episode and 10 cases of healthy controls using Ficoll density gradient centrifugation, were incubated with MSCs pretreated with Hemin, ZnPP and mock respectively.RESULTS AND CONCLUSION: The expression of HO-1 in MSCs can be induced and inhibited in vitro. The higher concentration of Hemin added to MSCs, the higher expression of HO-1 mRNA was tested (P < 0.05). With the increasing concentration of ZnPP added to MSCs, the expression of HO-1 mRNA was becoming lower (P < 0.05). The proportion of CD4 +CD25+CD127low/-regulatory T cells in CD4+ T cells could be up-regulated by MSCs (P < 0.01) and also by MSCs with induction of HO-1 expression (P < 0.01). While MSCs with inhibition of HO-1 expression could down-regulate the proportion of CD4 +CD25+CD127low/-regulatory T cells in CD4 + T cells (P < 0.01). The expression of HO-1 partially contributed to the up-regulation of CD4+CD25+CD127low/- regulatory T cells mediated by MSCs in asthma patients.%背景:上调CD4+CD25+CD127low/-调节性T 细胞是目前治疗哮喘的新靶点.骨髓间充质干细胞在体外可上调正常人外周血的调节性T 细胞,但机制尚未明确.目的:观察血红素加氧酶1 对间充质干细胞

  6. Biological features of intrahepatic CD4+CD25+ T cells in the naturally tolerance of rat liver transplantation

    Institute of Scientific and Technical Information of China (English)

    LU Ling; ZHANG Feng; PU Liyong; YAO Aihua; YU Yue; SUN Beicheng; LI Guoqiang

    2007-01-01

    The biological features of intrahepatic CD4+CD25+ T regulatory cells in the naturally tolerance of rat liver transplantation were explored.Orthotopic liver transplantation was performed in two allogeneic rat strain combinations,one with fatal immunosuppression despite a complete major histocompatibility complex mismatch.The subjects were divided into three groups according to different donors and recipients [Tolerance group:LEW-to-DA;Rejection group:DA-to-LEW;Syngegnic group(control group):DAto-DA].The proportion of intrahepatic CD4+CD25+ T cells from three groups was determined by flow cytometry(FCM)in different time.The intrahepaitc CD4+CD25+ T cells were isolated by magnetic activated cell sorting(MACS)method and identified by FCM.The Foxp3 mRNA was detected by reverse transcriptase polymerase chain reaction(RT-PCR).And their suppression on the proliferation of CD4+CD25- T effector cells was analyzed by cell proliferation assay in vitro.Beginning immediately after transplantation,the proportion of Treg cells increased over time in both allogeneic groups but was significantly greater in the Rejection group.The proportion of Treg cells declined after day 5,and such reduction was more dramatic in the Rejection group than in the Tolerance group.Animals in the Tolerance group showed a second increase in the proportion after day 14.Intrahepatic CD4+CD25+T cells isolated from spontaneous tolerance models inhibited the proliferation of mixed lymphocyte reaction.The purity of CD4+CD25+ T cells sorted by MACS was 86%-93%.The CD4+CD25+ T cells could specifically express the Foxp3 gene compared with CD4+CD25- T cells.In vitro,the spleen cells from LEW rats can irritate the proliferation of CD4+CD25+ T cells more obviously than the syngegnic spleen cells.CD4+CD25+ Tr cells could suppress the proliferation of CD4+CD25- T cells,but the inhibition was reversed by exogenous IL-2(200 U/mL).The CD4+CD25+ T regulatory cells specifically express the Foxp3 gene,which may play an

  7. Change of CD4+CD25+Foxp3+Treg cells and Foxp3 expression in a mouse model of asthma and the effects of Icariin%哮喘小鼠CD4+CD25+Foxp3+调节T细胞和Foxp3蛋白的变化及淫羊藿苷干预作用

    Institute of Scientific and Technical Information of China (English)

    金华良; 董竞成; 罗清莉; 厉蓓; 吴金峰; 吕玉宝; 杜文静; 徐海林; 王根发; 王镓

    2012-01-01

    目的:研究哮喘小鼠CD4+CD25+Foxp3+调节T(Treg)细胞和Foxp3蛋白的变化,探讨淫羊藿苷对哮喘干预机制.方法:将BALB/c小鼠32只随机分为正常对照组、哮喘模型组、淫羊藿苷组、地塞米松阳性对照组,每组8只.哮喘模型制备用卵白蛋白(OVA)致敏大鼠并雾化吸入刺激,治疗组采用相应药物灌胃给药,对照组用等量生理盐水代替.最后一次激发24小时后,采用Buxco肺功能仪有创法检测小鼠气道反应性;HE染色评价观察小鼠肺组织病理形态学改变;流式细胞术检测脾脏CD4+CD25+Foxp3+Treg细胞比例及Foxp3蛋白表达量;免疫印迹法测定肺组织Foxp3蛋白表达量;ELISA法测定血清及肺泡灌洗液(BALF)IL-10水平.结果:与正常对照组比较:模型组气道阻力及气道炎症指数显著增加(P 0.05);脾Foxp3蛋白表达显著下降(P 0.05);与模型组比较:淫羊藿苷组气道阻力和气道炎症明显减轻(P 0.05),但淫羊藿苷可进一步上调哮喘小鼠肺组织Foxp3蛋白表达量(P 0.05). The expression of Foxp3 was significantly decreased in spleen, but was remarkably increased in lung tissues (P < 0. 05) . In addition, the level of IL-10 in serum of model group was significantly decreased ( P < 0. 05 ) . Icariin significantly attenuated airway hyperresponsiveness and inflammation, and further markedly enhanced expression of Foxp3 in lung tissues and IL-10 levels in serum, as compared lo OVA-exposed mice. Conclusion: The expression of Foxp3 protein is decreased within spleen CD4+ CD25 + Foxp3 + Treg cells in asthma. Icariin can increase the Foxp3 expression in lung tissue and levels of serum IL-10, which may contribute lo its antiaslhmalic effecls.

  8. The frequency of CD127low expressing CD4+CD25high T regulatory cells is inversely correlated with human T lymphotrophic virus type-1 (HTLV-1 proviral load in HTLV-1-infection and HTLV-1-associated myelopathy/tropical spastic paraparesis

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    Chieia Marco

    2008-07-01

    Full Text Available Abstract Background CD4+CD25high regulatory T (TReg cells modulate antigen-specific T cell responses, and can suppress anti-viral immunity. In HTLV-1 infection, a selective decrease in the function of TReg cell mediated HTLV-1-tax inhibition of FOXP3 expression has been described. The purpose of this study was to assess the frequency and phenotype of TReg cells in HTLV-1 asymptomatic carriers and in HTLV-1-associated neurological disease (HAM/TSP patients, and to correlate with measures of T cell activation. Results We were able to confirm that HTLV-I drives activation, spontaneous IFNγ production, and proliferation of CD4+ T cells. We also observed a significantly lower proportion of CTLA-4+ TReg cells (CD4+CD25high T cells in subjects with HAM/TSP patients compared to healthy controls. Ki-67 expression was negatively correlated to the frequency of CTLA-4+ TReg cells in HAM/TSP only, although Ki-67 expression was inversely correlated with the percentage of CD127low TReg cells in healthy control subjects. Finally, the proportion of CD127low TReg cells correlated inversely with HTLV-1 proviral load. Conclusion Taken together, the results suggest that TReg cells may be subverted in HAM/TSP patients, which could explain the marked cellular activation, spontaneous cytokine production, and proliferation of CD4+ T cells, in particular those expressing the CD25highCD127low phenotype. TReg cells represent a potential target for therapeutic intervention for patients with HTLV-1-related neurological diseases.

  9. Effects of PQL on levels of immunal proteins, C3, C4 and expressions of CD4, CD8, and CD25 on lymphocytes of peripherial blood in patients with breast carcinoma undergoing radiotherapy

    International Nuclear Information System (INIS)

    Objective: To evaluate the effects of panaxatriol in quinquefolium leaf (PQL) on immune functions in the patients with breast carcinoma undergoing radiotherapy. Methods: Thirty patients with breast carcinoma undergoing radiotherapy were randomly divided into 2 groups: panaxatriol treatment group (n=17) and control group (n=13). All patients in panaxatriol treatment group were administrated with PQL as well as radiotherapy, no administration in control group. After radiotherapy with 40 Gy, the white cells count and IgG, IgA, IgM, C3, and C4 values, and the percentages of CD4+, CD8+, CD25+ and lymphocyte transformation rate of peripheral blood in the patients were examined. Results: There were (4.02 ± 0.67) x 109 ·L-1 and (6.17 ± 1.20) x 109 ·L-1 white blood cells, respectively, in control and PQL groups. The lymphocyte transformation rate of peripheral blood were 7966 ± 1562 in the control group, while 18035 ± 1577 in PQL group, which was significantly higher than that in control group (P+ and CD25+ T lymphocytes in PQL group were significantly higher than those in control group (P<0.05, P<0.001). Conclusion: The PQL could enhance the cellular immune function in patients with breast carcinoma undergoing radiotherapy. (authors)

  10. Characteristics and PD-1 expression of peripheral CD4+CD127loCD25hiFoxP3+ Treg cells in chronic HCV infected-patients

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    Zhang Ting

    2011-06-01

    Full Text Available Abstract Background Both regulatory T cells (Tregs and PD-1/PD-L1 pathway were critically involved in HCV viral persistence. However, the association between them was not well investigated. Herein, we aimed to investigate the distributional profiles of Tregs subsets and association between PD-1 expression on these subsets and development of HCV long-term persistence. Methods CD45RA and CD27 were employed to separate peripheral Tregs as naïve/central memory/effector memory/effector subsets. The phenotypic characteristics and PD-1 expression of Tregs were studied by flow cytometry. Results In the present study, the majority of Tregs was identified as central memory phenotype in chronic hepatitis C patients compared with nearly equal contribution of naïve and central memory subsets in healthy individuals. PD-1 expression was elevated in all CD4+ T cell subset in chronic HCV infected patients, including Tregs. Of note, higher level of PD-1 expression was found on TEM- and effector-Treg than naïve- and TCM-Tregs subsets. The ratio of TEM-Tregs/naive-Tregs and TEM-Tregs/TCM-Tregs regarding to PD-1 MFI were significantly lower in CHC patients compared to controls. Conclusions Our study indicated that distinctive characteristics of PD-1 expression on Tregs in HCV infection suggests associated with impaired adaptive immunity as well as viral long-term persistence. The cross talk between Treg cells and PD-1 induced inhibition in chronic HCV infection deserved further exploration for HCV infection associated immune pathogenesis.

  11. Role of CD4+CD25+ regulation cells and expressing of FOXP3 in the pathogenesis of children with asthma%哮喘患儿CD4+CD25+调节性T细胞及FOXP3的表达

    Institute of Scientific and Technical Information of China (English)

    史彧; 褚莉莉; 张兰芳; 赵德育

    2009-01-01

    目的:研究CD4+CD25+调节性T细胞在哮喘儿童外周血中的比例改变,并探讨其临床意义.方法:采用细胞内染色的流式细胞术检测急性发作期哮喘患儿外周血CD4+CD25+Treg及其调控基因FOXP3的表达,并与健康对照组进行比较.结果:急性发作期哮喘患儿外周血CD4和CD25双阳性细胞所占比例与健康对照组无明显差异(P>0.05),而CD4+CD25+FOXP3调节性T细胞在外周血中的比例明显低于健康对照组(P<0.01).结论:哮喘患儿外周血CD4+CD25+FOXP3调节性T细胞在外周血中的比例明显减少,可能与哮喘的发病机制有关.

  12. 哮喘患儿CD4+CD25+T细胞FoxP3表达的意义%Significance of Expression of Intracellular FoxP3 in CD4+CD25+T Cells in Children with Asthma

    Institute of Scientific and Technical Information of China (English)

    陈跃华; 朱华芳; 范婷婷

    2007-01-01

    目的 研究表达FoxP3的CD4+CD25+调节性T细胞在哮喘儿童外周血中的比例改变,探讨其在发病机制中的意义.方法 采用细胞内染色的流式细胞术及定量PCR方法,分别在蛋白质和mRNA水平检测哮喘患儿外周血FoxP3表达,并与健康对照组进行比较.结果 哮喘患儿CD4和CD25双阳性细胞所占比例与健康对照组比较无明显差异,而CD4+CD25high和 CD4+CD25+FoxP3+细胞明显低于对照组(Pa<0.05).FoxP3 mRNA表达水平与蛋白质表达水平变化一致.结论 哮喘患儿CD4+CD25+FoxP3+调节性T细胞明显减少,可能与哮喘的发病机制有关.

  13. Depletion of CD4+CD25+ regulatory T cells can promote local immunity to suppress tumor growth in benzo[a]pyrene-induced forestomach carcinoma

    Institute of Scientific and Technical Information of China (English)

    Yi-Ling Chen; Jung-Hua Fang; Ming-Derg Lai; Yan-Shen Shan

    2008-01-01

    AIM: To elucidate the distribution of CD4+CD25+ regulatory T cells (Tregs) in different lymphoid tissues and its local enhancement on tumor growth before and after depletion of CD4+CD25+ Tregs.METHODS: Female ICR mice were gavaged with benzo[a]pyrene (BaP) to induce forestomach carcinoma. CD4+CD25+ Tregs were intraperitoneally depleted with monoclonal antibody PC61. These mice were divided into BaP-only, BaP+IgG, BaP+PC61, and control groups. The forestomach of mice was dissected for histological analysis, and tunnel test was performed for apoptosis of tumor cells. CD4+CD25+ Tregs were sorted from different lymphoid tissues and expression of Foxp3, IL-10, and chemokine receptors was analyzed by flow cytometry, semi-quantitative and real-time polymerase chain reaction.RESULTS: The mice gavaged with only BaP showed increased forestomach papilloma and carcinoma at wk 16 and 32. The proportion of CD4+CD25+ Tregs was significantly higher in peri-stomach regional lymph nodes than in other lymphoid tissues. These CD4+CD25+ Tregs in regional lymph nodes expressed higher levels of Foxp3 and IL-10, enriched in the CD62L-subset, and CCR1 and CCR5 chemokine receptors. In mice gavaged with BaP+PC61, the number of tumor nodules and tumor volume decreased significantly with massive infiltrating cells and apoptosis of tumor cells. In the draining regional lymph nodes, the number of CD4+CD25+ Tregs also decreased significantly.CONCLUSION: Inducible and activated CD4+CD25+ Tregs in the draining regional lymph nodes suppress host local immunity during tumor growth. Depletion of CD4+CD25+ Tregs can promote host local immunity to suppress tumor growth.

  14. CD4+CD25+调节性T细胞及Foxp3表达与支气管哮喘发病的相关性研究%Study of Relationship between CD4+ CD25+ Regulator T Cells as well as Expression of Foxp3 and Bronchial Asthma

    Institute of Scientific and Technical Information of China (English)

    刘春花; 刘恩梅

    2006-01-01

    CD4+CD25+调节性T细胞(CD4+CD25+Tr)在胸腺内发育,迁移到外周组织发挥功能,通过对自身抗原反应T细胞活化和增殖起负调节作用,保持自身稳定,抑制自身免疫性疾病以及哮喘和炎症性肠病等炎症性疾病的发生,并调控器官移植耐受和肿瘤免疫.近年发现Foxp3是CD4+CD25+Tr成熟、分化以及发挥功能的调节基因.作为小儿最常见的慢性疾病,支气管哮喘的免疫学发病机制尚不完全清楚,CD4+CD25+Tr与Foxp3表达在哮喘发病中的作用已引起人们的广泛关注,其研究结果将为进一步探讨哮喘的发病机制和寻找有效的治疗方法提供理论依据.

  15. 支气管哮喘患者外周血CD4+ CD25+调节性T细胞水平及Foxp3 mRNA表达的分析%CD4+ CD25+ regulatory T cells and expressions of forkhead/winged helix transcription factor ( Foxp 3 ) mRNA in peripheral blood of patients with asthma

    Institute of Scientific and Technical Information of China (English)

    薛克营; 周咏明; 熊盛道; 熊维宁

    2006-01-01

    目的 观察支气管哮喘(哮喘)患者外周血单个核细胞(PBMCs)中CD4+ CD25+调节性T细胞(Treg)水平及叉状头/翅膀状螺旋转录因子(Foxp3)mRNA表达的变化,探讨CD4+ CD25+ Treg在哮喘发病中的作用.方法采用流式细胞仪检测78例哮喘患者(急性发作期组30例,慢性持续期组25例,缓解期组23例)和29例健康志愿者(正常对照组)PBMCs中CD4+ CD25+ Treg的比例;反转录-聚合酶链反应(RT-PCR)检测PBMCs中Foxp3 mRNA的表达.结果 急性发作期组和慢性持续期组PBMCs中CD4+ CD25+ Treg的比例及Foxp3 mRNA的表达明显低于缓解期组和正常对照组(P<0.05);缓解期组CD4+ CD25+ Treg的比例及Foxp3 mRNA的表达虽亦低于正常对照组,但差异无统计学意义(P>0.05);急性发作期组CD4+ CD25+ Treg的比例及Foxp3 mRNA的表达低于慢性持续期组(P<0.05).结论 哮喘患者外周血中具有免疫抑制活性的CD4+ CD25+ Treg数量减少,功能下降,可能参与哮喘的发生和发展.

  16. The expression of tumor necrosis factor-a induced protein 8 like-2 in CD4+ CD25+ regulatory T cells%肿瘤坏死因子-α诱导蛋白-8样分子2在调节性T细胞中的表达

    Institute of Scientific and Technical Information of China (English)

    栾樱译; 姚咏明

    2011-01-01

    目的:观察肿瘤坏死因子-α诱导蛋白-8样分子2(TIPE2)在CD4CD25调节性T细胞(CD4CD25Treg)中的表达.方法:免疫磁珠法分离正常BALB/C小鼠脾脏CD4CD25Tregs,流式细胞术鉴定CD4CD25Treg的纯度.激光共聚焦荧光法检测Treg细胞中TIPE2的分布,并进行初步定位;进一步采用逆转录一聚合酶链反应(RT-PCR)和Western blot技术分别从基因和蛋白水平检测Treg细胞中TIPE2表达.结果:免疫磁珠分选法得到的CD4CD25Tregs纯度在92%以上,台盼蓝染色显示细胞活性大于97%.Western blot证实Treg细胞中存在清晰TIPE2条带,分子质量为21 kD;采用RT-PCR技术在Treg细胞中检测到147 bp大小的特异性TIPE2目的基因条带.结论:TIPE2可表达于小鼠CD4CD25Treg细胞.%Objective: To investigate the expression of tumor necrosis factor-α induced protein 8 like2 ( TIPF2)in CD4+ CD25+ regulatory T cells ( CD4+ CD25+ Tregs). Methods: CD4+ CD25+ Tregs were isolated from the spleens of male BALB/C mice by magnetic beads, and the purity of these cells was determined by flow cytometry.The present study was designed to determine TIPE2 expression in Tregs by confocal microscopy analysis, Western blot and reverse transcription-polymerase chain reaction ( RT-PCR) analysis, respectively. Results: Purity of CD4+ CD25+ Tregs was greater than 92%. The expression of TIPF2 was detected by confocal microscopy, and it was a cytoplasmic protein expressed in CD4+ CD25+Tregs. To confirm the expression of TIPF2 , it was detected by Western blot analysis using specific TIPF2 antibody, and a clear band with a molecular mass of approximately 21 kD from CD4+ CD25+ Tregs was found. Moreover, to determine the gene expression of TIPF2 , total RNA was extracted from CD4+ CD25+ Tregs and RT-PCR was performed, a band of the size of 147 bp was noted as expected. Conclution: TIPF2 appears to be a cytoplasmic protein expressed in CD4+ CD25+ Tregs.

  17. Expression of PD-1 by CD4+CD25+CD127low Treg cells in the peripheral blood of lung cancer patients

    Directory of Open Access Journals (Sweden)

    Zhong AY

    2015-07-01

    Full Text Available AnYuan Zhong, Xue Pan, MinHua Shi Department of Respiratory Diseases, the Second Affiliated Hospital of Soochow University, Suzhou, People’s Republic of ChinaLung cancer is the most commonly diagnosed cancer and the leading cause of cancer death worldwide,1 with the majority of patients presenting with advanced disease.2 Treg cells diminish the activation and function of lymphocytes via cell-cell contact and secretion of soluble mediators.3 PD-1 is expressed on the surface of activated T and B cells and regulates their activation and proliferation.4 PD-L1 binds to the PD-1 receptor, leading to, among other responses, negative regulation of immune activity. Both Tregs and the PD-1/PD-L1 pathway play important roles in lung cancer pathogenesis;5,6 however, the association between these two factors remains poorly understood. Here, we examined PD-1 expression on Tregs, and compared these results with established clinical indicators of lung cancer. These analyses revealed significant expression of PD-1 on Tregs in lung cancer, which may be used to inform clinical diagnoses.

  18. Role of Foxp3 expression and CD+4CD+25 regulatory T cells on the pathogenesis of childhood asthma%Foxp3表达与CD+4CD+25调节性T细胞在儿童哮喘发病中的作用

    Institute of Scientific and Technical Information of China (English)

    罗征秀; 刘恩梅; 邓兵; 李欣; 陈坤华; 王莉佳; 黄英; 符州

    2006-01-01

    目的研究Foxp3基因表达与CD+4CD+25调节性T细胞在哮喘发病中的作用.方法以确诊哮喘的患儿为研究对象,急性发作期15例、缓解期15例,同期选10例正常儿童作对照,提取外周血单个核细胞(PBMC)进行CD4、CD25表面标志及血浆、培养上清液IL-4、IFN-γ、IL-10和TGF-β等细胞因子的ELISA检测,同时收集哮喘患儿和正常儿童的诱导痰,用RT-PCR方法检测PBMC及诱导痰中转录因子Foxp3-mRNA的表达.结果 PBMC CD+4CD+25T细胞百分率在哮喘急性发作期、缓解期分别为(10.1±2.1)%、(11.7±2.5)%,低于对照组的(15.5±2.7)%(P分别<0.01、<0.05);对照组PBMC在体外培养后CD+4CD+25细胞百分率显著升高,同培养前比较差异有统计学意义(P<0.01).PBMC Foxp3-mRNA表达水平(Foxp3/β-actin) 在哮喘急性发作期、缓解期分别为0.46±0.14 、0.50±0.19,低于对照组0.77±0.22,诱导痰Foxp3-mRNA表达水平哮喘患儿也低于对照组;PBMC在体外培养后对照组Foxp3-mRNA表达水平较培养前升高(P<0.05),而哮喘患儿培养前后Foxp3-mRNA表达水平无显著变化.血浆及培养上清液IFN-γ、TGF-β在哮喘急性发作期、缓解期低于对照组 (P<0.05),且IFN-γ、TGF-β与PBMC Foxp3-mRNA水平、CD+4CD+25细胞百分率呈正相关. 哮喘患儿血浆及培养上清液IL-4显著高于对照组,急性发作期血浆IL-10显著高于对照组,而缓解期与正常组无显著性差异;IL-4、IL-10与PBMC Foxp3-mRNA水平、CD+4CD+25细胞百分率无相关性.结论哮喘患儿的TGF-β分泌不足、Foxp3基因表达降低、CD+4CD+25调节性T细胞数量减少及分化发育障碍可能在儿童哮喘的发病中起重要作用.

  19. The Expression of CD25 and FOXP3 in Mouse Asthma and the Effect of Dexamethasone%CD25及FOXP3在支气管哮喘小鼠中的表达以及地塞米松的干预作用

    Institute of Scientific and Technical Information of China (English)

    马祥; 梁宗安; 毛辉; 刘雅; 王茂筠

    2010-01-01

    目的 分析哮喘小鼠模型肺组织及骨髓中CD25及FOXP3的表达,以及地塞米松的干预作用.方法 BALB/c小鼠随机分为正常对照、哮喘及地塞米松干预组,通过HE染色观察肺组织的病理变化,应用Western blot、RT-PCR方法检测肺组织FOXP3及CD25的表达,RT-PCR方法检测哮喘及地塞米松干预组肺及骨髓FOXP3 mRNA表达.结果 哮喘及地塞米松干预组肺组织FOXP3表达强于正常对照组(P0.05);哮喘及地塞米松干预组肺组织及骨髓细胞均有FOXP3 mRNA表达,且地塞米松组强于哮喘组(P<0.05).结论 哮喘小鼠肺内CD25及FOXP3表达增强,地塞米松会促进其表达;哮喘小鼠骨髓有F10XP3表达,地塞米松促进其表达.

  20. Downregulation of CD4+CD25+ regulatory T cells may underlie enhanced Th1 immunity caused by immunization with activated autologous T cells

    Institute of Scientific and Technical Information of China (English)

    Qi Cao; Dangsheng Li; Ningli Li; Li Wang; Fang Du; Huiming Sheng; Yan Zhang; Juanjuan Wu; Baihua Shen; Tianwei Shen; Jingwu Zhang

    2007-01-01

    Regulatory T cells (Treg) play important roles in immune system homeostasis, and may also be involved in tumor immunotolerance by suppressing Thl immune response which is involved in anti-tumor immunity. We have previously reported that immunization with attenuated activated autologous T cells leads to enhanced anti-tumor immunity and upregulated Thl responses in vivo. However, the underlying molecular mechanisms are not well understood. Here we show that Treg function was significantly downregulated in mice that received immunization of attenuated activated autologous T cells. We found that Foxp3 expression decreased in CD4+CD25+ T cells from the immunized mice. Moreover, CD4+CD25+Foxp3+ Treg obtained from immunized mice exhibited diminished immunosuppression ability compared to those from naive mice. Further analysis showed that the serum of immunized mice contains a high level of anti-CD25 antibody (about 30 ng/ml,/K0.01 vs controls). Consistent with a role of anti-CD25 response in the down-regulation of Treg, adoptive transfer of serum from immunized mice to naive mice led to a significant decrease in Treg population and function in recipient mice. The triggering of anti-CD25 response in immunized mice can be explained by the fact that CD25 was induced to a high level in the ConA activated autologous T cells used for immunization. Our results demonstrate for the first time that immunization with attenuated activated autologous T cells evokes anti-CD25 antibody production, which leads to impeded CD4+CD25+Foxp3+ Treg expansion and function in vivo. We suggest that dampened Treg function likely contributes to enhanced Thl response in immunized mice and is at least part of the mechanism underlying the boosted anti-tumor immunity.

  1. Healthy Preterm Newborns Show an Increased Frequency of CD4(+) CD25(high) CD127(low) FOXP3(+) Regulatory T Cells with a Naive Phenotype and High Expression of Gut-Homing Receptors.

    Science.gov (United States)

    Rennó, C; Nadaf, M I V; Zago, C A; Carneiro-Sampaio, M; Palmeira, P

    2016-06-01

    Treg cells are crucial to prevent immune dysregulation, but little is known about the frequency of these cells in neonates, particularly in very/moderate and late preterm newborns studied as separate groups. The CD4(+) CD25(hi) CD127(lo) FOXP3(+) Treg population was phenotypically characterized to assess maturation markers and gut-homing integrins by flow cytometry in the cord blood of healthy preterm newborns born at 30-33(6/7) gestation weeks (Group 1), at 34-36(6/7) gestation weeks (Group 2) and term newborns born at 37-41 gestation weeks (Group 3), compared to healthy adults. An inverse correlation of the Treg percentage and gestational age was found, with significantly higher frequencies in Group 1 compared to Groups 2 and 3 and in Group 2 compared to Group 3, and significantly higher Treg frequencies and numbers in the neonates compared to the adults. All of the newborns exhibited increased Treg frequencies with a naive phenotype compared to adults. Cytotoxic T-lymphocyte-associated protein 4 CTLA-4 expression in the naive Treg was decreased in both preterm groups compared with those from term newborns and adults, and in the memory Treg from Group 1 compared with the other groups. The frequencies of Treg expressing α4β7 and α4β1 integrins were higher in both preterm groups, but significantly different only in Group 1, when compared with those from the term newborns and the adults. In conclusion, although a high frequency of Treg is present in newborns, an immature phenotype with a higher expression of CD45RA and α4β7/α4β1 and a lower expression of CTLA-4 is found, particularly in the very preterm group. PMID:27007547

  2. Freeze and Thaw of CD4+CD25+Foxp3+ Regulatory T Cells Results in Loss of CD62L Expression and a Reduced Capacity to Protect against Graft-versus-Host Disease.

    Directory of Open Access Journals (Sweden)

    Mareike Florek

    Full Text Available The adoptive transfer of CD4+CD25+Foxp3+ regulatory T cells (Tregs in murine models of allogeneic hematopoietic cell transplantation (HCT has been shown to protect recipient mice from lethal acute graft-versus-host disease (GVHD and this approach is being actively investigated in human clinical trials. Here, we examined the effects of cryopreservation on Tregs. We found that freeze and thaw of murine and human Tregs is associated with reduced expression of L-selectin (CD62L, which was previously established to be an important factor that contributes to the in vivo protective effects of Tregs. Frozen and thawed murine Tregs showed a reduced capacity to bind to the CD62L binding partner MADCAM1 in vitro as well as an impaired homing to secondary lymphoid organs in vivo. Upon adoptive transfer frozen and thawed Tregs failed to protect against lethal GVHD compared with fresh Tregs in a murine model of allogeneic HCT across major histocompatibility barriers. In summary, the direct administration of adoptively transferred frozen and thawed Tregs adversely affects their immunosuppressive potential which is an important factor to consider in the clinical implementation of Treg immunotherapies.

  3. Change of CD4+ CD25+ regulatory T cell and expression of Foxp3 in a mouse model of asthma%哮喘小鼠CD4+CD25+调节性T细胞及Foxp3表达的变化

    Institute of Scientific and Technical Information of China (English)

    李茜; 沈华浩

    2008-01-01

    目的 探讨哮喘小鼠CD4+CD25+调节性T细胞数量的变化及相关转录因子Foxp3表达的变化.方法 24只5周龄C57BL/6小鼠随机分为哮喘模型组和对照组,每组12只,于第0天和第14天,哮喘组以腹腔注射致敏液[0.1%的鸡卵蛋白(OVA)0.1ml与等体积液态铝混合]0.2ml致敏,对照组注射等体积生理盐水,第24、25、26天开始雾化吸入1%的OVA(哮喘组)或生理盐水(对照组)进行激发,连续3 d,于最后1次激发后48 h处死小鼠.取脾脏,制备脾细胞悬液,流式细胞术检测脾脏CD4+CD25+T细胞占CD4+T细胞比例.取脾脏组织以RT-PCR和Western blot分别检测脾脏Foxp3 mRNA和蛋白的表达.数据结果以Levene法行方差齐性检验后,进行独立样本的t检验,以P<0.05为差异具有统计学意义.结果 脾脏CD4+CD25+T细胞占CD4+T细胞比例哮喘组为(7.03±2.19)%,对照组为(9.70±2.80)%,哮喘组低于对照组,两者相比差异有统计学意义(P=0.016);脾脏Foxp3 mRNA的表达水平哮喘组为(0.37±0.11),对照组为(0.237±0.118),哮喘组高于对照组,两者相比差异有统计学意义(P=0.009);脾脏Foxp3蛋白的表达水平哮喘组(0.692±0.171),对照组(0.515±0.135),哮喘组高于对照组,两者相比差异有统计学意义(P=0.01).结论 哮喘时抗原对CD4+CD25+T细胞的激活不足,可能在哮喘发病机制中起一定作用.

  4. Intestinal lamina propria retaining CD4+CD25+ regulatory T cells is a suppressive site of intestinal inflammation.

    Science.gov (United States)

    Makita, Shin; Kanai, Takanori; Nemoto, Yasuhiro; Totsuka, Teruji; Okamoto, Ryuichi; Tsuchiya, Kiichiro; Yamamoto, Masafumi; Kiyono, Hiroshi; Watanabe, Mamoru

    2007-04-15

    It is well known that immune responses in the intestine remain in a state of controlled inflammation, suggesting that not only does active suppression by regulatory T (T(REG)) cells play an important role in the normal intestinal homeostasis, but also that its dysregulation of immune response leads to the development of inflammatory bowel disease. In this study, we demonstrate that murine CD4(+)CD25(+) T cells residing in the intestinal lamina propria (LP) constitutively express CTLA-4, glucocorticoid-induced TNFR, and Foxp3 and suppress proliferation of responder CD4(+) T cells in vitro. Furthermore, cotransfer of intestinal LP CD4(+)CD25(+) T cells prevents the development of chronic colitis induced by adoptive transfer of CD4(+)CD45RB(high) T cells into SCID mice. When lymphotoxin (LT)alpha-deficient intercrossed Rag2 double knockout mice (LTalpha(-/-) x Rag2(-/-)), which lack mesenteric lymph nodes and Peyer's patches, are transferred with CD4(+)CD45RB(high) T cells, they develop severe wasting disease and chronic colitis despite the delayed kinetics as compared with the control LTalpha(+/+) x Rag2(-/-) mice transferred with CD4(+)CD45RB(high) T cells. Of note, when a mixture of splenic CD4(+)CD25(+) T(REG) cells and CD4(+)CD45RB(high) T cells are transferred into LTalpha(-/-) x Rag2(-/-) recipients, CD4(+)CD25(+) T(REG) cells migrate into the colon and prevent the development of colitis in LTalpha(-/-) x Rag2(-/-) recipients as well as in the control LTalpha(+/+) x Rag2(-/-) recipients. These results suggest that the intestinal LP harboring CD4(+)CD25(+) T(REG) cells contributes to the intestinal immune suppression. PMID:17404275

  5. The IL-6R α chain controls lung CD4+CD25+ Treg development and function during allergic airway inflammation in vivo

    OpenAIRE

    Doganci, Aysefa; Eigenbrod, Tatjana; Krug, Norbert; De Sanctis, George T.; Hausding, Michael; Erpenbeck, Veit J.; Haddad, El-Bdaoui; Schmitt, Edgar; Bopp, Tobias; Kallen, Karl-J.; Herz, Udo; Schmitt, Steffen; Luft, Cornelia; Hecht, Olaf; Jens M Hohlfeld

    2005-01-01

    The cytokine IL-6 acts via a specific receptor complex that consists of the membrane-bound IL-6 receptor (mIL-6R) or the soluble IL-6 receptor (sIL-6R) and glycoprotein 130 (gp130). In this study, we investigated the role of IL-6R components in asthma. We observed increased levels of sIL-6R in the airways of patients with allergic asthma as compared to those in controls. In addition, local blockade of the sIL-6R in a murine model of late-phase asthma after OVA sensitization by gp130–fraction ...

  6. Human mesenchymal stem cells elevate CD4+CD25+CD127low/- regulatory T cells of asthmatic patients via heme oxygenase-1.

    Science.gov (United States)

    Li, Jian-guo; Zhuan-sun, Yong-xun; Wen, Bing; Wu, Hao; Huang, Feng-ting; Ghimire, Hridaya bibhu; Ran, Pi-xin

    2013-09-01

    Up-regulation of CD4+CD25+CD127low/- regulatory T cells (Tregs) is a new target in the treatment of asthma. Human bone marrow mesenchymal stem cells can up-regulate CD4+CD25+CD127low/- regulatory T cells in vitro, meanwhile, heme oxygenase-1 (HO-1) plays an important role in the development and maintenance of CD4+CD25+ regulatory T cells. However the mechanism has not yet been adequately understood. Hence, we wondered what effect of Heme Oxygenase-1 made on regulation of CD4+CD25+CD127low/- regulatory T cells mediated by mesenchymal stem cells. Peripheral blood mononuclear cells isolated from asthmatic patients and healthy controls were co-cultured with human bone marrow mesenchymal stem cells which were pretreated with Hemin (the revulsive of Heme Oxygenase-1), Protoporphyrin Ⅸ zinc (the inhibitor of Heme Oxygenase-1) and saline. The expression of Heme Oxygenase-1 in MSCs was enhanced by Hemin and inhibited by Protoporphyrin  zinc in vitro. Overexpression of Heme Oxygenase-1 elevated the proportion of CD4+CD25+CD127low/- regulatory T cells in CD4+ T cells, meanwhile, inhibition of Heme Oxygenase-1 decreased the proportion of CD4+CD25+CD127low/- regulatory T cells in CD4+ T cells as compared with mesenchymal stem cells alone. Taken together, these data demonstrated that Heme Oxygenase-1 contributed to the up-regulation of CD4+CD25+CD127low/- regulatory T cells mediated by mesenchymal stem cells in asthma.  PMID:23893806

  7. Association of PD-1 expression on CD4~+CD25~(nt/hi)CD127~(lo) regulatory T cells with disease progression in HIV-1 infected patients%HIV-1感染者CD4~+CD25~(nt/hi)CD127~(lo)调节性T细胞PD-1表达水平与疾病进展的关系

    Institute of Scientific and Technical Information of China (English)

    曹清华; 薛以乐; 王盈

    2009-01-01

    目的:阐明HIV-1感染者外周血中具有CD4~+CD25~(nt/hi)CD127~(lo)特征的调节性T细胞(Treg)表面PD-1的表达水平与疾病进展的关系.方法:选取108名未经治疗的不同进展期的HIV-1感染者和27名健康人对照, 采集静脉血, 用Ficoll-Hypaque密度梯度离心法分离获得PBMC, 加入PerCP-CD4抗体、 FITC-CD25抗体、 PE-CD127抗体和APC-PD-1抗体, 经细胞表面四色染色、流式细胞术(FCM)分析Treg表面PD-1的表达;另将50 L全血加入Trucount绝对计数管, 采用Multitest CD3/CD8/CD45/CD4试剂盒检测CD4+T细胞绝对数;分离静脉血血浆, NucliSens EasyQ测定血浆HIV-1病毒载量;实验数据采用SPSS14.0 统计学软件分析处理.结果:HIV-1感染者Treg表面PD-1表达水平显著高于健康人(5.33%±2.24% vs 1.72%±0.65%, P<0.01);AIDS期(7.87%±2.23%)明显高于进展期(5.21%±1.72%, P<0.05)和新近感染者(3.22%±1.01%, P<0.05);HIV-1感染者Treg表面PD-1表达水平与血浆中的HIV-1病毒载量和CD4~+T细胞绝对数密切相关.结论:首次证实HIV-1感染者外周血中Treg表面PD-1表达增加, 且表达水平与病程进展相关.该结果为进一步揭示HIV-1感染中Treg的效应机制、探索新的免疫治疗方案提供了理论及实验依据.%AIM: To investigate whether Programmed death-1 (PD-1) expression on peripheral CD4~+CD25~(nt/hi)CD127~(lo) regulatory T cells (Treg) was associated with disease progression in HIV-1-infected patients. METHODS: Peripheral blood from 108 HIV-1-infected patients in distinct disease progression statuses and 27 healthy individuals were collected in the present investigation. PBMCs were isolated by centrifugation on Ficoll-Hypaque, followed by staining with anti-CD4-PerCP, anti-CD25-FITC, anti-CD127-PE and anti-PD-1-APC. PD-1 expression on Treg was analyzed by four-color staining flow cytometry. CD4~+T cell absolute counts were determined using Multitest CD3/CD4/CD8/CD45 kit and plasma viral loads were detected on Nucli

  8. The expression of CD25, CD11b, MHC-II, SWC1, SWC7, CD45RA and CD45RC on porcine chi delta T lymphocytes isolated from different organs

    Czech Academy of Sciences Publication Activity Database

    Štěpánová, Kateřina; Šinkora, Marek

    Malden: WILEY-BLACKWELL, 2011. s. 59-59. ISSN 0019-2805. [Annual Congress of the British Society for Immunology. 05.12.2011-08.12.2011, Liverpool] R&D Projects: GA ČR GA524/07/0087; GA ČR GAP502/10/0038; GA MŠk ME09089 Institutional research plan: CEZ:AV0Z50200510 Keywords : germ free * CD25 Subject RIV: EC - Immunology

  9. CD4(+)CD25 (+)CD127 (low/-) T cells: a more specific Treg population in human peripheral blood.

    Science.gov (United States)

    Yu, Ning; Li, Xiaomei; Song, Weiya; Li, Dongmei; Yu, Daliang; Zeng, Xiaofeng; Li, Mengtao; Leng, Xiaomei; Li, Xiangpei

    2012-12-01

    The quantitative identification and enrichment of viable regulatory T cells (Treg) requires reliable surface markers that are selectively expressed on Treg. Foxp3 is the accepted marker of nTreg, but it cannot be used to isolate cells for functional studies. In this study, we compared four staining profiles of Treg, including CD4(+)CD25(high) T cells, CD4(+)CD39(+) T cells, CD4(+)CD73(+) T cells, and CD4(+)CD25(+)CD127(low/-) T cells. We found that CD4(+)CD25(+)CD127(low/-) T cells expressed the highest level of Foxp3 and had the strongest correlation with CD4(+)CD25(+)Foxp3(+) T cells, the accepted identifying characteristics for "real" nTreg cells. Moreover, functional data showed that CD4(+)CD25(+)CD127(low/-) T cells could effectively suppress the proliferation of CD4(+)CD25(-) T cells, suggesting that compared with the other three populations, CD4(+)CD25(+)CD127(low/-) T cells best fit the definition of naturally occurring regulatory T cells in human peripheral blood. Finally, we showed that CD4(+)CD25(+)CD127(low/-) can be used to quantitate Treg cells in individuals with systemic lupus erythematosus supporting the use of CD4(+)CD25(+)CD127(low/-) to identify human Treg cells. PMID:22752562

  10. Effect of methylprednisolone on CD4 + CD25 + T regulator cells in peripheral blood with asthmatic patients in vitro%甲泼尼龙对哮喘患者外周血CD4+CD25+调节性T细胞影响的体外研究

    Institute of Scientific and Technical Information of China (English)

    鞠云飞; 孙立锋; 胡华; 杨燕; 滕格玲

    2011-01-01

    目的 观察哮喘患者外周血中CD4+ CD25+调节性T细胞(Treg)功能状态及甲泼尼龙对其的影响.方法 清晨取静脉血5 mL,常规分离外周血单个核细胞,分为健康组、哮喘组及1×10-7 mol·L-1甲泼尼龙哮喘干预组,刺激培养48 h,用ELISA法测定IL - 10及TGF - β1的浓度;流式细胞仪检测CD4+ CD25+Treg的比例及细胞内Foxp3表达,用SPSS 17.0进行分析.结果 哮喘组,外周血CD4+ CD25+ Treg细胞比例降低,CD4+ CD25+ Foxp3+ Treg细胞减少,TGF -β1分泌减少,IL - 10分泌有增高趋势,但与健康组比较无统计学意义;甲泼尼龙可以明显增加哮喘急性发作期患者CD4+ CD25+ Treg比例,Foxp3表达明显增强;但未能增加TGF -β1及IL-10的分泌.结论 哮喘患者CD4+ CD25+ Treg功能低下,可能是哮喘发病机制之一;甲泼尼龙可能通过上调CD4+ CD25+ Treg数量,起到控制哮喘急性发作的作用.%Objective To observe the state of CD4+ CD25 + T regulator ( Treg ) cells and the effect of methylprednisolone ( MP) on it in peripheral blood with asthmatic patients. Methods Thirty patients with asthma and 20 healthy controls who visited the Chest Hospital of Shandong Province from Jan, 2010 to Dec, 2010 were included. Five mL fasting blood samples were collected and peripheral blood mononuclear cells ( PBMCs) were isolated using Ficoll - Hypaque density gradient centrifugation. PBMCs were incubated with phytohemagglutinin ( PHA) . The samples were divided into three groups, including healthy control, asthma group, 1 x 10-7 mol · L-1 MP group. After 48 h incubation, supernatant was harvested to determine levels of IL - 10 and TGF - β1 by ELISA. Intracellular 3 - colour flow cytometry were used to assess the expression of Foxp3. All data were analyzed using SPSS 17.0. Results The ratio of CD4+ CD25 + Treg cells and CD4 + CD25 + Foxp3 + Treg cells declined in asthmatic patients compared with healthy control. The secretion of TGF - β1 weaked . The secretion of

  11. CD4+CD25+ Treg cell and Foxp3 expression play key roles in the pathogenesis of cervical carcinoma%宫颈癌患者外周血CD4+CD25+调节性T细胞及其Foxp3表达的临床意义

    Institute of Scientific and Technical Information of China (English)

    徐华; 王丹波

    2015-01-01

    目的 评价宫颈癌患者外周血CD4+CD25+调节性T细胞(Regulatery T cell,Treg)及其Foxp3表达的临床意义.方法 收集中国医科大学附属盛京医院自2012年1月至2013年6月手术治疗的早期宫颈癌患者(FIGO分期≤Ⅱ期)34例,高级别宫颈上皮内瘤变(CIN2/3)患者30例以及20例健康女性作为正常对照.流式细胞技术检测各研究组手术前、后及对照组外周血Treg及Foxp3+ Treg细胞占CD4+T细胞比例,应用t检验和单因素方差法进行统计学分析.结果 比较宫颈癌、CIN2/3及对照组外周血Treg及Foxp3+ Treg细胞所占CD4+T细胞比例,各组间差异有统计学意义(P<0.05);术后早期(7 d),宫颈癌及CIN2/3患者外周血Foxp3+ Treg细胞所占比例较术前显著下降(P<0.05),但Treg细胞所占比例无显著性变化(P>0.05);术后恢复期(1个月),宫颈癌及CIN2/3患者外周血Foxp3+ Treg细胞所占比例较术后早期无显著变化,但Treg细胞所占比例较术后早期显著性降低(P<0.05).结论 宫颈癌患者外周血Treg细胞及其Foxp3的表达均与病变程度相关;Treg细胞Foxp3的表达具有不稳定性;宫颈癌肿瘤微环境可能是维持Treg细胞分化增殖及Foxp3稳定表达的外在客观条件.

  12. CD4+CD25+Treg细胞与支气管哮喘%CD4+ CD25+ Treg cells and bronchial asthma

    Institute of Scientific and Technical Information of China (English)

    鞠云飞; 孙立锋; 胡华

    2011-01-01

    The main function of CD4+ CD25+ Treg cells are immunological anergy and inhibition,which is essential to the maintenance of immunological tolerance in the host.CD4+ CD25+ Treg cells produce inhibitory cytokines (TGF-β and IL-10),express membrane molecules (CTLA-4,GITR,etc) and Foxp3.There are abnormal in function and quantity of CD4+ CD25+ Treg cells of peripheral blood from asthmatic patients,which maybe one of the pathogenesis of asthma.Glucocorticoids can inhibit the airway inflamation of asthma by impacting CD4+ CD25+ Treg cells.%CD4+ CD25+ Treg细胞的主要作用表现为免疫无能性和免疫抑制性,是外周免疫耐受形成机制的主要组成部分.其主要作用机制为分泌抑制性细胞因子(IL-10和TGF-β)、表达细胞表面分子(CTLA-4、GITR等)及Foxp3等.支气管哮喘患者外周血CD4+ CD25+ Treg功能及数量存在异常,这可能是支气管哮喘发病机制之一.糖皮质激素可以通过影响CD4+ CD25+ Treg的状态起到抑制支气管哮喘气道炎症的作用.

  13. Different thresholds of T cell activation regulate FIV infection of CD4+CD25+ and CD4+CD25- cells

    International Nuclear Information System (INIS)

    Cellular activation plays an important role in retroviral replication. Previously, we have shown that CD4+CD25+ T cells by the virtue of their partially activated phenotype represent ideal candidates for a productive feline immunodeficiency virus (FIV) infection. In the present study, we extended our previous observations with regard to FIV replication in CD4+CD25+ and CD4+CD25- cells under different stimulation conditions. Both CD4+CD25+ and CD4+CD25- cells remain latently infected in the absence of IL-2 or concanvalinA (ConA), respectively; harboring a replication competent provirus capable of reactivation several days post-infection. While CD4+CD25+ cells require low levels of exogenous IL-2 and virus inputs for an efficient FIV replication, CD4+CD25- T cells can only be productively infected in the presence of either high concentrations of IL-2 or high virus titers, even in the absence of mitogenic stimulation. Interestingly, while high virus input activates CD4+CD25- cells to replicate FIV, it induces apoptosis in a high percentage of CD4+CD25+ T cells. High IL-2 concentrations but not high virus inputs lead to surface upregulation of CD25 and significant cellular proliferation in CD4+CD25- cells. These results suggest that CD4+CD25+ and CD4+CD25- T cells have different activation requirements which can be modulated by both viral and cytokine stimuli to reach threshold activation levels in order to harbor a productive FIV infection. This holds implications in vivo for CD4+CD25+ and CD4+CD25- cells to serve as potential reservoirs of a productive and latent FIV infection

  14. Anticancer immunotherapy by CTLA-4 blockade: obligatory contribution of IL-2 receptors and negative prognostic impact of soluble CD25.

    Science.gov (United States)

    Hannani, Dalil; Vétizou, Marie; Enot, David; Rusakiewicz, Sylvie; Chaput, Nathalie; Klatzmann, David; Desbois, Melanie; Jacquelot, Nicolas; Vimond, Nadège; Chouaib, Salem; Mateus, Christine; Allison, James P; Ribas, Antoni; Wolchok, Jedd D; Yuan, Jianda; Wong, Philip; Postow, Michael; Mackiewicz, Andrzej; Mackiewicz, Jacek; Schadendorff, Dirk; Jaeger, Dirk; Zörnig, Inka; Hassel, Jessica; Korman, Alan J; Bahjat, Keith; Maio, Michele; Calabro, Luana; Teng, Michele Wl; Smyth, Mark J; Eggermont, Alexander; Robert, Caroline; Kroemer, Guido; Zitvogel, Laurence

    2015-02-01

    The cytotoxic T lymphocyte antigen-4 (CTLA-4)-blocking antibody ipilimumab induces immune-mediated long-term control of metastatic melanoma in a fraction of patients. Although ipilimumab undoubtedly exerts its therapeutic effects via immunostimulation, thus far clinically useful, immunologically relevant biomarkers that predict treatment efficiency have been elusive. Here, we show that neutralization of IL-2 or blocking the α and β subunits of the IL-2 receptor (CD25 and CD122, respectively) abolished the antitumor effects and the accompanying improvement of the ratio of intratumoral T effector versus regulatory cells (Tregs), which were otherwise induced by CTLA-4 blockade in preclinical mouse models. CTLA-4 blockade led to the reduction of a suppressive CD4(+) T cell subset expressing Lag3, ICOS, IL-10 and Egr2 with a concomitant rise in IL-2-producing effector cells that lost FoxP3 expression and accumulated in regressing tumors. While recombinant IL-2 improved the therapeutic efficacy of CTLA-4 blockade, the decoy IL-2 receptor α (IL-2Rα, sCD25) inhibited the anticancer effects of CTLA-4 blockade. In 262 metastatic melanoma patients receiving ipilimumab, baseline serum concentrations of sCD25 represented an independent indicator of overall survival, with high levels predicting resistance to therapy. Altogether, these results unravel a role for IL-2 and IL-2 receptors in the anticancer activity of CTLA-4 blockade. Importantly, our study provides the first immunologically relevant biomarker, namely elevated serum sCD25, that predicts resistance to CTLA-4 blockade in patients with melanoma. PMID:25582080

  15. The Detection and Significance of CD4+CD25+CD127lo/-Tregs in Peripheral Blood CD4+T Cells of Healthy and Asthma Patients during Exacerbation and Remission%急性发作期和缓解期哮喘患者外周血CD4+CD25+CD127lo/-Treg的检测和意义

    Institute of Scientific and Technical Information of China (English)

    上官文姬; 沈惠风; 王利民; 叶人诵

    2011-01-01

    目的:通过检测健康人、急性发作期和缓解期哮喘患者外周血CD4+CD25+CD127lo/-Treg细胞的表达水平,探讨该细胞表达水平的变化与哮喘患者病情严重程度的关系.方法:采集急性发作期、缓解期哮喘患者及健康人外周抗凝血,用流式细胞术检测其外周血中CD4+CD25+CD127lo/-Treg细胞占CD4+T细胞的比例.结果:哮喘急性发作组和缓解组患者外周血中CD4+CD25+CD127lo/-Treg细胞占CD4+T细胞的百分比低于健康对照组(P<0.05);哮喘急性发作期组CD4+CD25+CD127lo/-Treg细胞占CD4+T细胞的百分比低于哮喘缓解组(P<0.05).结论:CD4+CD25+ CD127lo/-Treg细胞数量减少可能参与了哮喘的发病过程.%Objective:To investigate the relationship between the expression levels of CD4 + CD25 + CD127lo/-Tregs in peripheral blood of asthma patients during exacerbation or remission. Methods: A total of 2 ml peripheral blood were collected in healthy or asthma patients during exacerbation and remission. The percentage of CD4+ CD25 + CDl27lo/-Tregs in CD4+ T cells by flow cytometry were detected. Results: A significant decrease of the percentage of CD4+ CD25+ CD127lo/- Tregs in CD4+ T cells was observed in exacerbation and remission groups compared with control group (P<0.05). This was also detected in exacerbation group compared with remission group (P<0.05). Conclusions: The CD4+ CD25+ CD127lo/-Tregs play a key role in the pathogenesis of asthma and its decrease,which may lead to immune dysfunction, may be involved in the mechanisms of asthma.

  16. Regulatory CD4+CD25+ T Cells Dampen Inflammatory Disease in Murine Mycoplasma Pneumonia and Promote IL-17 and IFN-γ Responses

    Science.gov (United States)

    Odeh, Adam N.; Simecka, Jerry W.

    2016-01-01

    Mycoplasmas cause respiratory diseases characterized by persistent infection and chronic airway inflammation. Mycoplasma lung disease is immunopathologic, with CD4+ Th cells determining both disease severity and resistance to infection. Th2 cell responses promote immunopathology, while Th1 cells confer resistance to infection. However, regulatory CD4+ T cells may also have a role in the pathogenesis of mycoplasma respiratory diseases. We hypothesized Treg cells control the severity of the inflammatory lesions and may also promote persistence of infection. To examine this, BALB/c mice were depleted of CD25+ cells, and had increased disease severity due to Mycoplasma pulmonis infection. Increases in mycoplasma antibody responses and lymphocyte infiltration into lungs also occurred after CD25+ cell depletion. CD4+CD25+ regulatory T cells promoted IFN-γ and IL-17 mycoplasma-specific CD4+ T cell responses in vitro and in vivo, while dampening IL-13+ Th responses. Neither IL-10 nor TGF-ß expression was detected in CD4+CD25+ T cells from lymph nodes. Thus, a regulatory T cell population plays an important role in controlling damaging immune responses in mycoplasma respiratory disease but does not contribute to persistence of infection. It appears that a regulatory T cell population preferentially dampens Th2 cell-mediated inflammatory responses to mycoplasma through a mechanism independent of IL-10 or TGF-ß characteristic of “classic” Treg cells. PMID:27175511

  17. TL1A increases expression of CD25, LFA-1, CD134 and CD154, and induces IL-22 and GM-CSF production from effector CD4 T-cells

    DEFF Research Database (Denmark)

    Reichwald, Kirsten; Jørgensen, Tina Z.; Skov, Søren

    2014-01-01

    Elevated levels of the cytokine TL1A is associated with several autoimmune diseases e.g. rheumatoid arthritis and inflammatory bowel disease. However, the exact role of TL1A remains elusive. In this study, we investigated the function of TL1A in a pro-inflammatory setting. We show that TL1A toget...... of CD25 (IL-2Rα) and CD11a (α-chain of LFA-1) on CD4 T-cells, likely governing increased IL-2/IL-15 sensitivity and cell-cell contact. Along with this, TL1A co-stimulation caused a specific induction of IL-22 and GM-CSF from the activated T-cells. These results substantially contribute...

  18. RA8, A human anti-CD25 antibody against human treg cells

    Energy Technology Data Exchange (ETDEWEB)

    Arias, Robyn; Flanagan, Meg; Miller, Keith D.; Nien, Yu-Chih; Hu, Peisheng; Gray, Dixon; Khawli, Leslie A.; Epstein, Alan L.

    2007-06-01

    Although anti-CD25 antibodies exist for clinical use in patients, there is a need for the development of a human Treg antibody that will abrogate the immunosuppressive function of this small but critical T cell subtype. Based upon mounting evidence that the level of Treg cells in the tumor microenvironment correlates with clinical prognosis and stage in man, it appears that Treg cells play an important role in the tumor's ability to overcome host immune responses. In mice, the rat anti-mouse CD25 antibody PC61 causes depletion of CD25-bearing Treg cells both peripherally in lymphatic tissues and in the tumor microenvironment, without inducing symptoms of autoimmunity. A similar antibody, though with the ability to delete Treg cells specifically, would be an important new tool for reversing tumor escape associated with Treg immunosuppression in man. To begin to generate such a reagent, we now describe the development of a human anti-CD25 antibody using a novel yeast display library. The target antigen CD25-Fc was constructed and used for five rounds of selection using a non-immune yeast display library that contained as many as 109 single chain variable fragments (scFv). Two unique clones with low KD values (RA4 and RA8) were then selected to construct fully human anti-CD25 antibodies (IgG1/kappa) for stable expression. One antibody, RA8, showed excellent binding to human CD25+ cell lines and to human Treg cells and appears to be an excellent candidate for the generation of a human reagent that may be used in man for the immunotherapy of cancer.

  19. Allo-PBSCT患者CD4+CD25+调节性T细胞的体外研究%Study on post-allogeneic peripheral blood stem cell transplantation patients'CD4 + CD25 + regulatory T cells in vitro

    Institute of Scientific and Technical Information of China (English)

    翟海龙; 赖永榕

    2011-01-01

    Objective To investigate the proliferation reaction of CD4+ CD25+ Tregs in the stimulating of costimulato-ry signal, lymphocyte reactions mixed with CD4+ CD25- T cells of CD4+ CD25+ Tregs, and cytokine secretion state of the two cells in allogeneic peripheral blood stem cell transplantation ( Allo-PBSCT) patients. Methods CD4+ CD2S+ Tregs and CD4+ CD25- T cells from peripheral blood obtained from 36 patients who had undergone Allogeneic peripheral blood stem cell transplantation (Allo-PBSCT), 7 healthy volunteers as control, were isolated with magnetic cells sorting separation. Then CD4+ CD25+ Tregs and CD4+ CD25+ Tregs + CD4+ CD25- T cells were cultered for 72 hours, stimulated by an-ti-CD3-mAbs and anti-CD28-mAbs. After that the cultures added with CCK-8 solution were incubated for 1 hour. Then OD450 were detected by ELISA. IL-10, TGF-β and IFN-γ from the two above cell cultures were detected by ELISA method. Results OD450 values of CD4+ CD25+ Tregs were both extremely lower than that of CD4+ CD25- T cells and CD4+ CD25+ Tregs + CD4+ CD25- T cells( P < 0.01). IL-10, TGF-p and IFN-γ secreted by CD4 + CD25+ Tregs in vitro from patients with and without GVHD were signigicantly lower than that of CD4+ CD25- T cells( P < 0.01 ). The 3 cytokines secreted by CD4+ CD25- Tregs + CD4+ CD25- T cells group were also signigicantly lower than that of CD4+ CD25- T cells( P <0.05 ). The cytokines secretory of Allo-PBSCT group was similar with that of control group. Conclusions If the suppressive function of CD4+ CD25+ Tregs are utilized, incidence of GVHD post- Allo-PBSCT may decrease.%目的 探讨异基因外周血干细胞移植(Allo-PBSCT)患者外周血CD4+ CD25+调节性T细胞(Tregs)在协同刺激信号作用下的增殖反应、与CD4+ CD25 -T细胞混合淋巴细胞反应及上述两种培养细胞的细胞因子分泌情况.方法 对36例Allo-PBSCT患者离体CD4+ CD25+ Tregs在抗CD3-mAbs和抗CD28-mAbs的刺激下行CD4+CD25 +Tregs培养和CD4+ CD25+ Tregs、CD4

  20. Effects of estrogen on CD4+CD25+ regulatory T cell in peripheral blood during pregnancy

    Institute of Scientific and Technical Information of China (English)

    Yuan-Huan Xiong; Zhen Yuan; Li He

    2013-01-01

    Objective:To investigate the effects of estrogen (E2) level on regulatory T cells (Treg) in peripheral blood during pregnancy. Methods:A total of 30 healthy non-pregnant women were selected as control group, 90 pregnant women of early, middle and late pregnancy and 30 postpartum women at 1 month after parturition were selected as experimental groups including early pregnancy group, middle pregnancy group and late pregnancy group;the proportions of CD4+CD25+Treg and CD4+CD25+CD127-Treg among CD4+T cells were detected by flow cytometry;the serum estrogen content in peripheral blood was detected by electrochemical immune luminescence method. Results: E2 level was coincident with the change of Tregs number during pregnancy. The estrogen content in peripheral blood increased gradually from early pregnancy to late pregnancy, then decreased significantly after parturition, and the level at 1 month after parturition down to the level in non-pregnancy group (P>0.05);the level of E2 in pregnancy groups were significantly higher than those in non-pregnancy group (P0.05);the proportions in middle and late pregnancy groups were significantly higher than those in early pregnancy group (P0.05). There was correlation between Tregs number with estrogen level during pregnancy. The proportion of CD4+CD25+ Treg and CD4+CD25+CD127- Treg were positively correlated with estrogen level. Conclusions:High proportion of CD4+CD25+Treg and CD4+CD25+CD127-Treg is closely related to the high level of E2 during pregnancy. It suggested that high level of estrogen may induce an increase of CD4+CD25+Treg in peripheral blood, and then influence the immune function of pregnant women. The results of this experiment might play an important role of estrogen in immune-modulation during pregnancy.

  1. Endogenous IRBP can be dispensable for generation of natural CD4+CD25+ regulatory T cells that protect from IRBP-induced retinal autoimmunity

    OpenAIRE

    Grajewski, Rafael S.; Silver, Phyllis B.; Agarwal, Rajeev K.; Shao-bo SU; Chan, Chi-Chao; Liou, Gregory I.; Caspi, Rachel R.

    2006-01-01

    Susceptibility to experimental autoimmune uveitis (EAU), a model for human uveitis induced in mice with the retinal antigen interphotoreceptor retinoid-binding protein (IRBP), is controlled by “natural” CD4+CD25+ regulatory T (T reg) cells. To examine whether endogenous expression of IRBP is necessary to generate these T reg cells, we studied responses of IRBP knockout (KO) versus wild-type (WT) mice. Unexpectedly, not only WT but also IRBP KO mice immunized with a uveitogenic regimen of IRBP...

  2. Role of regulatory CD4+CD25+ Foxp3 T cells in bronchial asthma in Egyptian children.

    Science.gov (United States)

    Bakr, Salwa I; Mahran, Manal Z; Soliman, Dina A

    2013-01-01

    CD4+CD25+high Foxp3 regulatory T (Treg) cells are known to play a key role in balancing immune response to maintain peripheral tolerance against harmless antigens or allergens. Defective immunological suppression by CD4+CD25+high Foxp3 Treg cells can be a cause of the inflammation that leads to an allergic condition such as asthma. The aims of the study are to (1) determine CD4+CD25+high Foxp3 Treg cells frequency in the peripheral blood of children with and without asthma; and (2) investigate the association between CD4+CD25+high Foxp3 Treg cells frequency with disease severity and corticosteroid therapy. Sixty asthmatic children with varying disease severity (20 mild, 20 moderate and 20 severe) were enrolled in the study. Severe asthmatic children were further subdivided into two groups, one on corticosteroid therapy and the other was not on corticosteroid. Twenty age and sex matched healthy children were enrolled as controls. Number of circulating CD4+CD25+high Foxp3 Tregs were measured using flow cytometry. Our finding demonstrates that children with asthma had a significant decrease of CD4+CD25high Foxp3 Treg cells and Tregs/T effectors ratio in peripheral blood compared to children without asthma. Patients with moderate asthma demonstrated lower frequency of CD4+CD25+high Foxp3 Treg cells compared to mild and severe asthmatic patients. Those on corticosteroid therapy revealed significant increase in CD4+CD25+high Foxp3 Treg cells and decrease in T effectors. It is concluded that asthmatic children have decreased number of CD4+CD25+high Foxp3 Treg cells leading to increase in effectors cells which mediate inflammation in the airways. Corticosteroid therapy plays a role in elevating number of CD4+CD25+high Foxp3 Treg cells and maintaining its suppressor function. PMID:24617045

  3. Use of CD25 as an immunohistochemical marker for acquired ocular toxoplasmosis

    Directory of Open Access Journals (Sweden)

    Cristina Miyamoto

    2010-10-01

    Full Text Available PURPOSE: Toxoplasmosis is the most common cause of posterior infectious uveitis worldwide. It is often impossible to determine its congenital or acquired nature. Interleukin-2 (IL-2 in peripheral blood has been described as a possible marker for acquired toxoplasmosis. The purpose of this study is to evaluate the histopathological characteristics of ocular toxoplasmosis cases using CD25 as a marker for the expression of interleukin-2. METHODS: Ten formalin-fixed, paraffin-embedded enucleated globes from ten immunocompetent patients with clinical diagnosis of toxoplasmosis were evaluated. Four patients had the acquired form of ocular toxoplasmosis (positive IgM while six were IgM negative and IgG positive for toxoplasmosis. Histopathological slides were reviewed for the extension of the retinal necrosis, number of toxo cysts, the granulomatous inflammatory reaction, the presence of T and B cells within the choroid and the IL-2 expression. Immunohistochemistry using monoclonal antibodies was performed to observe the expression of CD4, CD8, CD20, CD25, and CD68. RESULTS: The histopathological evaluation disclosed no differences between acquired and the other ocular toxoplasmosis cases regarding the characteristics studied. However, CD25 showed a higher expression of IL-2 on the 4 acquired cases of ocular toxoplasmosis compared to the remainders. CONCLUSIONS: To the best of our knowledge, this is the first report showing that the use of CD25 as a marker for interleukin-2 could differentiate acquired ocular toxoplasmosis.

  4. Glucocorticoid induced TNFR-related protein (GITR as marker of human regulatory T cells: expansion of the GITR+CD25- cell subset in patients with systemic lupus erythematosus

    Directory of Open Access Journals (Sweden)

    E. Bartoloni Bocci

    2011-06-01

    Full Text Available Objectives: Regulatory T cells (TREG represent a T cell subset able to modulate immune response by suppressing autoreactive T-lymphocytes. The evidence of a reduced number and an impaired function of this cell population in autoimmune/ inflammatory chronic diseases led to the hypothesis of its involvement in the pathogenesis of these disorders. Glucocorticoid-induced TNFR-related protein (GITR is a well known marker of murine TREG cells, but little is known in humans. The aim of this study was to investigate the characteristics of TREG cells in systemic lupus erythematosus (SLE and the potential role of GITR as marker of human TREG. Methods: Nineteen SLE patients and 15 sex- and age-matched normal controls (NC were enrolled. CD4+ T cells were magnetic sorted from peripheral blood by negative selection. Cell phenotype was analyzed through flow-cytometry using primary and secondary antibodies and real time polymerase-chain reaction (PCR using TaqMan probes. Results: The CD25highGITRhigh subset was significantly decreased in SLE patients with respect to NC (0.37±0.21% vs 0.72±0.19%; p<0.05. On the opposite, the CD25-GITRhigh cell population was expanded in the peripheral blood of SLE patients (3.5±2.25 vs 0.70±0.32%, p<0.01. Interestingly, FoxP3 at mRNA level was expressed in both CD25- GITRhigh and CD25highGITRhigh cells, suggesting that both cell subsets have regulatory activity. Conclusions: CD4+CD25-GITRhigh cells are increased in SLE as compared to NC. The expression of high level of GITR, but not CD25, on FoxP3+ cells appears to point to a regulatory phenotype of this peculiar T cell subset.

  5. CD4+CD25−Foxp3+ T cells play a role in tuberculous hydrothorax rather than malignant hydrothorax

    OpenAIRE

    Tang, Ying; Peng, Li-Ping; Qin, Gui-Xiang; Sun, Jing-Ting; Xu, Li-Jun; Jiang, Yan-Fang

    2015-01-01

    Background Foxp3+ T cells regulate inflammation and tumorigenesis. However, little is known about the role of different subsets of Foxp3+ T cells in malignant or tuberculous hydrothorax. Methods The numbers of CD4+CD25+Foxp3+, CD4+CD25−Foxp3+ T cells and the levels of some inflammatory cytokines in patients with tuberculous hydrothorax, malignant hydrothorax, and healthy controls (HCs) were examined by flow cytometry and ELISA. The potential association between the numbers of different subset...

  6. 肥大细胞体外诱生CD4~+CD25~+Foxp3~+调节性T细胞的作用%Mast cells induce CD4~+CD25~+Foxp3~+ regulatory T cells in vitro

    Institute of Scientific and Technical Information of China (English)

    张维娜; 周巧丹; 雒真龙; 陈忠华; 吴轲; 周鸿敏; 何文涛; 高英; 汪理; 林星光; 方泽民; 蔡兰军

    2009-01-01

    探索小鼠骨髓源性肥大细胞能否在体外诱生CD4~+CD25~+Foxp3~+调节性T细胞.从小鼠股骨获取骨髓细胞,加入完全RPMI 1640培养基(含IL-3和SCF各10 ng/m1)诱导4周.用甲苯胺蓝染色法观察肥大细胞异染颗粒;流式检测CD117和FcεRIα双阳性细胞比例.将肥大细胞与同基因来源的T细胞按不同比例(1,2、1:1、2:1)置于48孔培养板中培养,作为三个实验组;未加肥大细胞的单独T细胞组作对照组.四个组均加入CD3抗体和CD28抗体各2μg/ml,IL-2 1 000U/ml,5 d后流式检测Foxp3表达情况.RT-PCR,免疫组化法检测肥大细胞中TGF-β1的表达.与对照组相比,实验组Foxp3表达均升高(对照组:3.37%±0.40%;实验组为:8.23%±0.80%、10.87%±1.25%、13.63%±0.55%).RT-PCR和免疫组化均检出TGF-β1表达.肥大细胞能在体外诱导T细胞转化为CD4~+CD25~+Foxp3~+调节性T细胞,可能与肥大细胞表达TGF-β1有关.%To detect whether mouse mast cells can induce CD4~+ CD25~+ Foxp3+ regulatory T cells (Treg) in vitro. Bone marrow cells obtained from C57BL/6 (H-2b) mice were cultured with IL-3 (10 ng/ml) and SCF (10 ng/ml) for 4 weeks. The purity of bone marrow mast cells (BMMCs) was tested by flow cytometry. The expression of TGF-β1 in BMMCs was tested by RT-PCR and immunohistochemistry. Then the BMMCs were co-cultured with T cells of C57BL/6 mice at 1 : 2, 1 : 1 and 2 : 1 ratios in the presence of anti-CD3 antibody(2μg/ml), anti-CD28 antibody(2μg/ml) and IL-2 (1 000 U/ml). The percentages of CD4~+ CD25~+ Foxp3+ T cells in the co-cultured system were compared among different groups by flow cytometry on day 5. It was found that the percentages of CD4~+ CD25~+ Foxp3+ cells were significantly higher in the ratio of BMMC/T 2 : 1 group(13.63 ± 0.55%), the 1 : 1 group(10.87% ± 1.25%)and 1 : 2 group(8. 23% ± 0.80%) than in the control group(3.37% ± 0.40%). The expression of TGF-β1 was determined in the mouse BMMCs by RT-PCR and immunohistochemistry. Our find

  7. CD4+CD25+调节性T细胞及其分子标记物与支气管哮喘%CD4+CD25+regulatory T cell and its molecular marker with bronchial asthma

    Institute of Scientific and Technical Information of China (English)

    马祥; 毛辉; 梁宗安

    2009-01-01

    CD4+CD25+regulatory T cells are a kind of lymphocytes characterized by immune inhibition and immune inability,FOXP3 is a specific molecular marker of CD4+CD25+regulatory T cells,and critical for the generation,peripheral expression and function of CD4+CD25+regulatory T cells.In recent years,a series of studies showed that CD4+CD25+regulatory T cells interfere with the development and progression of bronchial asthma,the intervention of regulatory T cell and its relative genes may offer a novel therapeutic strategy for bronchial asthma.%CD4+CD25+调节性T细胞是一类以免疫抑制和免疫无能为特征的淋巴细胞群,FOXP3是CD4+CD25+调节性T细胞一个特征性的分子标志物,并且对CD4+CD25+调节性T细胞的发育、外周表达和功能维持有着关键性的作用.近年来,多项研究显示CD4+CD25+调节性T细胞参与并影响了支气管哮喘的发生、发展过程,对调节性T细胞或其相关基因的干预也许会成为支气管哮喘治疗的新方向.

  8. CD4(+CD25(-Nrp1(+ T cells synergize with rapamycin to prevent murine cardiac allorejection in immunocompetent recipients.

    Directory of Open Access Journals (Sweden)

    Qing Yuan

    Full Text Available Besides CD4(+CD25(+Foxp3(+ regulatory T cells (Tregs, other immunosuppressive T cells also participated in the regulation of immune tolerance. Reportedly, neuropilin-1 (Nrp1 might be one of the molecules by which regulatory cells exert their suppressive effects. Indeed, CD4(+CD25(-Nrp1(+ T cells exhibit potent suppressive function in autoimmune inflammatory responses. Here we investigated the specific role of CD4(+CD25(-Nrp1(+ T cells in the setting of the transplant immune response. Through MLR assays, we found that CD4(+CD25(-Nrp1(+ T cells suppressed the proliferation of naive CD4(+CD25(- T cells activated by allogeneic antigen-stimulation. Adoptive transfer of CD4(+CD25(-Nrp1(+ T cells synergized with rapamycin to induce long-term graft survival in fully MHC-mismatched murine heart transplantation, which was associated with decreased IFN-γ, IL-17 and increased IL-10, TGF-β, Foxp3 and Nrp1 expression in the grafts. Importantly, our data indicated that CD4(+CD25(-Nrp1(+ T cell transfer augments the accumulation of Tregs in the recipient, and creates conditions that favored induction of hyporesponsiveness of the T effector cells. In conclusion, this translational study indicates the possible therapeutic potential of CD4(+CD25(-Nrp1(+ T cells in preventing allorejection. CD4(+Nrp1(+ T cells might therefore be used in bulk as a population of immunosuppressive cells with more beneficial properties concerning ex vivo isolation as compared to Foxp3(+ Tregs.

  9. Influences of Inhaled Corticosteroids on CD4+ CD25+ Regulatory T Cells and Foxp3 mRNA of Asthmatic Patients%吸入激素对哮喘患者外周血CD4+CD25+调节性T细胞水平及Foxp3mRNA表达的影响

    Institute of Scientific and Technical Information of China (English)

    薛克营; 柯明耀; 姜燕; 陈玲玲; 吴雪梅; 谢红旗

    2011-01-01

    目的 观察支气管哮喘患者外周血CD4 CD25调节性T细胞(CD4CD25Treg)、Foxp3 mRNA的水平及吸入糖皮质激素对其的影响.方法 流式细胞仪检测非急性发作期哮喘患者吸入激素前、后和健康志愿者(正常对照组)外周血单个核细胞(PBMCs)中CD4CD25Treg的比例,RT-PCR检测Foxp3 mRNA表达变化,肺功能仪检测第1秒用力呼气容积占预计值百分比(FEV.%pred)和呼气峰流量(PEF).结果 哮喘组治疗前CD4CD25Treg水平及Foxp3 mRNA的表达显著低于正常对照组(PCD25Treg水平及Foxp3 mRNA的表达较治疗前显著升高(P%pred、PEF显著低于正常对照组(P%pred、PEF较治疗前显著增加(PCD25Treg数量减少,功能下降,参与了哮喘的发病过程;吸入激素可以上调哮喘患者外周血CD4CD25Treg及Foxp3 mRNA的水平,改善哮喘患者的肺功能.%Objective To investigate the percentage of CD4+ CD25+ Treg cells and expression of Foxp3 mRNA in asthmatic patients and the impacts of inhaled steroids.Methods The percentages of CD4 + CID25 + Treg cells was assayed by flow cytometry and the expression of Foxp3 mRNA was detected by RT-PCR in peripheral blood mononuclear cells from the patients with chronic persistent asthma before and after steroids inhalation in comparison with healthy control.The forced expired volum in one second/predicted value( FEV1 % pred)and peak expired flow (PEF) were measured by spirometry.Results The level of CD4 + CD25 + Treg cells and the expression of Foxp3 mRNA were lower in asthmatics before steroids treatment than those in control ( P < 0.05 ) which were increased significantly after steroids treatment ( P < 0.05 ).FEV1 % pred and PEF were declined significantly than those in control but improved markedly after treatment (P < 0.05 ).Conclusions The insufficiency of amount and function of immue-suppressive CD4 + CD25 + Treg cells may play a role in the pathogenesis of asthma.Inhaled steroids can improve the lung function of asthmatics

  10. Frequently Increased but Functionally Impaired CD4+CD25+ Regulatory T Cells in Patients with Oral Lichen Planus.

    Science.gov (United States)

    Zhou, Leilei; Cao, Tianyi; Wang, Yufeng; Yao, Hui; Du, Guanhuan; Chen, Guangjie; Niu, Xiaoyin; Tang, Guoyao

    2016-06-01

    Oral lichen planus (OLP) is a T cell-mediated chronic inflammatory mucosal disease, and CD4(+)CD25(+) regulatory T cells (Tregs) are considered involved in the pathogenesis of OLP. In this study, to investigate whether there are intrinsic factors that might cause functional changes in Tregs in this disease, we evaluated the frequency of Tregs in peripheral blood and oral lesions and the expression levels of function-related transcription factors, forkhead/winged-helix transcription factor box P3 (FOXP3), transforming growth factor β (TGF-β), interleukin 10 (IL-10), and TGF-β receptors (TβRI and TβRII) mRNAs in Tregs of patients with oral lichen planus (OLP). We also investigated the frequency of pro-inflammatory cytokines (IFN-γ and IL-17A) producing Foxp3(+) regulatory cells. Increased proportions of Tregs were found in OLP patients. The expression of FOXP3 on mRNA and protein level was elevated in the Tregs of OLP. The expression of TGF-β was lower both on the mRNA and serum level, whereas the expression of IL-10 showed no significant difference between the OLP patients and normal controls. The percentages of CD4(+)FOXP3(+)IL-17(+) T cells were significantly higher than that of normal controls, whereas the percentages of CD4(+)FOXP3(+)IFN-γ(+) T cells did not differ significantly. Furthermore, impaired suppressive function of CD4(+)CD25(+) T cells was demonstrated in OLP patients by in vitro proliferation assay. These data indicate that Tregs in OLP are frequently expanded but functionally deficient. This could explain, at least in part, why the increased Tregs in OLP fail to control the pathogenesis and development of this autoimmune disease. PMID:27106476

  11. Percentage of CD4+, CD8+, and CD25+ T lymphocytes in peripheral blood of pigs in the course of experimental burns and necrectomy

    Directory of Open Access Journals (Sweden)

    Aleksiewicz Roman

    2015-09-01

    Full Text Available The aim of the study was the evaluation of changes in the percentage profile of CD4+, CD8+, and CD25+ T lymphocytes, and their predictive value with respect to the course of experimental skin burns and early necrectomy in pigs. Thirty Large White Landrace pigs of both genders, weighing 50 kg (±2 kg, were used. Burns to their skin were performed with the use of a computer-controlled heating plate, applied to the animal’s body and heated to 2000°C, using 2.5 kg pressure for 10 s. It produced a burn of 30% (±2% of body surface with a range of damage between II b° and III°. In animals of each experimental group fascial necrectomy was performed, according to the testing module. Blood from experimental and non-treated control animals was collected from the external jugular vein before the beginning of the experiment (hour 0 and at 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, 144, 156, 168, and 180 h of the experiment. An immune response profile was evaluated using flow cytometry analysis of the level and expression dynamics of CD4+, CD8+, and CD25+ particles on the surface of T lymphocytes. The study demonstrated that experimentally-induced burns in pigs caused cell-mediated immune response reflected in the changes in the percentage of CD4+, CD8+, and CD25+ T lymphocytes, and that early necrectomy in burnt pigs acted in a protective manner for the organism, based on the immunological index values. The study also proved that the dynamics of cell-mediated immunological response intensification determined on the basis of the percentage of CD4+, CD8+, and CD25+ T lymphocytes is conditioned by the size of the burnt surface and the time of necrectomy procedure.

  12. Analysis of CD4+CD25+ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma

    Institute of Scientific and Technical Information of China (English)

    XUE Keying; ZHOU Yongming; XIONG Shengdao; XIONG Weining; TANG Tao

    2007-01-01

    The changes of CD4+CD25+ regulatory T cells (CD4+CD25+ Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible roles of CD4+CD25+ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4+CD25+ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4+CD25+ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups (P<0.05). Although the CD4+CD25+ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them (P>0.05). As compared with persistent group, exacerbation group had lower CD4+CD25+ Treg ratio and Foxp3 mRNA (P<0.05). It was indicated that the decrease of CD4+CD25+Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.

  13. Impaired NK cells' activity and increased numbers of CD4 + CD25+ regulatory T cells in multidrug-resistant Mycobacterium tuberculosis patients.

    Science.gov (United States)

    Fan, Renhua; Xiang, Yangen; Yang, Li; Liu, Yanke; Chen, Pingsheng; Wang, Lei; Feng, Wenjun; Yin, Ke; Fu, Manjiao; Xu, Yixin; Wu, Jialin

    2016-05-01

    Multidrug-resistant tuberculosis (MDR-TB) often causes persistent infection and chemotherapy failure, which brings heavy burden of society and family. Many immune cell subsets and regulatory mechanisms may operate throughout the various stages of infection. The presence of regulatory T cells (Tregs) is thought to be an important mechanism that TB successfully evades the immune system. Tregs play a central role in the prevention of autoimmunity and in the control of immune responses. The role of Tregs in MDR-TB infection and persistence is inadequately documented. The current study was designed to determine whether CD4 + CD25+ regulatory T cells may modulate innate immunity (such as NK cells) against human tuberculosis. Our results indicated that the numbers of CD4 + CD25+ Treg cells increased in MDR-TB patients' blood, and the cytokine production of IL-10 increased from MDR-patients compared with healthy subjects, along with the lower activity and low CD69 expression of NK cells in patients. These results suggested that immunity to MDR-TB patients induced circulating CD4 + CD25+ T regulatory cells expansion, which may be related to the persistence of Mycobacterium tuberculosis (M. tb) infection, and to the balance between effectors immune responses and suppression immune responses. PMID:27156613

  14. Chemokines involved in protection from colitis by CD4+CD25+ regulatory T cells

    DEFF Research Database (Denmark)

    Kristensen, Nanna Ny; Brudzewsky, Dan; Gad, Monika;

    2006-01-01

    /chemokine receptor-specific gene expression profiling system of 67 genes, the authors have determined the expression profile of chemokine and chemokine receptor genes in the rectum of colitic mice and in mice that have been protected fromcolitis by CD4CD25 regulatory T cells. In mice protected from colitis, the...... authors found down regulation of the mRNA expression of the inflammatory chemokine receptors CCR1 and CXCR3 and their ligands CXCL9, CXCL10, CCL5, and CCL7. Also the transcripts for CCR9, CCL25, CCL17, and CXCL1 are found down regulated in protected compared with colitic animals. In addition, the authors...

  15. Research progress of CD+4 CD+25 Foxp3 Treg cells in the hematological malignancies%调节性T细胞在血液系统恶性肿瘤中的研究进展

    Institute of Scientific and Technical Information of China (English)

    邵亮; 张铀

    2008-01-01

    胸腺产生的CD+4 CD25调节性T细胞(Treg细胞)被认为在控制自身免疫、防止移植物排斥反应、抑制抗感染自身免疫以及抑制异基因免疫反应中发挥重要作用.最近有报道此类细胞有抑制机体抗肿瘤的作用.Foxp3是Treg细胞最具特异性的蛋白分子标志物.阐述了血液系统恶性肿瘤患者CD+4 CD+25 Foxp3 Treg的表达以及其对免疫治疗的潜在作用.%Thymus derived CD+4; CD+25 regulatory T cells (Treg) are thought to be specific T cells that play an important role in controlling autoimmunity, preventing transplant rejection, restraining anti-infectious immune response, suppressing allogeneic immune respons. More recently, these cells are reported to have the ability of suppressing antitumor immune response. Foxp3 are the most specific protein of Treg. In this review, we will discuss the expression of CD+4 CD+25 Treg in patients with hematological malignancies and its implication for immunotherapy.

  16. CD4+CD25+ regulatory T cell depletion modulates anxiety and depression-like behaviors in mice.

    Directory of Open Access Journals (Sweden)

    Soo-Jeong Kim

    Full Text Available Stress has been shown to suppress immune function and increase susceptibility to inflammatory disease and psychiatric disease. CD4(+CD25(+ regulatory T (Treg cells are prominent in immune regulation. This study was conducted to determine if anti-CD25 antibody (Ab mediated depletion of Treg cells in mice susceptibility to stress-induced development of depression-like behaviors, as well as immunological and neurochemical activity. To accomplish this, an elevated plus-maze test (EPM, tail suspension test (TST, and forced swim test (FST were used to examine depression-like behaviors upon chronic immobilization stress. Immune imbalance status was observed based on analysis of serum cytokines using a mouse cytometric bead array in conjunction with flow cytometry and changes in the levels of serotonin (5-HT and dopamine (DA in the brain were measured by high performance liquid chromatography (HPLC. The time spent in the open arms of the EPM decreased significantly and the immobility time in the FST increased significantly in the anti-CD25 Ab-treated group when compared with the non stressed wild-type group. In addition, interlukin-6 (IL-6, tumor necrosis factor-á (TNF-á, interlukin-2 (IL-2, interferon-gamma (IFN-γ, interlukin-4 (IL-4 and interlukin-17A (IL-17A concentrations were significantly upregulated in the stressed anti-CD25 Ab-treated group when compared with the non stressed wild-type group. Furthermore, the non stressed anti-CD25 Ab-treated group displayed decreased 5-HT levels within the hippocampus when compared with the non stressed wild-type group. These results suggest that CD4(+CD25(+ Treg cell depletion modulated alterations in depressive behavior, cytokine and monoaminergic activity. Therefore, controlling CD4(+CD25(+ Treg cell function during stress may be a potent therapeutic strategy for the treatment of depression-like symptoms.

  17. Pretreatment With Inactivated Bacillus Calmette-Guerin Increases CD4+CD25+ Regulatory T Cell Function and Decreases Functional and Structural Effects of Asthma Induction in a Rat Asthma Model.

    Science.gov (United States)

    Gong, Ping; Li, Yun; Tan, Yu-Pin; Li, Hong

    2016-04-01

    Bacillus Calmette-Guerin (BCG) has been shown to have therapeutic effects on asthma through CD4+CD25+ regulatory T cells (Tregs). We sought to assess pretreatment with inactivated BCG on CD4+CD25+ Tregs and its functional and structural effects in rat asthma model. The rat asthma model was established using ovalbumin (OVA) sensitization and challenge. Ten rats were pretreated with BCG prior to OVA and received continued BCG injections during OVA challenge (BCG+OVA group), 10 rats were treated with OVA alone (OVA group), and 10 rats were treated with saline (control group). After 9 weeks, histamine dihydrochloride effect on airway resistance was measured. Number of CD4+CD25+ Tregs was measured by flow cytometry, expression of Foxp3 and CTLA-4 mRNA was measured, and serum TGF-β levels were determined. Differential cell count in bronchoalveolar lavage fluid (BALF) was determined, and lung tissue was processed and stained with hematoxylin and eosin, Masson's trichrome, and alcine blue and periodic acid Schiff's reaction to evaluate inflammatory cell infiltration, collagen deposition, and presence of goblet cells, respectively. BCG treatment led to an increase in CD4+CD25+ Tregs, as well as an increase in Foxp3 and CTLA-4 expression and serum TGF-β levels. In addition, we observed a decrease in histamine dihydrochloride-induced airway resistance, a decrease in inflammatory leukocytes in BALF, and a decrease in airway remodeling indicators in BCG+OVA-treated rats compared with OVA-treated rats. Intradermally injected inactivated BCG has the potential to improve airway inflammation, airway resistance, and airway remodeling through a mechanism that may involve CD4+CD25+ Tregs. PMID:26495900

  18. 白癜风患者外周血CD4+CD25+调节性T细胞的检测%Detection of peripheral CD4+CD25+ regulatory T lymphocytes in patients with vitiligo

    Institute of Scientific and Technical Information of China (English)

    白明辉; 王竞; 涂彩霞; 张蕴颖; 刘敏; 李国艳; 钟良瑞

    2009-01-01

    Objective To determine the level of peripheral CD4+CD25+ regulatory T lymphocytes in patients with vitiligo at different stages and to study its relationship with the development of vitiligo. Methods Blood samples were collected from 34 outpatients with vitiligo, including 19 cases of progressive vitiligo and 15 cases of stable vitiligo, as well as from 20 normal human controls. Flow cytometry was used to detect the levels of peripheral CD4+ and CD4+CD25+ T lymphocytes in these samples. Results Compared with the controls, the percentage of CD4+CD25+ regulatory T lymphoeytes in peripheral lymphocytes was significantly lower in patients with progressive vitiligo than those in patients with stable vitiligo and normal human con-trois [(2.43±0.30)% vs (3.49±0.39)% and (3.34±0.24)%, both P <0.05], but no significant difference was found between patients with stable vitiligo and normal human controls (P>0.05). A significantly nega-tive correlation was observed between the percentage of CD4+CD25+ regulatory T lymphocytes and lesion area in patients with progressive vitiligo (r = -0.48, P <0.05), but not in patients with stable vitiligo (P >0.05). There was no significant correlation between the course of disease and the percentage of peripheral CD4+CD25+ regulatory T lymphocytes in patients with progressive vitiligo or stable vitiligo (both P > 0.05). Conclusion There is an abnormal proportion of peripheral CD4+CD25+ regulatory T lymphocytes in patients with vitiligo, which may be related to the development of vitiligo.%目的 检测不同病期白癜风患者外周血CD4+CD25+调节性T细胞水平,探讨其与白癜风发病的关系.方法 白癜风患者34例,进展期19例,稳定期15例.通过流式细胞仪对不同病期白癜风患者外周血CD4+、CD4+CD25+T细胞水平进行检测,并与20例正常人比较.结果 进展期患者外周血中CD4+CD25+调节性T细胞占外周血淋巴细胞的表达率低于正常对照组(P<0.05);稳定期患者与正

  19. CD4 + CD25 + T cells protect against experimentally induced asthma and alter pulmonary dendritic cell phenotype and function

    OpenAIRE

    Lewkowich, Ian P.; Herman, Nancy S.; Schleifer, Kathleen W.; Dance, Matthew P.; Chen, Brian L.; Dienger, Krista M.; Sproles, Alyssa A.; Shah, Jaimin S.; Köhl, Jörg; Belkaid, Yasmine; Wills-Karp, Marsha

    2005-01-01

    The role of natural CD4 + CD25 + regulatory T (T reg) cells in the control of allergic asthma remains poorly understood. We explore the impact of T reg cell depletion on the allergic response in mice susceptible (A/J) or comparatively resistant (C3H) to the development of allergen-induced airway hyperresponsiveness (AHR). In C3H mice, anti-CD25–mediated T reg cell depletion before house dust mite treatment increased several features of the allergic diathesis (AHR, eosinophilia, and IgE), whic...

  20. 抗ICOS抗体体外对哮喘大鼠血液和淋巴液CD4+CD25+Foxp3+调节性T细胞数量及其功能影响%Influence of anti-ICOS antibody on quantity and function of CD4 + CD25 + Foxp3 + Treg from lymph and blood of rats with bronchial asthma

    Institute of Scientific and Technical Information of China (English)

    赵磊; 冯学斌; 王跃嗣; 崔晴晴; 曹茵茵

    2012-01-01

    AIM: To investigate the influences of anti-ICOS antibody (anti-ICOSAb) on quantity and function of CD4+ CD25+ Foxp3+ Treg cells from lymph and peripheral blood of rats with bronchial asthma. METHODS: The mononuclear cells ( MNC) from lymph and blood were co-cultured with anti-ICOSAb, and then the percentage of CD4+ CD25+ Foxp3+ Treg cells were analyzed by flow cytometer (FCM) and the levels of IL-10 and TGF-pi in supernatants were determined by ELISA. RESULTS; The MNC were collected from lymph and blood at 0, 24 and 48 h after the last challenge, respectively, and the cells were cultured for 96 h in vitro. The percentage of CD4+ CD25 + Foxp3+ Treg cells in the MNC from lymph was significantly higher than that from blood in each group (P < 0.05); The percentage of CD4 + CD25 Foxp3+ Treg cells in the MNC from lymph and blood in asthma group was significantly lower compare with the normal control group (P < 0.05); The percentage of CD4+ CD25 + Foxp3+ Treg cells in the MNC from lymph and blood in the anti-ICOSAb group obviously decreased compare with the asthma group ( P < 0. 05). At 0 h after the last challenge, the level of IL-10 in the supernatant of MNC from lymph and blood in the anti-ICOSAb group were significantly lower than that of the control and asthma groups (P < 0. 05), while there were no significant differences of TGF-pi expression in the supernatant of MNC from lymph and blood in each group at different time points. CON-CLUSION: Blocking the ICOS/ICOSL signaling pathway by anti-ICOSAb could exacerbate the deficiency of CRT CD25+ Foxp3+Treg cells from lymph and Wood in bronchial asthmatic rat, meanwhile inhibit the CD4 + CD25 + Foxp3+Treg cells secreting IL-10 at 0 h after the last challenge, but have no significant effect on the secretion of TGF-β1.%目的:研究抗ICOS抗体对哮喘大鼠外周血和淋巴液来源CD4+ CD25+ Foxp3+调节性T细胞(Treg)数量及其功能的影响.方法:抗ICOS抗体处理血液和淋巴液中单个核细胞(MNC

  1. A study of the effects of Schistosoma japonicum soluble egg antigen on CD4+CD25+regulatory T cells in patients with asthma%CD4+CD25+调节性T细胞在血吸虫可溶性虫卵抗原影响哮喘中的作用研究

    Institute of Scientific and Technical Information of China (English)

    朱云娟; 刘佩梅; 杨秀珍; 刘霞; 吴增强; 纪伟华; 安桂珍; 沈悦云; 刘金霞; 李健

    2011-01-01

    Objective To study the relationship between Schistosoma japonicum soluble egg antigen (SEA) and asthma and the effects of SEA on CD4+ CD25+ regulatory T cells (CD4+ CD25+ Treg) and expression of the Foxp3 gene. Methods BALB/C mice were each injected with 50 μg SEA peritoneally and through the foot pad once a week for 4 weeks. In the control group, all injections were with normal saline. Then asthma was induced with ovalbumin (OVA) in all mice. After mice were sacrificed, the lungs were subjected to pathologic examination; the bronchoalveolar lavage fluid (BALF) was collected and different cells were classified and counted after smearing and staining. Spleen cells were separated and the percentage of CD4+ CD25+ Treg out of total CD4+ T cells was determined using flow cytometry. Total spleen RNA was prepared and synthesized into cDNA through reverse transcription; cDNA was subjected to PCR amplification to determine the level of Foxp3 mRNA expression. Results Mild pulmonary inflammation was observed in the SEA immunization group, whereas severe inflammation was observed in the control group. Staining of the BALF revealed that the SEA immunization group had a much lower BALF cell density than did the control group. In the SEA immunization group, the percentage of eosinophils out of total cells was(2. 22± 1. 52)% while it was (19. 93±4. 08)% in the control group. The difference between the 2 groups was statistically significant (P<0. 05). Flow cytometry revealed that the percentage of CD4+ CD25+ Treg out of total CD4+ T cells was (32. 24±2. 19) % in the SEA immunization group while it was (27. 41±2. 87) % in the control group. The difference between the 2 groups was statistically significant (P<0. 05). The level of Foxp3 mRNA expression was higher than that in the control group as well. Conclusion SEA inhibits the development of asthma to some extent and it seems influence immune regulation through its effect on CD4+CD25+Treg.%目的 研究血吸虫可溶性虫

  2. Differential influence of the tumour-specific non-human sialic acid containing GM3 ganglioside on CD4+CD25- effector and naturally occurring CD4+CD25+ regulatory T cells function.

    Science.gov (United States)

    de León, Joel; Fernández, Audry; Clavell, Marilyn; Labrada, Mayrel; Bebelagua, Yanin; Mesa, Circe; Fernández, Luis E

    2008-04-01

    Increasing evidences suggest that the aberrant expression of certain gangliosides on malignant cells could affect host's anti-tumour-specific immune responses. We have recently documented the relevance of the N-glycolylated variant of GM3 ganglioside (NGcGM3), a tumour-specific non-human sialic acid containing ganglioside, for tumour progression. However, evidences about the implication of host's immunity in NGcGM3-promoted cancer progression had not been obtained previously. In this work, we compared tumour growth of X63 myeloma cells pre-treated or not with an inhibitor of the glucosylceramide synthase enzyme, in wild or CD4+ T cell-depleted BALB/c mice. Results clearly showed a relationship between the agonistic effect of NGcGM3 in tumour growth and the presence of CD4+ T lymphocytes. For the first time, a description of a ganglioside-differential effect over purified CD4+CD25- and naturally occurring regulatory CD4+CD25+ T cells is provided. While NGcGM3 similarly down-modulated the CD4 expression in both cell populations, the inhibitory capacity of the CD4+CD25+ lymphocytes and their proliferation, induced by an anti-CD3 mAb and IL2, were not modified. In a different fashion, a reduction in proliferative capacity and a noteworthy secretion of anti-inflammatory cytokines were detected when CD4+CD25- T cells were cultured in the presence of NGcGM3. Considering the relevance of dendritic cells (DC) on primary activation of T cells, the effect of NGcGM3 over DC differentiation and TLR4-mediated maturation was also assessed. Our results indicate that NGcGM3 contributes to cancer progression mainly by influencing DC and CD4+CD25- T lymphocyte functions, rather than increasing the inhibitory capacity of naturally occurring regulatory T cells. PMID:18310617

  3. Detection and Significance of CD4+CD25+CD127dim Regulatory T Cells in Individuals with Severe Aplastic Anemia

    Directory of Open Access Journals (Sweden)

    Weiwei Qi

    2015-09-01

    Full Text Available Objective: To investigate the relationship between CD4+CD25+CD127dim regulatory T cells (Tregs and immune imbalance in acquired severe aplastic anemia (SAA. Materials and Methods: The quantity of CD4+CD25+CD127dim Tregs in 44 SAA patients and 23 normal controls was measured by flow cytometry. Correlations between Tregs and T cell subsets, dendritic cell (DC subsets, granulocyte counts, and percentage of reticulocytes (RET% were analyzed. Results: The percentage of CD4+CD25+CD127dim Tregs in peripheral blood lymphocytes (PBLs of untreated patients was lower than in recovery patients and normal controls (0.83±0.44% vs. 2.91±1.24% and 2.18±0.55%, respectively, p<0.05. The percentage of CD4+CD25+CD127dim Tregs in CD4+ T lymphocytes of recovery patients was higher than that of untreated patients and normal controls (9.39±3.51% vs. 7.61±5.3% and 6.83±1.4%, respectively, p<0.05. The percentage of CD4+ T lymphocytes in PBLs of untreated patients was lower than in recovery patients and normal controls (13.55±7.37% vs. 31.82±8.43% and 32.12±5.88%, respectively, p<0.05. T cell subset (CD4+/CD8+ ratio was 0.41±0.24 in untreated patients, which was lower than in recovery patients (1.2±0.4 and normal controls (1.11±0.23 (p<0.05. DC subset (myeloid DC/plasmacytoid DC ratio, DC1/DC2 ratio was 3.08±0.72 in untreated patients, which was higher than in recovery patients (1.61±0.49 and normal controls (1.39±0.36 (p<0.05. The percentage of CD4+CD25+CD127dim Tregs in PBLs was positively associated with T cell subset (r=0.955, p<0.01 and negatively associated with DC subset (r=-0.765, p<0.01. There were significant positive correlations between CD4+CD25+CD127dim Tregs/PBL and granulocyte counts and RET% (r=0.739 and r=0.749, respectively, p<0.01. Conclusion: The decrease of CD4+CD25+CD127dim Tregs in SAA patients may cause excessive functioning of T lymphocytes and thus lead to hematopoiesis failure in SAA.

  4. Where CD4 + CD25 + T reg cells impinge on autoimmune diabetes

    OpenAIRE

    Chen, Zhibin; Herman, Ann E.; Matos, Michael; Mathis, Diane; Benoist, Christophe

    2005-01-01

    Foxp3 is required for the generation and activity of CD4 + CD25 + regulatory T (T reg) cells, which are important controllers of autoimmunity, including type-1 diabetes. To determine where T reg cells affect the diabetogenic cascade, we crossed the Foxp3 scurfy mutation, which eliminates T reg cells, with the BDC2.5 T cell receptor (TCR) transgenic mouse line. In this model, the absence of T reg cells did not augment the initial activation or phenotypic characteristics of effector T cells in ...

  5. 56 Increased Frequency of CD4+ CD25+ FOXP3- in Allergic Conjunctivitis Patients

    OpenAIRE

    Galicia-Carreón, Jorge; Alonso-Sánchez, Miguel E.; Robles-Contreras, Atzin; Hong, Enrique; Chávez, Raul; Jiménez-Martínez, Maria C.

    2012-01-01

    Background Allergic conjunctivitis (AC) is one of the most common eye disorders in clinical practice. It has been shown that AC is a disorder mediated by Th2 lymphocytes producing IL-4 and IL-5, where the eye damage is caused by a type I hypersensitivity. It has been suggested in asthma and rhinitis that T regulatory cells (Tregs) CD4+ CD25+ FOXP3+ have been involved in control allergic status, favoring an optimal microenvironment with immunosuppressive cytokines (IL-10, TGF-β). However is un...

  6. Interplay of T Helper 17 Cells with CD4+CD25high FOXP3+ Tregs in Regulation of Allergic Asthma in Pediatric Patients

    Directory of Open Access Journals (Sweden)

    Amit Agarwal

    2014-01-01

    Full Text Available Background. There is evidence that Tregs are important to prevent allergic diseases like asthma but limited literature exists on role of TH17 cells in allergic diseases. Methods. Fifty children with asthma and respiratory allergy (study group and twenty healthy children (control group were recruited in this study. Total IgE levels and pulmonary function tests were assessed. The expression of Tregs and cytokines was determined by flow cytometry. Results. The average level of total IgE in study group (316.8 ± 189.8 IU/mL was significantly higher than controls (50 ± 17.5 IU/mL, P<0.0001. The frequency of TH17 cells and culture supernatant level of IL-17 in study group (12.09 ± 8.67 pg/mL was significantly higher than control group (2.01 ± 1.27 pg/mL, P<0.001. Alternatively, the frequency of FOXP3 level was significantly lower in study group [(49.00 ± 13.47%] than in control group [(95.91 ± 2.63%] and CD4+CD25+FOXP3+ to CD4+CD25+ ratio was also significantly decreased in study group [(6.33 ± 2.18%] compared to control group [(38.61 ± 11.04%]. The total serum IgE level is negatively correlated with FOXP3 level (r=-0.5273, P<0.0001. The FOXP3 expression is negatively correlated with the IL-17 levels (r=-0.5631, P<0.0001 and IL-4 levels (r=-0.2836, P=0.0460. Conclusions. Imbalance in TH17/Tregs, elevated IL-17, and IL-4 response and downregulation of FOXP3 were associated with allergic asthma.

  7. Exploration of the Partial Different Roles of CD4 + CD25 + Tregs and CD4 + CD25HighTregs in Sero-resistance Syphilitic Patients%梅毒血清固定患者CD4+CD25+Treg细胞和CD4+CD25High Treg细胞功能的差异探讨

    Institute of Scientific and Technical Information of China (English)

    张明海; 赵建斌

    2013-01-01

    目的 探讨梅毒血清固定患者外周血CD4+ CD25+ Treg细胞和CD4+ CD25HighTreg细胞功能是否发生变化.方法 采用流式细胞术(FCM)检测28例梅毒血清固定患者和28例梅毒转阴患者外周血CD4+CD25+ Treg细胞和CD4+ CD25High Treg细胞中FOXP3和CTLA-4的水平;采用免疫磁珠细胞分选技术(MACS)和Realtime-PCR技术检测CD4+ CD25+ Treg细胞的FOXP3和CTLA-4mRNA水平.结果 梅毒血清固定组外周血CD4+ CD25+Treg细胞功能增强,而梅毒血清转阴组外周血CD4+ CD25HighTreg细胞功能稳定.结论 CD4+CD25+Treg和CD4+ CD25HighTreg细胞在梅毒血清固定形成中作用的方式或途径存在部分差异.

  8. 90Y-daclizumab, an anti-CD25 monoclonal antibody, provided responses in 50% of patients with relapsed Hodgkin’s lymphoma

    Science.gov (United States)

    Janik, John E.; Morris, John C.; O’Mahony, Deirdre; Pittaluga, Stefania; Jaffe, Elaine S.; Redon, Christophe E.; Bonner, William M.; Brechbiel, Martin W.; Paik, Chang H.; Whatley, Millie; Chen, Clara; Lee, Jae-Ho; Fleisher, Thomas A.; Brown, Maggie; White, Jeffrey D.; Stewart, Donn M.; Fioravanti, Suzanne; Lee, Cathryn C.; Goldman, Carolyn K.; Bryant, Bonita R.; Junghans, Richard P.; Carrasquillo, Jorge A.; Worthy, Tat’Yana; Corcoran, Erin; Conlon, Kevin C.; Waldmann, Thomas A.

    2015-01-01

    Despite significant advances in the treatment of Hodgkin’s lymphoma (HL), a significant proportion of patients will not respond or will subsequently relapse. We identified CD25, the IL-2 receptor alpha subunit, as a favorable target for systemic radioimmunotherapy of HL. The scientific basis for the clinical trial was that, although most normal cells with exception of Treg cells do not express CD25, it is expressed by a minority of Reed–Sternberg cells and by most polyclonal T cells rosetting around Reed–Sternberg cells. Forty-six patients with refractory and relapsed HL were evaluated with up to seven i.v. infusions of the radiolabeled anti-CD25 antibody 90Y-daclizumab. 90Y provides strong β emissions that kill tumor cells at a distance by a crossfire effect. In 46 evaluable HL patients treated with 90Y-daclizumab there were 14 complete responses and nine partial responses; 14 patients had stable disease, and nine progressed. Responses were observed both in patients whose Reed–Sternberg cells expressed CD25 and in those whose neoplastic cells were CD25− provided that associated rosetting T cells expressed CD25. As assessed using phosphorylated H2AX (γ-H2AX) as a bioindicator of the effects of radiation exposure, predominantly nonmalignant cells in the tumor microenvironment manifested DNA damage, as reflected by increased expression of γ-H2AX. Toxicities were transient bone-marrow suppression and myelodysplastic syndrome in six patients who had not been evaluated with bone-marrow karyotype analyses before therapy. In conclusion, repeated 90Y-daclizumab infusions directed predominantly toward nonmalignant T cells rosetting around Reed–Sternberg cells provided meaningful therapy for select HL patients. PMID:26438866

  9. 90Y-daclizumab, an anti-CD25 monoclonal antibody, provided responses in 50% of patients with relapsed Hodgkin's lymphoma.

    Science.gov (United States)

    Janik, John E; Morris, John C; O'Mahony, Deirdre; Pittaluga, Stefania; Jaffe, Elaine S; Redon, Christophe E; Bonner, William M; Brechbiel, Martin W; Paik, Chang H; Whatley, Millie; Chen, Clara; Lee, Jae-Ho; Fleisher, Thomas A; Brown, Maggie; White, Jeffrey D; Stewart, Donn M; Fioravanti, Suzanne; Lee, Cathryn C; Goldman, Carolyn K; Bryant, Bonita R; Junghans, Richard P; Carrasquillo, Jorge A; Worthy, Tat'Yana; Corcoran, Erin; Conlon, Kevin C; Waldmann, Thomas A

    2015-10-20

    Despite significant advances in the treatment of Hodgkin's lymphoma (HL), a significant proportion of patients will not respond or will subsequently relapse. We identified CD25, the IL-2 receptor alpha subunit, as a favorable target for systemic radioimmunotherapy of HL. The scientific basis for the clinical trial was that, although most normal cells with exception of Treg cells do not express CD25, it is expressed by a minority of Reed-Sternberg cells and by most polyclonal T cells rosetting around Reed-Sternberg cells. Forty-six patients with refractory and relapsed HL were evaluated with up to seven i.v. infusions of the radiolabeled anti-CD25 antibody (90)Y-daclizumab. (90)Y provides strong β emissions that kill tumor cells at a distance by a crossfire effect. In 46 evaluable HL patients treated with (90)Y-daclizumab there were 14 complete responses and nine partial responses; 14 patients had stable disease, and nine progressed. Responses were observed both in patients whose Reed-Sternberg cells expressed CD25 and in those whose neoplastic cells were CD25(-) provided that associated rosetting T cells expressed CD25. As assessed using phosphorylated H2AX (γ-H2AX) as a bioindicator of the effects of radiation exposure, predominantly nonmalignant cells in the tumor microenvironment manifested DNA damage, as reflected by increased expression of γ-H2AX. Toxicities were transient bone-marrow suppression and myelodysplastic syndrome in six patients who had not been evaluated with bone-marrow karyotype analyses before therapy. In conclusion, repeated (90)Y-daclizumab infusions directed predominantly toward nonmalignant T cells rosetting around Reed-Sternberg cells provided meaningful therapy for select HL patients. PMID:26438866

  10. Changes of Th17 cells and CD4+CD25+ regulatory T cells in peripheral blood of asthmatic children and their relationship with the situation of asthma.%哮喘患儿外周血Th17细胞CD4+CD25+调节性细胞变化及其与病情相关分析

    Institute of Scientific and Technical Information of China (English)

    马秋莉; 彭韶; 梁鹏; 李会娟; 张曼

    2011-01-01

    Objective To observe the levels of Thl7 cells, CD4+CD25+ regulatory T cells in the asthmatic children and relationship between the two types of cells and children' s condition. Methods Flow cytometry was used to detect the percentages of the Thl7 cells and CD4+CD25+ regulatory T cells in the peripheral blood of acute asthma children (asthma group, n - 60) .alleviated period asthma children (n = 30) and healthy children (healthy control group, n = 30).Acute asthma children were divided into 3 groups: mild, moderate and severe asthmatic patients. Results Compared with the healthy control group (1.02% ± 0.28%) and alleviated period asthma children (1.65% ± 0.38%), the numbers of CD4+ cells (Thl7) expressing IL-17 in peripheral blood of acute asthma(2.24% ± 1.02%) were increased, and the differences were statistically significant (P<0.05). The levels of CD4+CD25+T cells in peripheral blood of acute asthma and alleviated period asthma children (5.37% ± 0.80% ; 6.05% ± 0.87%) were significantly lower than those of healthy children (7.11% ± 0.89%) (P < 0.05). Thl7 cells were positively correlated with the course of childhood asthma(r = 0.649, P < 0.05).CD4+CD25+ regulatory T cells were negatively correlated with the course of childhood asthma (r =-0.599, P < 0.05). Conclusion The immunization response of Thl7 cells in peripheral blood of asthmatic children is strengthened, but the immune function of CD4+CD25+regulatory T cells is decresed. The severity of asthma is closely related to the inbal-ance of Th71ATreg cellular immunity.%目的 探讨支气管哮喘患儿外周血中辅助T细胞(Th) 17细胞和CD4+ CD25+调节性T细胞(Treg)的变化与儿童哮喘病情的相关性.方法 收集2009年月5月至2010年4月于郑州大学第一附属医院就诊的患儿,均为首次确诊哮喘或规范吸入激素停用>3个月后复发及近1个月内无明显感染者.采用流式细胞仪测定患儿外周血中Th17细胞及CD4+CD25+Treg比例的变化.结果 Th

  11. Inhibition of CD4+CD25+ regulatory T-cell function by calcineurin-dependent interleukin-2 production

    OpenAIRE

    Zeiser, Robert; Nguyen, Vu H; Beilhack, Andreas; Buess, Martin; Schulz, Stephan; Baker, Jeanette; Contag, Christopher H.; Negrin, Robert S.

    2006-01-01

    CD4+CD25+ regulatory T (Treg) cells control immunologic tolerance and antitumor immune responses. Therefore, in vivo modification of Treg function by immunosuppressant drugs has broad implications for transplantation biology, autoimmunity, and vaccination strategies. In vivo bioluminescence imaging demonstrated reduced early proliferation of donor-derived luciferase-labeled conventional T cells in animals treated with Treg cells after major histocompatibility complex mismatch bone marrow tran...

  12. 丹参注射液联合地塞米松对哮喘大鼠CD4+CD25+调节性T细胞的影响%Influence of danshen injection combined with dexamethasone on CD4+CD25+ regulatory T cells of asthmatic rats

    Institute of Scientific and Technical Information of China (English)

    薛克营; 程立; 王成国; 李威; 石明

    2008-01-01

    目的 探讨丹参注射液联合地塞米松(DXM)抑制哮喘气道炎症的免疫学机制.方法 50只Wistar大鼠随机分成正常对照(NC)组、哮喘组、丹参组、DXM组、联合用药组,计数支气管肺泡灌洗液(BALF)中细胞总数并分类,HE染色行肺组织病理学检查,流式细胞仪检测外周血单个核细胞(PBMCs)中CD4+CD25+调节性T细胞(CD4+CD25+Treg)比例,ELISA检测BALF中IL-4、IL-5含量.结果 与哮喘组比较,药物干预组细胞总数、中性粒细胞(Neu)、淋巴细胞(Lym)、嗜酸性粒细胞(Eos)百分率下降(P<0.05,P<0.01),联合用药组下降程度大于丹参组、DXM组(P<0.05).哮喘组呈显著炎症变化,丹参组呈中度炎症变化,DXM组呈轻度炎症变化,联合用药组无炎症改变.与哮喘组比较,药物干预组CD4+CD25+Treg/CD4+T升高(P<0.05),IL4、IL-5含量下降(P<0.05),联合用药组CD4+CD25+ Tree/CD4+T升高程度和IL-4、IL-5下降程度大于丹参组和DXM组(P<0.05).结论 丹参注射液具有抑制哮喘大鼠气道炎症的作用,和DXM联合应用后,抑制作用更加明显,该作用可能和促进CD4+CD25+Treg产生,进而抑制IL-4、IL-5产生,纠正Th1/Th2失衡,最终减轻气道炎症有关.%Objective To investigate the immunological mechanism of inhibitory effect of Danshen injection combined with dexamethasone(DXM) on asthmatic airway inflammation.Methods 50 Wistar rats were randomly divided into normal control(NC),asthma,Danshen,DXM and Danshen+DXM group.Cytology study of Bronchoalveolar lavage fluid(BALF) was conducted.Pathology of lung tissue was done through HE.Flow eytometry was used to detect CD4+CD25+ regulatory T Cells(CD4+CD25+ Treg) ratio in peripheral blood mononuclear cells(PBMCs).IL-4 and IL-5 levels in BALF were detected by ELISA.Results Total cells number,percentage of lymphocytes,neutrophils and eosinophils(Eos) in BALF of the three treated groups were lower than that in asthma group(P<0.05,P<0.01),particularly in Danshen+DXM group

  13. Aberrant Expression of Novel Cytokine IL-38 and Regulatory T Lymphocytes in Childhood Asthma

    OpenAIRE

    Man Chu; Ida M.T. Chu; Edmund C.M. Yung; Christopher W. K. Lam; Ting F. Leung; Wong, Gary W.K.; Wong, Chun K

    2016-01-01

    We investigated the expression of novel anti-inflammatory interleukin (IL)-38 and regulatory T (Treg) lymphocytes in childhood asthma patients. The protein and mRNA expression level of IL-38, periostin, peripheral CD4+CD25+CD134+ T lymphocytes as well as CD4+CD25highFoxP3+ and CD4+CD25highCD127− Treg lymphocytes from 40 asthmatic patients and 20 normal control (NC) subjects were studied using ELISA, qPCR and flow cytometry. Serum and supernatant cytokines/chemokines were determined by multipl...

  14. Circulating CD4~+CD25~+ and CD8~+CD28~- T regulate cells in multiple myeloma%多发性骨髓瘤患者外周血CD4~+CD25~+和CD8~+CD28~-调节性T细胞研究

    Institute of Scientific and Technical Information of China (English)

    贾丽; 谢晓宝; 邱国强; 钱新瑜; 周民; 肖溶

    2009-01-01

    Objective: The study was designed to evaluate the changes and significance of circulating CD4~+CD25~+ and CD8~+CD28~- regulatory T cells (Tregs) in patients with multiple myeloma (MM).Methods:CD4~+CD25~+ and CD8~+CD28~-Tregs in peripheral blood of 38 patients with MM and of 20 healthy doners were measured by flow cytometry.Serum albumin and β_2-MG in patients with MM were measured using bromocresol green method,transmission turbidimetry respectively.Results:Compared to those of the controls,the proportions of CD4~+CD25~(+/high),CD4~+CD25~(high) CD127~(low) and CD8~+CD28~-Treg cells in newly diagnosed MM patients were elevated.Furthermore,the proportions of CD4~+CD25~(high) and CD4~+CD25~(high)CD127~(low) Tregs in each clinical stage were elevated when compared to those of the controls.The number of the Tregs were increasing with clinical stages and were significantly higher in stage Ⅲ MM than in stageⅠ MM;In stageⅡand Ⅲ MM,there were also elevated proportions of CD8~+CD28~- Tregs,increasing with clinical stages.However,there were no differences when compared between stage Ⅰ MM and the controls;Both the proportions of CD4~+CD25~(+/high) and CD4~+CD25~(high)CD127~(low) Tregs in active MM were not different from stable MM,although all of them were higher than those of controls.The proportion of CD8~+CD28~- Tregs was higher in active MM than in stable MM and controls,but there were no differences when compared between active and stable MM.The proportions of both CD4~+CD25~(high) Tregs and CD4~+CD25~(high)CD127~(low)Tregs had negative correlation with the levels of serum albumin.Conclusion:MM patients have elevated levels of circulating CD4~+CD25~+ and CD8~+CD28~-Tregs,which may be an important mechanism of MM immune evasion,and may be associated with clinical stages,disease progression and prognosis of MM to some extent.%目的:探讨CD4~+CD25~+和CD8~+CD28~-调节性T细胞(Tregs)在多发性骨髓瘤(MM)患者外周血中的变化及意义.方

  15. 白三烯受体拮抗剂对哮喘气道重塑及Th17细胞/CD4+CD25+调节性T细胞表达的影响%Effects of leukotriene receptor antagonists on airway remodeling and Th17 cells/CD4+CD25+regulatory T cells expresson in asthma

    Institute of Scientific and Technical Information of China (English)

    李丽(综述); 李敏(审校)

    2014-01-01

    支气管哮喘的气道重塑是气道炎症反复作用的结果,白三烯是气道重塑中的重要炎症介质之一。影响气道重塑的因素较多,近年来Th17细胞和CD4+CD25+调节性T细胞(CD4+CD25+treg细胞)在气道重塑中的作用日益受到重视。白三烯受体拮抗剂是治疗哮喘的有效药物,能在一定程度上抑制气道重塑,但其作用机制及对Th17细胞/CD4+CD25+treg细胞表达的影响机制尚不十分清楚。因此,阐明Th17细胞/CD4+CD25+treg细胞平衡在气道重塑中的表达变化、白三烯受体拮抗剂干预气道重塑的具体作用途径和生物效应及对Th17细胞/CD4+CD25+treg细胞表达的影响,将为以后哮喘患儿的预防和治疗提供新的靶点。%The airway remodeling of bronchial asthma is the result of repeated airway inlfammation. Its occurrence is a complex process involving many cytokines, inlfammatory mediators and associated cellular components, of which leukotrienes are important mediators of inlfammation in the airway remodeling. Many factors inlfuence Airway remodeling. In recent years, effects of Th17 cells and CD4+CD25+regulatory T cells (CD4+CD25+treg cells) on airway remodeling is growing in importance. Leukotriene receptor antagonist is an effective drug in the treatment of asthma and can suppress airway remodeling. But the exact mechanisms and its impact on the proportion of Th17 cells/CD4+CD25+treg cells is not yet clear. Therefore, the clariifcation of the changes of Th17 cells/CD4+CD25+treg cells expression in airway remodeling and the speciifc pathways, biological effects, inlfuence of the proportion of Th17 cells/CD4+CD25+treg cells expression after leukotriene receptor antagonist intervene can provide a new target for prevention and the treatment of asthma in the future.

  16. Effect of Lactobacillus salivarius on Th1/Th2 cytokines and the number of spleen CD4+ CD25+ Foxp3+ Treg in asthma Balb/c mouse

    OpenAIRE

    Yun, Xiang; SHANG, YUNXIAO; Li, Miao

    2015-01-01

    Background: Bronchial asthma is a chronic airway inflammatory disease that involves T lymphocytes. Methods: In order to explore the effect of Lactobacillus salivarius on Th1/Th2 cytokines and the number of spleen CD4+ CD25+ Foxp3+ Treg in asthma Balb/c mouse, we constructed acute asthma model with ovalbumin to observe the mouse behavior change in Balb/c mice. The expression of GATA-3 mRNA and T-bet mRNA was measured by real-time PCR. The proportion of CD4+ CD25+ Foxp3+ Treg/CD4+ was determine...

  17. Anti-CD25 monoclonal antibody Fc variants differentially impact regulatory T cells and immune homeostasis.

    Science.gov (United States)

    Huss, David J; Pellerin, Alex F; Collette, Brian P; Kannan, Arun K; Peng, Liaomin; Datta, Abhishek; Wipke, Brian T; Fontenot, Jason D

    2016-07-01

    Interleukin-2 (IL-2) is a critical regulator of immune homeostasis through its non-redundant role in regulatory T (Treg) cell biology. There is major interest in therapeutic modulation of the IL-2 pathway to promote immune activation in the context of tumour immunotherapy or to enhance immune suppression in the context of transplantation, autoimmunity and inflammatory diseases. Antibody-mediated targeting of the high-affinity IL-2 receptor α chain (IL-2Rα or CD25) offers a direct mechanism to target IL-2 biology and is being actively explored in the clinic. In mouse models, the rat anti-mouse CD25 clone PC61 has been used extensively to investigate the biology of IL-2 and Treg cells; however, there has been controversy and conflicting data on the exact in vivo mechanistic function of PC61. Engineering antibodies to alter Fc/Fc receptor interactions can significantly alter their in vivo function. In this study, we re-engineered the heavy chain constant region of an anti-CD25 monoclonal antibody to generate variants with highly divergent Fc effector function. Using these anti-CD25 Fc variants in multiple mouse models, we investigated the in vivo impact of CD25 blockade versus depletion of CD25(+) Treg cells on immune homeostasis. We report that immune homeostasis can be maintained during CD25 blockade but aberrant T-cell activation prevails when CD25(+) Treg cells are actively depleted. These results clarify the impact of PC61 on Treg cell biology and reveal an important distinction between CD25 blockade and depletion of CD25(+) Treg cells. These findings should inform therapeutic manipulation of the IL-2 pathway by targeting the high-affinity IL-2R. PMID:27012310

  18. Evaluation of CD4+CD25+ regulatory T cells in patients with hepatocellular carcinoma and liver cirrhosis

    Directory of Open Access Journals (Sweden)

    Amal Abdel Aleem, ** Eman A Abdel Rahman and ***Abdel Aty M. Elgonimy

    2011-04-01

    Full Text Available The emergence of a Tumor results from the disruption of cell growth regulation as well as from failure of the host to provoke a sufficient immunological anti-tumor response. Regulatory T cells CD4+CD25+ (Tregs play an important role in maintaining peripheral self-tolerance, thus preventing autoimmune pathologies. However, in certain situations Tregs can impair effective immunity to some pathogens and tumor cells. Hepatocellular carcinoma (HCC is one of the leading causes of cancer-related death in the world, and in developed countries it is expected to continue to increase because of the epidemic of chronic hepatitis C virus (HCV infection. Previous studies also showed that Tregs infiltrating HCC tumors were an indicator of poor prognosis. Aim: of this study was to evaluate CD4+CD25+ Treg cells in patients with HCC and liver cirrhosis and their correlation with liver tumor markers and grading. Patients and Methods: The study included 30 patients, 15 patients had HCC (group I and 15 were cirrhotic patients (group II. In addition, 10 healthy subjects were used as controls. All patients were subjected to clinical examination and laboratory investigations including liver function tests, hepatitis B markers (HBs Ag, HBeAg and HBc-Ab and HCV antibodies were detected by ELISA. and Bilharzial Abs by indirect hemagglutination test. CD4+CD25+ Tcells were quantified in the blood by flow cytometry, alpha feto protein by Cobas e 411, To evaluate HCC grading ,abdominal sonography, C.T.and liver biopsy were done. Patients were categorized into mildely differentiated (grad 1, moderately differentiated (grad 11 and poorly differentiated (grad 111. Results: There were significant increased in serum AFP, and CD4+CD25+% in patients with HCC, and in patients with liver cirrhosis when compared to control group (p<0.05, and highly significant increased in AFP, and CD4+CD25+ % in patients with HCC when compared to patients with liver cirrhosis (p<0.001. In HCC patients

  19. TRAIL对肿瘤侵润CD4+CD25+Treg的调节作用%Regulation of TRAIL on tumor infiltrating CD4+CD25+Treg

    Institute of Scientific and Technical Information of China (English)

    袁海芹; 刁智娟; 周剑锁; 刘彦信; 史娟; 郑德先

    2011-01-01

    0bjective:To investigate the regulation of TRAIL on tumor infiltrating CD 4+ CD25 +Treg cells.Methods:The inpact of TRAL on CCL22 secretion of tumor cells was detected by ELISA .Recam binant soluble TRAL was adminsttated into subcutaneous solid 4T1 tumor and tumor volum e was measured .Tumor infiltrating lym phocytes w ere isolated and assayed by flow cytam etry to evaluate the change of CD4+ CD25+ Treg cells in umor.Results:rsTRAIL increased CCL22 secretion into supematant of tumor cell 4Tl and B16 cells.TRAIL treat ment did notinhibit the s .C.4T1 tumor growth ,but turmor infiltrating CD 4+ CD25+ Treg increased obviously .Conclusion :Augmention in CCL22 secretion of 4T1 cancer cells might recruit Tregs,therefore, leading to turmor infiltrating CD 4+CD 25+Treg increase .This study provides novel data tor the physiological function research of TRAIL and cancer therapy application .%目的:探讨TRAIL对肿瘤侵润CD4+CD25+Treg细胞的调节作用.方法:ELISA检测TRAIL对肿瘤细胞分泌CCL22的影响;建立对TRAIL耐受的4T1肿瘤细胞皮下实体瘤模型,瘤内给予重组TRAIL蛋白,检测肿瘤体积的变化;分离肿瘤侵润的淋巴细胞,采用流式细胞术检测瘤内CD4+CD25+Treg细胞的变化.结果:TRAIL引起肿瘤细胞4T1和B16培养上清中CCL22水平增加;TRAIL治疗组与对照组相比,对TRAIL耐受的4T1移植瘤体积没有明显变化,但TRAIL治疗组的肿瘤侵润CD4+CD25+Treg细胞显著增加.结论:TRAIL引起肿瘤细胞分泌CCL22,可因此诱导CD4+CD25+Treg细胞趋化至肿瘤部位导致肿瘤侵润的CD4+CD25+Treg比例增加,为TRAIL的生理功能和在肿瘤治疗中的应用提供了新的资料.

  20. CD25 shedding by human natural occurring CD4+CD25+ regulatory T cells does not inhibit the action of IL-2

    DEFF Research Database (Denmark)

    Pedersen, Anders Elm; Lauritsen, Jens Peter Holst

    2009-01-01

    Regulatory T (Treg) cells are important for the maintenance of peripheral tolerance and inhibition of pathogenic T-cell responses. Therefore, they are important for the limitation of chronic inflammation but can also be deleterious by e.g. limiting antitumour immune responses. Natural occurring...... Tregs are known to inhibit CD4+ T cell in a contact-dependent manner, but at the same time, various suppressive factors are secreted. We, here, demonstrate that human naturally occurring CD4+CD25+ Tregs are able to shed large amounts of soluble CD25 upon activation. Secretion of sCD25 could add to the...... inhibitory effect of Tregs as such secretion in other settings has been proposed to act as a sink for local IL-2. However, we here demonstrate that supernatant from human Tregs containing high concentration of sCD25 does not inhibit proliferation of CD4+CD25(-) T cells or inhibit the action of IL-2 in an in...

  1. CD4+CD25+Treg细胞与支气管哮喘%CD4+CD25+Treg cells and bronchial asthma

    Institute of Scientific and Technical Information of China (English)

    鞠云飞; 孙立锋; 胡华

    2011-01-01

    CD4+CD25+Treg细胞的主要作用表现为免疫无能性和免疫抑制性,是外周免疫耐受形成机制的主要组成部分.其主要作用机制为分泌抑制性细胞因子(IL-10和TGF-β)、表达细胞表面分子(CTLA-4、GITR等)及Foxp3等.支气管哮喘患者外周血CD4+CD25+Treg功能及数量存在异常,这可能是支气管哮喘发病机制之一.糖皮质激素可以通过影响CD4+CD25+Treg的状态起到抑制支气管哮喘气道炎症的作用.

  2. Progress of CD4 + CD25 + regulatory T cells in pathogenesis of idiopathic thrombocytopenic purpura%CD4+CD25+调节性T细胞在特发性血小板减少性紫癜发病机制中的作用

    Institute of Scientific and Technical Information of China (English)

    刘小群

    2011-01-01

    CD4 + CD25 + regulatory T cells(Treg) are thought to be a subgroup of mature CD4 + T cells.Forkhead winged helix transcription factor-3 (Foxp3)is specifically expressed on them and plays a key role in their development and function. CD4 + CD25 + Treg cells can maintain the stabilization of internal environment by two principal pathways to suppress the immunological function: the direct suppression of the target cells by cell-contact and the secretion of suppressor cytokines. At present, it has been considered that decreased number and dysfunction of CD4+ CD25+ Treg cells are closely related to pathogenesis of autoimmune disease. Recent findings show that CD4+ CD25+ Treg cells play an important role in pathogenesis of idiopathic thrombocytopenic purpura.%CD4+ CD25+调节性T细胞(regulatory T cell,Treg)是一种成熟的CD4+T细胞亚群,而叉头翼状螺旋转录因子3(forkhead winged helix transcription factor-3,Foxp3)特异性表达于该细胞上,对其发育和功能起关键作用。CD4+ CD25+ Treg细胞主要通过直接接触和分泌抑制性细胞因子两大途径发挥免疫抑制功能,维持机体内环境的稳定。目前认为其数目减少和功能障碍与自身免疫性疾病的发生密切相关。近年来研究表明CD4+ CD25+ Treg细胞在特发性血小板减少性紫癜的发病中起重要作用。

  3. Murine CD4+CD25- cells activated in vitro with PMA/ionomycin and anti-CD3 acquire regulatory function and ameliorate experimental colitis in vivo

    Directory of Open Access Journals (Sweden)

    Majowicz Anna

    2012-12-01

    Full Text Available Abstract Background Induced regulatory T (iTreg lymphocytes show promise for application in the treatment of allergic, autoimmune and inflammatory disorders. iTreg cells demonstrate advantages over natural Treg (nTreg cells in terms of increased number of starting population and greater potential to proliferate. Different activation methods to generate iTreg cells result in iTreg cells that are heterogeneous in phenotype and mechanisms of suppression. Therefore it is of interest to explore new techniques to generate iTreg cells and to determine their physiological relevance. Methods Using phorbol myristate acetate (PMA/ionomycin and anti-CD3 activation of CD4+CD25- cells we generated in vitro functional CD4+CD25+ iTreg (TregPMA cells. Functionality of the generated TregPMA cells was tested in vivo in a mouse model of inflammatory bowel disease (IBD - CD45RB transfer colitis model. Results TregPMA cells expressed regulatory markers and proved to ameliorate the disease phenotype in murine CD45RB transfer colitis model. The body weight loss and disease activity scores for TregPMA treated mice were reduced when compared to diseased control group. Histological assessment of colon sections confirmed amelioration of the disease phenotype. Additionally, cytokine analysis showed decreased levels of proinflammatory colonic and plasma IL-6, colonic IL-1 β and higher levels of colonic IL-17 when compared to diseased control group. Conclusions This study identifies a new method to generate in vitro iTreg cells (TregPMA cells which physiological efficacy has been demonstrated in vivo.

  4. 哮喘患儿外周血CD4+CD25+调节性T细胞的测定及临床意义%The level of CD4+CD25+regulatory T cells and its clinical significance in children with asthma

    Institute of Scientific and Technical Information of China (English)

    湛洁谊; 卢慧敏; 林穗玲

    2014-01-01

    Objective To explore the proportion change of CD4+CD25+regulatory T cells (Treg) in peripheral blood of children with asthma and to analyze its significance. Methods A total of 150 asthmatic children were divided into three groups according to their clinical features50 subjects in acute asthma attack group, 50 subjects in clinical re-mission of asthma group and 50 subjects in cough variant asthma group,meanwhile, 50 healthy children were enrolled in the control group. The levels of CD4+CD25+Treg in peripheral blood of all children were detected by flow cytometer. Results The CD4+CD25+Treg level in acute attack group were lowest of the four groups (P0.05). Conclusion The CD4+CD25+regulatory T cells may be involved in the pathogenesis of asthma,the level of CD4+CD25+regulatory T cells correlated with the severity of asthma in children.%目的:探讨CD4+CD25+调节性T细胞在哮喘儿童外周血中的比例改变,并探讨其临床意义。方法150例哮喘患儿按临床表现分为急性发作组(50例)、临床缓解期组(50例)和咳嗽变异性哮喘组(50例),另选择50名健康儿童为正常对照组。应用流式细胞仪检测上述各组外周血CD4+CD25+调节性T细胞占CD4+T细胞的百分比。结果急性发作期组患儿外周血CD4+CD25+调节性T细胞水平较缓解组、咳嗽变异性哮喘组及健康对照组明显下降(P0.05)。结论CD4+CD25+调节性T细胞可能参与了哮喘的发生与发展,哮喘的严重程度可能与CD4+CD25+调节性T细胞的水平相关。

  5. CDK6-mediated repression of CD25 is required for induction and maintenance of Notch1-induced T-cell acute lymphoblastic leukemia.

    Science.gov (United States)

    Jena, N; Sheng, J; Hu, J K; Li, W; Zhou, W; Lee, G; Tsichlis, N; Pathak, A; Brown, N; Deshpande, A; Luo, C; Hu, G F; Hinds, P W; Van Etten, R A; Hu, M G

    2016-05-01

    T-cell acute lymphoblastic leukemia (T-ALL) is a high-risk subset of acute leukemia, characterized by frequent activation of Notch1 or AKT signaling, where new therapeutic approaches are needed. We showed previously that cyclin-dependent kinase 6 (CDK6) is required for thymic lymphoblastic lymphoma induced by activated AKT. Here, we show CDK6 is required for initiation and maintenance of Notch-induced T-ALL. In a mouse retroviral model, hematopoietic stem/progenitor cells lacking CDK6 protein or expressing kinase-inactive (K43M) CDK6 are resistant to induction of T-ALL by activated Notch, whereas those expressing INK4-insensitive (R31C) CDK6 are permissive. Pharmacologic inhibition of CDK6 kinase induces CD25 and RUNX1 expression, cell cycle arrest and apoptosis in mouse and human T-ALL. Ablation of Cd25 in a K43M background restores Notch-induced T leukemogenesis, with disease that is resistant to CDK6 inhibitors in vivo. These data support a model whereby CDK6-mediated suppression of CD25 is required for initiation of T-ALL by activated Notch1, and CD25 induction mediates the therapeutic response to CDK6 inhibition in established T-ALL. These results both validate CDK6 as a molecular target for therapy of this subset of T-ALL and suggest that CD25 expression could serve as a biomarker for responsiveness of T-ALL to CDK4/6 inhibitor therapy. PMID:26707936

  6. Ribavirin Does Not Impair the Suppressive Activity of Foxp3+CD4+CD25+ Regulatory T Cells

    OpenAIRE

    Lee, Jeewon; CHOI, YOON SEOK; Shin, Eui-Cheol

    2013-01-01

    Ribavirin is an antiviral drug used in combination with pegylated interferon-α (IFN-α) for the treatment of hepatitis C virus (HCV) infection. Recently, ribavirin was reported to inhibit the suppressive activity of regulatory T (Treg) cells. In the present study, we re-evaluated the effect of ribavirin on Foxp3+CD4+CD25+ Treg cells from normal donors. First, we examined the expression of CTLA-4 and CD39, which are known to play a role in the suppressive function of Treg cells. We found that r...

  7. 体外培养扩增CD4+CD25+调节性T细胞的实验观察%Proliferation of CD4+CD25+ regulatory T cells of rat by different cytokines in vitro

    Institute of Scientific and Technical Information of China (English)

    王子函; 朱继业; 李涛; 冷希圣

    2008-01-01

    Objective To evaluate the effects of cytokines on tlle proliferation and function of CD4+CD25+ regulatory T cell(Treg).Mellaotis Tregs were isolated from na(i)ve C57BL/6 mice spleen and lymph nodes.Mature dendritic cells(mDC)were isolated from DBA/2 mice,co-cultured with Tregs,and divided into 4 groups with or without interleukin-2(IL-2),interleukin-4(IL-4),and interleukin-15(IL-15)added into the culture fluid.Fluorescence.activated cell sorting(FACS)was used to detect the Treg proliferation and apoptosis with CFSE and annexin-V staining.The co-cuhure increased Tregs were divided into 5 groups:CFSE labeled naive CIM+ CD25-T cells,self-proliferated Treg,Treg mixedly cultured with IL-2 mDC,and Teff,Treg mixedly cultured with IL-4,mDC,and Teff,and Treg mixedly cultured with IL-15,mDC,and Teff,a control group included Teff co-cultured with mDC.FACS was used 5 d later to evalugte the suppressive function of the Treg Oil the Teff. The expression of Foxp3,indicating the phenotype of Treg was detected.Results FASC showed that the values of precursor frequency(PF)of the Tregs stimulated by IL-2,IL-4,and IL-15 were 31.3%,28.9%,and 34.5%respectively,all significantly higher than that of the control group(14.5% all P<0.05),and the values of proliferation index(PI)of the Tregs stimulated by IL-2,IL-4,and IL-15 were 1.9,1.7,and 1.8 respectively,all significantly higher than that of the control group(1.5,all P<0.05).,The apoptotic rates of the Tregs stimulated by IL-2,IL-4,and IL-15 were 12.8%,11.4%,and 12.7% respectively,all signifieanfly lower tllan that of the control group(28.9%,P<0.05).The Foxp3 expression rate of the Tregs stimulated by IL-2,IL-4,and IL-15was 91.75%.Conclusion IL-2.IL-4.and IL-15 in the in vitro culture of Treg stimulate the Treg proliferation,reduce tlleir apoptosis,and maintain their suppressive function.The proliferated Tregs still maintain their phenotype,highly expressing Foxp3.%目的 观察细胞因子体外培养扩增CD4+CD25+调节性T细

  8. Induction of CD4+CD25+Foxp3+regulatory T cell response by glatiramer acetate in type 1 diabetes

    Institute of Scientific and Technical Information of China (English)

    Guoliang Cui; Yuebo Zhang; Zhenwei Gong; Jingwu Z Zhang; Ying Qin Zang

    2009-01-01

    Glatiramer acetate (GA) is an immunomodulatory peptide drug used to treat multiple sclerosis. Its treatment ef-fect has been expanded to other autoimmune conditions such as uveoretinitis, inflammatory bowel disease, graft re-jection and hepatic fibrosis. Here, we report that GA was effective in altering the clinical course of diabetes in cyclo-phosphamide (CY)-potentiated non-obese diabetic (CY-NOD) mice. Treatment with GA significantly reduced the dia-betic rate in the mice and ameliorated insulitis, which coincided with increased CD4+CD25+Foxp3+T cell response in treated mice. GA treatment led to increased expression of transcription factor Foxp3 and elevated production of interleukin-4 (IL-4) both in vivo and in vitro. It was evident that the effect of GA on up-regulation of Foxp3 was me-diated partially through IL-4. IL-4 was found to maintain Foxp3 expression and regulatory function of CD4+CD25+ regulatory T cells (Tregs). This study provides new evidence that GA has treatment potential for type 1 diabetes through the induction of Tregs and that increased IL-4 production is partially responsible for the enhanced Treg's function in GA treatment.

  9. Protective Effect of CXCR3+CD4+CD25+Foxp3+ Regulatory T Cells in Renal Ischemia-Reperfusion Injury

    Directory of Open Access Journals (Sweden)

    Cao Jun

    2015-01-01

    Full Text Available Regulatory T cells (Tregs suppress excessive immune responses and are potential therapeutic targets in autoimmune disease and organ transplantation rejection. However, their role in renal ischemia-reperfusion injury (IRI is unclear. Levels of Tregs and expression of CXCR3 in Tregs were analyzed to investigate their function in the early phase of renal IRI. Mice were randomly divided into Sham, IRI, and anti-CD25 (PC61 + IRI groups. The PC61 + IRI group was established by i.p. injection of PC61 monoclonal antibody (mAb to deplete Tregs before renal ischemia. CD4+CD25+Foxp3+ Tregs and CXCR3 on Tregs were analyzed by flow cytometry. Blood urea nitrogen (BUN, serum creatinine (Scr levels, and tubular necrosis scores, all measures of kidney injury, were greater in the IRI group than in the Sham group. Numbers of Tregs were increased at 72 h after reperfusion in kidney. PC61 mAb preconditioning decreased the numbers of Tregs and aggravated kidney injury. There was no expression of CXCR3 on Tregs in normal kidney, while it expanded at 72 h after reperfusion and inversely correlated with BUN, Scr, and kidney histology score. This indicated that recruitment of Tregs into the kidney was related to the recovery of renal function after IRI and CXCR3 might be involved in the migration of Tregs.

  10. 应用iDC从G-CSF动员的外周血中诱导产生CD4+CD25+调节性T细胞%Characteristics of induced CD4+CD25+ regulatory T lymphocytes in G-CSF mobilized peripheral blood by iCD

    Institute of Scientific and Technical Information of China (English)

    苏虹; 王忱诚; 王保龙; 彭美玲

    2012-01-01

    -CSF mobilized and un-mobilized peripheral blood. Then flow cytometry (FCM) and mixed lymphocyte reaction ( MLR) were used respectively to test the expression of CD25+ , Foxp 3 and suppres-sive function of these iTregs The differences of the rate and the suppressive effect were compared among different groups. Results After treated with iDC , there was significant difference in the expression of CD25 + between different groups with CD4 + CD25- T cells isolated from G-CSF mobilized and un-mobilized groups , which were(90. 4 ± 5. 3)% and (76. 8 ±4. 1)% ,respectively(P <0. 01). The expressions of Foxp3 of the above groups were (64. 1 ± 2.1')% and ( 59. 5 ± 3. 2 ) % respectively , the difference was also significant ( P < 0. 05 ) . The iTregs came from G-CSF mobilized peripheral blood induced by iDC had more suppressive effect compared with the unmobilized group in vitro, the difference was significant ( P < 0. 01). Conclusion The induced CD4+ CD25 + Tregs in G-CSF mobilized peripheral blood by iDC have more suppressive effect· in vitro It indicates that G-CSF mobilized peripheral blood can serve as an important source of CD4+CD25 + Tregs, which has potential to be used as an immunotherapy agent for autoimmune diseases.

  11. The relative values of CD8+CD25+Foxp3brigh Treg cells correlate with selected lung function parameters in asthma.

    Science.gov (United States)

    Eusebio, M; Kuna, P; Kraszula, L; Kupczyk, M; Pietruczuk, M

    2015-06-01

    The study aimed to detect CD8(+)CD25(+)FoxP3(brigh) Tregs and investigate their possible association with selected lung function values. CD8(+)CD25(+)FoxP3(brigh) Tregs were detected by flow cytometry in the peripheral blood of 25 patients with severe asthma (SA), 25 patients with mild-to-moderate asthma (MA), and 25 age-matched healthy donors (NC). The percentages of CD8(+)CD25(+)FoxP3(brigh) Tregs of the patients with severe (3.4 ± 4.55), and mild-to-moderate asthma (7.5 ± 8.15), were markedly lower than those of controls (12.1 ± 13.2). The mean forced expiratory volume in 1 s (FEV1) % predicted value in severe asthma subpopulation was significantly lower (67.05 ± 15.98%) when compared with that of mild-to-moderate asthma subgroup (87.71 ± 16.12%). Interestingly, the percentages of CD8(+)CD25(+)FoxP3(brigh) Tregs correlate with mean peak expiratory flow (PEF)% predicted values in severe (r = 0.7, P asthma. In contrast, this parameter was positively correlated with FEV1% predicted values in the severe asthmatics only (r = 0.71, P Tregs and selected lung function parameters, suggesting that this parameter has potential as a marker for inflammation and airflow obstruction. PMID:25921629

  12. CD4(+), CD25(+), FOXP3 (+) T Regulatory Cell Levels in Obese, Asthmatic, Asthmatic Obese, and Healthy Children.

    Science.gov (United States)

    Donma, Metin; Karasu, Erkut; Ozdilek, Burcu; Turgut, Burhan; Topcu, Birol; Nalbantoglu, Burcin; Donma, Orkide

    2015-08-01

    The aim of this prospective case control study is to determine CD4(+), CD25(+), and FoxP3(+) T regulatory cells (Tregs) and T helper cells (Ths) in obese, asthmatic, asthmatic obese, and healthy children. Obese (n = 40), asthmatic (n = 40), asthmatic obese (n = 40), and healthy children (n = 40) were included in this study. Blood samples collected from children were marked with CD4, CD25, ve Foxp3 in order to detect Tregs and Ths by flow cytometric method. Statistical analyses were performed. p ≤ 0.05 was chosen as meaningful threshold. Tregs exhibiting anti-inflammatory nature were significantly lower in obese (0.16 %; p ≤ 0.001), asthmatic (0.25 %; p ≤ 0.01), and asthmatic obese (0.29 %; p ≤ 0.05) groups than control group (0.38 %). Ths were counted higher in asthma group than control (p ≤ 0.01) and obese (p ≤ 0.001) groups. T cell immunity plays important roles in chronic inflammatory diseases such as obesity and asthma pathogeneses. Decreased numbers of Tregs found in obese, asthmatic, and asthmatic obese children might represent a challenge of these cells. PMID:25655390

  13. Neonatal bacillus Calmette-Guerin vaccination inhibits de novo allergic inflammatory response in mice via alteration of CD4+CD25+T-regulatory cells

    Institute of Scientific and Technical Information of China (English)

    Qian LI; Hua-hao SHEN

    2009-01-01

    Aim: The hygiene hypothesis suggests a lack of bacterial infections would favor the development of allergic diseases. My-cobacterium bovis bacille Calmette-Guerin (BCG) infection can inhibit allergen-induced asthma reactions, but the underly-hag mechanism of this infection on the immunological responses is unclear. T-regulatory (Treg) cells are thought to play a role as a crucial immunoregulatory cells that are capable of regulating adaptive immune responses. We conducted this study to investigate whether the protective effect of the BCG vaccination on allergic pulmonary inflammation is associated with the alteration of CD4+CD25+ Treg cells in a murine asthma model and the mechanisms of Treg cells. Methods: Newborn C57BL/6 mice were vaccinated 3 times with BCG on d 0, 7, and 14 and subsequently sensitized and challenged with ovalbumin. Eosinophil infiltration was investigated. The frequencies of spleen CD4+CD25+ Treg cells and the expression of specific transcriptional factor Foxp3 were assayed. The cytotoxic lymphocyte associated antigen (CTLA)-4 expression and cytokine interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) levels were measured. Results: We showed that treatment of mice with BCG inhibited de novo allergic inflammatory response in a mouse model of asthma. BCG treatments are associated with the increase of CD4+CD25+ Treg cells and Foxp3 expression, accompanied by an increased CTLA-4 expression and cytokine IL-10 and TGF-β levels (P<0.05). Conclusion: Neonatal BCG vaccinations ameliorate de novo local eosinophilic inflammation induced by allergen and in-crease the numbers of CD4+CD25+ Treg cells and Foxp3 expression. The cell-cell contact inhibition and regulatory cytokine production may be involved in the regulatory mechanism.

  14. Immunology Mechanism of CD4+ CD25 T Regulatory Cells Acting on Effector T Cells

    Institute of Scientific and Technical Information of China (English)

    FENGNing-han; WUHong-fei; WUJun; ZHANGWei; SUIYuan-gen; HEHou-guang; ZHANGChun-lei; ZHENGJun-song

    2004-01-01

    Objective: To detect the inhibiting co-stimulating molecule CTLA4 and cytokines secreted by Treg cells, and explore the immunology mechanism of T regulatory cells acting on effector T cells in co-cultured system(CCS) and separating-cultured system(SCS). Methods: Detecting the percentage of CTLA4 and CD28 expressed on the Treg ceils and effector T ceils, and then adding Treg cells to mixed lymphocyte reaction(MLR) system in CCS and TransWeil Milliceil-PCF SCS, at the same time, adding or not adding anti-IL-10 or anti-TGF.II1 to the reacting systems, examining the inhibitory capacity of Treg ceils exerting on the MLR. Results: Compared with effector T cells, Treg cells expressed higher level CTLA4 and secreted much more IL-10 and TGF-β(P<0.01). The inhibitory capacity of Treg cells co-cultured with effector T ceils is much stronger than that in separating cultured group(P<0.01). Moreover, the inhibiting rate of Treg ceils exerting on effector T ceils through secretin_g IL-10 was more powerful than that through secreting TGF-β1 (P<0.01). Coaclusion: Both ceil-to-ceil contact and cytokines secretion mechanisms are involved in CD4+ CD25+ Treg ceils operating function. However, the former is more important. Intresfingly, we for the first time pointfound that IL-10 plays more powerful roles than TGF-β1 in the cytokines secretion mechanism.

  15. Downregulation of IL-12 and a novel negative feedback system mediated by CD25+CD4+ T cells

    International Nuclear Information System (INIS)

    CD25+CD4+ regulatory T cells suppress immune responses and are believed to play roles in preventing autoimmune diseases. However, the mechanism(s) underlying the suppression and the regulation of their homeostasis remain to be elucidated. Here we show that these regulatory T cells downregulated CD25-CD4+ T-cell-mediated production of IL-12 from antigen-presenting cells, which can act as a growth factor for CD25-CD4+ T cells. We further found that CD25+CD4+ T cells, despite their well-documented 'anergic' nature, proliferate significantly in vitro only when CD25-CD4+ T cells are present. Notably, this proliferation was strongly dependent on IL-2 and relatively independent of IL-12. Thus, CD25+CD4+ T cells suppress CD25-CD4+ T-cell responses, at least in part, by inhibiting IL-12 production while they themselves can undergo proliferation with the mediation of CD25-CD4+ T cells in vitro. These results offer a novel negative feedback system involving a tripartite interaction among CD25+CD4+ and CD25-CD4+ T cells, and APCs that may contribute to the termination of immune responses

  16. Numerical status of CD4+CD25+FoxP3+ and CD8+CD28- regulatory T cells in multiple sclerosis

    Directory of Open Access Journals (Sweden)

    Ebrahim Kouchaki

    2014-04-01

    Full Text Available Objective(s: Regulatory T cells, including CD4+CD25+Fox3+ and CD8+CD28- cells play an important role in regulating the balance between immunity and tolerance. Since multiple sclerosis is an inflammatory autoimmune disease, regulatory T cells are considered to be involved in its pathogenesis. In this study, we investigated the circulatory numbers of the two mentioned types of regulatory T cells and also their association with different clinical characteristics in 84 multiple sclerosis patients. Materials and Methods: 84 patients with multiple sclerosis and 75 normal individuals were studied. Demographic and clinical information of all participants were collected via questionnaire and clinical examination as well as MRI. The peripheral blood frequency of two different subgroups of regulatory T cells (CD4+ CD25+Foxp3+ and CD8+CD28- cells were analyzed by flow cytometry using anti-human antibodies conjugated with CD4-FITC / CD25-PE/Foxp3-PE-Cy5, CD3-PE/CD8a-PE-Cy5/CD28-FITC. Results: The frequency of CD4+CD25+Foxp3+ cells in multiple sclerosis patients was significantly less than that in healthy controls (P=0.006 and in mild forms less than that in sever forms (P=0.003. There was not any correlation between the frequency of regulatory T cells and different clinical variables. Conclusion: Our results showed that the number of CD4+CD25+Foxp3+ cells decreases significantly in multiple sclerosis patients, which probably shows the regulatory role of these cells in multiple sclerosis.

  17. CD4+CD25+调节性T细胞对哮喘大鼠免疫功能影响%Effect of CD4+CD25+Regulatory T Cells on the Immunologic Function in Rats with Asthma

    Institute of Scientific and Technical Information of China (English)

    薛克营; 金卫国; 王成国; 程立; 杨中卫; 王正艳

    2009-01-01

    目的:观察CD4+CD25+调节性T细胞(CD4+CD25+Treg)对CD4+CD25-T细胞增殖和Th1/Th2细胞因子分泌的影响,探讨其在哮喘气道炎症中的作用机制.方法:将哮喘大鼠CD4+CD25-T细胞分别与卵白蛋白(OVA)免疫耐受大鼠CD4+Cd25+Treg细胞和哮喘大鼠CD4+CD25+Treg细胞联合培养,3H胸腺嘧啶核苷(3H-TdR)掺入法测量细胞增殖情况,ELISA检测细胞IL-4、IL-5和IFN-γ含量.结果:OVA耐受大鼠CD4+CD25+Treg细胞能抑制CD4+CD25-T细胞增殖和Th2细胞因子分泌(P<0.05);哮喘大鼠CD4+CD25+Treg细胞可明显抑制IFN-γ的分泌(P<0.05).结论:OVA免疫耐受大鼠CD4+CD25+Treg细胞可能通过抑制哮喘大鼠CD4+CD25-T细胞增殖和影响Th1/Th2平衡发挥作用,哮喘大鼠CD4+CD25+Treg细胞存在功能异常,可能与哮喘的发病有关.

  18. Effect of different doses of rapamycin (RAPA) on Kunming-strain mouse CD4 + CD25 + Treg cells proliferations%不同剂量雷帕霉素对小鼠体内CD4+CD25+Treg细胞的影响

    Institute of Scientific and Technical Information of China (English)

    彭磊磊; 葛圣林; 张成鑫

    2011-01-01

    目的 研究不同剂量雷帕霉素对小鼠体内Treg细胞的影响.方法 将SPF级昆明系小鼠60只随机分为对照组(A)和实验组(B、C、D),B、C、D三组分别灌胃雷帕霉素1、2、3 mg·kg-1,A组每天予以无菌水灌胃,共3周.3周后,无菌条件下心脏采血,EDTA抗凝,分离脾脏,制备单细胞悬液,采用流式细胞仪检测小鼠外周血和脾脏中CD4+CD25+调节性T细胞水平(CD4+CD25+Treg细胞占CD4+ T细胞的百分比).结果 实验组(B、C、D)小鼠外周血和脾细胞中CD4+CD25+Treg细胞水平分别为(9.62±1.43)%、(13.76±1.97)%、(15.41±2.45)%和(12.23±4.56)%、(23.03±6.18)%、(25.17±6.42)%,对照组(A)小鼠外周血和脾细胞中CD4+CD25+Treg细胞水平分别为(3.52±0.65)%和(6.53±3.01)%,无论是在外周血还是脾细胞中,B、C、D组CD4+CD25+Treg细胞水平明显高于A组(P0.05).结论 雷帕霉素能够诱导昆明系小鼠体内CD4+CD25+Treg细胞增殖,其使用剂量可以影响CD4+CD25+Treg细胞的增殖程度.%Aim To investigate how rapamycin (RAPA) at different doses levels induce Kunming-strain mouse CD4 + CD25 + Treg cells proliferations. Methods 60 Kunming-strain mice at the age of 8 weeks were divided into a control group (A) and three experimental groups (B, C,D). The mice in groups B,C and D were fed RAPA 1 ,2 and 3 mg · kg -1 intragastric administration. The mice in group A were given sterile water as the control group. After three weeks, under sterile conditions by collecting the peripheral blood and then seperating the splenocytes (EDTA anticoagulant) ,we were able to generate a single-cell suspension. The level of CD4 + CD25 + Treg cells in the mouse peripheral blood and splenocytes were detected by flow cytometer. (The ratio of CD4 + CD25 + Treg cells to CD4 + CD25 Treg cells). Results The CD4 + CD25 + Treg cells in the mouse peripheral blood and splenocytes of the experimental groups (B, C, D) were (9.62± 1.43)% ,(13.76 ± 1.97)% ,(15.41 ±2.45)% and (12.23 ±4

  19. Conversion of Peripheral CD4+CD25− Naive T Cells to CD4+CD25+ Regulatory T Cells by TGF-β Induction of Transcription Factor Foxp3

    OpenAIRE

    Chen, Wanjun; Jin, Wenwen; Hardegen, Neil; Lei, Ke-Jian; Li, Li; Marinos, Nancy; McGrady, George; Wahl, Sharon M.

    2003-01-01

    CD4+CD25+ regulatory T cells (Treg) are instrumental in the maintenance of immunological tolerance. One critical question is whether Treg can only be generated in the thymus or can differentiate from peripheral CD4+CD25− naive T cells. In this paper, we present novel evidence that conversion of naive peripheral CD4+CD25− T cells into anergic/suppressor cells that are CD25+, CD45RB−/low and intracellular CTLA-4+ can be achieved through costimulation with T cell receptors (TCRs) and transformin...

  20. An experimental study of the relationship between CD4+CD25+T regulatory cells and the Th1/Th2 imbalance in asthmatic mice%哮喘小鼠调节性T细胞与Th1/Th2因子失衡关系的实验研究

    Institute of Scientific and Technical Information of China (English)

    綦梅伶; 路明; 武怡; 吴铭

    2011-01-01

    Objective To investigate the effect of the variations of CD4 +CD25 + T regulatory cell counts in the peripheral blood in mouse model of asthma on the expressions of serum Th1/Th2 - related cytokines, IFN - γ/ and IL - 4,and to explore their roles in the pathogenesis of bronchial asthma. Methods Twenty healthy female BALB/c mice were randomized into two groups ( n = 10 each ). The mice in the model group were subjected to intraperitoneal injection of OVA/AIC OH )3 on days 0, 7 and 14 respectively, followed by challenge with inhalation of nebulized OVA daily from days 21 to 26. The control group underwent treatment with normal saline instead of OVA for intraperitoneal injection during primary sensitization and aerosol inhalation during secondary induction. The animals were sacrificed on day 27. The histopathological changes of the murine pulmonary tissues were observed by microscopy. The variations in CD4+ CD25 + T+ regulatory cell counts in the blood and expression levels of the cytokines 1L -4 and IFN - γ/ in the serum were determined by flow cytometry and ELISA, respectively. Results The counts of CD4+CD25 +T regulatory cells in the peripheral blood.their proportion in CD4 + T cells and the serum IFN - γ level were significantly lower ( P <0. 01 ) and serum 1L -4 level was significantly higher ( P <0. 01 ) in the model group than those in the control group. Conclusion Asthmatic mice show an evident Th1/Th2 imbalance, with the hyperfunction of the Th2 , and its ruechanism is related to the decreased inhibition of CD4 + CD25 + T regulatory cells.%目的 研究哮喘小鼠外周血CD4+CD25+Treg细胞数量改变与血清Th1/Th2相关细胞因子IFN-γ和IL-4水平的关系,探讨其在支气管哮喘发病中的作用.方法 健康雌性BALB/c小鼠20只,随机分为2组(每组10只):①模型组于第0、7、14天予OVA/Al(OH)3腹腔注射,第21~26天每天以OVA雾化吸入激发.②对照组则以生理盐

  1. 激素抵抗性哮喘患者外周血CD4+CD25+Foxp3+调节性T细胞及白细胞介素10、转化生长因子β1水平变化及其临床意义%The clinical significance of CD4+ CD25+ Foxp3+ regulatory T cell and interleukin-10,transforming growth factor-β1 in patients with steroid-resistant asthma

    Institute of Scientific and Technical Information of China (English)

    王善飞; 赵平; 许庆元

    2013-01-01

    was detected by flow cytometry and serum IL-10 and TGF-β1 levels were detected by enzyme-linked immunosorbent assay.Results The percentage of CD4 + CD25 +Foxp3 +Treg in CD4+ T cell and its absolute value in SRA group and SSA group were 0.0225 ± 0.0063,(1.09 ± 0.23) × 107/L and 0.0345 ± 0.0094,(1.35 ± 0.14) × 107/L,they were significandy lower than those in control group [0.0537 ± 0.0128,(2.06 ± 0.27) × 107/L],and SRA group was significandy lower than SSA group,there were statistical differences (P < 0.05).The levels of serum TGF-β1 in SRA group and SSA group were significantly lower than those in control group [(138.12 ± 23.26),(176.25 ± 40.37) ng/L vs.(281.22 ±47.15) ng/L],there was statistical difference (P <0.05).The levels of serum IL-10 in SRA group was significantly lower than that in control group [(516.43 ± 86.33)ng/L vs.(763.02 ± 90.19) ng/L],there was statistical difference (P < 0.05).There was no statistical difference in the level of serum IL-10 between SSA group and control group (P > 0.05).The levels of serum IL-10 and TGF-β 1 in SRA group were significantly lower than those in SSA group,there were statistical differences (P < 0.05).The levels of serum IL-10 and TGF-β1 had positive correlation with CD4 + CD25 + Foxp3 +Treg in SRA group and SSA group (P < 0.01).Conclusion The interaction among CD4+ CD25+ Foxp3 +Treg,IL-10 and TGF-β1 may play an important role in the SRA occurrence and development,while by increasing peripheral blood CD4 + CD25 + Foxp3 +Treg number and stabilizing its function can increase IL-10 and TGF-β1 expression,which may be an important way to treat SRA.

  2. Cortical control of facial expression.

    Science.gov (United States)

    Müri, René M

    2016-06-01

    The present Review deals with the motor control of facial expressions in humans. Facial expressions are a central part of human communication. Emotional face expressions have a crucial role in human nonverbal behavior, allowing a rapid transfer of information between individuals. Facial expressions can be either voluntarily or emotionally controlled. Recent studies in nonhuman primates and humans have revealed that the motor control of facial expressions has a distributed neural representation. At least five cortical regions on the medial and lateral aspects of each hemisphere are involved: the primary motor cortex, the ventral lateral premotor cortex, the supplementary motor area on the medial wall, and the rostral and caudal cingulate cortex. The results of studies in humans and nonhuman primates suggest that the innervation of the face is bilaterally controlled for the upper part and mainly contralaterally controlled for the lower part. Furthermore, the primary motor cortex, the ventral lateral premotor cortex, and the supplementary motor area are essential for the voluntary control of facial expressions. In contrast, the cingulate cortical areas are important for emotional expression, because they receive input from different structures of the limbic system. J. Comp. Neurol. 524:1578-1585, 2016. © 2015 Wiley Periodicals, Inc. PMID:26418049

  3. CD8{sup +}CD25{sup +} T cells reduce atherosclerosis in apoE(−/−) mice

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Jianchang; Dimayuga, Paul C.; Zhao, Xiaoning; Yano, Juliana; Lio, Wai Man; Trinidad, Portia; Honjo, Tomoyuki; Cercek, Bojan; Shah, Prediman K.; Chyu, Kuang-Yuh, E-mail: Chyuk@cshs.org

    2014-01-17

    Highlights: •The role of a sub-population of CD8{sup +} T cells with suppressor functions was investigated in atherosclerosis. •CD8{sup +}CD25{sup +} T cells from adult apoE(−/−) mice had phenotype characteristics of T suppressor cells. •These CD8{sup +}CD25{sup +} T cells reduced CD4{sup +} T cell proliferation and CD8{sup +} cytotoxic activity in vitro. •Adoptive transfer of CD8{sup +}CD25{sup +} T cells significantly reduced atherosclerosis. •CD8{sup +}CD25{sup +} T cells have a suppressive function in atherosclerosis. -- Abstract: Background: It is increasingly evident that CD8{sup +} T cells are involved in atherosclerosis but the specific subtypes have yet to be defined. CD8{sup +}CD25{sup +} T cells exert suppressive effects on immune signaling and modulate experimental autoimmune disorders but their role in atherosclerosis remains to be determined. The phenotype and functional role of CD8{sup +}CD25{sup +} T cells in experimental atherosclerosis were investigated in this study. Methods and results: CD8{sup +}CD25{sup +} T cells were observed in atherosclerotic plaques of apoE(−/−) mice fed hypercholesterolemic diet. Characterization by flow cytometric analysis and functional evaluation using a CFSE-based proliferation assays revealed a suppressive phenotype and function of splenic CD8{sup +}CD25{sup +} T cells from apoE(−/−) mice. Depletion of CD8{sup +}CD25{sup +} from total CD8{sup +} T cells rendered higher cytolytic activity of the remaining CD8{sup +}CD25{sup −} T cells. Adoptive transfer of CD8{sup +}CD25{sup +} T cells into apoE(−/−) mice suppressed the proliferation of splenic CD4{sup +} T cells and significantly reduced atherosclerosis in recipient mice. Conclusions: Our study has identified an athero-protective role for CD8{sup +}CD25{sup +} T cells in experimental atherosclerosis.

  4. Electroacupuncture Attenuates Ovalbumin-Induced Allergic Asthma via Modulating CD4+CD25+ Regulatory T Cells

    Directory of Open Access Journals (Sweden)

    Youngjoo Kwon

    2012-01-01

    Full Text Available A mouse pulmonary hypersensitivity experimental model that mimics human asthma was developed, and electroacupuncture (EA treatment was shown to reduce allergic inflammatory processes. In addition, we also assessed whether the beneficial effects of EA on allergic asthma could be correlated with CD4+CD25+Foxp3+ regulatory T cells (Treg. Cellular profiles and histopathologic analysis demonstrated that peribronchial and perivascular inflammatory cell infiltrates were significantly decreased in the EA-treated groups when compared to the OVA and anti-CD25 Ab-injected (Treg depletion groups. Furthermore, total BAL cells were reduced in the EA groups when compared to other groups. Interestingly, the population of CD4+CD25+Foxp3+Tregs in pneumonocytes increased in EA-treated group when compared to OVA and Treg depletion groups. These results imply that EA stimulation at ST 36 may affect CD4+CD25+Foxp3+ Treg in an OVA-induced experimental model and may enhance Treg function by suppressing other T cells and limiting the immune response.

  5. Reactivity of naive CD4+CD25- T cells against gut microflora in healthy mice

    DEFF Research Database (Denmark)

    Gad, Monika; Lundsgaard, Dorthe; Kjellev, Stine;

    2006-01-01

    We have previously shown that conventional as well as germ-free CD4+ T cells depleted of CD25+ cells from the gut-associated lymphoid tissue and the periphery proliferate specifically in response to enterobacterial antigen exposure whereas unfractionated CD4+ T cells are not reactive under these ...

  6. Serum 25-hydroxyvitamin D and CD4+CD25(+high) FoxP3+ Regulatory T cell as Predictors of Severity of Bronchial Asthma in Children.

    Science.gov (United States)

    Ismail, Ahlam M; Aly, Sanaa S; Fayed, Hanan M; Ahmed, Samar S

    2015-01-01

    Bronchial asthma (BA) is one of the common chronic diseases of childhood. Vitamin D deficiency has been associated with BA. Suppressor regulatory T cells (Treg) are important for the induction, maintenance of immunological tolerance to allergens. This study assessed serum 25-hydroxyvitamin D (vitamin D) and the percentages of CD4+CD25+(high) Foxp3+ Treg, in peripheral blood, as predictors of asthma severity and level of clinical control. The study enrolled 72 children divided equally between asthmatic children (AC) and age and sex matched controls. Diagnostic criteria and level of asthma severity followed the Global Initiative for Asthma guidelines. Serum vitamin D was determined by an immunoassay and the percentages of CD4+CD25+ig Foxp3+ Treg by flow cytometry. Serum vitamin D level and percentage of CD4+CD25+(high) Treg were lower in AC compared to controls (P asthma compared to mild and moderate forms (P = 0.008) and in uncontrolled attacks compared to partially or completely controlled children. No difference in percentage of Treg in relation to asthma severity and clinical control was observed. Since AC has decreased serum vitamin D with inverse relationship between its levels and asthma severity, we conclude that it can be used to predict severity of asthma. PMID:26415368

  7. CD4+CD25+Foxp3+调节性T细胞与IL-33在儿童哮喘发病机制中的作用%Roles of CD4+CD25+Foxp3+ regulatory T cells and IL-33 in the pathogenesis of asthma in children

    Institute of Scientific and Technical Information of China (English)

    潘珍珍; 李羚; 郭赟; 贺建

    2014-01-01

    ObjectiveTo study the roles of CD4+CD25+Foxp3+ regulatory T cells (Treg) and IL-33 in the pathogenesis of asthma in children.MethodsFlow cytometry was used to detect peripheral blood CD4+CD25+Foxp3+Treg proportion in CD4+T lymphocytes in.45 children with asthma, 50 children with wheezing caused by respiratory syncytial virus infection and 40 healthy children. Serum levels of IFN-γ, IL-4, IL-5 and IL-33 were measured using ELISA.ResultsThe level of peripheral blood CD4+CD25+Foxp3+Treg in the asthma group was significantly lower than in the wheezing and control groups (P<0.05). In contrast, serum levels of IL-33 in the asthma group was signiifcantly higher than in the wheezing and control groups (P<0.05). Peripheral blood CD4+CD25+Foxp3+Treg level was negatively correlated with serum IL-33 level in the asthma group(r=-0.156,P<0.01). ConclusionsCD4+CD25+Foxp3+Treg may interact with IL-33 in the pathogenesis of childhood asthma.%目的:探讨CD4+CD25+Foxp3+调节性T细胞(Treg)与IL-33在儿童哮喘发病中的作用。方法采用流式细胞仪检测45例哮喘患儿(哮喘组)、50例呼吸道合胞病毒感染喘息患儿(喘息组)及40例健康儿童(对照组)外周血CD4+CD25+Foxp3+Treg细胞百分比,采用ELISA法检测各组外周血血清IFN-γ、IL-4、IL-5及IL-33浓度,进行比较分析。结果哮喘组患儿体内CD4+CD25+Foxp3+Treg 水平较喘息组及对照组均降低(P<0.05);哮喘组患儿体内IL-33水平较喘息组及对照组均升高(P<0.05),哮喘组患儿体内CD4+CD25+Foxp3+Treg与IL-33呈负相关(r=-0.156,P<0.01)。结论在哮喘患儿发病机制中,CD4+CD25+Foxp3+Treg与IL-33可能存在相互作用。                                                                             [中国当代儿科杂志,2014,16(12):1211-1214

  8. Effect of bacillus calmette-guerin on CD+4 CD+25 CDlo127 treg in asthmatic mouse%卡介苗对哮喘小鼠CD+4CD+25CDlo127Treg表达的影响

    Institute of Scientific and Technical Information of China (English)

    葛荣领; 张建华

    2011-01-01

    目的 探讨卡介苗干预对哮喘小鼠调节性T细胞(Treg)及T淋巴细胞凋亡的影响.方法 27只小鼠随机分为3组:哮喘组,干预组,对照组.哮喘组小鼠用卵清蛋白(OVA)、氢氧化铝腹腔注射致敏,OVA雾化吸入激发,干预组在致敏前BCG干预3次,对照组生理盐水代替OVA及氢氧化铝.流式细胞术检测外周血(记为培养前)及植物血凝素(PHA)刺激培养后的外周血单个核细胞中Treg的表达,Comet法检测T淋巴细胞凋亡率.结果 培养前:哮喘组与对照组比较Treg显著降低,干预组与哮喘组比较显著升高.培养后:哮喘组与对照组比较Treg显著降低,干预组与哮喘组比较显著升高.同组小鼠Treg培养前后的比较:哮喘组、干预组表达无改变,对照组表达明显升高.T淋巴细胞凋亡率,哮喘组明显低于对照组,干预组明显高于哮喘组.结论 哮喘小鼠存在Treg数量小足和分化障碍,BCG干预能促进其数量上升,并促进T淋巴细胞凋亡.%Objective To explore the effect of BCG intervention on expression of CD/ CD+4 CD+25 Treg and T lymphocyte apopto-sis in asthmatic mouse. Methods 27 mice were randomly divided into 3 groups, the asthma model group, the treatment group and the normal control group. The mice were sensitized by ovalbumin and AL( OH) 3 with intraperitoneal injection, challenged with atomization inhalation. The treatment group were treated 3 times with BCG subcutaneous injection before sensitization. The normal control group were treated with saline water taking the place of ovalbumin and AL( OH) 3. Treg in peripheral blood and PBMC were detected by flow cytome-try, PBMC were stimulated and cultured by PHA. Counted the apoptosis ratio of T lymphocyte by Comet Assay. Results In peripheral blood: Compared the asthmatic group with the control group, Treg decreased remarkably; Compared the treatment group with the asthmatic group, it increased obviously. In PBMC: Compared the asthmatic group with the

  9. The level of CD4+CD25nt/hiCD127lo regulatory T cells and its clinical significance in children with asthma%哮喘患儿外周血 CD4+CD25nt/hi CD127lo调节性T细胞的测定及临床意义

    Institute of Scientific and Technical Information of China (English)

    湛洁谊; 卢慧敏; 林穗玲

    2016-01-01

    Objective To explore the proportion change of CD4+ CD25nt/hi CD127lo regulatory T cells(Treg)in pe-ripheral blood of children withasthma and to analyze its significance.Methods 150 asthmatic children were divided into three groups according to their clinicalfeatures:50 subjects in acute asthma attack group,50 subjects in clinical remis-sion of asthma group and 50 subjects in cough variant asthma group,meanwhile,50 healthy children were enrolled in the control group.The levels of CD4+ CD25nt/hi CD127lo Treg in peripheral blood of all children were detected by flow cy-tometer.Results The CD4+ CD25nt/hi CD127lo Treg level in acute attack group were lowest of the four groups (P 0.05).Conclusion The CD4+ CD25nt/hi CD127lo regulatory T cells may be involved in the pathogenesis of asthma,The level of CD4+ CD25nt/hi CD127lo regulatory T cells correlated with the severity of asthma in children.%目的:探讨 CD4+ CD25nt/hi CD127lo 调节性 T 细胞在哮喘儿童外周血中的比例改变,并探讨其临床意义。方法150例哮喘患儿按临床表现分为急性发作组(50例)、临床缓解期组(50例)和咳嗽变异性哮喘组(50例),另选择50名健康儿童为正常对照组。应用流式细胞仪检测上述各组外周血 CD4+ CD25nt/hi CD127lo 调节性 T 细胞占 CD4+ T细胞的百分比。结果急性发作期组患儿外周血 CD4+ CD25nt/hi CD127lo 调节性 T 细胞水平较缓解组、咳嗽变异性哮喘组及健康对照组明显下降(P <0.05),而缓解期组及咳嗽变异性哮喘组与健康对照组比较差异无统计学意义(P >0.05)。结论CD4+ CD25nt/hi CD127lo 调节性 T 细胞可能参与了哮喘的发生与发展,哮喘的严重程度可能与 CD4+CD25nt/hi CD127lo 调节性 T 细胞的水平相关。

  10. Electroacupuncture Attenuates Ovalbumin-Induced Allergic Asthma via Modulating CD4+CD25+ Regulatory T Cells

    OpenAIRE

    Youngjoo Kwon; Sung-Hwa Sohn; Gihyun Lee; Youngeun Kim; Hyejung Lee; Minkyu Shin; Hyunsu Bae

    2012-01-01

    A mouse pulmonary hypersensitivity experimental model that mimics human asthma was developed, and electroacupuncture (EA) treatment was shown to reduce allergic inflammatory processes. In addition, we also assessed whether the beneficial effects of EA on allergic asthma could be correlated with CD4+CD25+Foxp3+ regulatory T cells (Treg). Cellular profiles and histopathologic analysis demonstrated that peribronchial and perivascular inflammatory cell infiltrates were significantly decreased in ...

  11. CD4(+)CD25(hi)Foxp3(+) Cells Exacerbate Bleomycin-Induced Pulmonary Fibrosis.

    Science.gov (United States)

    Birjandi, Shirin Z; Palchevskiy, Vyacheslav; Xue, Ying Ying; Nunez, Stefanie; Kern, Rita; Weigt, S Sam; Lynch, Joseph P; Chatila, Talal A; Belperio, John A

    2016-08-01

    Idiopathic pulmonary fibrosis is a fatal lung disease with a median survival of 2 to 5 years. A decade of studies has downplayed inflammation contributing to its pathogenesis. However, these studies preceded the discovery of regulatory T cells (Tregs) and all of their functions. On the basis of human studies demonstrating Tregs can decrease graft-versus-host disease and vasculitides, there is consideration of their use to treat idiopathic pulmonary fibrosis. We hypothesized that Treg therapy would attenuate the fibroplasia involved in a preclinical murine model of pulmonary fibrosis. IL-2 complex was used in vivo to expand CD4(+)CD25(hi)Foxp3(+) cells in the lung during intratracheal bleomycin challenge; however, this unexpectedly led to an increase in lung fibrosis. More important, this increase in fibrosis was a lymphocyte-dependent process. We corroborated these results using a CD4(+)CD25(hi)Foxp3(+) cellular-based therapy. Mechanistically, we demonstrated that CD4(+)CD25(hi)Foxp3(+) cells undergo alterations during bleomycin challenge and the IL-2 complex had no effect on profibrotic (eg, transforming growth factor-β) or type 17 immune response cytokines; however, there was a marked down-regulation of the type 1 and augmentation of the type 2 immune response cytokines from the lungs. Collectively, our animal studies show that a specific lung injury can induce Treg alterations, which can augment pulmonary fibrosis. PMID:27317904

  12. CD4+CD25+调节性T细胞在支气管哮喘中的研究进展%The researches on CD4+CD25+ Treg in bronchial asthma

    Institute of Scientific and Technical Information of China (English)

    李营营; 冯学斌

    2007-01-01

    CD4+CD25+调节性T细胞(CD4+CD25+Treg)是一种表型和功能特异的T细胞亚群,它在抑制自身免疫性疾病的发生,诱导机体免疫耐受机制中发挥着重要作用.研究CD4+CD25+Treg在支气管哮喘中的功能机制,对阐明哮喘的发病机制及探索疾病免疫治疗方面有重要意义.

  13. Detection of CD4+/CD8+T Lymphocyte Ratio and CD4+CD25+ Treg in Peripheral Blood of Patients with Sporadic Vitiligo%散发型白癜风患者外周血CD4+/CD8+T细胞比值及CD4+CD25+T细胞的检测

    Institute of Scientific and Technical Information of China (English)

    叶蓉; 马刚; 胡小平; 彭曦

    2012-01-01

    目的 检测散发型白癜风患者外周血CD4+/CD8+T细胞比值及CD4+CD25+调节性T细胞水平,探讨其与散发型白癜风发病的关系.方法 散发型白癜风患者29例,男13例,女16例.通过流式细胞仪对散发型白癜风患者外周血CD4+/CD8+T细胞比值及CD4+CD25+调节性T细胞水平进行检测,并与20例健康人相比较.结果 与健康对照组相比,散发型白癜风患者外周血中CD4+/CD8+T细胞比值的差异无统计学意义(P>0.05),而CD4+CD25+调节性T细胞水平明显减少,差异有统计学意义(P<0.05),但在不同病程的患者中CD4+CD25+调节性T细胞数量的差异无统计学意义(P>0.05).结论 散发型白癜风患者外周血中存在CD4+CD25+调节性T细胞水平下降,可能与散发型白癜风的发生发展有一定关系.%Objective To detect the CD4+/CD8+ T lymphocyte ratio and the CD4+CD25+ Treg level in peripheral blood of patients with sporadic vitiligo, and to investigate its role in the pathogenesis of sporadic vitiligo. Methods Peripheral blood samples were taken from 29 outpatients with sporadic vitiligo, including 13 males and 16 females. The CD4+/CD8+ T lymphocyte ratio and the CD4+CD25+ Treg level was detected in peripheral blood of patients with sporadic vitiligo by flow cytometry, as well as controlled samples from 20 healthy human. Results There was no difference on the ratio of CD4+/CD8+ T lymphocyte between the patients with sporadic vitiligo and healthy people (P>0.05). Compared to the controlled group, the proportion of CD4+CD25+ Treg was significantly lower in sporadic vitiligo patients(P0.05). Conclusion The level of CD4+CD25+ Treg is lower in peripheral blood of sporadic vitiligo patients, which might play a role in the pathogenesis and development of sporadic vitiligo.

  14. 老年脓毒症休克患者CD4+CD25+调节性T细胞的变化及对预后的影响%Effects of changes of CD4+ CD25+ regulatory T cells on prognosis in elderly patients with septic shock

    Institute of Scientific and Technical Information of China (English)

    呼邦传; 孙仁华; 徐云祥; 杨向红; 李茜; 韩芳

    2010-01-01

    the 1st day (1.76 % ±0.31% vs. 1.68 %±0.24 %, P>0.05)and the 4th day (1.94%±0.32% vs. 1.82% ±0.28%, P>0.05). However, compared with the survivors, non-survivors had a higher percentage of CD4+ CD25+ FoxP3/CD4+ T cells (2.65%±0.28% vs. 1.79%±0.27%, P<0.01) at the 7-10th day of septic shock after being diagnosed.Furthermore, from the 4th day to the 7-10th day, the expressions of monocyte HLA-DR in the nonsurvivors were significantly lower than in the survivors (P<0. 01), and they were inversely correlated with the percentage of CD4+ CD25+ FoxP3/CD4+ T cells at the 4th day (r=-0.39, P=0.023) and the 7-10th day (r= -0. 58, P<0. 01) respectively. The multiple logistic regression analysis showed that the percentages of CD4+ CD25+ FoxP3/CD4+ T cells (OR = 3.47, 95% CI: 1.33-10.0) and HLA-DR (OR= 0. 27, 95% CI: 0.14-0.73) were independent predictors of 28-day mortality rate.Conclusions Persistent higher percentage of CD4+ CD25+ Treg cells in the elderly patients with septic shock indicates that the patients are under the states of immunosuppression and have a higher risk ofmortality in intensive care unit at admission.

  15. 抗CD3单克隆抗体对人CD4+CD25+T淋巴细胞凋亡和自噬的影响%Apoptosis and autophagy induced by anti-CD3 monoclonal antibody in human peripheral blood CD4+CD25+T lymphocytes

    Institute of Scientific and Technical Information of China (English)

    邬伟明; 洪国斌; 徐晓华

    2009-01-01

    目的 观察抗CD3单克隆抗体对分离培养的人外周血CD4+CD25+T淋巴细胞自噬、凋亡及其分泌的代表性因子转化生长因子β(TGF-β)的影响.方法 采用密度梯度离心法及尼龙棉柱法分离32例健康者外周血T淋巴细胞,磁性细胞分离器(MACS)分离得到CD4+CD25+T淋巴细胞,分别利用电镜及流式细胞仪观察、检测各组(抗CD3单克隆抗体1 mg/L组、IgG1同型抗体对照组)干预72 h后细胞的凋亡率、自噬率,用ELISA法检测细胞培养上清液中细胞因子TGF-β的水平.结果 抗CD3单克隆抗体组CD4+CD25+T淋巴细胞自噬率、凋亡率及TGF-β水平均增加(均P0.05).结论 抗CD3单克隆抗体可促进CD4+CD25+T淋巴细胞凋亡和自噬及TGF-β分泌,且自噬与凋亡间相互独立.%Objective To observe the autophagy,apoptosis rate and TGF-β secretion in human peripheral blood CD4+CD25+T lymphocytes induced by anti-CD3 monoclonal antibody.Methods Peripheral blood T lymphocyte of 32 healthy adults were separated by Percoll (1.073 g/ml) and harvested by using nylon column.Then,CD4+CD25+T lymphocytes were isolated by (magnetic cell sorting MACS).The cultured cells were divided into different groups (anti-CD3 mAb group 1 mg/L,IgG1 isotype control group).The apoptosis and autophagy rate of the cells cultured for 72 h were analyzed by electron microscope and flow cytometry.The cell culture supernatants were collected after 72 h and analyzed for levels of TGF-β by ELISA.Results The autophagy,apoptosis rate and TGF-β secretion in human peripheral CD4+CD25+T lymphocytes of the health group were increased by anti-CD3 mAb(all P<0.01);There was no significant statistic difference in correlation between apoptosis and autophagy of T lmyphocytes.Conclusions Anti-CD3 mAb could increase the autophagy rate,apoptosis rate and TGF-β secretion of peripheral blood CD4+CD25+Tregs,and there was no significant statistic difference between autophagy and apoptosis of CD4+CD25+Tregs.

  16. The potential role of cell surface complement regulators and circulating CD4+ CD25+ T-cells in the development of autoimmune myasthenia gravis

    OpenAIRE

    Hamdoon, Mohamed Nasreldin Thabit; Fattouh, Mona; El-din, Asmaa Nasr; Elnady, Hassan M.

    2016-01-01

    Introduction CD4+CD25+ regulatory T-lymphocytes (T-regs) and regulators of complement activity (RCA) involving CD55 and CD59 play an important role in the prevention of autoimmune diseases. However, their role in the pathogenesis of human autoimmune myasthenia gravis (MG) remains unclear. This study aimed to determine the frequency of peripheral blood T-regs and CD4+ T-helper (T-helper) cells and the red blood cells (RBCs) level of expression of CD55 and CD59 in MG patients. Methods Fourteen ...

  17. Increased cytotoxicity of CD4+ invariant NKT cells against CD4+CD25hiCD127lo/− regulatory T cells in allergic asthma

    OpenAIRE

    Nguyen, Khoa D.; Vanichsarn, Chris; Nadeau, Kari C.

    2008-01-01

    CD4+CD25hiCD127lo/− regulatory T cells (Treg) have been implicated in the resolution of asthma-associated inflammation while the opposite role of CD4+ invariant NKT (iNKT) cells has been the subject of recent investigations. Studies here focused on mechanisms of interaction between CD4+ iNKT cells and Treg to further explore their roles in allergic asthma (AA). Flow cytometry analysis revealed a significant increase in the expression of the natural cytotoxicity receptors NKp30 and NKp46 by CD...

  18. XCL1 Enhances Regulatory Activities of CD4+CD25highCD127low/− T Cells in Human Allergic Asthma1

    OpenAIRE

    Nguyen, Khoa D.; Fohner, Alison; Booker, Jerome D.; Dong, Chen; Krensky, Alan M.; Nadeau, Kari C.

    2008-01-01

    Chemokine-mediated recruitment of regulatory cell subsets to the airway during inflammation and enhancement of their activities are potential strategies for therapeutic development in allergic asthma (AA). In this study, we aim to explore the role of XCL1, a chemokine associated with immune suppression and allergy, on CD4+CD25highCD127low/− regulatory T cell (Treg) function in AA. Flow cytometry and PCR analysis showed a reduction in XCL1 and XCR1 expression in AA Treg compared with healthy c...

  19. Differences of CD4+CD25+ regulatory T cells between food allergy mice and normal mice%食物过敏与正常小鼠CD4+CD25+调节性T细胞的差异性研究

    Institute of Scientific and Technical Information of China (English)

    程茜; 陈玉梅

    2013-01-01

    Objective:To compare the quantities and functions of CD4+CD25+ regulatory T(Treg)cells between food allergy mice model and normal mice model. Methods: SPF Balb/c female mice of 4-6 weeks on egg-free diet were randomly divided into ovalbumin (OVA) group(n=8) and normal control group(n=8). Quantitative changes of CD4+CD25+ Treg cells in the spleen mononuclear cell suspension were analyzed by flow cytometry. Levels of serum interleukin-lO(IL-lO) and transforming growth factor-β1(TGF-βl) in OVA mice and normal control mice were detected by enzyme-linked immunosorbent assay (ELISA). Results: Percentage of CD4+ CD25+ Treg cells in OVA group was lower than that in normal control group (P<0.05 ), and percentage of forkhead transcription factor3+(Foxp3 + )CD4+CD25 + Treg cells in OVA group was significantly lower in OVA group than in normal control group(P<0.01). Levels of IL-10 and TGF-β1 were lower in OVA group than in normal control group(P<0.05)(P<0.01). Conclusions:There are differences in quantities and functions of Treg cells between food allergy and normal mice.%目的:比较食物过敏(food allergy,FA)小鼠与正常小鼠模型CD4+CD25+调节性T(regulatory T,Treg)细胞数量及功能差异.方法:4~6周龄SPF级无鸡蛋喂养Balb/c雌鼠随机分为2组,每组8只.分别为卵清蛋白(ovalbumin,OVA)致敏组和正常对照组.采用流式细胞术分析其脾脏单个核细胞悬液中CD4+CD25+ Treg细胞的数量变化;酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测OVA致敏组与正常对照组血清白介素10(interleukin-10,IL-10)和转化生长因子-β1 (transforming growth factor-β1,TGF-β1)水平.结果:OVA组CD4+CD25+T淋巴细胞的百分比低于正常对照组(P<0.05),并且叉状头转录因子3阳性(forkhead transcription factor3+,Foxp3+)CD4+CD25+Treg细胞的百分比显著低于正常对照组(P<0.01);OVA组IL-10低于正常对照组(P<0.05),TGF-β1水平显著低于正常对照组(P<0.01).结论:FA

  20. Nanoscale Relationship Between CD4 and CD25 of T Cells Visualized with NSOM/QD-Based Dual-Color Imaging System

    Science.gov (United States)

    Fan, Jinping; Lu, Xiaoxu; Liu, Shengde; Zhong, Liyun

    2015-10-01

    In this study, by using of near-field scanning optical microscopy (NSOM)/immune-labeling quantum dot (QD)-based dual-color imaging system, we achieved the direct visualization of nanoscale profiles for distribution and organization of CD4 and CD25 molecules in T cells. A novel and interesting finding was that though CD25 clustering as nanodomains were observed on the surface of CD4+CD25high regulatory T cells, these CD25 nanodomains were not co-localized with CD4 nanodomains. This result presented that the formation of these CD25 nanodomains on the surface of CD4+CD25high T cells were not associated with the response of T cell receptor (TCR)/CD3-dependent signal transduction. In contrast, on the surface of CD4+CD25low T cells, CD25 molecules distributed randomly without forming nanodomains while CD4 clustering as nanodomains can be observed; on the surface of CD8+CD25+ T cells, CD25 clustering as nanodomains and co-localization with CD8 nanodomains were observed. Collectively, above these results exhibited that TCR/CD3-based microdomains were indeed required for TCR/CD3-mediated T cells activation and enhanced the immune activity of CD4+CD25low T cells or CD8+CD25+ T cells. In particular, it was found that the formation of CD25 nanodomains and their segregation from TCR/CD3 microdomains were the intrinsic capability of CD4+CD25high T cells, suggesting this specific imaging feature of CD25 should be greatly associated with the regulatory activity of CD4+CD25high T cells. Importantly, this novel NSOM/QD-based dual-color imaging system will provide a useful tool for the research of distribution-function relationship of cell-surface molecules.

  1. Immunity to experimental Salmonella typhimurium infections in rats. Transfer of immunity with primed CD4+CD25high and CD4+CD25low T lymphocytes

    DEFF Research Database (Denmark)

    Thygesen, P; Brandt, L; Jørgensen, T;

    1994-01-01

    The protective effect of primed CD4+ T lymphocytes against a lethal dose of 10(8) viable Salmonella typhimurium was studied in Lewis rats. Primed CD4+ T lymphocytes were obtained by inoculating Lewis rats with a non-lethal dose of 10(6) viable S. typhimurium. Four weeks after the infection, spleen...... fluorescence-activated cell sorter. Untreated Lewis rats were injected with 10(4) different primed CD4+ T-cell populations 24 h prior to the lethal dose of 10(8) viable S. typhimurium. Blood samples were drawn from the orbital plexus 1, 2, 3, and 4 weeks after the infection, and analysed for specific IgM and...... levels were not correlated with protection against S. typhimurium infections, although it showed that a higher and more persistent level of specific IgG antibodies was produced in animals receiving the CD4+CD25high fraction. It is concluded that 10(4) primed CD4+ T lymphocytes can induce immunity in...

  2. Inhibiting effect of Astragalus polysaccharides on the functions of CD4+CD25highTreg cells in the tumor microenvironment of human hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    LI Qiang; BAO Jian-min; LI Xiao-li; ZHANG Ti; SHEN Xiao-hong

    2012-01-01

    Background Astragalus polysaccharides (APS),the main active extract from Astragalus membranaceus (a traditional Chinese medicinal herb),is associated with a variety of immunomodulatory activities.The purpose of the present study was to examine the effect of APS on the function of Treg cells in the tumor microenvironment of human hepatocellular carcinoma (HCC) and to identify the pharmacologic mechanism of APS responsible for the anti-chemotactic activity in CD4+CD25highTreg cells in tumor site of HCC.Methods The prevalence of Treg in fresh tissue samples from 31 patients with HCC after radicalhepatectomy was detected.CD4,CD25 and CD127 were selected as Treg cell makers to phenotype cell populations.The expression of FOXp3 mRNA was also analyzed.The migration and proliferation of Treg cells were observed.Interleukin (IL)-4,IL-10,IFN-γ and SDF-1 in cell supernatant were detected.For all tests,functions of Treg cells were evaluated after treatment with APS.Results APS can inhibit the growth and proliferation of CD4+CD25+Treg cells in vitro in a dose- and time-dependent manner.APS may inhibit CD4+CD25+Treg cells through restoring the cytokine imbalance and reducing the expression of FOXp3 in local HCC microenvironments.SDF-1 played an important role in there recruitment of Treg cells into the tumor microenvironment of HCC.APS might have inhibiting effects on Treg cell migration by blocking SDF-1 or its receptor through the CXCR4/CXCL12 pathway.Conclusions The increase in numbers of tumor associated Treg cells might play a role in modulation of the immune response against HCC.APS can restore the cytokine balance in the tumor micro environment and suppress the expression of FOXp3 mRNA to inhibit the immune suppressive effects of Treg cells.The application of APS in the tumor microenvironment might act to enhance the anti-tumor effects of the immunotherapy-based methods,and consequently to increase the survival rate in HCC.

  3. Characterization of protective human CD4CD25 FOXP3 regulatory T cells generated with IL-2, TGF-β and retinoic acid.

    Directory of Open Access Journals (Sweden)

    Ling Lu

    Full Text Available BACKGROUND: Protective CD4+CD25+ regulatory T cells bearing the Forkhead Foxp3 transcription factor can now be divided into three subsets: Endogenous thymus-derived cells, those induced in the periphery, and another subset induced ex-vivo with pharmacological amounts of IL-2 and TGF-β. Unfortunately, endogenous CD4+CD25+ regulatory T cells are unstable and can be converted to effector cells by pro-inflammatory cytokines. Although protective Foxp3+CD4+CD25+ cells resistant to proinflammatory cytokines have been generated in mice, in humans this result has been elusive. Our objective, therefore, was to induce human naïve CD4+ cells to become stable, functional CD25+ Foxp3+ regulatory cells that were also resistant to the inhibitory effects of proinflammatory cytokines. METHODOLOGY/PRINCIPAL FINDINGS: The addition of the vitamin A metabolite, all-trans retinoic acid (atRA to human naïve CD4+ cells suboptimally activated with IL-2 and TGF-β enhanced and stabilized FOXP3 expression, and accelerated their maturation to protective regulatory T cells. AtRA, by itself, accelerated conversion of naïve to mature cells but did not induce FOXP3 or suppressive activity. The combination of atRA and TGF-β enabled CD4+CD45RA+ cells to express a phenotype and trafficking receptors similar to natural Tregs. AtRA/TGF-β-induced CD4+ regs were anergic and low producers of IL-2. They had potent in vitro suppressive activity and protected immunodeficient mice from a human-anti-mouse GVHD as well as expanded endogenous Tregs. However, treatment of endogenous Tregs with IL-1β and IL-6 decreased FOXP3 expression and diminished their protective effects in vivo while atRA-induced iTregs were resistant to these inhibitory effects. CONCLUSIONS/SIGNIFICANCE: We have developed a methodology that induces human CD4(+ cells to rapidly become stable, fully functional suppressor cells that are also resistant to proinflammatory cytokines. This methodology offers a practical

  4. The level and significance of CD4 + CD25 + Foxp3 + regulation T cells in leprosy%CD4+CD25+Foxp3+调节性T细胞在麻风病中的水平和意义

    Institute of Scientific and Technical Information of China (English)

    李彩霞; 徐元品; 邹子宏; 周晓鸿

    2013-01-01

    Objective: To detect the expression and significance of CD4+ CD25 + Foxp3 + regulation T cells in leprosy. Methods; The leprosy patients were 51, the cured leprosy people were 26, the ENL were 5, and the normal persons were 56. We used the flow cytometry to test the Tregs in four groups above mentioned. And use the SPSS 19.0 to analysis the results. Results;The percentage of Tregs in leprosy patients, cured people, ENL patients and normal persons were (17.626 ±8.1977)% ,(38.442 ±4.7618)% ,(6. 380 ± 1.5482) % and (9.998 ± 1.7062) %. The Tregs in cured people and leprosy were higher than normal and the Tregs in cured people were higher than leprosy patients, (P < 0.05); The Tregs in ENL were lower than normal; Among leprosy patients(P < 0.05 ) , the Tregs in PB was higher than MB, (P<0.05). Conclusion;Tregs may break the immune tolerance and related to the leprosy.%目的:探讨CD4+ CD25+ Foxp3+调节性T细胞(Treg)在麻风病发病中的作用.方法:采用流式细胞仪检测51例现症麻风患者,5例发生Ⅱ型麻风反应(ENL)的患者,26例治愈麻风患者及56例正常体检者外周血CD4+ CD25+ Foxp3+调节性T细胞(Treg)占CD4+的百分比.结果:现症组、治愈组、Ⅱ型麻风反应(ENL)组及正常对照组外周血Treg的水平分别为:(17.626±8.1977)%、(38.442±4.761 8)%、(6.380±1.548 2)%和(9.998±1.706 2)%.治愈组与现症组麻风患者的Treg高于正常对照组,并且治愈组的Treg比现症组高(P<0.05),ENL组的Treg比正常对照组低(P<0.05);在现症麻风病人中,少菌型患者(PB)及多菌型患者(MB) Treg的百分比分别为:(29.629±7.999 5)%和(15.709±6.4809)%,均高于正常对照组(9.998±1.706 2)% (P <0.05),并且少菌型患者(PB)的Treg高于多菌型患者(MB) (P <0.05).结论:治愈组与现症组麻风患者的Treg均高于正常对照组,发生Ⅱ型麻风反患者(ENL组)的Treg比正常对照组低,Treg可能打破了外周免疫耐受,参与了麻风病的发生发展.

  5. 支气管哮喘患儿CD4+ CD25+ Foxp3+ IL-10+ T淋巴细胞水平变化和意义%Changes and significances of CD4 + CD25 + FOXP3 + IL-10 + T cells in children with asthma

    Institute of Scientific and Technical Information of China (English)

    王炜; 李芳; 段国威; 吴晓玲; 宋青

    2013-01-01

    目的:探讨对尘螨过敏的轻度持续支气管哮喘患儿淋巴细胞增殖情况及CD4+ CD25+ Foxp3+ IL-10+T淋巴细胞水平变化.方法:选择30例对尘螨过敏的轻度持续支气管哮喘患儿作为实验组,同时选择30例健康儿童作为对照组.分离所有入选者的外周血淋巴细胞,给予户尘螨浸出液刺激48 h后检测淋巴细胞增殖的刺激指数以及CD4+ CD25+ Foxp3+ IL-10+T淋巴细胞水平变化.结果:支气管哮喘患儿外周血淋巴细胞接受户尘螨浸出液刺激后淋巴细胞增殖的刺激指数显著高于对照组(P<0.01),支气管哮喘患儿CD4+ CD25+ Foxp3+ IL-10+T淋巴细胞水平显著低于对照组(P<0.01).结论:支气管哮喘患儿接受过敏原刺激后发生明显的增殖反应,支气管哮喘患儿存在CD4+ CD25+ Foxp3+ IL-10+T淋巴细胞功能障碍.%Objective: To explore the proliferation of T lymphocytes and the changes of CD4 + CD25 + Foxp3+ IL - 10 + T lymphocytes in children allergic to house dust mite with mild persistent asthma. Methods: Thirty children allergic to house dust mite ( HDM) with mild persistent asthma were selected as experimental group and 30 healthy children were selected as control group. Peripheral blood mononu-clear cells (PBMCs) were isolated from the subjects. After 48 hours of in vitro stimulation with HDM extracts, the stimulation indexes (SI) of lymphocytes and the changes of CD4 + CD25 + Foxp3+ IL - 10+ T lymphocytes were measured by flow cytometry. Results: SI of lymphocytes increased significantly in asthma group compared with control group, there was statistically significant difference ( P < 0. 001) . The level of CD4+ CD25+ Foxp3+ IL — 10+ T lymphocytes in asthma group was statistically significantly lower than that in control group (P < 0.01) . Conclusion: T lymphocyte proliferation obviously can be observed in asthmatic children when they were stimulated by ,HDM extract. There is functional insufficiency of CD4+ CD25+ Foxp3+ IL - 10+ T

  6. Detection of interleukin-10 and transforming growth factor-β1 in the culture supernatant of CD4+CD25+ T cells from patients with alopecia areata%斑秃患者外周血CD4+CD25+T细胞培养上清液白介素10和转化生长因子β1检测

    Institute of Scientific and Technical Information of China (English)

    马新华; 邵文俊; 金宛宛; 高宇

    2014-01-01

    Objective To evaluate the potential association of CD4+CD25+ T cells with alopecia areata.Methods Totally,this study enrolled 23 patients with progressive alopecia areata,25 patients with stable alopecia areata,and 25 healthy controls.Peripheral blood was isolated from these subjects followed by isolation of CD4+ CD25+ regulatory T cells,which were then cuhured with the presence of anti-CD3 and-CD28 monoclonal antibodies for four days.Subsequently,enzyme-linked immunosorbent assay was performed to measure the levels of interleukin (IL)-10 and transforming growth factor (TGF)-β1 in the culture supematant of these T cells.Results The levels of IL-10 and TGF-β1 were (31.68 ± 6.78) pg/ml and (32.29 ± 6.8) pg/ml respectively in the culture supernatant of CD4+CD25+ regulatory T cells from patients with progressive alopecia areata,significantly lower than those from the healthy controls ((57.34 ± 14.15) pg/ml and (57.43 ± 15.16) pg/ml,both P < 0.05) and patients with stable alopecia areata ((52.56 ± 13.02) pg/ml and (61.75 ± 14.10) pg/ml,both P < 0.05).However,no significant difference was observed in the supernatant levels of IL-10 or TGF-β1 between the healthy controls and patients with stable alopecia areata.Conclusions The secretion of IL-10 and TGF-β1 by CD4+CD25+ T cells is decreased in patients with progressive alopecia areata,which may contribute to the pathogenesis of alopecia areata.%目的 探讨CD4+CD25+T细胞与斑秃发病之间的关系.方法 收集了3组研究对象,其中健康对照组25例、稳定期斑秃患者25例、进展期斑秃患者23例.抽取所有对象外周血,提取CD4+CD25+T细胞,培养4d,收集培养上清液,ELISA法检测上清液IL-10和TGF-β1水平.结果 进展期斑秃患者外周血CD4+CD25+T细胞培养的IL-10和TGF-β1分别为(31.68±6.78) pg/ml和(32.29±6.80) pg/ml,明显低于健康对照组(57.34±14.15) pg/ml、(57.43±15.16) pg/ml和稳定期斑秃患者(52.56±13.02) pg/ml和(61.75±14.10) pg

  7. Naturally Occurring Self-Reactive CD4+CD25+ Regulatory T Cells: Universal Immune Code

    Institute of Scientific and Technical Information of China (English)

    Nafiseh Pakravan; Agheel Tabar Molla Hassan; Zuhair Muhammad Hassan

    2007-01-01

    Naturally occurring thymus-arisen CD4+CD25+ regulatory T (Treg) cells are considered to play a central role in self-tolerance. Precise signals that promote the development of Treg cells remain elusive, but considerable evidence suggests that costimulatory molecules, cytokines, the nature of the TCR and the niche or the context in which the T cell encounters antigen in the thymus play important roles. Analysis of TCR from Treg cells has demonstrated that a large proportion of this population has a higher avidity to self-antigen in comparison with TCR from CD4+CD25- cells and that peripheral antigen is required for their development, maintenance, or expansion. Treg cells have been shown to undergo expansion in the periphery, likely regulated by the presence of self-antigen. Many studies have shown that the involvement of Treg cells in the tolerance induction is antigen-specific, even with MHC-mismatched,in transplantation/graft versus host disease (GVHD), autoimmunity, cancer, and pregnancy. Theses studies concluded a vital role for self-reactive Treg cells in maintenance of the body integrity. Based on those studies, we hypothesize that self-reactive Treg cells are shared among all healthy individuals and recognize same self-antigens and their TCR encodes for few dominant antigens of each organ which defines the healthy self. These dominant self antigens can be regarded as "universal immune code".

  8. Plasmodium falciparum-mediated induction of human CD25Foxp3 CD4 T cells is independent of direct TCR stimulation and requires IL-2, IL-10 and TGFbeta.

    Directory of Open Access Journals (Sweden)

    Anja Scholzen

    2009-08-01

    Full Text Available CD4(+CD25(+Foxp3(+ regulatory T cells (Tregs regulate disease-associated immunity and excessive inflammatory responses, and numbers of CD4(+CD25(+Foxp3(+ Tregs are increased during malaria infection. The mechanisms governing their generation, however, remain to be elucidated. In this study we investigated the role of commonly accepted factors for Foxp3 induction, TCR stimulation and cytokines such as IL-2, TGFbeta and IL-10, in the generation of human CD4(+CD25(+Foxp3(+ T cells by the malaria parasite Plasmodium falciparum. Using a co-culture system of malaria-infected red blood cells (iRBCs and peripheral blood mononuclear cells from healthy individuals, we found that two populations of Foxp3(hi and Foxp3(int CD4(+CD25(hi T cells with a typical Treg phenotype (CTLA-4(+, CD127(low, CD39(+, ICOS(+, TNFRII(+ were induced. Pro-inflammatory cytokine production was confined to the Foxp3(int subset (IFNgamma, IL-4 and IL-17 and inversely correlated with high relative levels of Foxp3(hi cells, consistent with Foxp3(hi CD4 T cell-mediated inhibition of parasite-induced effector cytokine T cell responses. Both Foxp3(hi and Foxp3(int cells were derived primarily from proliferating CD4(+CD25(- T cells with a further significant contribution from CD25(+Foxp3(+ natural Treg cells to the generation of the Foxp3(hi subset. Generation of Foxp3(hi, but not Foxp3(int, cells specifically required TGFbeta1 and IL-10. Add-back experiments showed that monocytes expressing increased levels of co-stimulatory molecules were sufficient for iRBC-mediated induction of Foxp3 in CD4 T cells. Foxp3 induction was driven by IL-2 from CD4 T cells stimulated in an MHC class II-dependent manner. However, transwell separation experiments showed that direct contact of monocytes with the cells that acquire Foxp3 expression was not required. This novel TCR-independent and therefore antigen-non specific mechanism for by-stander CD4(+CD25(hiFoxp3(+ cell induction is likely to reflect a

  9. Th17细胞与Treg细胞在支气管哮喘发病机制中的研究进展%Th17 cell and CD4 + CD25 + Treg in iathogenesis of asthma

    Institute of Scientific and Technical Information of China (English)

    姚斌

    2012-01-01

    近年来,众多研究发现支气管哮喘的发病机制已不能单纯用Th1/Th2平衡理论来解释,CD4+ CD25+调节性T细胞和Th17细胞及其细胞因子IL-10、IL-17、转化生长因子-β等与支气管哮喘发病明显相关.由于Th17细胞与CD4+ CD25+调节性T细胞在功能上相互拮抗,而在分化上密切相关,因此这两种细胞的免疫失衡也是支气管哮喘发病的重要原因.糖皮质激素可通过维甲酸相关孤核受体γt信号途径降低IL-17的表达,还可以通过诱导转录因子Foxp3的表达调控CD4+ CD25+调节性T细胞的分化和功能.%Many studies have suggested that pathogenesis of asthma could no longer be interpreted merely by “Th1/Th2 balance” theory.CD4 + CD25 + Treg and Th17 cells,as well as their cytokines such as IL-10,transforming growth factor-β,and IL-17,account for asthma.CD4 + CD25 + Treg and Th17 are functionally antagonistic to each other,and also go with each other during their differentiation.Therefore,immunity-unbalance of CD4 + CD25 + Treg and Th17 is one of the most important factors that triggers asthma.Glucocorticoid has been shown to down regulate IL-17 expression by retinoic acid receptors γt signaling pathway,and regulate differentiation and function of CD4 + CD25 + Treg by inducing expression of transcription factor Foxp3,all of these are immuno-mechanisms of glucocorticoid in asthma treatment.

  10. CD4+CD25+Foxp3+ T regulatory cells, Th1 (CCR5, IL-2, IFN-γ) and Th2 (CCR4, IL-4, Il-13) type chemokine receptors and intracellular cytokines in children with common variable immunodeficiency.

    Science.gov (United States)

    Kutukculer, Necil; Azarsiz, Elif; Aksu, Guzide; Karaca, Neslihan Edeer

    2016-06-01

    Common variable immunodeficiency (CVID) is a heterogeneous group of primary antibody deficiencies characterized by decreased serum immunoglobulin G along with a decrease in serum IgA and/or IgM, defective specific antibody production, and recurrent bacterial infections. Abnormal lymphocyte trafficking, dysregulated cellular responses to chemokines, and uncontrolled T cell polarization may be involved in the pathogenesis and may help to understand the clinical complications. We evaluated T helper cell subsets (chemokine receptors CCR4, CCR5, and CCR7), expressions on T lymphocytes, intracellular cytokines - IL-2, IL-4, IL-13, IFN- γ-on CD4(+) T cells, and expression of CD4(+)CD25(+)Foxp3(+) regulatory T cells of 20 CVID patients and 26 healthy controls. Autoimmune clinical findings and other complications were also determined. Percentages and absolute numbers of CD4(+)CD25(+) Foxp3(+) cells did not show any significant difference between CVID cases and healthy controls nor between severe and moderate disease patients. The only significant difference regarding Th1 and Th2 type intracellular cytokines was the decreased absolute numbers of CD3(+)CD4(+)IL4(+) cells in CVID cases. There were some findings about T helper cell type dominance in CVID patients such as positive correlation between hepatomegaly and high IL-2 and IFN-γ in CD3(+)CD4(+) cells and very high expression of CCR5 (Th1) on CD3(+)CD4(+) cells in patients with granuloma. Th1 (CCR5) and Th2 (CCR4) type chemokine receptors did not show any dominance in CVID cases. However, frequencies of CCR7 expressing CD3(+) T cells, CD3(+)CD4(+) T helper cells and CD3(+)CD8(+) T cytotoxic cells were significantly lower in severe CVID patients. In addition, presence of autoimmune clinical findings was negatively correlated with CCR7(+) cells. As CCR7 is a key mediator balancing immunity and tolerance in the immune system, the abnormality of this mediator may contribute to the profound immune dysregulation seen in CVID

  11. CD4+CD25+ regulatory T cells: I. Phenotype and physiology

    DEFF Research Database (Denmark)

    Holm, Thomas Lindebo; Nielsen, Janne; Claesson, Mogens H

    2004-01-01

    it has become increasingly clear that regulatory CD4+CD25+ T cells (Treg cells) play an important role in the maintenance of immunological self-tolerance, and that this cell subset exerts its function by suppressing the proliferation or function of autoreactive T cells. Based on human and murine...... observations, this review presents a characterization of the phenotype and functions of the Treg cells in vitro and in vivo. An overview of the surface molecules associated with and the cytokines produced by the Treg cells is given and the origin, activation requirements and mode of action of the Treg cells...... are discussed. Finally, we address the possibility that Treg cells may play a central role in immune homeostasis, regulating not only autoimmune responses, but also immune responses toward foreign antigens....

  12. CD4+CD25+ regulatory T cells: II. Origin, disease models and clinical aspects

    DEFF Research Database (Denmark)

    Nielsen, Janne; Holm, Thomas Lindebo; Claesson, Mogens H

    2004-01-01

    Autoimmune diseases afflict approximately 5% of the population and reflect a failure in the immune system to discriminate between self and non-self resulting in the breakdown of self-tolerance. Regulatory CD4+CD25+ T cells (Treg cells) have been shown to play an important role in the maintenance of...... immune homeostasis and self-tolerance by counteracting the development and effector functions of potentially autoreactive T cells. We have in the previous APMIS review described the phenotype and physiology of Treg cells. The present overview deals with the thymic origin of Treg cells and their role in...... disease models such as autoimmune gastritis and inflammatory bowel disease. Finally, we will consider some aspects of the therapeutic potential of Treg cells....

  13. CD4+CD25+T调节细胞及Foxp3在哮喘中的研究进展%Research Progress of CD4+CD25+T Regulatory Cells and Foxp3 in Children with Asthma

    Institute of Scientific and Technical Information of China (English)

    王娜; 孔宪明; 曹兰芳

    2006-01-01

    CD4+CD25+T调节细胞是具有免疫调节(或抑制)作用的T细胞,其主要功能是抑制自身反应性T细胞的活化和增殖,参与自身免疫调节,维持自身免疫耐受,在儿童哮喘等疾病中发挥重要作用.Foxp3已作为CD4+CD25+T调节细胞的特异性标志.本文就CD4+CD25+T调节细胞及Foxp3的生物学特性、效应机制及其与儿童哮喘发病的关系作一综述.

  14. CD25 signaling regulates the function and stability of peripheral Foxp3+ regulatory T cells derived from the spleen and lymph nodes of mice.

    Science.gov (United States)

    Wang, Kunpeng; Gu, Jian; Ni, Xuhao; Ding, Zheng; Wang, Qi; Zhou, Haoming; Zheng, SongGuo; Li, Bin; Lu, Ling

    2016-08-01

    Regulatory T cells (Tregs) play a critical role in sustaining immune tolerance and maintaining immune balance to alloantigen after transplatation. However, the functions of peripheral Tregs in different organs have not been fully characterized. Here, we showed that spleen-derived Tregs exhibited higher expression of Foxp3, greater suppressive capacity, and lower levels of IL-17A secretion than lymph node-derived Tregs in vitro in the presence or absence of inflammatory cytokines, such as IL-6. We found a higher percentage of CD25(bright) Tregs among spleen-derived Tregs than among lymph node-derived Tregs. Additionally, in vivo experiments demonstrated that adoptive transfer of spleen-derived Tregs, but not lymph node-derived Tregs, alleviated ischemia-reperfusion injury. These results reveal novel functions of Tregs derived from peripheral organs. In particular, spleen-derived Tregs, primarily consisting of CD25(bright) cells, may provide a more significant contribution to the suppression of immune-mediated autoimmune and inflammatory disease. PMID:27344615

  15. 不同剂量粉尘螨提取液对哮喘小鼠CD4+CD25+调节性T细胞的影响%Effect of Dermatophagoides farinae extract at various dosages on CD4+CD25+ regulatory T cells of mice with asthma

    Institute of Scientific and Technical Information of China (English)

    吴雪郡; 黄英; 韩洁; 王模奎; 王莹; 王莉佳

    2012-01-01

    Objective To investigate the effect of Dermatophagoides farinae (Derf) extract at various dosages on CD4+CD25+ regulatory T cells (Tregs) of mice with asthma and optimize the maintenance dosage of the extract for immune therapy. Methods Mouse model of anaphylactic asthma was established by injecting i. p. CS7BL / 6 mice with the extract of Derf. Thirty-two model mice were divided into one control and three test groups. The mice in test groups 1, 2 and 3 were injected i. p. with Der f at low (100μg/ mouse), moderate (1 mg/mouse) and high (2 mg/mouse) dosages respectively, while those in control groups with physiological saline. The contents of CD4+CD25+ T cells and Foxp3+CD4+CD25+Tregs in splenocytes of mice were determined by flow cytometry, while the IL-10 and TGF-β1 levels in sera by ELISA. Results The percentages of CD4+CD25+T cells in CD4+T cells and the percentages of Foxp3+CD4+CD25+Tregs in CD4+CD25+ T cells as well as serum IL-10 and TGF-β1 levels were significantly lower in control group and test group 1 than in test groups 2 and 3 (each P 0. 05). Conclusion The optimal maintenance dosage of Der f extract for specific immune therapy of asthma in mice was 1 mg/ mouse.%目的 探讨不同剂量粉尘螨提取液对哮喘小鼠CD4+CD25调节性T细胞(Regulatory T cells,Tregs)的影响,确定粉尘螨提取液免疫治疗的最佳维持剂量.方法 用粉尘螨提取液经腹腔注射C57BL/6小鼠,建立过敏性哮喘模型,将32只哮喘小鼠随机分为4组:生理盐水组以及粉尘螨低(100 μg/只)、中(1 mg/只)和高(2 mg/只)剂量组,治疗完成后,采用流式细胞术检测小鼠脾细胞中CD4+CD25+T细胞及Foxp3+C D4+C D25 +Tregs的含量,ELISA法检测小鼠血清中细胞因子IL-10、TGF-β1的水平.结果 生理盐水组、粉尘螨低剂量组小鼠脾细胞中CD4+CD25+T细胞占CD4+T细胞的百分比均明显低于粉尘螨中剂量组和高剂量组(P均<0.05),而中剂量组与高剂量组比

  16. Th17/CD4+CD25+Treg细胞与自身免疫病

    Institute of Scientific and Technical Information of China (English)

    王琴; 崔向军

    2011-01-01

    CD4+CD25+调节T细胞(CD4+CD25+Treg)是调节性T细胞的一个重要亚类,Treg可以通过IL-2、TGF-β1等促炎症细胞因子调节Th17/Treg平衡,调控Thl7细胞介导的炎症反应、自身免疫反应。本文将Thl7细胞/CD4+CD25+Treg细胞与自身免疫性疾病的关系作一综述。

  17. Aberrant Expression of Novel Cytokine IL-38 and Regulatory T Lymphocytes in Childhood Asthma

    Directory of Open Access Journals (Sweden)

    Man Chu

    2016-07-01

    Full Text Available We investigated the expression of novel anti-inflammatory interleukin (IL-38 and regulatory T (Treg lymphocytes in childhood asthma patients. The protein and mRNA expression level of IL-38, periostin, peripheral CD4+CD25+CD134+ T lymphocytes as well as CD4+CD25highFoxP3+ and CD4+CD25highCD127− Treg lymphocytes from 40 asthmatic patients and 20 normal control (NC subjects were studied using ELISA, qPCR and flow cytometry. Serum and supernatant cytokines/chemokines were determined by multiplex assay. Serum IL-38, IL-5, IL-17, IL-6, interferon-γ, periostin, IL-1β and IL-13 concentrations were significantly higher in asthmatic patients with or without steroid treatment than those in controls (all p < 0.05. The percentages of both CD4+CD25highFoxP3+ and CD4+CD25highCD127− Treg lymphocytes were markedly decreased in asthmatic patients with and without steroid treatment than those in controls (all p < 0.05. The elevated IL-38 concentration negatively correlated with the percentage of Treg lymphocytes in asthmatic patients with high level (>40 ng/mL of periostin (p < 0.05. Although the comparable mRNA levels of IL-38 and its receptor IL-36R were found between patients and controls, the mRNA level of IL-38 positively correlated with IL-36R and negatively correlated with IL-10 in all asthmatic patients (both p < 0.05. The percentage of CD4+CD25+CD134+ activated T lymphocytes was also significantly higher in asthmatic patients with steroid treatment than those in controls (p < 0.05. This cross-sectional study demonstrated that the overexpression of circulating IL-38 may play a role in the immunopathogenesis in asthma.

  18. 激素抵抗性哮喘患者外周血CD4+CD25+Foxp3+调节性T细胞及IL-10、TGF-β1的变化及意义%Changes and clinical significance of CD4+ CD25+Foxp3+ regulatory T cell and IL-10,TGF-β1 in steroid-resistant asthma patients of peripheral blood

    Institute of Scientific and Technical Information of China (English)

    钱一龙; 赵振中; 朱建俊; 谢中华; 王珠美

    2013-01-01

    目的 观察激素抵抗性哮喘(SRA)患者外周血CD4+ CD25+ Foxp3+调节性T细胞(Treg)及白介素10(IL-10)、转化生长因子β1(TGF-β1)的变化,分析其在SRA发病机制中的作用.方法 采用流式细胞术检测40例SRA患者(激素抵抗组)外周血单个核细胞CD4+ CD25+ Foxp3+ Treg数目,并计算CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的百分比;酶联免疫吸附试验(ELISA)法检测其血清IL-10、TGF-β1水平,并与激素敏感性患者(激素敏感组,46例)及正常体检者(正常组,30例)进行对比.结果 激素抵抗组患者外周血CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的百分比、CD4+ CD25+Foxp3+ Treg绝对值及血清IL-10、TGF-β1水平均明显低于激素敏感组与正常组(P<0.01,P<0.05);激素敏感组患者外周血CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的百分比、CD4+ CD25+ Foxp3+Treg绝对值及血清TGF-β1水平明显低于正常组(P<0.01,P<0.05),血清IL-10无明显差异(P>0.05);CD4+ CD25+ Foxp3+ Treg/CD4+T及CD4+ CD25+ Foxp3+ Treg绝对数均与血清IL-10、TGF-β1水平呈明显正相关(P<0.01).结论 SRA患者外周血CD4+ CD25+ Foxp3+ Treg数目减少及IL-10、TGF-β1含量减低可能与SRA的发生、发展有关.%Objective To investigate the changes of CD4+ CD25+ Foxp3+ regulatory T cell and IL-10,TGF-β1 in steroid-resistant asthma patients of peripheral blood,to study their significance.Methods The CD4+ CD25+ Foxp3+ regulatory T cell in 40 cases steroid-resistant asthma patients were detected by flow cytometry.The levels of serum IL-10 and TGF-β1 were detected by ELISA.The control groups were 30 individuals having general physical examination and 46 patients with steroid-sensitive patients.Results The proportion and number of CD4+ CD25+ Foxp3+ regulatory T cell and the levels of serum IL-10,TGF-β1 in steroid-resistant asthma group of Peripheral Blood were decreased significantly than steroid-sensitive group and healthy control group (P <0.01,P <0

  19. LAP TGF-Beta Subset of CD4+CD25+CD127− Treg Cells is Increased and Overexpresses LAP TGF-Beta in Lung Adenocarcinoma Patients

    Directory of Open Access Journals (Sweden)

    Lorenzo Islas-Vazquez

    2015-01-01

    Full Text Available Lung cancer is the leading cause of cancer death worldwide. Adenocarcinoma, the most commonly diagnosed histologic type of lung cancer, is associated with smoking. Cigarette smoke promotes inflammation on the airways, which might be mediated by Th17 cells. This inflammatory environment may contribute to tumor development. In contrast, some reports indicate that tumors may induce immunosuppressive Treg cells to dampen immune reactivity, supporting tumor growth and progression. Thus, we aimed to analyze whether chronic inflammation or immunosuppression predominates at the systemic level in lung adenocarcinoma patients, and several cytokines and Th17 and Treg cells were studied. Higher proportions of IL-17-producing CD4+ T-cells were found in smoking control subjects and in lung adenocarcinoma patients compared to nonsmoking control subjects. In addition, lung adenocarcinoma patients increased both plasma concentrations of IL-2, IL-4, IL-6, and IL-10, and proportions of Latency Associated Peptide (LAP TGF-β subset of CD4+CD25+CD127− Treg cells, which overexpressed LAP TGF-β. This knowledge may lead to the development of immunotherapies that could inhibit the suppressor activity mediated by the LAP TGF-β subset of CD4+CD25+CD127− Treg cells to promote reactivity of immune cells against lung adenocarcinoma cells.

  20. LAP TGF-Beta Subset of CD4+CD25+CD127− Treg Cells is Increased and Overexpresses LAP TGF-Beta in Lung Adenocarcinoma Patients

    Science.gov (United States)

    Islas-Vazquez, Lorenzo; Prado-Garcia, Heriberto; Aguilar-Cazares, Dolores; Meneses-Flores, Manuel; Galicia-Velasco, Miriam; Romero-Garcia, Susana; Camacho-Mendoza, Catalina; Lopez-Gonzalez, Jose Sullivan

    2015-01-01

    Lung cancer is the leading cause of cancer death worldwide. Adenocarcinoma, the most commonly diagnosed histologic type of lung cancer, is associated with smoking. Cigarette smoke promotes inflammation on the airways, which might be mediated by Th17 cells. This inflammatory environment may contribute to tumor development. In contrast, some reports indicate that tumors may induce immunosuppressive Treg cells to dampen immune reactivity, supporting tumor growth and progression. Thus, we aimed to analyze whether chronic inflammation or immunosuppression predominates at the systemic level in lung adenocarcinoma patients, and several cytokines and Th17 and Treg cells were studied. Higher proportions of IL-17-producing CD4+ T-cells were found in smoking control subjects and in lung adenocarcinoma patients compared to nonsmoking control subjects. In addition, lung adenocarcinoma patients increased both plasma concentrations of IL-2, IL-4, IL-6, and IL-10, and proportions of Latency Associated Peptide (LAP) TGF-β subset of CD4+CD25+CD127− Treg cells, which overexpressed LAP TGF-β. This knowledge may lead to the development of immunotherapies that could inhibit the suppressor activity mediated by the LAP TGF-β subset of CD4+CD25+CD127− Treg cells to promote reactivity of immune cells against lung adenocarcinoma cells. PMID:26582240

  1. Increased frequency and compromised function of T regulatory cells in systemic sclerosis (SSc is related to a diminished CD69 and TGFbeta expression.

    Directory of Open Access Journals (Sweden)

    Timothy R D J Radstake

    Full Text Available BACKGROUND: Regulatory T cells (Tregs are essential in the control of tolerance. Evidence implicates Tregs in human autoimmune conditions. Here we investigated their role in systemic sclerosis (SSc. METHODS/PRINCIPAL FINDINGS: Patients were subdivided as having limited cutaneous SSc (lcSSc, n = 20 or diffuse cutaneous SSc (dcSSc, n = 48. Further subdivision was made between early dcSSc (n = 24 and late dcSSc (n = 24 based upon the duration of disease. 26 controls were studied for comparison. CD3+ cells were isolated using FACS and subsequently studied for the expression of CD4, CD8, CD25, FoxP3, CD127, CD62L, GITR, CD69 using flow cytometry. T cell suppression assays were performed using sorted CD4CD25(highCD127(- and CD4CD25(lowCD127(high and CD3(+ cells. Suppressive function was correlated with CD69 surface expression and TGFbeta secretion/expression. The frequency of CD4(+CD25(+ and CD25(highFoxP3(highCD127(neg T cells was highly increased in all SSc subgroups. Although the expression of CD25 and GITR was comparable between groups, expression of CD62L and CD69 was dramatically lower in SSc patients, which correlated with a diminished suppressive function. Co-incubation of Tregs from healthy donors with plasma from SSc patients fully abrogated suppressive activity. Activation of Tregs from healthy donors or SSc patients with PHA significantly up regulated CD69 expression that could be inhibited by SSc plasma. CONCLUSIONS/SIGNIFICANCE: These results indicate that soluble factors in SSc plasma inhibit Treg function specifically that is associated with altered Treg CD69 and TGFbeta expression. These data suggest that a defective Treg function may underlie the immune dysfunction in systemic sclerosis.

  2. Human and Mouse CD8+CD25+FOXP3+ Regulatory T Cells at Steady State and during Interleukin-2 Therapy

    OpenAIRE

    Churlaud, Guillaume; Pitoiset, Fabien; Jebbawi, Fadi; Lorenzon, Roberta; Bellier, Bertrand; Rosenzwajg, Michelle; Klatzmann, David

    2015-01-01

    International audience In addition to CD4+ regulatory T cells (Tregs), CD8+ suppressor T cells are emerging as an important subset of regulatory T cells. Diverse populations of CD8+ T cells with suppressive activities have been described. Among them, a small population of CD8+CD25+FOXP3+ T cells is found both in mice and humans. In contrast to thymic-derived CD4+CD25+FOXP3+ Tregs, their origin and their role in the pathophysiology of autoimmune diseases (AIDs) are less understood. We repor...

  3. Role of Circulating CD4+ CD25high Foxp3+ Regulatory T-Cells in Paediatric Asthma

    OpenAIRE

    Ensaf Khalil Mohammed*, Zeinab Farag Asheiba

    2011-01-01

    Background: The role of T-Helper 2 (Th2) cells in the pathogenesis of allergy and asthma has been well described. However, the immunologic mechanisms that down modulate and protect against the development of these disorders are poorly characterized. A spectrum of CD4+ T cells, including, FOXP3-positive CD4+CD25+ T regulatory cells (Tregs) might play a critical role in regulating these diseases. Objective: To investigate the role of CD4+CD25high FoxP3 Tregs in the pathogenesis of pediatric ast...

  4. CD4+CD25+调节性T细胞在支气管哮喘中的研究进展%Changes and effects of CD4+CD25+ regulatory T cells in pathogenesis of asthma

    Institute of Scientific and Technical Information of China (English)

    宋丽; 施森; 李敏

    2007-01-01

    近年来,关于CD4+CD25+调节性T细胞在支气管哮喘发病中作用的研究有相当大的进展,支气管哮喘的发病机制已不能单纯用Th1/Th2平衡理论来解释,CD4+CD25+调节性T细胞越来越受到重视,它通过细胞接触和分泌细胞因子发挥免疫抑制功能,维持自身免疫平衡,控制支气管哮喘的发展,Foxp3、IL-6等在其功能调控中发挥重要作用.支气管哮喘中,有关CD4+CD25+调节性T细胞作用机制及数量变化的观点不一,糖皮质激素通过增强CD4+CD25+调节性T细胞功能发挥治疗作用.

  5. CD4+CD25+调节性T细胞在心脏疾病中的研究进展%The Research Progress of CD4 +CD25 + Regulatory T Cells in Heart Diseases

    Institute of Scientific and Technical Information of China (English)

    牛瑞兵

    2011-01-01

    CD4+ CD25+ regulatory T cells( Treg ) are a special subgroup of T cells in vivo with an immunoregulatory function. In recent years, massive domestic and oversea research indicated that Treg is closely related with various cardiovascular disease's occurrence and development. Therefore, understanding of Treg and its immunosuppression's mechanism in the heart disease is helpful to improve the immunotherapy effect for heart diseases. Here is to make a summary of CD4+ CD25 + Treg' production, features, functional mechanism and its correlation with heart diseases.%CD4 +CD25 +调节性T细胞(Treg)是体内具有免疫调节功能的一类特殊T细胞亚群.近年来,国内外的大量研究表明Treg与很多种心血管疾病的发生和进展密切相关.因此,认识Treg及其在心脏疾病中免疫抑制的机制,有助于提高心脏疾病的免疫治疗效果.现就CD4 +CD25 +Treg的产生、特征、作用机制及其与心脏疾病的联系予以综述.

  6. 佛波醇酯加离子霉素诱导脐血和成人外周血CD4+CD25+T细胞分泌IL-2的相关机制研究%Mechanisms underlying the induction of IL- 2 secretion by PDB plus ionomycin in CD4 + CD25 + T cells from cord blood and adult peripheral blood

    Institute of Scientific and Technical Information of China (English)

    肇静娴; 曾耀英; 李海仙; 曾祥凤; 季煜华; 何贤辉

    2006-01-01

    目的:以佛波醇酯加离子霉素作为刺激剂,验证CD4+CD25+调节性T细胞本身并不存在分泌IL-2障碍;同时通过对脐血和成人外周血的比较性研究,了解脐血CD4+CD25+T细胞的成熟度.方法:以autoMACS从足月婴儿脐血(CB)和成人外周血(PB)分选CD4+CD25+和CD4+CD25-T细胞,以PDB+ionomycin作为刺激剂,培养45h后流式细胞术检测各组细胞表达CD69和CD25水平,并以Luminex多重细胞因子检测技术检测培养上清中7种细胞因子的浓度.结果:经PDB+ionomycin刺激后,CB、PB的CD4+CD25+和CD4+CD25-T细胞均发生增殖,但在培养45h后CD4+CD25+T细胞均出现细胞状态变差或死亡倾向.CB、PB的CD4+CD25+T细胞活化后CD25分子表达进一步上调,高于CD25-细胞活化后的CD25分子密度.经PDB+ionomycin刺激后,PB CD4+CD25+和CD4+CD25-T细胞均分泌高水平的IFN-γ、IL-2和TNF-α,但CD25+细胞分泌IL-5、IL-4和IL-10水平远远高于CD25-细胞;CBCD4+CD25+和CD4+CD25-T细胞亦分泌高水平的IL-2和TNF-α,但IFN-γ水平远远低于PB,基本不分泌IL-5、IL-4和IL-10.结论:CD4+CD25+T细胞本身并不存在合成和分泌IL-2障碍,其可能具有与传统T细胞不同的T细胞受体信息转导模式;脐血CD4+CD25+T细胞功能尚未完全成熟.

  7. Shen-Qi-Jie-Yu-Fang exerts effects on a rat model of postpartum depression by regulating inflammatory cytokines and CD4+CD25+ regulatory T cells

    Science.gov (United States)

    Li, Jingya; Zhao, Ruizhen; Li, Xiaoli; Sun, Wenjun; Qu, Miao; Tang, Qisheng; Yang, Xinke; Zhang, Shujing

    2016-01-01

    Background Shen-Qi-Jie-Yu-Fang (SJF) is composed of eight Chinese medicinal herbs. It is widely used in traditional Chinese medicine for treating postpartum depression (PPD). Previous studies have shown that SJF treats PPD through the neuroendocrine mechanism. Aim To further investigate the effect of SJF on the immune system, including the inflammatory response system and CD4+CD25+ regulatory T (Treg) cells. Materials and methods Sprague Dawley rats were used to create an animal model of PPD by inducing hormone-simulated pregnancy followed by hormone withdrawal. After hormone withdrawal, the PPD rats were treated with SJF or fluoxetine for 1, 2, and 4 weeks. Levels of Treg cells in peripheral blood were measured by flow cytometry analysis. Serum interleukin (IL)-1β and IL-6 were evaluated by enzyme-linked immunosorbent assay, and gene and protein expressions of IL-1RI, IL-6Rα, and gp130 in the hippocampus were observed by reverse-transcription polymerase chain reaction and Western blot. Results Serum IL-1β in PPD rats increased at 2 weeks and declined from then on, while serum IL-6 increased at 1, 2, and 4 weeks. Both IL-1β and IL-6 were downregulated by SJF and fluoxetine. Changes in gene and protein expressions of IL-1RI and gp130 in PPD rats were consistent with changes in serum IL-1β, and were able to be regulated by SJF and fluoxetine. The levels of Treg cells were negatively correlated with serum IL-1β and IL-6, and were decreased in PPD rats. The levels of Treg cells were increased by SJF and fluoxetine. Conclusion Dysfunction of proinflammatory cytokines and Tregs in different stages of PPD was attenuated by SJF and fluoxetine through the modulation of serum concentrations of IL-1β and IL-6, expressions of IL-1RI, and gp130 in the hippocampus, and CD4+CD25+ Treg cells in peripheral blood. PMID:27143890

  8. Cerebral ischemia increases bone marrow CD4+CD25+FoxP3+ regulatory T cells in mice via signals from sympathetic nervous system.

    Science.gov (United States)

    Wang, Jianping; Yu, Lie; Jiang, Chao; Fu, Xiaojie; Liu, Xi; Wang, Menghan; Ou, Chunying; Cui, Xiaobing; Zhou, Chengguang; Wang, Jian

    2015-01-01

    Recent evidence has shown that an increase in CD4(+)CD25(+)FoxP3(+) regulatory T (Treg) cells may contribute to stroke-induced immunosuppression. However, the molecular mechanisms that underlie this increase in Treg cells remain unclear. Here, we used a transient middle cerebral artery occlusion model in mice and specific pathway inhibitors to demonstrate that stroke activates the sympathetic nervous system, which was abolished by 6-OHDA. The consequent activation of β2-adrenergic receptor (AR) signaling increased prostaglandin E2 (PGE2) level in bone marrow. β2-AR antagonist prevented the upregulation of PGE2. PGE2, which acts on prostaglandin E receptor subtype 4 (EP4), upregulated the expression of receptor activator for NF-κB ligand (RANKL) in CD4(+) T cells and mediated the increase in Treg cells in bone marrow. Treatment of MCAO mice with RANKL antagonist OPG inhibited the increase in percent of bone marrow Treg cells. PGE2 also elevated the expression of indoleamine 2,3 dioxygenase in CD11C(+) dendritic cells and promoted the development of functional Treg cells. The effect was neutralized by treatment with indomethacin. Concurrently, stroke reduced production of stromal cell-derived factor-1 (SDF-1) via β3-AR signals in bone marrow but increased the expression of C-X-C chemokine receptor (CXCR) 4 in Treg and other bone marrow cells. Treatment of MCAO mice with β3-AR antagonist SR-59230A reduced the percent of Treg cells in peripheral blood after stroke. The disruption of the CXCR4-SDF-1 axis may facilitate mobilization of Treg cells and other CXCR4(+) cells into peripheral blood. This mechanism could account for the increase in Treg cells, hematopoietic stem cells, and progenitor cells in peripheral blood after stroke. We conclude that cerebral ischemia can increase bone marrow CD4(+)CD25(+)FoxP3(+) regulatory T cells via signals from the sympathetic nervous system. PMID:25110149

  9. Incomplete depletion and rapid regeneration of Foxp3+ regulatory T cells following anti-CD25 treatment in malaria-infected mice

    OpenAIRE

    Couper, Kevin N.; Blount, Daniel G.; de Souza, J. Brian; Suffia, Isabelle; Belkaid, Yasmine; Riley, Eleanor M.

    2007-01-01

    Investigation of the role of regulatory T cells (Treg) in model systems is facilitated by their depletion using anti-CD25 antibodies, but there has been considerable debate about the effectiveness of this strategy. Here, we have compared the depletion and repopulation of CD4+CD25+Foxp3+ Treg in uninfected and malaria-infected mice using 7D4 and/or PC61 anti-CD25 antibodies. We find that numbers and percentages of CD25hi cells, but not Foxp3+ cells, are transiently reduced after 7D4 treatment ...

  10. Cutting edge: TNFR-shedding by CD4+CD25+ regulatory T cells inhibits the induction of inflammatory mediators.

    NARCIS (Netherlands)

    Mierlo, G.J. van; Scherer, H.U.; Hameetman, M.; Morgan, M.E.; Flierman, R.; Huizinga, T.W.J.; Toes, R.E.

    2008-01-01

    CD4+CD25+ regulatory T (Treg) cells play an essential role in maintaining tolerance to self and nonself. In several models of T cell-mediated (auto) immunity, Treg cells exert protective effects by the inhibition of pathogenic T cell responses. In addition, Treg cells can modulate T cell-independent

  11. CD4~+ CD25~+调节性T细胞对哮喘大鼠气道炎症的影响%The effects of CD4~+ CD25~+ regulatory T cells on the airway inflammation of asthmatic rats

    Institute of Scientific and Technical Information of China (English)

    薛克营; 王成国; 程立; 杨中卫; 王正艳; 李威; 石明; 唐友勇

    2009-01-01

    目的 观察CD4~+CD25~+调节性T细胞(CD4~+CD25~+Treg)对哮喘大鼠气道炎症的影响.方法 将卵白蛋白(OVA)免疫耐受大鼠CD4+CD25~+Treg细胞过继转移给哮喘大鼠,然后观察支气管肺泡灌洗液(BALF)中细胞计数及分类,ELISA检测BALF中IL-4、IL-5和IFN-γ及血清OVA特异性IgE含量,HE染色观察肺组织的病理改变.结果 与哮喘组比较,过继转移CD4~+CD25~+ Treg细胞后哮喘大鼠BALF中细胞总数、中性粒细胞和淋巴细胞百分率降低(P<0.05),嗜酸性粒细胞(Eos)百分率明显降低(P<0.01);BALF中IL-4和IL-5含量明显降低,IFN-γ含量明显升高,血清OVA特异性IgE含量明显降低(P<0.05);气道炎症明显减轻.结论 过继转移OVA免疫耐受大鼠CD4~+CD25~+ Treg细胞可以明显抑制哮喘的慢性气道炎症.%Objective To observe the effects of CD4~+ CD25~+ regulatory T cells ( CD4~+ CD25~+ Treg) on the airway inflammation of asthmatic rats. Methods CD4~+ CD25~+ Treg of OVA- immune tolerance rats were transferred to asthmatic rats. Then bronchoalveolar lavage fluid (BALF) was collected, and cytology study was conducted. The IL-4, IL-5, IFN-γ and OVA-specific serum IgE level in BALF were determined by ELISA. The lung tissue was obtained, and histologieal analysis was done through H. E. Results Total cells number, the percentage of lymphocytes and neutrophils in BALF, the IL-4 and IL-5 BALF levels and the OVA-specific serum IgE level of adoptive transfer group were decreased ( P < 0.05 ) , and the percentage of eosinophils ( Eos) was significantly lower than that of asthma group ( P < 0.01) , while its BALF IFN-γ level was higher than that of asthma group( P <0. 05). Compared with that of asthma group, peribronchiole inflammatory of treated group was alleviated. Conclusion CD4 ~+ CD25~+ Treg of OVA- immune tolerance rats transferred to asthmatic rats can significantly alleviate the airway inflammation of asthmatic rats.

  12. Changes of CD4+CD25+Regulatory T Cells in Patients with Acute Coronary Syndrome and the Effects of Atorvastatin

    Institute of Scientific and Technical Information of China (English)

    HU Zhenping; LI Dazhu; HU Yingfeng; YANG Keping

    2007-01-01

    The function of CD4+CD25+regulatory T lymphocytes (Treg) in patients with acute coronary syndrome (ACS) and the effects of atorvastatin were investigated. Forty-eight patients with ACS were randomly divided into two groups: group C receiving conventional therapy (n=24), and group C+A receiving conventional therapy+atorvastatin (10 mg/day, n=24). T lymphocytes from ACS patients (before and 2 weeks after the treatment) or 18 healthy subjects were separated and the flow cytometry was used to measure the percentage of Treg. The inhibitory ability of Treg on effector T cells was determined by mixed lymphocyte reaction (MLR). ELISA was used to measure the serum levels of cytokines (IL-10, TGF-β1 and IFN-γ) before and after treatment. The results showed that as compared with normal control group, Treg percentage was decreased significantly (P<0.01), the in- hibitory ability of Treg on the T lymphocytes proliferation was reduced (P<0.01), IFN-γ, levels were increased and IL-10 and TGF-β1 levels were lowered in ACS patients. After treatment with atorvas- tatin, Treg percentage and the inhibitory ability of Treg on T lymphocytes proliferation were signifi- cantly increased in ACS patients. Serum IFN-γ, was decreased significantly, while IL-10 and TGF-β1 were elevated significantly as compared with the non-atorvastatin group. The number of Treg was positively correlated with serum TGF-β1, but negatively with serum IFN-γ and CRP. It was concluded that ACS was associated with decreased number and defected function of Treg, which may play an important role in initiating immune-inflammatory response in ACS. The inhibitory ef- fects of atorvastatin on inflammation in ACS may be due to its beneficial effects on Treg and restora- tion of immune homeostasis.

  13. Induction of FOXP3 expression in naive human CD4+FOXP3− T cells by T-cell receptor stimulation is transforming growth factor-β–dependent but does not confer a regulatory phenotype

    OpenAIRE

    Tran, Dat. Q.; Ramsey, Heather; Shevach, Ethan M.

    2007-01-01

    Thymic-derived natural T-regulatory cells (nTregs) are important for the induction of self-tolerance and the control of autoimmunity. Murine CD4+CD25−Foxp3− cells can be induced to express Foxp3 after T-cell receptor (TCR) activation in the presence of transforming growth factor β (TGFβ) and are phenotypically similar to nTregs. Some studies have suggested that TCR stimulation of human CD4+CD25− cells results in the induction of transient expression of FOXP3, but that the induced cells lack a...

  14. Overexpression of CD45RA isoforms in carriers of the C77G mutation leads to hyporeactivity of CD4+CD25highFoxp3+ regulatory T cells.

    Science.gov (United States)

    Pokoyski, C; Lienen, T; Rother, S; Schock, E; Plege-Fleck, A; Geffers, R; Schwinzer, R

    2015-12-01

    Disorders in regulatory T-cell (T(reg)) function can result in the breakdown of immunological self-tolerance. Thus, the identification of mechanisms controlling the activity of T(reg) is of great relevance. We used T(reg) from individuals carrying the C77G polymorphism as models to study the role of CD45 molecules in humans. C77G prevents splicing of CD45 exon A thereby leading to an aberrant expression pattern of CD45 isoforms in affected individuals. Resting and in vitro expanded/activated CD4(+)CD25(high)Foxp3(+) T(reg) from carriers of C77G strongly expressed CD45RA isoforms whereas these isoforms were almost absent in cells from individuals with wild-type CD45. C77G T(reg) showed diminished upregulation of activation markers, lower phosphorylation of p56(lck)(Y505) and a reduced proliferative potential when stimulated with anti-TcR or anti-TcR plus CD28 mAb suggesting decreased responsiveness to activating stimuli. In addition, the capacity to suppress proliferation of conventional CD4(+) T cells was impaired in C77G T(reg). Furthermore, microarray studies revealed distinct gene expression patterns in T(reg) from C77G carriers. These data suggest that the changes in CD45 isoform combination resulting from the C77G mutation alter the responsiveness of T(reg) to TcR-mediated signaling. Targeting CD45 isoform expression might be a useful approach to modulate T(reg) function. PMID:26355564

  15. Allergen-responsive CD4+CD25+ Regulatory T Cells in Children who Have Outgrown Cow's Milk Allergy

    OpenAIRE

    Karlsson, Malin R.; Rugtveit, Jarle; Brandtzaeg, Per

    2004-01-01

    Cow's milk allergy in children is often of short duration, which makes this disorder an interesting clinical model for studies of tolerance to dietary antigens. Here, we studied T cell responses in 21 initially allergic children who, after a milk-free period of >2 mo, had cow's milk reintroduced to their diet. Children who outgrew their allergy (tolerant children) had higher frequencies of circulating CD4+CD25+ T cells and decreased in vitro proliferative responses to bovine β-lactoglobulin i...

  16. Translational control of gene expression and disease

    NARCIS (Netherlands)

    Calkhoven, Cornelis F; Müller, Christine; Leutz, Achim

    2002-01-01

    In the past decade, translational control has been shown to be crucial in the regulation of gene expression. Research in this field has progressed rapidly, revealing new control mechanisms and adding constantly to the list of translationally regulated genes. There is accumulating evidence that trans

  17. Dynamic changes in percentages of CD4+CD25+regulatory T cells and Th17 cells in process of airway remodeling in mouse model of asthma%哮喘小鼠气道重塑过程中CD4+CD25+调节性T细胞和Th17细胞表达的动态变化

    Institute of Scientific and Technical Information of China (English)

    娄春艳; 李敏; 李丽

    2015-01-01

    Objective To study the dynamic changes in Th17 cells and CD4+CD25+regulatory T cells (Treg) in the spleen and to analyze their relationship with airway remodeling. Methods A total of 48 female speciifc pathogen-free Balb/c mice were randomly divided into control and asthmatic groups. To establish the asthmatic airway remodeling model, the mice were sensitized to ovalbumin (OVA) through intraperitoneal injection of OVA and aluminum hydroxide suspension and challenged by inhalation of aerosol OVA. The matched control group was treated with normal saline instead. In 24 hours after 2-week, 4-week, and 8-week aerosol inhalation, 8 mice were randomly selected from each group and sacriifced. Then histopathological examination of the left lung was performed to measure the degree of airway remodeling. The percentages of Th17 and CD4+CD25+Treg cells in total CD4+cells from the spleen were determined by lfow cytometry. Results In the asthmatic group, the ratios of total bronchial wall area to bronchial basement membrane perimeter (WAt/Pbm) and bronchial smooth muscle area to bronchial basement membrane perimeter (WAm/Pbm) signiifcantly increased as the challenge proceeds (P<0.01). The percentage of Th17 cells derived from the cell suspension of the spleen gradually increased and it was positively correlated with the degree of asthmatic airway remodeling (P<0.01). The percentage of CD4+CD25+Treg cells from the suspension gradually decreased and it was negatively correlated with the degree of asthmatic airway remodeling (P<0.01). Conclusions In mice with asthma, as the challenge proceeds, the airway remodeling becomes more severe, the percentage of Th17 cells increases, and the percentage of CD4+CD25+Treg cells decreases. The immunological imbalance is possibly one of the important factors inducing airway remodeling.%目的:探讨哮喘小鼠Th17细胞和CD4+CD25+调节性T细胞(Treg)在脾组织中表达水平的变化规律及与气道重塑的关系

  18. The presence of B7-H4+ macrophages and CD25+CD4+ and FOXP3+ regulatory T cells in the microenvironment of nasal polyps - a preliminary report.

    Directory of Open Access Journals (Sweden)

    P Strek

    2011-04-01

    Full Text Available The nasal polyp (NP seems to represent the end-stage of longstanding inflammation in patients with chronic rhinosinusitis. The aim of our study has been to evaluate the presence of two regulatory cell populations in the microenvironment of NP: CD4+CD25high Foxp3+ (Treg cells and B7-H4-expressing macrophages. Treg cells are actively able to inhibit T lymphocytes, while the population of B7-H4-expressing macrophages has recently been described as characterized by a regulatory function similar to that of Treg cells. For our study, we evaluated 14 NP tissue samples. The samples were divided into two main groups, eosinophilic (NP and lymphocytic (NP, according to the predominant type of immune cell infiltration. The presence of Treg cells and B7-H4 positive macrophages in the samples was analyzed by FACS. Treg cells and B7-H4-expressing macrophages were identified in all the examined nasal polyps. The percentages of both Treg cells and of B7H4 positive cells found in the eosinophilic nasal polyps were higher than those found in the lymphocytic nasal polyps. Treg cells and B7H4+ macrophage subpopulations were present in the NP microenvironment and the alterations in their percentages were related to a distinct pattern of immune cell infiltration.

  19. Seasonal influences of winter and summer on CD4+CD25+Treg in SD rats of RA kidney deficiency%冬夏季节肾虚型痹证大鼠CD4+CD25+Treg表达变化

    Institute of Scientific and Technical Information of China (English)

    张淼; 王彤; 陈怀民; 陈彦钦; 邓杨春

    2013-01-01

    目的:以中医学“天人相应”的整体调控思想为指导,以“肾应冬”为切入点,从褪黑素高位调节的角度,通过动物实验探讨肾虚型痹证CD4+CD25+调节性T细胞(Treg)表达的变化.方法:采用胶原诱导的关节炎(CIA)大鼠模型,以冬至、夏至两个时间点,测定各组大鼠血清中CD4+CD25+ Treg/CD4+ Treg变化.结果:与正常组大鼠相比,CIA模型组、手术组及伪手术组CD4+CD25+Treg/CD4+Treg水平均明显降低,并且差异具有统计学意义(P<0.05,P<0.01).说明痹证可导致Treg水平降低.在季节性比较中,冬季正常组、CIA模型组及伪手术组大鼠Treg水平明显低于夏季组,且差异具有统计学意义(P<0.05,P<0.01).结论:Treg水平的改变具有自然节律性,人体冬天的免疫能力低于夏天,正与冬季阳气藏于内的中医理论相一致.%Objective:From the Chinese medicine point of view,man and nature should be in correspondence.Following its overall regulation as guidance and putting‘kidney should be winter'as an entry point,the essay investigates the relationship between RA (a Chinese medicine category) Kidney deficiency CD4+CD25+ Treg and the season changes through animal experiments.Methods:Adopting collagen Ⅱ-induced arthritis,CIA rat model which was the closest to the human RA clinical features,in the Winter Solstice and Summer solstice,the observation on the arthritis index of each rats group and the level change evaluationfor the CD4+ CD25+ Treg/CD4+ Treg.Results:Compared with normal group rat,the CD4+CD25+ Treg/CD4+Treg level of the model of rheumatoid arthritis,CIA group,operation group,and the sham-operation group was significantly decreased and the difference had statistics significance (P<0.05,P<0.01).It proved that RA could lead to the decrease of the treg level.In the season comparison,the treg level of the winter normal group,the model of rheumatoid arthritis,CIA group and the sham operation group was lower than the summer

  20. B cell–deficient NOD.H-2h4 mice have CD4+CD25+ T regulatory cells that inhibit the development of spontaneous autoimmune thyroiditis

    OpenAIRE

    Yu, Shiguang; Maiti, Prasanta K; Dyson, Melissa; Jain, Renu; Braley-Mullen, Helen

    2006-01-01

    Wild-type (WT) NOD.H-2h4 mice develop spontaneous autoimmune thyroiditis (SAT) when given 0.05% NaI in their drinking water, whereas B cell–deficient NOD.H-2h4 mice are SAT resistant. To test the hypothesis that resistance of B cell–deficient mice to SAT was due to the activity of regulatory CD4+CD25+ T (T reg) cells activated if autoantigen was initially presented on non–B cells, CD25+ T reg cells were transiently depleted in vivo using anti-CD25. B cell–deficient NOD.H-2h4 mice given three ...

  1. Characterization of Protective Human CD4+CD25+ FOXP3+ Regulatory T Cells Generated with IL-2, TGF-β and Retinoic Acid

    OpenAIRE

    Lu, Ling; Zhou, Xiaohui; Wang, Julie; Zheng, Song Guo; Horwitz, David A.

    2010-01-01

    Background Protective CD4+CD25+ regulatory T cells bearing the Forkhead Foxp3 transcription factor can now be divided into three subsets: Endogenous thymus-derived cells, those induced in the periphery, and another subset induced ex-vivo with pharmacological amounts of IL-2 and TGF-β. Unfortunately, endogenous CD4+CD25+ regulatory T cells are unstable and can be converted to effector cells by pro-inflammatory cytokines. Although protective Foxp3+CD4+CD25+ cells resistant to proinflammatory cy...

  2. Detection and significance of CD4+ CD25+ CD127low/-Treg cells in patients with SLE%系统性红斑狼疮患者外周血CD4+CD25+CD127low/-调节性T细胞的检测及意义

    Institute of Scientific and Technical Information of China (English)

    韦月梅; 邹洪才; 崔俊; 孔建忠; 田安国; 葛建英

    2013-01-01

    Objective To investigate the feasibility of application of CD4+ CD25+ CD127low/- as an Treg cells new marker in patients with systemic lupus erythematosus (SLE). Methods The proportions of CD4+CD25+CD127low-/and CD4+CD25+ FoxP3+Treg cells in peripheral blood of SLE patients(group A) and healthy people(group B) were determined by flow cytometry. The correlation between CD4+ CD25+ CD127low/- Treg cells and CD4+ CD25+ FoxP3+ Treg cells was analyzed. Results The proportions of CD4+CD25+CD127low/- Treg cells and CD4+ CD25+ FoxP3+Treg cells in group A were significantly lower than those in group B [(3. 31 + 0. 82)% and (2. 28 + 0. 47)% vs. (6. 07 + 1. 59)% and (5. 01 + 1. 09)%](P<0. 01). The proportion of CD4+ CD25+ CD127 low/- Treg cells was positively correlated to that of CD4+ CD25+ FoxP3+ T cells in both groups(r=0. 713 and r=0. 709, P<0. 01). Conclusion The surface marker CD4+ CD25+ CD127low/- can be used to identify Treg cells. The decreases of CD4+CD25+CD127low/- Treg cells may play an important role in the pathogenesis of SLE.%目的 探讨用膜表面标志CD4+ CD25+ CD127low/-作为检测调节性T(Treg)细胞标记的可行性,并探讨其在系统性红斑狼疮(SLE)中的临床意义.方法 用流式细胞术检测SLE组及健康对照组外周血CD4+ CD25+ CD127low/-Treg细胞及CD4+ CD25+ FoxP3+ Treg细胞的比例,并分析两组CD4+ CD25+ CD127low/-Treg细胞与CD4+ CD25+ FoxP3+ Treg细胞比例之间的相关性.结果 SLE组外周血CD4+ CD25+ CD127low/-Treg细胞比例为(3.31±0.82)%CD4+ CD25+ FoxP3+ Treg细胞比例为(2.28±0.47)%,均显著低于健康对照组的(6.07±1.59)%和(5.01±1.09)%(P<0.01).SLE组及健康对照组外周血CD4+ CD25+ CD127low/-Treg细胞比例与CD4+ CD25+FoxP3+ Treg细胞比例之间呈显著正相关(r=0.713、r=0.709,P<0.01).结论 膜表面标志CD4+ CD25+ CD127low/-可以用来鉴定Treg细胞;SLE患者外周血CD4+ CD25+ CD127low/-Treg细胞的显著减少可能与SLE的发病有关.

  3. Shen-Qi-Jie-Yu-Fang exerts effects on a rat model of postpartum depression by regulating inflammatory cytokines and CD4+CD25+ regulatory T cells

    Directory of Open Access Journals (Sweden)

    Li JY

    2016-04-01

    Full Text Available Jingya Li,1,* Ruizhen Zhao,1,* Xiaoli Li,1 Wenjun Sun,1 Miao Qu,1 Qisheng Tang,1 Xinke Yang,1 Shujing Zhang2 1Third Affiliated Hospital, 2School of Basic Medical Sciences, Beijing University of Chinese Medicine, Beijing, People’s Republic of China *These authors contributed equally to this work Background: Shen-Qi-Jie-Yu-Fang (SJF is composed of eight Chinese medicinal herbs. It is widely used in traditional Chinese medicine for treating postpartum depression (PPD. Previous studies have shown that SJF treats PPD through the neuroendocrine mechanism. Aim: To further investigate the effect of SJF on the immune system, including the inflammatory response system and CD4+CD25+ regulatory T (Treg cells. Materials and methods: Sprague Dawley rats were used to create an animal model of PPD by inducing hormone-simulated pregnancy followed by hormone withdrawal. After hormone withdrawal, the PPD rats were treated with SJF or fluoxetine for 1, 2, and 4 weeks. Levels of Treg cells in peripheral blood were measured by flow cytometry analysis. Serum interleukin (IL-1β and IL-6 were evaluated by enzyme-linked immunosorbent assay, and gene and protein expressions of IL-1RI, IL-6Rα, and gp130 in the hippocampus were observed by reverse-transcription polymerase chain reaction and Western blot. Results: Serum IL-1β in PPD rats increased at 2 weeks and declined from then on, while serum IL-6 increased at 1, 2, and 4 weeks. Both IL-1β and IL-6 were downregulated by SJF and fluoxetine. Changes in gene and protein expressions of IL-1RI and gp130 in PPD rats were consistent with changes in serum IL-1β, and were able to be regulated by SJF and fluoxetine. The levels of Treg cells were negatively correlated with serum IL-1β and IL-6, and were decreased in PPD rats. The levels of Treg cells were increased by SJF and fluoxetine. Conclusion: Dysfunction of proinflammatory cytokines and Tregs in different stages of PPD was attenuated by SJF and fluoxetine through

  4. Central muscarinic cholinergic activation alters interaction between splenic dendritic cell and CD4+CD25- T cells in experimental colitis.

    Directory of Open Access Journals (Sweden)

    Peris Munyaka

    Full Text Available BACKGROUND: The cholinergic anti-inflammatory pathway (CAP is based on vagus nerve (VN activity that regulates macrophage and dendritic cell responses in the spleen through alpha-7 nicotinic acetylcholine receptor (a7nAChR signaling. Inflammatory bowel disease (IBD patients present dysautonomia with decreased vagus nerve activity, dendritic cell and T cell over-activation. The aim of this study was to investigate whether central activation of the CAP alters the function of dendritic cells (DCs and sequential CD4+/CD25-T cell activation in the context of experimental colitis. METHODS: The dinitrobenzene sulfonic acid model of experimental colitis in C57BL/6 mice was used. Central, intracerebroventricular infusion of the M1 muscarinic acetylcholine receptor agonist McN-A-343 was used to activate CAP and vagus nerve and/or splenic nerve transection were performed. In addition, the role of α7nAChR signaling and the NF-kB pathway was studied. Serum amyloid protein (SAP-A, colonic tissue cytokines, IL-12p70 and IL-23 in isolated splenic DCs, and cytokines levels in DC-CD4+CD25-T cell co-culture were determined. RESULTS: McN-A-343 treatment reduced colonic inflammation associated with decreased pro-inflammatory Th1/Th17 colonic and splenic cytokine secretion. Splenic DCs cytokine release was modulated through α7nAChR and the NF-kB signaling pathways. Cholinergic activation resulted in decreased CD4+CD25-T cell priming. The anti-inflammatory efficacy of central cholinergic activation was abolished in mice with vagotomy or splenic neurectomy. CONCLUSIONS: Suppression of splenic immune cell activation and altered interaction between DCs and T cells are important aspects of the beneficial effect of brain activation of the CAP in experimental colitis. These findings may lead to improved therapeutic strategies in the treatment of IBD.

  5. CD4+ CD25+调节性T细胞及其分子标记物与支气管哮喘

    Institute of Scientific and Technical Information of China (English)

    马祥; 毛辉; 梁宗安

    2009-01-01

    CD4+ CD25+调节性T细胞是一类以免疫抑制和免疫无能为特征的淋巴细胞群,FOXP3是CD4+CD25+调节性T细胞一个特征性的分子标志物,并且对CD4+CD25+调节性T细胞的发育、外周表达和功能维持有着关键性的作用.近年来,多项研究显示CD4+ CD25+调节性T细胞参与并影响了支气管哮喘的发生、发展过程,对调节性T细胞或其相关基因的干预也许会成为支气管哮喘治疗的新方向.

  6. Essential role for interleukin-2 for CD4+CD25+ T regulatory cell development during the neonatal period

    OpenAIRE

    Bayer, Allison L.; Yu, Aixin; Adeegbe, Dennis; Malek, Thomas R.

    2005-01-01

    Although many aspects of CD4+CD25+ T regulatory (Treg) cell development remain largely unknown, signaling through the IL-2R represents one feature for the production of Treg cells. Therefore, the present study was undertaken to further define early developmental steps in the production of Treg cells, including a more precise view on the role of interleukin (IL)-2 in this process. After adoptive transfer of wild-type Treg cells into neonatal IL-2Rβ−/− mice, only a small fraction of donor Treg ...

  7. Distinct roles of CTLA-4 and TGF-b in CD4+ CD25+ regulatory T cell function

    OpenAIRE

    Tang, Qizhi Z; Bluestone, Jeffrey A.

    2004-01-01

    Both CTLA-4 and TGF- have been implicated in suppression by CD4+CD25+ regulatory T cells (Treg). In this study, the relationship between CTLA-4 and TGF- in Treg function was examined. Blocking CTLA-4 on wild-type Treg abrogated their suppressive activity in vitro, whereas neutralizing TGF- had no effect, supporting a TGF--independent role for CTLA-4 in Treg-mediated suppression in vitro. In CTLA-4-deficient mice, Treg development and homeostasis was normal. Moreover, Treg from CTLA-4-deficien...

  8. Fermented fish oil suppresses T helper 1/2 cell response in a mouse model of atopic dermatitis via generation of CD4+CD25+Foxp3+ T cells

    Directory of Open Access Journals (Sweden)

    Han Sang-Chul

    2012-08-01

    Full Text Available Abstract Background Allergic skin inflammation such as atopic dermatitis (AD, which is characterized by pruritus and inflammation, is regulated partly through the activity of regulatory T cells (Tregs. Tregs play key roles in the immune response by preventing or suppressing the differentiation, proliferation and function of various immune cells, including CD4+ T cells. Recent studies report that fermentation has a tremendous capacity to transform chemical structures or create new substances, and the omega-3 polyunsaturated fatty acids (n-3 PUFAs in fish oil can reduce inflammation in allergic patients. The beneficial effects of natural fish oil (NFO have been described in many diseases, but the mechanism by which fermented fish oil (FFO modulates the immune system and the allergic response is poorly understood. In this study, we produced FFO and tested its ability to suppress the allergic inflammatory response and to activate CD4+CD25+Foxp3+ Tregs. Results The ability of FFO and NFO to modulate the immune system was investigated using a mouse model of AD. Administration of FFO or NFO in the drinking water alleviated the allergic inflammation in the skin, and FFO was more effective than NFO. FFO treatment did increase the expression of the immune-suppressive cytokines TGF-β and IL-10. In addition, ingestion of FFO increased Foxp3 expression and the number of CD4+CD25+Foxp3+ Tregs compared with NFO. Conclusions These results suggest that the anti-allergic effect of FFO is associated with enrichment of CD4+CD25+ Foxp3+ T cells at the inflamed sites and that FFO may be effective in treating the allergic symptoms of AD.

  9. 银屑病患者外周血CD4+CD25+调节性T细胞水平的研究

    Institute of Scientific and Technical Information of China (English)

    陈丽芳; 陈小敏; 杨秀丽; 史维平; 秦小卫; 郝树媛

    2007-01-01

    调节性T细胞(regulatory T cell,Treg)是一组具有免疫调节功能的T细胞亚群.对于维持机体内环境的稳定有着重要的作用。根据CD4+CD25+Treg来源的不同将其分为固有CD4+CD25+Treg和适应性CD4+CD25+Treg。固有CD4+CD25+Treg是由胸腺T细胞自然分化发育而来的一个主要Treg亚群,而适应性CD4+CD25+Treg是在特定的免疫环境中,经抗原刺激后成熟的T细胞。我们测定银屑病患者外周血中CD4+CD25+Treg,分析研究其与银屑病的相关性。

  10. Changes in Th1 cells and CD4+CD25+Treg cells in non-obese diabetic mice at early stage of diabetes

    Directory of Open Access Journals (Sweden)

    Hong-jun WANG

    2013-11-01

    Full Text Available Objective To investigate the changes in Th1 cells and CD4+CD25+Treg cells in non-obese diabetic (NOD mice at early stage of diabetes, and to evaluate the significance of these changes. Methods Four week- (group A, 8 week- (group B and 16 week-old (group C female NOD mice (8 each were used in present study. The spleen, thymus and pancreas were harvested. Th1 and CD4+CD25+Treg cells in spleen were determined by flow cytometer, and the ratios of Th1/CD4+T, CD4+CD25+Treg/CD4+T and Th1/CD4+CD25+Treg were calculated. Subsequently, CD4–CD8–T, CD4+CD8+T, CD4–CD8+T and CD4+CD8–T cells in thymus were determined by flow cytometer, and the ratio of CD25+Treg/CD4+CD8–T was calculated. The histopathological changes in pancreas were also evaluated by HE staining and immunohistochemistry staining. Results The proportion of Th1 cells in spleen and the ratios of Th1/CD4+T and Th1/CD4+CD25+Treg were higher significantly in group C than in group A and B. However, no significant differences were found in the proportion of spleen CD4+CD25+Treg cells and the ratio of CD4+CD25+Treg/CD4+T among the three groups. Compared with group A, no obvious changes were found in thymus CD4–CD8–T, CD4+CD8+T, CD4–CD8+T and CD4+CD8–T cells in group B and C, but the ratio of thymus CD25+Treg/CD4+CD8–T increased significantly in group B and C. Lymphocytic infiltration was observed in pancreatic islets of group B and C as shown with HE staining, but Foxp3+T cells were not seen in pancreatic islets by immunohistochemistry. Conclusion Th1 cells are gradually increased at early stage of diabetes in NOD mice, but CD4+CD25+Treg cells are relatively default. These changes may play an important role in the progress of diabetes. DOI: 10.11855/j.issn.0577-7402.2013.11.004

  11. Population-level control of gene expression

    Science.gov (United States)

    Nevozhay, Dmitry; Adams, Rhys; van Itallie, Elizabeth; Bennett, Matthew; Balazsi, Gabor

    2011-03-01

    Gene expression is the process that translates genetic information into proteins, that determine the way cells live, function and even die. It was demonstrated that cells with identical genomes exposed to the same environment can differ in their protein composition and therefore phenotypes. Protein levels can vary between cells due to the stochastic nature of intracellular biochemical events, indicating that the genotype-phenotype connection is not deterministic at the cellular level. We asked whether genomes could encode isogenic cell populations more reliably than single cells. To address this question, we built two gene circuits to control three cell population-level characteristics: gene expression mean, coefficient of variation and non-genetic memory of previous expression states. Indeed, we found that these population-level characteristics were more predictable than the gene expression of single cells in a well-controlled environment. This research was supported by the NIH Director's New Innovator Award 1DP2 OD006481-01 and Welch Foundation Grant C-1729.

  12. Control-flow checking via regular expressions

    OpenAIRE

    Benso, Alfredo; Di Natale, Giorgio; Di Carlo, Stefano; Prinetto, Paolo Ernesto; Tagliaferri, Luca

    2001-01-01

    The present paper explains a new approach to program control flow checking. The check has been inserted at source-code level using a signature methodology based on regular expressions. The signature checking is performed without a dedicated watchdog processor but resorting to inter-process communication (IPC) facilities offered by most of the modern operating systems. The proposed approach allows very low memory overhead and trade-off between fault latency and program execution time overhead

  13. Stromal CD4/CD25 positive T-cells are a strong and independent prognostic factor in non-small cell lung cancer patients, especially with adenocarcinomas.

    Science.gov (United States)

    Kayser, Gian; Schulte-Uentrop, Luzie; Sienel, Wulf; Werner, Martin; Fisch, Paul; Passlick, Bernward; Zur Hausen, Axel; Stremmel, Christian

    2012-06-01

    Within the concert of immune reactions against tumour cells cytotoxic and regulatory T-cells are of utmost importance. Several studies revealed contradictory results on this issue. We therefore focused on functional expression patterns and localization of tumour-infiltrating T-lymphocytes in non-small cell lung cancer (NSCLC) and their impact on patient's survival. 232 curatively operated NSCLC patients were included. After histological reevaluation and construction of tissue-multi-arrays immunohistochemical doublestains for CD3/CD8 and CD4/CD25 were performed to evaluate the total number of T-cells and their subsets of cytotoxic and activated T-cells. Additionally, the localization of the lymphocytes was included in the analysis. Hereby, T-cells within the tumour stroma were regarded as stromal, those among cancer cells as intraepithelial. The number of lymphocytes differed significantly between the histological subtypes being most prominent in large cell carcinomas. Survival analysis showed that high numbers of stromal T-lymphocytes are of beneficial prognostic influence in NSCLC patients. This also proved to be an independent prognostic factor in adenocarcinomas. Thus, in a large and well characterized cohort of NSCLC this is the first study to determine the prognostic value of stromal T-lymphocytes, as these are an independent prognosticator in NSCLC especially in adenocarcinomas whereas intraepithelial T-cells are not. PMID:22300751

  14. CD4+CD25+FoxP3+ Regulatory Tregs inhibit fibrocyte recruitment and fibrosis via suppression of FGF-9 production in the TGF-β1 exposed murine lung.

    Science.gov (United States)

    Peng, Xueyan; Moore, Meagan W; Peng, Hong; Sun, Huanxing; Gan, Ye; Homer, Robert J; Herzog, Erica L

    2014-01-01

    Pulmonary fibrosis is a difficult to treat, often fatal disease whose pathogenesis involves dysregulated TGF-β1 signaling. CD4+CD25+FoxP3+ Regulatory T cells ("Tregs") exert important effects on host tolerance and arise from naïve CD4+ lymphocytes in response to TGF-β1. However, the precise contribution of Tregs to experimentally induced murine lung fibrosis remains unclear. We sought to better understand the role of Tregs in this context. Using a model of fibrosis caused by lung specific, doxycycline inducible overexpression of the bioactive form of the human TGF-β1 gene we find that Tregs accumulate in the lung parenchyma within 5 days of transgene activation and that this enhancement persists to at least 14 days. Anti-CD25 Antibody mediated depletion of Tregs causes increased accumulation of soluble collagen and of intrapulmonary CD45+Col Iα1 fibrocytes. These effects are accompanied by enhanced local concentrations of the classical inflammatory mediators CD40L, TNF-α, and IL-1α, along with the neuroimmune molecule fibroblast growth factor 9 (FGF-9, also known as "glial activating factor"). FGF-9 expression localizes to parenchymal cells and alveolar macrophages in this model and antibody mediated neutralization of FGF-9 results in attenuated detection of intrapulmonary collagen and fibrocytes without affecting Treg quantities. These data indicate that CD4+CD25+FoxP3+ Tregs attenuate TGF-β1 induced lung fibrosis and fibrocyte accumulation in part via suppression of FGF-9. PMID:24904415

  15. CD4+CD25+FoxP3+ Regulatory Tregs inhibit fibrocyte recruitment and fibrosis via suppression of FGF-9 production in the TGF-b1 exposed murine lung

    Directory of Open Access Journals (Sweden)

    EricaHerzog

    2014-05-01

    Full Text Available Pulmonary fibrosis is a difficult to treat, often fatal disease whose pathogenesis involves dysregulated TGF-b1 signaling. CD4+CD25+FoxP3+ Regulatory T cells (“Tregs” exert important effects on host tolerance and arise from naïve CD4+ lymphocytes in response to TGF-b1. However the precise contribution of Tregs to experimentally induced murine lung fibrosis remains unclear. We sought to better understand the role of Tregs in this context. Using a model of fibrosis caused by lung specific inducible overexpression of the bioactive form of the human TGF-b1 gene we find that Tregs accumulate in the lung parenchyma within five days of transgene activation and that this enhancement persists to at least fourteen days. Anti-CD25 Antibody mediated depletion of Tregs causes increased detection of soluble collagen and of intrapulmonary CD45+CD34+Col Ia1 fibrocytes. These findings are accompanied by enhanced local concentrations of the classical inflammatory mediators CD40L, TNF-a, and IL-1a, along with the neuroimmune molecule fibroblast growth factor 9 (FGF-9, also known as “glial activating factor”. FGF-9 expression localizes to structural cells and alveolar macrophages in this model and antibody mediated neutralization of FGF-9 results in attenuated detection of intrapulmonary collagen and fibrocytes without affecting Treg quantities. These data indicate that CD4+CD25+FoxP3+ Tregs attenuate TGF-b1 induced lung fibrosis and fibrocyte accumulation in part via suppression of FGF-9.

  16. 重组疫苗E.coli LLO/OVA对小鼠CD4+CD25+Treg细胞调节作用的研究%The study for recombinant E.coli LLO/OVA regulating the function of CD4+CD25+Treg cells in mice

    Institute of Scientific and Technical Information of China (English)

    徐曼; 蒋小卫; 米粲

    2009-01-01

    目的:探讨重组E.coli LLO/OVA对小鼠CD4+CD25+Treg细胞的调节作用.方法:E.coli LLO/OVA和E.coli OVA分别免疫小鼠后,磁珠分离脾脏CD11c、CD4+CD25+Treg和CD4+CD25-T细胞,比较两组CD11c细胞对CD4+CD25+Treg细胞分泌IL-10的影响,以及CD4+CD25+Treg细胞抑制CD4+CD25-T细胞增殖的作用;流式细胞术分析荷瘤小鼠OVA特异性CD8+T细胞比率,观察去除CD4+CD25+Treg细胞前后两组黑色素瘤B16-OVA荷瘤小鼠肺转移情况.结果:E.coli LLO/OVA免疫组小鼠脾脏CD4+CD25+Treg细胞产生IL-10水平明显低于E.coli OVA免疫组小鼠(P<0.05),CD4+CD25+Treg细胞对CD4+CD25-T 细胞增殖的抑制作用明显减弱(P<0.05),且OVA特异性CD8+T细胞数量明显增多(P<0.05).在去除CD4+CD25+Treg细胞前后,E.coli LLO/OVA免疫的荷瘤小鼠肺转移结节数无明显减少(P>0.05).结论:重组E.coli LLO/OVA可通过下调小鼠CD4+CD25+Treg细胞数量、抑制其功能而促进机体特异性抗肿瘤免疫.

  17. CD4+CD25+ T lymphocytes and regulation of the immune system: perspectives for a pathophysiological understanding of sepsis.

    Science.gov (United States)

    Siqueira-Batista, Rodrigo; Gomes, Andréia Patrícia; Azevedo, Sarah Fumian Milward; Vitorino, Rodrigo Roger; Mendonça, Eduardo Gomes de; Sousa, Flávio Oliveira de; Oliveira, Alcione de Paiva; Cerqueira, Fábio Ribeiro; Paula, Sérgio Oliveira de; Oliveira, Maria Goreti de Almeida

    2012-09-01

    The systemic inflammatory response represents the core pathogenic event of sepsis, underlying clinical manifestations and laboratory findings in patients. Numerous studies have shown that CD4+CD25+ T lymphocytes, also known as regulatory T lymphocytes (Treg), participate in the development of sepsis due to their ability to suppress the immune response. The present article discusses the role of Treg lymphocytes in sepsis based on a specific search strategy (Latin American and Caribbean Health Sciences / Literatura Latino-americana e do Caribe em Ciências da Saúde - LILACS, PubMed, and Scientific Electronic Library Online - SciELO) focusing on two main topics: the participation of Treg cells in inflammation and immunity as well as perspectives in the computational physiological investigation of sepsis. PMID:23917832

  18. Peripheral Dendritic Cells and CD4+CD25+Foxp3+ Regulatory T Cells in the First Trimester of Normal Pregnancy and in Women with Recurrent Miscarriage.

    Directory of Open Access Journals (Sweden)

    Maciej Kwiatek

    Full Text Available The development of pregnancy is possible due to initiation of immune response in the body of the mother resulting in immune tolerance. Miscarriage may be caused by the impaired maternal immune response to paternal alloantigens located on the surface of trophoblast and fetal cells. The aim of the study was to compare the population of circulating dendritic cells (DCs and CD4+CD25+Foxp3+ regulatory T cells (TREGs in the first trimester of a normal pregnancy and in women with recurrent miscarriage and an attempt to determine the relationship between these cells and the role they may play in human reproductive failures. The study was conducted in a group of 33 first trimester pregnant women with recurrent miscarriage and in a group of 20 healthy pregnant women in the first trimester of normal pregnancy. Among mononuclear cells isolated from peripheral blood, the populations of DCs and TREGs were assessed by flow cytometry. The percentage of myeloid DCs and lymphoid DCs showed no significant difference between study and control group. Older maternal age and obesity significantly reduced the pool of circulating myeloid and lymphoid DCs (R=-0.39, p=0.02. In miscarriages the percentage of circulating TREGs was significantly lower compared to normal pregnancies (p=0.003. Among the analysed factors the percentage of TREGs was the most sensitive and the most specific parameter which correlated with the pregnancy loss. The reduction in the population of circulating TREGs suggests immunoregulatory mechanisms disorder in a pregnancy complicated by miscarriage.

  19. CD4+CD25+Foxp3+ regulatory T cells depletion may attenuate the development of silica-induced lung fibrosis in mice.

    Directory of Open Access Journals (Sweden)

    Fangwei Liu

    Full Text Available BACKGROUND: Silicosis is an occupational lung disease caused by inhalation of silica dust characterized by lung inflammation and fibrosis. Previous study showed that Th1 and Th2 cytokines are involved in silicosis, but Th1/Th2 polarization during the development of silicosis is still a matter of debate. Regulatory T cells (Treg cells represent a crucial role in modulation of immune homeostasis by regulating Th1/Th2 polarization, but their possible implication in silicosis remains to be explored. METHODOLOGY/PRINCIPAL FINDINGS: To evaluate the implication of Treg cells in the development of silicosis, we generated the Treg-depleted mice model by administration of anti-CD25 mAbs and mice were exposed to silica by intratracheal instillation to establish experimental model of silica-induced lung fibrosis. The pathologic examinations show that the Treg-depleted mice are susceptive to severer inflammation in the early stage, with enhanced infiltration of inflammatory cells. Also, depletion of Treg cells causes a delay of the progress of silica-induced lung fibrosis in mice model. Further study of mRNA expression of cytokines reveals that depletion of Tregs leads to the increased production of Th1-cytokines and decreased production of Th2-cytokine. The Flow Cytometry and realtime PCR study show that Treg cells exert the modulation function both directly by expressing CTLA-4 at the inflammatory stage, and indirectly by secreting increasing amount of IL-10 and TGF-β during the fibrotic stage in silica-induced lung fibrosis. CONCLUSION/SIGNIFICANCE: Our study suggests that depletion of Tregs may attenuate the progress of silica-induced lung fibrosis and enhance Th1 response and decelerate Th1/Th2 balance toward a Th2 phenotype in silica-induced lung fibrosis. The regulatory function of Treg cells may depend on direct mechanism and indirect mechanism during the inflammatory stage of silicosis.

  20. P38 MAP Kinase Signaling Is Required for the Conversion of CD4+CD25− T Cells into iTreg

    OpenAIRE

    Samuel Huber; Jörg Schrader; Gerhard Fritz; Katrin Presser; Steffen Schmitt; Ari Waisman; Stefan Lüth; Manfred Blessing; Johannes Herkel; Christoph Schramm

    2008-01-01

    CD4+CD25+ regulatory T cells (Treg) are important mediators of immune tolerance. A subset of Treg can be generated in the periphery by TGF-beta dependent conversion of conventional CD4+CD25- T cells into induced Treg (iTreg). In chronic viral infection or malignancy, such induced iTreg, which limit the depletion of aberrant or infected cells, may be of pathogenic relevance. To identify potential targets for therapeutic intervention, we investigated the TGF-beta signaling in Treg. In contrast ...

  1. High proportions of FOXP3+CD25high T cells in neonates are positively associated with allergic sensitization later in childhood

    OpenAIRE

    Strömbeck, A; Rabe, H.; Lundell, A-C; Andersson, K.; Johansen, S; Adlerberth, I; Wold, A E; Hesselmar, B; Rudin, A

    2014-01-01

    Background The role of FOXP3+ regulatory T cells in the prevention against sensitization and allergy development is controversial. Objective We followed 65 newborn Swedish children from farming and non-farming families from birth to 3 years of age and investigated the relation between CD4+ T cell subsets in blood samples and development of sensitization and allergic disease. Methods The proportions of FOXP3+CD25high, CTLA-4+CD25+, CD45RO+, HLA-DR+, CCR4+ or α4β7+ within the CD4+ T cell popula...

  2. EAE大鼠胸腺CD4+CD25+Foxp3+Treg细胞的动态变化及α-硫辛酸的干预作用%The variation of CD4+ CD25+ Foxp3+T regulative cells of thymus in different courses of EAE group and the effection of alpha lipoic acid

    Institute of Scientific and Technical Information of China (English)

    王燕燕; 蔺辉前; 檀国军; 郭书英; 张建娥

    2011-01-01

    目的 探讨实验性变态反应性脑脊髓炎(EAE)大鼠不同病程中胸腺CD4+ CD25+ Foxp3+Treg细胞变化情况及α-硫辛酸对EAE大鼠胸腺的干预作用.方法 取不同时期对照组、自然病程EAE组及α-硫辛酸EAE组大鼠的胸腺组织做流式细胞学,动态检测CD4+ CD25+ Foxp3+Treg细胞的变化情况.结果 EAE组大鼠急性期、复发期CD4+ CD25+ Foxp3+Treg细胞较同时期对照组明显减少(P 0. 05 ). Conclusion CD4 + CD25 + Foxp3 + Treg cells may play a role in the occurrence of EAE. There is significant relation between the development of EAE and CD4 + CD25 + Foxp3 + Treg cells. The function of ALA may not play through CD4 + CD25 + Foxp3 +Treg cells in immune adjustment at EAE. As the age added,the thymus may not be the main immune organ.

  3. 离体CD4+CD25+调节性T细胞的扩增和混合淋巴细胞反应研究

    Institute of Scientific and Technical Information of China (English)

    翟海龙

    2011-01-01

    为研究健康人外周血CD4+CD25+调节性T细胞(CD4+CD25+regulatory Tcells,CD4+CD25+Tregs)在协同刺激信号作用下的扩增反应及与CD4+CD25-T细胞混合淋巴细胞反应,采用免疫磁珠法分离CD4+CD25+Tregs和CD4+CD25-T细胞,在抗CD3-mAbs和抗CD28-mAbs的刺激下行CD4+CD25+Tregs培养和CD4+CD25+Tregs+CD4+CD25-T细胞混合淋巴细胞培养72h。然后加入CCK-8溶液孵育1h,用酶标仪检测OD4so值。结果为:CIM+CD25+Tregs组OD。50值极显著性地低于CI)4+CD25-T细胞组(P〈0.01)。CIM+CD25+Tregs与CD4+CD25-T细胞混合组0D450值也极显著性地低于CIM+CD25-T细胞组(P〈0.01)。CD4+CD25+Tregs表现出无反应性特征,还可抑制CD4+CD25-T细胞扩增,因此,CD4+CD25+Tregs不只是很惰性的免疫细胞,而是对保持免疫耐受发挥了积极的调节作用。

  4. CD4+/CD25+T细胞与旋毛虫急性感染后大鼠的肠道免疫活化%The Study of CD 4 +/CD 25 +T Cell in the Pathogensis of Post-infectious Irritable Bowel Syndrome Induced By Acute Trichinella Infection

    Institute of Scientific and Technical Information of China (English)

    蔺蓉; 丁震; 马欢; 钱伟; 侯晓华

    2011-01-01

    Objective To investigate the change of blood Tregulatory cell ( Treg) after different time point of acute trichinella infection. Methods Male sprague-dawley rats were divided 4 groups in terms of sampling time;control group and PI-IBS group(2 weeks,4 weeks and 8 weeks after acute trichinella infection). Flow cytometry was used to detect the percent of Treg both in blood and laminal propria lymphocyte ( LPL). Results ( 1) The intestinal pathologic scores in 2 and 4 weeks groups are higher than that in control group;the scores in 8,12 weeks groups recover from the intestinal inflammation. (2)2 weeks after trichinella spirals infection,there is a significant decrease of CD 4 * CD 25 VCD 4 * both in peripheral blood and LPL, which is almost abolished in 8 weeks group. Conclusion There is declination of blood Treg and LPL-Treg percent after the early stage of acute trichinella infection,and this might involved in the pathogenesis of post-infections irritable bowel syndrome,%目的 研究不同时间点旋毛虫肠道急性感染后外周血及肠道黏膜内调节T细胞(CD 4+/CD 25+ Treg)的变化规律.方法 研究分为旋毛虫急性感染组(感染后2周,4周及8周)、对照组(与感染组相应年龄及饲养环境).测定肠道病理损伤程度,外周血中及肠道黏膜固有层淋巴细胞中CD 4 +/CD 25+T细胞占CD 4+淋巴细胞的百分比.结果 (1)模型动物感染后2周肠道各部位炎症评分明显高于正常组(回肠末端2.85±0.22,近端结肠2.66±0.27,远端结肠2.43±0.19,P<0.05);4周组炎症评分较2周组降低,但仍高于正常组;至感染后8周大致恢复正常.(2)感染2周后外周血Treg百分比明显下降(3.91±0.15 vs 2.28±0.13,P<0.01),下降比例41.7%.造模后4周,外周血Treg百分比仍低于正常对照组(3.91±0.15 vs 3.01±0.14,P<0.01).而至模型诱导成功的8周后,血Treg的比例(CD4+ CD25+/CD4+)趋于正常(3,48±0.15,0.01<P<0.05).(3)在急性旋毛虫感染2周后LPL- Treg

  5. 小鼠乳腺癌模型中CD4~+CD25~(bright)CCR6~+Treg的检测及其意义%The detection and its significance of CD4~+ CD25~(bright) CCR6~+ Treg regulatory T cells in murine mammary carcinoma model

    Institute of Scientific and Technical Information of China (English)

    徐林; 徐薇; 蒋正刚; 熊思东

    2010-01-01

    为检测CD4~+ CD25~(bright) CCR~(6+) Treg在小鼠乳腺癌实验动物模型中的分布,并探讨其意义.采用FACS检测正常小鼠和4T1荷瘤小鼠中CD4~+ CD25~(bright)Treg的记忆分子CCR6的表达水平,同时检测CD4~+ CD25~(bright)Treg的CCR6~+和CCR6~-两个亚群的Foxp3表达情况;用增殖抑制实验观察了两个亚群分别对CD4~+CD25 T细胞增殖的抑制作用;用FACS检测CD4~+ CD25~(bright)CCR6~+ Treg在正常小鼠和4T1荷瘤小鼠中PBMC、LN和TIL中的分布情况.结果:4T1荷瘤小鼠中CD4~+ CD25~(bright)Treg的记忆分子CCR6的表达水平较正常小鼠增加;CD4~+ CD25~(bright)Treg的CCR6~+和CCR6~-两个亚群均高表达Foxp3,均能在体外有效抑制CD4~+ CD25 T细胞的增殖;与正常对照相比,CD4~+ CD25~(bright)CCR6~+ Treg在4T1荷瘤模型的引流淋巴结中比例明显增加,并在肿瘤局部存在显著的富集.上述结果提示在肿瘤免疫中存在CD4~+ CD25~(bright)CCR6~+ Treg,其具有效应/记忆样表型,并在肿瘤局部有明显的富集,这可能是肿瘤长期免疫逃逸的重要机制.

  6. CD4+/CD25+ regulatory cells inhibit activation of tumor-primed CD4+ T cells with IFN-gamma-dependent antiangiogenic activity, as well as long-lasting tumor immunity elicited by peptide vaccination

    OpenAIRE

    Casares, N. (Noelia); L. Arribillaga; Sarobe, P.; Dotor, J. (Javier); Lopez-Diaz-de-Cerio, A. (Ascensión); Melero, I; Lasarte, J.J. (Juan José)

    2003-01-01

    CD25(+) regulatory T (T reg) cells suppress the activation/proliferation of other CD4(+) or CD8(+) T cells in vitro. Also, down-regulation of CD25(+) T reg cells enhance antitumor immune responses. In this study, we show that depletion of CD25(+) T reg cells allows the host to induce both CD4(+) and CD8(+) antitumoral responses following tumor challenge. Simultaneous depletion of CD25(+) and CD8(+) cells, as well as adoptive transfer experiments, revealed that tumor-specific CD4(+) T cells, w...

  7. Interaction of indoleamine 2, 3-dioxygenase and CD4 + CD25 + Foxp3 + regulatory T cell in asthmatic mice%IDO与Treg在支气管哮喘小鼠中的相互作用及其意义

    Institute of Scientific and Technical Information of China (English)

    周丽蓉; 张劼; 罗永艾

    2013-01-01

    Objective To explore the interaction and the role of indoleamine 2,3-dioxygenase (IDO) and CD4 + CD25 + Foxp3 + regulatory T cell (Treg) in a mice model of allergic bronchial asthma.Methods BALB/c mice were sensitized and challenged by ovalbumin (OVA).Penh were measured to evaluate the airway responsiveness by noninvasive lung functional instrument.Bronchoalveolar lavage cytology was analyzed.IFN-γ,IL-4 and IL-10 in BALF were detected by enzyme-linked immunosorbent assay (ELISA).The mRNA expression of IDO and Foxp3 was measured by real-time fluorescence-based quantitative PCR.The protein expression of IDO was detected by immunohistochemistry.The percentage of Treg in CD4 + cells was assessed by flow cytometry.Results The airway responsiveness,the total cell number,the eosinophils and IL-4 in BALF of the asthmatic group significantly increased as compared with the control group (P < 0.01).The levels of IFN-γand IL-10 in BALF,the mRNA expression of IDO and Foxp3,the protein expression of IDO,and the percentage of Treg in CD4 + cells in the asthmatic group were significantly lower than those in the control group (P <0.01).The mRNA expression of IDO and Foxp3 was positively correlated with each other (r =0.819,0.807,P <0.05).The protein expression of IDO was positively correlated with the percentage of Treg in CD4 +cells (r =0.783,0.765,P < 0.05).Conclusions IDO and Treg reciprocally regulate each other,which surmounts immune tolerance and induces asthma.Therefore,IDO and Treg may play important roles in asthma.%目的 探讨吲哚胺2,3双加氧酶(indoleamine 2,3-dioxygense,IDO)与CD4+ CD25+ Foxp3+调节性T细胞(Treg)之间的相关性及在支气管哮喘发病机制中的作用.方法 BALB/c小鼠用随机数字表法分成对照组和哮喘组,每组8只.哮喘组以鸡卵清蛋白(ovalbumin,OVA)致敏,激发小鼠建立哮喘模型,无创肺功能仪检测气道反应性,支气管肺泡灌洗液(BALF)进行细胞学分析,ELISA检测BALF

  8. In vitro effects of mesenchymal stem cells on secreting function of T lymphocytes and CD4~+CD25~+T cells from patients with immune thrombo-cytopenia

    Institute of Scientific and Technical Information of China (English)

    赵霞

    2014-01-01

    Objective To analyze in vitro the effect of mesenchymal stem cells(MSCs)on secreting cytokines by T lymphocytes and ratio of CD4+CD25+T cells from patients with immune thrombocytopenia(ITP).Methods Human bone marrow-derived MSCs were isolated by Ficoll Hypaque and cultured for proliferating to passage cells.Allogeneic T lymphocytes

  9. Reconstitution of Scid mice with CD4+CD25- T cells leads to rapid colitis: an improved model for pharmacologic testing

    DEFF Research Database (Denmark)

    Kjellev, Stine; Lundsgaard, Dorthe; Poulsen, Steen Seier;

    2006-01-01

    . Purification of CD4+CD25- T cells is a simple procedure, and does not require flow-cytometric sorting. Fecal consistency score and colonic weight:length ratio are readily measurable and consistent disease parameters. This model is thus highly suitable for pharmacological testing of intervention strategies....

  10. Diminished frequency and function of CD4(+) CD25(high) regulatory T cells associated with active uveitis in Vogt-Koyanagi-Harada syndrome

    NARCIS (Netherlands)

    Chen, L.; Yang, P.Z.; Zhou, H.Y.; He, H.; Ren, X.R.; Chi, W.; Wang, L.; Kijlstra, A.

    2008-01-01

    PURPOSE. CD4(+)CD25(high) regulatory T (Treg) cells have been shown to be involved in the pathogenesis of autoimmune diseases. Vogt-Koyanagi-Harada (VKH) syndrome is an organ-specific autoimmune disease. This study was designed to phenotypically and functionally characterize peripheral blood CD4(+)C

  11. Apoptosis induced by low-dose aminophylline in asthmatic peripheral blood CD4+CD25+T regulatory cells%小剂量氨茶碱对支气管哮喘患者外周血CD4+CD25+调节性T细胞凋亡的影响

    Institute of Scientific and Technical Information of China (English)

    梁瑞韵; 伍卫; 黄瑾; 江山平; 张蔚

    2009-01-01

    目的 观察小剂量氨茶碱对分离培养的健康人和支气管哮喘(简称哮喘)患者外周血CD4+CD25+调节性T细胞(T regulatory cells,Treg)凋亡的影响.方法 经密度梯度离心法、尼龙棉柱法、磁珠分离法分离出健康人和哮喘患者外周血CD4+CD25+Treg,分小剂量氨茶碱(1.13 mg/L)、及空白组培养72 h后,用流式细胞仪检测凋亡率变化.结果 ①健康人外周血CD4+CD25+Treg的纯度为77.4%~92.3%,哮喘患者CD4+CD25+Treg的纯度为75.2%~93.8%.②CD4+CD25+Treg占外周血CD4+T细胞的比例在健康组为4.12%~7.98%,在哮喘组为4.51%~8.68%.两者差异无统计学意义.③小剂量氨茶碱均可以诱导健康组及哮喘组外周血CD4+CD25+Treg凋亡率增加(P<0.05).结论 小剂量氨茶碱(1.13 mg/L)可能通过促进CD4+CD25+Treg凋亡来发挥免疫调节作用.

  12. Ozone and allergen exposure during postnatal development alters the frequency and airway distribution of CD25+ cells in infant rhesus monkeys

    International Nuclear Information System (INIS)

    The epidemiologic link between air pollutant exposure and asthma has been supported by experimental findings, but the mechanisms are not understood. In this study, we evaluated the impact of combined ozone and house dust mite (HDM) exposure on the immunophenotype of peripheral blood and airway lymphocytes from rhesus macaque monkeys during the postnatal period of development. Starting at 30 days of age, monkeys were exposed to 11 cycles of filtered air, ozone, HDM aerosol, or ozone + HDM aerosol. Each cycle consisted of ozone delivered at 0.5 ppm for 5 days (8 h/day), followed by 9 days of filtered air; animals received HDM aerosol during the last 3 days of each ozone exposure period. Between 2-3 months of age, animals co-exposed to ozone + HDM exhibited a decline in total circulating leukocyte numbers and increased total circulating lymphocyte frequency. At 3 months of age, blood CD4+/CD25+ lymphocytes were increased with ozone + HDM. At 6 months of age, CD4+/CD25+ and CD8+/CD25+ lymphocyte populations increased in both blood and lavage of ozone + HDM animals. Overall volume of CD25+ cells within airway mucosa increased with HDM exposure. Ozone did not have an additive effect on volume of mucosal CD25+ cells in HDM-exposed animals, but did alter the anatomical distribution of this cell type throughout the proximal and distal airways. We conclude that a window of postnatal development is sensitive to air pollutant and allergen exposure, resulting in immunomodulation of peripheral blood and airway lymphocyte frequency and trafficking

  13. Aberrant Expression of Novel Cytokine IL-38 and Regulatory T Lymphocytes in Childhood Asthma.

    Science.gov (United States)

    Chu, Man; Chu, Ida M T; Yung, Edmund C M; Lam, Christopher W K; Leung, Ting F; Wong, Gary W K; Wong, Chun K

    2016-01-01

    We investigated the expression of novel anti-inflammatory interleukin (IL)-38 and regulatory T (Treg) lymphocytes in childhood asthma patients. The protein and mRNA expression level of IL-38, periostin, peripheral CD4⁺CD25⁺CD134⁺ T lymphocytes as well as CD4⁺CD25(high)FoxP3⁺ and CD4⁺CD25(high)CD127(-) Treg lymphocytes from 40 asthmatic patients and 20 normal control (NC) subjects were studied using ELISA, qPCR and flow cytometry. Serum and supernatant cytokines/chemokines were determined by multiplex assay. Serum IL-38, IL-5, IL-17, IL-6, interferon-γ, periostin, IL-1β and IL-13 concentrations were significantly higher in asthmatic patients with or without steroid treatment than those in controls (all p Treg lymphocytes were markedly decreased in asthmatic patients with and without steroid treatment than those in controls (all p Treg lymphocytes in asthmatic patients with high level (>40 ng/mL) of periostin (p asthma. PMID:27438823

  14. CD4+ CD25+ Foxp3+ T regulatory cells with limited T cell receptor diversity in control of autoimmunity1

    OpenAIRE

    Adeegbe, Dennis; Matsutani, Takaji; Yang, Jing; Altman, Norman H; Malek, Thomas R.

    2009-01-01

    The importance of high TCR diversity of Treg cells for self-tolerance is poorly understood. To address this issue, TCR diversity was measured for Treg cells after transfer into IL-2Rβ-/- mice, which develop lethal autoimmunity due to failed production of Treg cells. Here we show that high TCR diversity of pre-transferred Treg cells led to selection of therapeutic Treg cells with lower TCR diversity that prevented autoimmunity. Pre-transferred Treg cells with lower diversity led to selection o...

  15. CD4+ CD25+ Foxp3+调节性T细胞在成人过敏性哮喘急性发作期和缓解期外周血中的表达%The Level of CD4+ CD25+ Foxp3+ Tregs in Peripheral Blood of Allergic Asthma Patients with Acute Stage or Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    陈敏; 樊黎; 吴晓立

    2012-01-01

    目的:探讨成人过敏性哮喘急性发作期和缓解期外周血中CD4+ CD25+ Foxp3+ 调节性T 细胞的表达情况及与病情的相关性.方法:收集健康人、过敏性哮喘急性发作期和缓解期患者外周血,流式细胞术测定CD4+ CD25+ Foxp3+ Treg 的比例.结果:CD4+ CD25+ Foxp3+ Treg 细胞在三组间的差异有统计学意义.哮喘缓解期组、哮喘急性期组较健康对照组该细胞比例均明显下降,哮喘急性期组较哮喘缓解期组更低.CD4+ CD25+ Foxp3+ Treg 细胞数量与哮喘病情严重程度呈负相关.结论:CD4+ CD25+ Foxp3+ 调节性T 细胞可能在过敏性哮喘的发病过程中起重要作用,它的降低导致的免疫缺陷可能与哮喘的发生相关.

  16. 脐血CD4+CD25+CD127low调节T细胞及淋巴细胞亚群分析%Study on CD4+CD25+CD127low Regulatory T Cells and Lymphocytes Subsets in Umbilical Cord Blood

    Institute of Scientific and Technical Information of China (English)

    张劼; 陈军浩

    2012-01-01

    目的 检测新生儿脐带血淋巴细胞亚群和调节性T细胞比率,了解脐带血免疫学的特征.方法 使用Sysmex XE2100血细胞分析仪分别计数脐带血、新生儿母亲及对照组外周血淋巴细胞;采用流式细胞术分别检测其CD3+T细胞、CD19+B细胞、CD3- CD16+56+ NK细胞、CD3+ CD4+细胞、CD3+ CD8+细胞占淋巴细胞百分比,以及CD4+ CD25+CD127low调节性T细胞占CD4+细胞的百分比.结果 脐带血、新生儿母亲及对照组淋巴细胞计数分别为(3.68±1.07)×109/L,(1.42±0.44)×109/L和(2.06±0.88)×109/L;B淋巴细胞为:15.71%±3.89%,11.13%±3.79%和9.69%±2.22%;CD4+T细胞为:50.27%±9.08%,37.25%±7.13%和34.65%±7.17%;调节性T细胞为:6.94%±1.09%,5.09%±0.95%和4.8%1±0.99%.上述检测结果脐带血均显著高于母亲及对照组,P<0.01,母亲与对照组差异无统计学显著性意义P>0.05.三组间CD3+T细胞(69.64%±9.97%,74.83%±5.91%和69.41%±5.42%)和NK(11.36%±7.93%,10.48%±6.78%和16.31%±4.69%)细胞无显著性差异,P>0.05.脐带血中CD8+T细胞低于母亲及对照组(19.38%±6.62%,32.39%±2.08%和31.16%±1.87%),P<0.01.结论 脐血中高水平的CD4+ CD25+ CD127low调节性T细胞和低水平的CD8+T细胞有助于保持脐带血的低免疫状态.%Objective The proportions of lymphocytes subsets and regulatory T cells in umbilical cord blood were analyzed to explore the characteristic of umbilical cord blood in immunology. Methods To count lymphocytes by Sysmex XE2100 hema-tocyte cytometer. The percentages of CD3 + T cells,CD19+ B cells,CD3- CD16 + 56+ NK cells,CD3+ CD4+ cells,CD3+ CD8+ cells,and CD4+CD25+CD127low regulatory T cells among the umbilical cord bloods and the peripheral bloods from their mothers or control group were determined by flow cytometry. Results The quantity of lymphocytes in cord blood was significantly higher compared with mothers or with control group (3. 68

  17. Optogenetic Control of Gene Expression in Drosophila.

    Directory of Open Access Journals (Sweden)

    Yick-Bun Chan

    Full Text Available To study the molecular mechanism of complex biological systems, it is important to be able to artificially manipulate gene expression in desired target sites with high precision. Based on the light dependent binding of cryptochrome 2 and a cryptochrome interacting bHLH protein, we developed a split lexA transcriptional activation system for use in Drosophila that allows regulation of gene expression in vivo using blue light or two-photon excitation. We show that this system offers high spatiotemporal resolution by inducing gene expression in tissues at various developmental stages. In combination with two-photon excitation, gene expression can be manipulated at precise sites in embryos, potentially offering an important tool with which to examine developmental processes.

  18. 雾化吸入变应原对支气管哮喘外周血CD4+CD25+Foxp3+Treg的影响%Effects of Desensitization of Allergen Nebulization on Blood CD4 + CD25 + Foxp3 + Treg in the Prevention and Treatment of Asthma

    Institute of Scientific and Technical Information of China (English)

    李红华; 赵娟; 张景鸿

    2012-01-01

    Objective To evaluate the effect of allergen nebulization on the ratio of blood CD4 + CD25 + Foxp3 + Treg in asthma. Methods 40 patients with Der. p and Der. f allergy and newly diagnosed uncontrolled moderate persistent bronchial asthma were randomly divided into 2 groups; group A and group B (20 per group). The patients in group B were nebulized with specific allergen twice a week for 6 months. Both groups were treated with salmeterol xinafoate and fluticasone propionate powder. The percentage of CD4 + CD25 + Foxp3 + Treg cells in peripheral blood was determined by flow cytometry. ACT, airway responsiveness and lung function were performed before and after treatment. Results The negative conversion rates of Bronchial Provocation Test in group B were higher than group A significantly. The percentage of CD4 + CD25 + Foxp3 + Treg cells increased in group B when compared with group A(P <0. 05). Conclusion CD4 + CD25 + Foxp3 + T cells played an important immunomudulatory role in immunotherapy of allergen nebulization in treatment of asthma.%目的 探讨CD4+ CD25+ Foxp3+ Treg细胞在哮喘变应原雾化吸入减敏治疗的作用.方法 粉尘螨和屋尘螨点刺阳性的支气管哮喘患者随机分为A组(常规治疗)和B组(变应原吸入减敏),各20例.B组雾化吸入特异性变应原,A组雾化吸入以生理盐水代替,两组给予相同的基础治疗.治疗前、后用流式细胞术(FCM)检测外周血中CD4+CD25+ Foxp3+T细胞占CD4 +T细胞比例;进行哮喘控制评分和肺功能、气道反应性测定.结果 治疗后B组ACT评分高于A组,两组肺功能均有明显增加.B组支气管激发试验转阴率明显高于A组.B组外周血中CD4+ CD25+Foxp3+T细胞占CD4 +T细胞比例较A组明显升高(P<0.05).结论 CD4+ CD25+ Foxp3+T细胞在变应原雾化吸入减敏防治支气管哮喘中发挥免疫调节作用.

  19. 高迁移率族蛋白B1对小鼠调节性T细胞与CD4+CD25-T细胞相互作用的影响%Influence of high mobility group box-1 protein on the correlation between regulatory T cells and CD4+CD25-T cells of spleen in mice

    Institute of Scientific and Technical Information of China (English)

    张莹; 姚咏明; 于燕; 吴瑶; 盛志勇

    2008-01-01

    Objective To investigate the influence of high mobility group box-1 protein(HMGB1)on the immunosuppression function of splenic regulatory T cells(Tregs)and its potential regulatory mechanism underlying the effect on CD4+CD25-T cells in mice.nethods CD4+CD25+Tregs isolated from the spleens of male BALB/c mice by magnetic beads were seeded on 96-well(1×105 cells/well)cell culture plates coated with 1 μg/ml anti-CD3 and soluble CD28.After being stimulated with HMGB1 for different time and concentrations,the secretions of IL-2 and IL-10 were analyzed by ELISA.Tregs stimulated for 72 hours were cultured with CD4+CD25-T cells together.The suppressive activity of CD4+CD25+Treg to CD4+CD25-T cells was analyzed by MTT test.IL-2,IL-10,IL-4,and interferon(IFN)-γ in the cell suspensions were determined by ELISA.Resuits After stimulation with HMGB1,the suppressive activity of splenic Tregs in mice were significantly down-regulated at 72 hours,when the proportion of Tregs to CD4+CD25-T cells was 1:1.The secretion of IL-2 of Tregs stimulated by HMGB1 was not markedly changed(P>0.05).while a dose-dependent decrease between IL-10 induction and HMGB1 concentration was obviously(P<0.05).When CD4+CD25-T cells were cultured with stimulated Tregs,comparing with unstimulated-Treg group,levels of IL-2 and IFN-γ were elevated following the increased concentration of HMGB1(P<0.05 or P<0.01).Meanwhile the secretion of IL-4 and IL-10 significantly decreased when cultured with stimulated Tregs(P<0.05).Conclusions These data suggested that HMGB1 stimulation can result in significant down-regulation of immunosuppression of splenic Tregs in mice.HMGB1 might be a potential immunoregnlatory signal that influences the proliferation of effector T cells,secretion of IL-2 and cells-polarization by inhibiting CD4+CD25+Tregs activity.%目的 观察高迁移率族蛋白B1(HMGB1)对调节性T细胞(Treg)与CD4+CD25-T细胞相互作用的影响,并初步探讨其影

  20. The Relations of Parenting, Effortful Control, and Ego Control to Children's Emotional Expressivity.

    Science.gov (United States)

    Eisenberg, Nancy; Zhou, Qing; Losoya, Sandra H.; Fabes, Richard A.; Shepard, Stephanie A.; Murphy, Bridget, C.; Reiser, Mark; Guthrie, Ivanna K.; Cumberland, Amanda

    2003-01-01

    Examined longitudinal relations of observed parental warmth and positive expressivity and children's effortful control and ego control with children's high versus low emotional expressivity. Found that moderate child expressivity related to high effortful control. Children's ego overcontrol mediated relations between parental warmth/positive…

  1. The ex vivo Microenviroments in MLTC of Poorly Immunogenic Tumor Cells Facilitate Polarization of CD4+CD25+ Regulatory T Cells

    Institute of Scientific and Technical Information of China (English)

    Le Zhou; Hongyan Wang; Juxiang Xiao; Lusheng Si; Yili Wang

    2006-01-01

    CD4+CD25+ regulatory T (TR) cells play an important role in maintaining a balanced peripheral immune system.Recent studies have shown that TR cells may also play a key role in suppressing anti-tumor immune response. In order to investigate the tumor immune microenvironment and its influence on TR polarization, poorly immunogenic tumor cell line D5 (C57BL/6, H-2b), immunogenic tumor cell lines FBL3 (C57BL/6, H-2b) and H22 BALB/c, H-2d) were used to establish the syngeneic/allogeneic, poorly immunogenic/immunogenic mixed lymphocytes-tumor cell culture (MLTC). Our results revealed that the proportion of CD4+CD25+ T cells in MLTC of syngeneic primed splenocytes stimulated with D5 tumor cells was higher than that with H22 cells (0.43% vs 0.044%, and the similar results appeared in allogeneic splenocytes stimulated with D5 tumor cells (0.39% vs 0.04%).The splenocytes stimulated with supernatant from syngeneic MLTC of D5 tumor cells demonstrated higher proportion of CD4+CD25+ cells than that from allogeneic MLTC of D5 tumor cells, and the splenocytes stimulated with supernatant from syngeneic or allogeneic MLTC of H22 tumor cells generated lower proportion of CD4+CD25+ T cells than that of D5 tumor cells. The TGF-β1 and Th2-oriented cytokines (IL-4 and IL-10) were dominated in supernatants of syngeneic MLTC of poorly immunogenic tumor cells. Our results provided useful information for studying the mechanisms underlying tumor immune surveillance as well as for the tumor immunotherapy.

  2. Inhibitory effect of CD4~+CD25~+CCR6~+ regulatory T cells against CD8~+T cells in mouse mammary carcinoma model%CD4~+CD25~+CCR6~+调节性T细胞在小鼠乳腺癌模型中对CD8~+T细胞的抑制作用

    Institute of Scientific and Technical Information of China (English)

    徐林; 徐薇; 蒋正刚; 熊思东

    2009-01-01

    Objective:To observe the inhibitory effect of CD4~+CD25~+CCR6~+ regulatory T cells (CCR6~+ Tregs) against CD8~+T cells in vivo, and to investigate its relationship with tumor immune escape. Methods: Mouse mammary carcinoma models were established by inoculating mammary carcinoma 4T1 cells into nude mice. CCR6~+ Tregs were isolated by FACS, and the Foxp3 expression on CCR6~+ Tregs was further analyzed by FACS. 4T1 specific CD8~+T cells were labeled with CFSE after isolation by FACS, and then transferred into 4T1 bearing nude mice combined with or without CCR6~+ Tregs or CD4~+CD25~+CCR6-regulatory T cells (CCR6- Tregs). Tumor growth and survival of 4T1 bearing mice were observed. The proliferation, IFN-γ production, and granzyme B expression of CD8~+T cells were examined by FACS. Results: Both CCR6~+ Tregs and CCR6- Tregs expressed high levels of Foxp3. The tumors in CCR6~+ Tregs and CD8~+T cells co-transferred mice grew faster than those in CCR6- Tregs co-transferred and CD8~+T cell-transferred groups. The survival period of 4T1 bearing mice was significantly decreased in CCR6~+ Tregs co-transferred group (P<0.05). Furthermore, the proliferation, IFN-γ production and granzyme B expression of CD8~+ T cells were also dramatically decreased in CCR6~+ Tregs co-transferred group compared with those in CCR6- Tregs co-transferred and CD8~+ T cell-transferred groups (P<0.05). Conclusion: CCR6~+ Tregs can effectively inhibit the function of CD8~+ T cells, which might play an important role in tumor immune escape, tumor development and progress.%目的:观察CD4~+CD25~+CCR6~+调节性T细胞(简称CCR6~+Tregs)体内对CD8~+T细胞功能的抑制作用,并探讨其与肿瘤免疫逃逸的关系.方法:建立4T1乳腺癌细胞荷瘤裸鼠模型,FACS分选CCR6~+Tregs,检测其Foxp3的表达;FACS分选4T1特异性CD8~+T细胞,CFSE标记后分别与CCR6~+Tregs或CCR6-Tregs共同过继转输入4T1荷瘤裸鼠体内,观察荷瘤裸鼠肿瘤生长情况和小鼠存活

  3. Diminished CD4+/CD25+ T cell and increased IFN-gamma levels occur in dogs vaccinated with Leishmune in an endemic area for visceral leishmaniasis.

    Science.gov (United States)

    de Lima, Valéria Marçal Felix; Ikeda, Fabiana Augusta; Rossi, Cláudio N; Feitosa, Mary Marcondes; Vasconcelos, Rosemeride Oliveira; Nunes, Caris Maroni; Goto, Hiro

    2010-06-15

    The Leishmune vaccine has been used in endemic areas to prevent canine visceral leishmaniasis in Brazil, but cytokine production induced by vaccination has rarely been investigated in dogs. This study aimed to evaluate the immune response of dogs vaccinated with Leishmune FML vaccine (Fort Dodge) against total antigen of Leishmania (Leishmania) chagasi (TAg) and FML. Twenty healthy dogs from Araçatuba, São Paulo, Brazil, an endemic leishmaniasis area, received three consecutive subcutaneous injection of Leishmune vaccine at 21-day intervals. PBMC were isolated before and 10 days after completing vaccination and lymphoproliferative response and antibody production against FML or total promastigote antigen were tested. Cytokines IFN-gamma, IL-4 and TNF-alpha were measured in culture supernatant and CD4+/CD25+ and CD8+/CD25+ T cell presence was determined. Analysis of the data indicated that the vaccine conferred humoral responses (100%) against both antigens and cellular immunity to FML (85%) and total antigen (80%), the supernatant of cultured cells stimulated with TAg and FML showed an increase in IFN-gamma (P<0.05), and the vaccine reduced CD4+/CD25+ T cell presence compared to that observed before vaccination. These responses may constitute part of the immune mechanism induced by Leishmune. PMID:20132994

  4. Epigenetic control of antioxidant gene expression

    OpenAIRE

    Wild, Brigitte

    2015-01-01

    Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 29-10-2015 To respond to exogenous and endogenous stimuli, organisms have developed a variety of mechanisms to modulate the quantity, duration and the type of response to these stimuli. Of these mechanisms, one of the most important is the regulation of gene expression. This regulation of gene expression occurs at various levels but especially by th...

  5. 卡介苗多糖核酸对哮喘大鼠淋巴液和血液CD4+CD25+Foxp3+调节性T细胞的影响%The effect of BCG-PSN on CD4+CD25+Foxp3+Treg in blood and lymph of asthma rats

    Institute of Scientific and Technical Information of China (English)

    石涛; 冯学斌; 赵志旭; 张瑾锦

    2010-01-01

    目的:探讨卡介苗多糖核酸(BCG-PSN)对哮喘大鼠淋巴液和血液调节性T细胞数量及功能的影响.方法:将SD大鼠随机分为对照组、哮喘组和BCG-PSN组,分别收集不同时间点大鼠淋巴液和血液,采用流式细胞仪(FCM)检测CD4+CD25+Foxp3+调节性T细胞(CD4+CD25+Foxp3+Treg)百分率,酶联免疫吸附试验(ELISA)检测淋巴液和血浆白介素10(IL-10)和转录生长因子β1(TGF-β1)浓度.结果:各组在各时间点其淋巴液中CD4+CD25+Foxp3+Treg百分率、IL-10水平均较血液明显升高.哮喘组大鼠淋巴液和血液中CD4+CD25+Foxp3+Treg百分率、IL-10 、TGF-β1浓度均较对照组显著降低(P<0.05).BCG-PSN组淋巴液和血液中CD4+CD25+Foxp3+Treg百分率和IL-10水平较哮喘组明显升高(P<0.05),与对照组比较无显著性差异;而TGF-β水平在48小时较对照组和哮喘组明显升高(P<0.05).结论:哮喘大鼠淋巴液和血液存在明显CD4+CD25+Foxp3+Treg数量及功能不足.BCG-PSN可能通过增加哮喘大鼠外周血和淋巴液中CD4+CD25+Foxp3+Treg的数量及其产生IL-10和TGF-β水平,增强免疫抑制效应,从而发挥抑制哮喘炎症的作用.

  6. The role of CD4+CD25+ regulatory T cells in the pathogenesis of asthma in children%CD4+CD25+调节性T细胞在儿童哮喘发病机制中的作用初探

    Institute of Scientific and Technical Information of China (English)

    祖莹; 李成荣; 郑跃杰; 邓继岿; 付晓玲

    2006-01-01

    目的探讨哮喘急性发作患儿CD4+CD25+调节性T(Tr)细胞数量变化及影响其发育成熟的因素.方法观察20例哮喘患儿及相同数量同龄对照组.用流式细胞术检测外周血单个核细胞(PBMC)CD4、CD25表面标志及CD4+CD25+细胞内白细胞介素(IL-10)和转化生长因子受体(TGF-β)表达.用逆转录-聚合酶链反应(RT-PCR) 和荧光定量聚合酶链反应(Real-time PCR)检测PBMC Foxp3及SOCS1 mRNA表达.结果急性发作期哮喘患儿CD4+CD25+Tr细胞百分率(6.5%±1.9%)明显低于同龄对照组(12.0%±2.3%),P<0.01 ;CD4+CD25++IL-10及CD4+CD25++TGF-β分泌细胞亦明显低于对照组.Foxp3及SOCS1mRNA表达也显著降低( Foxp3:0.12±0.05 vs 1.71±0.58,P<0.001;SOCS1:0.38±0.19 vs 2.14±0.41,P<0.001).结论哮喘患儿CD4+CD25+Tr细胞明显降低可能参与哮喘发病,Foxp3及SOCS1表达降低可能是导致CD4+CD25+Tr细胞发育障碍的重要因素.

  7. Automatic Control of Gene Expression in Mammalian Cells.

    Science.gov (United States)

    Fracassi, Chiara; Postiglione, Lorena; Fiore, Gianfranco; di Bernardo, Diego

    2016-04-15

    Automatic control of gene expression in living cells is paramount importance to characterize both endogenous gene regulatory networks and synthetic circuits. In addition, such a technology can be used to maintain the expression of synthetic circuit components in an optimal range in order to ensure reliable performance. Here we present a microfluidics-based method to automatically control gene expression from the tetracycline-inducible promoter in mammalian cells in real time. Our approach is based on the negative-feedback control engineering paradigm. We validated our method in a monoclonal population of cells constitutively expressing a fluorescent reporter protein (d2EYFP) downstream of a minimal CMV promoter with seven tet-responsive operator motifs (CMV-TET). These cells also constitutively express the tetracycline transactivator protein (tTA). In cells grown in standard growth medium, tTA is able to bind the CMV-TET promoter, causing d2EYFP to be maximally expressed. Upon addition of tetracycline to the culture medium, tTA detaches from the CMV-TET promoter, thus preventing d2EYFP expression. We tested two different model-independent control algorithms (relay and proportional-integral (PI)) to force a monoclonal population of cells to express an intermediate level of d2EYFP equal to 50% of its maximum expression level for up to 3500 min. The control input is either tetracycline-rich or standard growth medium. We demonstrated that both the relay and PI controllers can regulate gene expression at the desired level, despite oscillations (dampened in the case of the PI controller) around the chosen set point. PMID:26414746

  8. Plasmodium falciparum–Mediated Induction of Human CD25hiFoxp3hi CD4 T Cells Is Independent of Direct TCR Stimulation and Requires IL-2, IL-10 and TGFβ

    OpenAIRE

    Scholzen, Anja; Mittag, Diana; Rogerson, Stephen J.; Cooke, Brian M.; Plebanski, Magdalena

    2009-01-01

    CD4+CD25+Foxp3+ regulatory T cells (Tregs) regulate disease-associated immunity and excessive inflammatory responses, and numbers of CD4+CD25+Foxp3+ Tregs are increased during malaria infection. The mechanisms governing their generation, however, remain to be elucidated. In this study we investigated the role of commonly accepted factors for Foxp3 induction, TCR stimulation and cytokines such as IL-2, TGFβ and IL-10, in the generation of human CD4+CD25+Foxp3+ T cells by the malaria parasite P...

  9. Statistical Quality Control of Microarray Gene Expression Data

    Directory of Open Access Journals (Sweden)

    Shen Lu

    2011-12-01

    Full Text Available This paper is about how to control the quality of microarray expression data. Since gene-expression microarrays have become almost as widely used as measurement tools in biological research, we survey microarray experimental data to see possibilities and problems to control microarray expression data. We use both variable measure and attribute measure to visualize microarray expression data. According to the attribute data's structure, we use control charts to visualize fold change and t-test attributes in order to find the root causes. Then, we build data mining prediction models to evaluate the output. According to the accuracy of the prediction model, we can prove control charts can effectively visualize root causes.

  10. Fel d 1-airway inflammation prevention and treatment by co-immunization vaccine via induction of CD4+CD25-Foxp3+ Treg cells

    OpenAIRE

    Pei, Yechun; Geng, Shuang; Liu, Lin; Yan, Fengxiang; Hong GUAN; Hou, Jian; Chen, Yongfu; Wang, Bin; An, Xiaorong

    2013-01-01

    Pet allergens are major causes for asthma and allergic rhinitis. Fel d 1 protein, a key pet allergen from domestic cat, can sensitize host and trigger asthma attack. In this study, we report that co-immunization with recombinant Fel d 1 protein (rFel d 1) plus plasmid DNA that contains Fe1 d 1 gene was effective in preventing and treating the natural Fel d 1 (nFel d 1) induced allergic airway inflammation in mice. A population of T regulatory cells (iTreg) exhibiting a CD4+CD25-Foxp3+ phenoty...

  11. Correlation of memory T cell responses against TRAP with protection from clinical malaria, and CD4 CD25 high T cells with susceptibility in Kenyans.

    Directory of Open Access Journals (Sweden)

    Stephen M Todryk

    Full Text Available BACKGROUND: Immunity to malaria develops naturally in endemic regions, but the protective immune mechanisms are poorly understood. Many vaccination strategies aim to induce T cells against diverse pre-erythrocytic antigens, but correlates of protection in the field have been limited. The objective of this study was to investigate cell-mediated immune correlates of protection in natural malaria. Memory T cells reactive against thrombospondin-related adhesive protein (TRAP and circumsporozoite (CS protein, major vaccine candidate antigens, were measured, as were frequencies of CD4(+ CD25(high T cells, which may suppress immunity, and CD56(+ NK cells and gammadelta T cells, which may be effectors or may modulate immunity. METHODOLOGY AND PRINCIPAL FINDINGS: 112 healthy volunteers living in rural Kenya were entered in the study. Memory T cells reactive against TRAP and CS were measured using a cultured IFNgamma ELISPOT approach, whilst CD4(+ CD25(high T cells, CD56(+ NK cells, and gammadelta T cells were measured by flow cytometry. We found that T cell responses against TRAP were established early in life (<5 years in contrast to CS, and cultured ELISPOT memory T cell responses did not correlate with ex-vivo IFNgamma ELISPOT effector responses. Data was examined for associations with risk of clinical malaria for a period of 300 days. Multivariate logistic analysis incorporating age and CS response showed that cultured memory T cell responses against TRAP were associated with a significantly reduced incidence of malaria (p = 0.028. This was not seen for CS responses. Higher numbers of CD4(+ CD25(high T cells, potentially regulatory T cells, were associated with a significantly increased risk of clinical malaria (p = 0.039. CONCLUSIONS: These data demonstrate a role for central memory T cells in natural malarial immunity and support current vaccination strategies aimed at inducing durable protective T cell responses against the TRAP antigen. They also

  12. CD4+CD25+Treg细胞在慢性荨麻疹发病中的作用

    Institute of Scientific and Technical Information of China (English)

    冯欢; 郭胤仕

    2011-01-01

    CD4+CD25+Treg细胞的功能紊乱在变态反应性疾病发生发展中的作用日益引起人们的关注.新近研究表明慢性荨麻疹是由于Treg细胞数量和(或)功能上的异常所引发.文章阐述了Treg细胞的来源、分类、作用机制及其与慢性荨麻疹之间的关系.

  13. 支气管哮喘患者外周血Th17、CD4+CD25+Treg细胞表达特征%The prevalence of blood Th17 and CD4+CD25+Treg cells in patients with bronchial asthma

    Institute of Scientific and Technical Information of China (English)

    施宇衡; 时国朝; 万欢英; 蒋黎华; 艾香艳; 朱海星; 汤葳

    2010-01-01

    目的:探讨外周血Th17和CD4+CD25+调节性T细胞(Treg)在支气管哮喘患者中的表达特征.方法:41例慢性持续期哮喘患者,分为间歇-轻度组(n=23)和中重度组(n=18),行肺功能检查和哮喘控制问卷(ACQ)调查,20例正常人作为对照.通过流式细胞术检测外周血Th17和CD4+CD25+Treg细胞的比例.ELISA检测血浆以及植物血凝素刺激24小时后外周血单个核细胞(PBMC)上清液中的IL-17、IL-10、TGF-β水平.结果:中重度哮喘组外周血Th17细胞比例及血浆IL-17水平高于间歇-轻度哮喘和正常人组,而外周血CD4+CD25+Foxp3+Treg细胞比例及血浆IL-10、TGF-β水平则降低.中重度哮喘组PBMC上清液中IL-17水平增高.哮喘患者FEV1(%预计值)与Th17细胞及血浆IL-17表达成负相关,与CD4+CD25+Treg表达成正相关.ACQ平均得分与Th17细胞和血浆IL-17表达成正相关,与外周血CD4+CD25+Treg表达成负相关.结论:中重度哮喘中外周血Th17细胞应答增强,而CD4+CD25+ Treg细胞缺乏,哮喘的严重程度及症状控制与外周血Th17/Treg免疫应答失衡密切相关.

  14. Practicing Qigong Baduanjin on drug addicts can reduce CD4+CD25+ regulatory T cells%习练健身气功八段锦降低戒毒人员CD4+CD25+调节性T细胞作用的研究

    Institute of Scientific and Technical Information of China (English)

    陈昌乐; 王艳; 李洁

    2014-01-01

    目的:通过比较习练健身气功八段锦前后,戒毒人员外周血中CD4+CD25+调节性T细胞(CD4+C D25+Treg)占CD4+T细胞的比例变化,研究健身气功八段锦提高戒毒人员免疫功能的分子机制.方法:80名戒毒人员随机分为健身气功八段锦习练组和对照组各40名.3个月后,流式细胞术法检测习练组和对照组外周血中CD4+CD25+T细胞、CD3+细胞、CD4+T细胞比例和CD4/CD8比值.免疫磁珠法分离并去除外周血中CD4+CD25+T细胞后,3H-TdR掺入法检测淋巴细胞增殖反应.结果:3个月后,习练组外周血中免疫抑制性CD4+CD25+T细胞比例明显低于对照组(P<0.05);而习练组外周血中CD3+细胞、CD4+T细胞、CD4/CD8细胞比例明显高于对照组(P<0.05).习练组外周血T淋巴细胞增殖反应明显高于对照组(P<0.05),免疫磁珠法去除CD4+CD25+T细胞后,对照组外周血T淋巴细胞增殖反应恢复至与习练组和健康对照组水平.结论:习练健身气功八段锦可降低戒毒患者外周血内CD4+CD25+Treg 比例,提高戒毒患者的免疫功能.

  15. 胃肠道癌肿患者外周血CD4+CD25+FOXP3+调节性T细胞的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    仇俊兰; 张惠; 曹斐; 吴惠莲; 皇甫照; 奉林

    2009-01-01

    目的:研究胃肠道癌肿患者外周血CD4+CD25+FOXP3+调节性T(Treg)细胞的表达,并探讨其临床意义。方法:通过免疫荧光术及流式细胞仪检测20例胃癌患者及20例结肠癌患者外周血CD4+CD25+FOXP3+Treg细胞、CD4+CD25+high Treg细胞、CD4+T细胞及CD4+ CTLA-4+T细胞。结果:胃癌组、结肠癌组与健康献血者比较外周血CD4+CD25+FOXP3+ Treg细胞、CD4+CD25+high Treg细胞及CD4+CTLA-4+T细胞显著增多,CD4+T细胞显著减少;胃癌、结肠癌患者之间其外周血中CD4+CD25+FOXP3+Treg细胞、CD4+CD25+high Treg细胞、CD4+T细胞及CD4+CTLA-4+T细胞无显著差异。结论:胃肠道癌肿患者外周血CD4+CD25+FOXP3+Treg细胞显著高于健康献血者,这可能与胃肠道癌肿患者的免疫抑制和肿瘤的进展相关。

  16. Different Responses of CD4+CD25+ Regulatory T Cells in Allergic Asthma Patients and Asymptomatic Sensitised People After Stimulation with Allergen in Vitro%CD4+CD25+调节性T细胞在过敏性哮喘和无症状的过敏者中表达的差异

    Institute of Scientific and Technical Information of China (English)

    王丽慧; 杨炯; 杨亦斌

    2011-01-01

    目的:比较过敏性哮喘患者、无症状的过敏者和不过敏的健康人外周血CD4+CD25+调节性T细胞(Treg)数量的差异及对特异性过敏原刺激的反应.方法:选取急性发作期的尘螨过敏性哮喘患者20例,无症状的尘螨过敏者24例,不过敏的健康人22例,分离外周血单个核细胞(PBMC)并进行CFSE染色,分别在不刺激和1mg/L尘螨抗原(rDer p 1)刺激情况下体外培养7 d,流式细胞仪检测CD4+CD25+Treg的比率和分裂指数;ELISA法检测各组细胞培养上清液和血清中白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)的水平.结果:过敏性哮喘患者CD4+CD25+Treg比率和IFN-γ基础水平均低于无症状的尘螨过敏者和不过敏的健康人,且过敏原刺激后增加不明显;但IL-4的基础水平最高,过敏原刺激后差异更显著.无论刺激与否,无症状的尘螨过敏者CD4+CD25+Treg比率均与不过敏的健康人的水平接近.但过敏原刺激后无症状的尘螨过敏者CD4+CD25+Treg比率高于过敏性哮喘患者,且存在高水平的IFN-γ反应.结论:接触过敏原后,过敏性哮喘患者和无症状的过敏者表现出不同的免疫反应模式.过敏性哮喘患者抗原特异性CD4+CD25+Treg反应不足,存在过高Th2反应.无症状过敏的发生可能与机体对特异性过敏原刺激产生高水平的CD4+CD25+Treg反应及高水平的IFN-γ反应有关.%Objective: To determine whether there are differences in the numbers and the responses of CD4+CD25+ regulatory T cells (Treg) in allergic asthmatics and people asymtomatic sensitised to specific allergen after specific allergen exposure. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from 20 patients sensitized to house dust mite with asthma, 24 asymptomatic subjects sensitized to house dust mite, and 22 non-allergic subjects. Cells were cultured without stimulation and with 1 mg/L antigen of dust mite rDer p 1. Frequencies and division index of CD4+ CD25+ Tregs were

  17. Chronic Heat Stress Inhibits Immune Responses to H5N1 Vaccination through Regulating CD4+CD25+Foxp3+ Tregs

    Directory of Open Access Journals (Sweden)

    Di Meng

    2013-01-01

    Full Text Available Chronic heat stress (CHS is known to have negative impacts on the immune responses in animals and increases their susceptibility to infections including the highly pathogenic avian influenza virus H5N1. However, the role of regulatory T cells (Tregs in CHS immunosuppression remains largely undefined. In this study, we demonstrated a novel mechanism by which CHS suppressed both Th1 and Th2 immune responses and dramatically decreased the protective efficacy of the formalin-inactivated H5N1 vaccine against H5N1 influenza virus infection. This suppression was found to be associated with the induced generation of CD4+CD25+FoxP3+ Tregs and the increased secretions of IL-10 and TGF-β in CD4+ T cells. Adoptive transfer of the induced Tregs also suppressed the protective efficacy of formalin-inactivated H5N1 virus immunization. Collectively, this study identifies a novel mechanism of CHS immunosuppression mediated by regulating CD4+CD25+Foxp3+ Tregs.

  18. 变应性鼻炎患者外周血CD4+CD25+Treg及Foxp3的检测及意义

    Institute of Scientific and Technical Information of China (English)

    王淑慧; 王桂琴

    2012-01-01

    目的 通过检测变应性鼻炎(AR)患者外周血中调节性T细胞(Treg)的含量,并与正常健康人外周血中Treg含量作比较,探讨Treg在AR患者发病过程中的作用机制.方法 30例AR患者为试验组(8周未用抗组胺、糖皮质激素),15例正常健康人为对照组,采用流式细胞术分别检测以上受试对象外周血中Treg的含量,结果以CD4+CD25+Treg、CD4+CD25highTreg及Foxp3占外周血T细胞的百分比表示.结果 AR患者外周血Treg含量明显低于正常人( P<0.05).结论 AR患者外周血Treg细胞比例明显减少,是变应性鼻炎的发病机制之一.

  19. Change and its significance of CD4+ CD25+ regulatory T cells in experimental murine mammary carcinoma model%小鼠乳腺癌实验动物模型中CD4+CD25+调节性T细胞的变化及意义

    Institute of Scientific and Technical Information of China (English)

    徐林; 蒋正刚; 李宝华; 熊思东

    2006-01-01

    目的 研究CD4+CD25+调节性T细胞在小鼠乳腺癌实验动物模型中的变化,并探讨其意义.方法 以4T1接种BALB/c小鼠建立小鼠乳腺癌动物模型;分别以接种后1周和4周为荷瘤早、晚期;用流式细胞术(FACS)检测小鼠CD4+CD25+Foxp3+调节性T细胞在肿瘤局部及引流淋巴结中的比例变化;通过特异性增殖和杀伤实验观察早、晚期肿瘤局部的免疫应答.结果 CD4+CD25+调节性T细胞在肿瘤浸润淋巴细胞中的比例晚期为7.36%±0.26%,高于早期4.47%±0.88%(P<0.05);在引流淋巴结中比例变化不大;肿瘤浸润淋巴细胞的抗原特异性增殖和杀伤能力明显减弱(P<0.05).结论 CD4+CD25+调节性T细胞在肿瘤局部存在富集,这可能是导致肿瘤局部免疫反应减弱的重要原因.

  20. CD4+CD25highCD127low Regulatory T Cells in Peripheral Blood Are Not an Independent Factor for Chronic Graft-versus-Host Disease after Allogeneic Stem Cell Transplantation

    Science.gov (United States)

    Perz, Jolanta B.; Gürel, Selma; Schonland, Stefan O.; Hegenbart, Ute; Ho, Anthony D.; Dreger, Peter

    2012-01-01

    Background. The therapeutic efficacy of allogeneic hemopoietic stem cell transplantation (HSCT) largely relies on the graft-versus-leukemia (GVL) effect. Uncontrolled graft-versus-host disease (GVHD) is a feared complication of HSCT. Regulatory T cells (Treg) are a subset of CD4+ T-helper cells believed to maintain tolerance after HSCT. It remains unclear whether low peripheral blood Treg have an impact on the risk for acute (aGVHD) and chronic GVHD (cGVHD). Methods. In this paper we enumerated the CD4+CD25highCD127low Treg in the peripheral blood of 84 patients after at least 150 days from HSCT and in 20 healthy age-matched controls. Results. Although similar mean lymphocyte counts were found in patients and controls, CD3+CD4+ T-cell counts were significantly lower in patients. Patients also had significantly lower Treg percentages among lymphocytes as compared to controls. Patients with cGVHD had even higher percentages of Treg if compared to patients without cGVHD. In multivariate analysis, Treg percentages were not an independent factor for cGVHD. Conclusions. This paper did not show a relation between deficient peripheral blood Treg and cGVHD, therefore cGVHD does not seem to occur as a result of peripheral Treg paucity. PMID:22666141

  1. HBV宫内感染新生儿外周血调节性T细胞表达%Detection and analysis of CD4 + CD25+ regulatory T cell in peripheral blood from newborns with HBV intrauterine infection

    Institute of Scientific and Technical Information of China (English)

    高怡; 郭健; 付振东; 郝海燕; 刘明慧; 汪波; 丰淑英; 王素萍

    2013-01-01

    目的 探讨CD4+ CD25+调节性T细胞(Treg)在乙肝病毒(HBV)宫内感染新生儿外周血中的表达和意义.方法 选择乙肝表面抗原(HBsAg)阳性孕妇及其分娩新生儿79例,用酶联免疫吸附试验检测孕妇和新生儿外周血HBV标志物,实时荧光定量PCR检测母亲和新生儿外周血HBV DNA含量,用流式细胞仪检测新生儿外周血CD4+ CD25+ Treg和CD4+ CD25+ Foxp3+ Treg水平.结果 宫内感染HBV新生儿外周血CD4+ CD25+Treg和CD4+ CD25+ Foxp3+ Treg占CD4+T细胞比例分别为(10.57±2.25)%和(1.67±0.37)%,均高于未感染新生儿,差异有统计学意义(P<0.05),随产妇HBV DNA载量增加,新生儿外周血CD4+ CD25+ Treg和CD4+CD25+ Foxp3+ Treg增加,CD4+ CD25+ Treg比例高于阴性组(P <0.05);CD4+CD25+ Treg和CD4+ CD25+Foxp3+ Treg均与产妇HBV DNA载量呈正相关(r=0.430、0.409,P<0.05).结论 Treg可能通过抑制机体细胞免疫反应影响乙肝病毒清除,新生儿HBV宫内感染可能与Treg表达上调有关.

  2. The vitamin D analog, TX527, promotes a human CD4+CD25highCD127low regulatory T cell profile and induces a migratory signature specific for homing to sites of inflammation.

    Science.gov (United States)

    Baeke, Femke; Korf, Hannelie; Overbergh, Lut; Verstuyf, Annemieke; Thorrez, Lieven; Van Lommel, Leentje; Waer, Mark; Schuit, Frans; Gysemans, Conny; Mathieu, Chantal

    2011-01-01

    The use of hypocalcemic vitamin D analogs is an appealing strategy to exploit the immunomodulatory actions of active vitamin D in vivo while circumventing its calcemic side effects. The functional modulation of dendritic cells by these molecules is regarded as the key mechanism underlying their ability to regulate T cell reactivity. In this article, we demonstrate the capacity of the vitamin D analog, TX527, to target T cells directly. Microarray analysis of purified human CD3(+) T cells, cultured in the presence of TX527, revealed differential expression of genes involved in T cell activation, proliferation, differentiation, and migratory capacity. Accordingly, functional analysis showed a TX527-mediated suppression of the T cell proliferative capacity and activation status, accompanied by decreased expression of effector cytokines (IFN-γ, IL-4, and IL-17). Furthermore, TX527 triggered the emergence of CD4(+)CD25(high)CD127(low) regulatory T cells featuring elevated levels of IL-10, CTLA-4, and OX40 and the functional capacity to suppress activation and proliferation of effector T cells. Moreover, the vitamin D analog profoundly altered the homing receptor profile of T cells and their migration toward chemokine ligands. Remarkably, TX527 not only modulated skin-homing receptors as illustrated for the parent compound, but also reduced the expression of lymphoid organ-homing receptors (CD62L, CCR7, and CXCR4) and uniquely promoted surface expression of inflammatory homing receptors (CCR5, CXCR3, and CXCR6) on T cells. We conclude that TX527 directly affects human T cell function, thereby inhibiting effector T cell reactivity while inducing regulatory T cell characteristics, and imprints them with a specific homing signature favoring migration to sites of inflammation. PMID:21131424

  3. Humoral immune response induced by an engineered cell-based neuroblastoma vaccine with or without CD25 blockade

    Institute of Scientific and Technical Information of China (English)

    Jin Zheng; Rimas Orentas; Xiaofei Yan; Hongli Liu

    2011-01-01

    Neuroblastoma is the most common extracranial solid cancer in childhood and it can develop in the nerve tissue of the adrenal gland, neck, chest, or spinal cord A number of tumor-associated antigens(TAAs), which can elicit humoral immunity, have been identified in cancer patients. To investigate the humoral immunity during neuroblastoma development, we treated A/J mice with an aggressive clone of neuroblastoma(AGN2a)cells, then vaccinated the mice with cells expressing AGN2a-CD80/CD137L under the condihons with or without regulatory T cell blockade. Strong humoral immunity was induced by AGN2a-CD80/CD137L immunization in the context of regulatory T cell blockade. Sera from treated mice were used to screen an AGN2a cDNA expression library for identifying TAAs by SEREX(serological analysis of recombinant cDNA expression libraries). Clones were identified by sequencing and comparative analysis of gene pools. Further investigation of these gene products revealed that most of them play a role in the neuronal differentiation, cell metabolism, and are highly expressed in other types of malignancy. Asz1(ankyrin repeat, SAM, and basic leucine zipper domaincontaining protein)was found in all tumor-bearing groups. These results implicated that these candidates identified from tumor-bearing mice may be neuroblastoma-associated antigens, which can be used as biomarkers in early diagnosis of neuroblastoma, whereas those identified from vaccinated mice may be the potential therapeutic targets.

  4. Effect of Astragalus on CD4+ CD25+ T cells and cytokines in the mice model of asthma%黄芪对哮喘小鼠CD4+ CD25+调节性T细胞及IL-4IFN-γ IL-10的影响

    Institute of Scientific and Technical Information of China (English)

    刘瑞文; 李春霞; 李志华; 王郡甫

    2011-01-01

    目的 探讨黄芪对哮喘小鼠CD4+ CD25+调节性T细胞及Ⅱ-4、IFN-γ、IL-10的影响.方法 将BALB/c小鼠30只随机分为对照组、哮喘组和黄芪组.以卵蛋白(OVA)致敏激发法制备小鼠哮喘模型.酶联免疫吸附试验( ELISA)法检测支气管肺泡灌洗液(BALF)中IL-4、IFN-γ、IL-10及血清中IL-10的含量;流式细胞术(FCM)、反转录聚合酶链反应(RT-PCR)分别检测小鼠脾脏中CD4+ CD25+调节性T细胞数量及FoxP3mRNA表达情况.结果 哮喘组小鼠BALF中IL-4含量明显高于对照组(P<0.01)而低于黄芪组(P<0.01).与对照组相比,哮喘组小鼠BALF中IFN-γ、IL-10、血清中IL-10含量及脾脏中CD4+ CD25+调节性T细胞数量、FoxP3 mRNA表达水平明显降低(P<0.01);而黄芪组的上述改变较哮喘组显著增加(P<0.05).结论 CD4+ CD25+调节性T细胞参与了支气管哮喘的发病过程,黄芪可通过上调CD4+ CD25+调节性T细胞、FoxP3mRNA的表达及增加IL-10含量减轻哮喘炎症.

  5. CD4+CD25+调节性T细胞在自身免疫性溶血性贫血患者中的变化及意义%Change and significance of CD4+CD25+ regulatory T cells in patients with autoimmune hemolytic anemia

    Institute of Scientific and Technical Information of China (English)

    黄东平; 何合胜

    2014-01-01

    Objective To investigate the change of regulatory T-cells (Treg) before and after therapy in patients with autoimmune hemolytic anemia (AIHA),and to study the role of Treg in AIHA.Methods Treg cells numbers was measured by flow cytometry.Results Before treatment,Treg cells in AIHA patients was (1.32 ± 0.87) %,which was significantly lower than (3.08 ± 0.96) % in the controls (t =-5.37,P < 0.01).After treatment,Treg cells in AIHA patients was significantly increased [(4.96 ± 1.13)%] (t =-16.94,P <0.01).Conclusion Treg cells decreased in AIHA patients.Glucocorticoid might play a role in AIHA treatment by up-regulating Treg cells number.%目的 探讨CD4+CD25+Foxp3+调节性T细胞(Treg)在自身免疫性溶血性贫血(AIHA)患者中的变化.方法 应用流式细胞术检测AIHA患者治疗前后外周血Treg的数量变化.结果 AIHA患者治疗前外周血中Treg细胞比例为(1.32±0.87)%,明显低于对照组(3.08±0.96)%(t=-5.37,P<0.01).治疗后Treg细胞比例为(4.96±1.13)%,明显高于治疗前(t=-16.94,P<0.01),且高于对照组(t=-4.96,P<0.01).结论 AIHA患者中Treg细胞数量下降,糖皮质激素通过上调Treg细胞数量可以控制自身免疫性溶血性贫血.

  6. Changes and clinical significance of CD4+CD25+Foxp3+ regulatory T cell and IL-10,TGF-β1 in steroid-resistant asthma patients of peripheral blood%激素抵抗性哮喘患者外周血CD4+CD25+Foxp3+调节性T细胞及IL-10、TGF-β1的变化及意义

    Institute of Scientific and Technical Information of China (English)

    钱一龙; 赵振中; 朱建俊; 谢中华; 王珠美

    2013-01-01

    目的 观察激素抵抗性哮喘(SRA)患者外周血CD4+CD25+Foxp3+调节性T细胞(Treg)及白介素10(IL-10)、转化生长因子β1(TGF-β1)的变化,分析其在SRA发病机制中的作用.方法 采用流式细胞术检测40例SRA患者(激素抵抗组)外周血单个核细胞CD4+CD25+Foxp3+ Treg数目,并计算CD4+CD25+Foxp3+ Treg占CD4+T淋巴细胞的百分比;酶联免疫吸附试验(ELISA)法检测其血清IL-10、TGF-β1水平,并与激素敏感性患者(激素敏感组,46例)及正常体检者(正常组,30例)进行对比.结果 激素抵抗组患者外周血CD4+CD25+Foxp3+Treg占CD4+T淋巴细胞的百分比、CD4+CD25+Foxp3+Treg绝对值及血清IL-10、TGF-β1水平均明显低于激素敏感组与正常组(P<0.01,P<0.05);激素敏感组患者外周血CD4+CD25+Foxp3+Treg占CD4+T淋巴细胞的百分比、CD4+CD25+Foxp3+Treg绝对值及血清TGF-β1水平明显低于正常组(P<0.01,P<0.05),血清IL-10无明显差异(P>0.05);CD4+CD25+Foxp3+Treg/CD4+T及CD4+CD25+Foxp3+Treg绝对数均与血清IL-10、TGF-β1水平呈明显正相关(P<0.01).结论 SRA患者外周血CD4+CD25+Foxp3+ Treg数目减少及IL-10、TGF-β1含量减低可能与SRA的发生、发展有关.

  7. Auditory stimulation of opera music induced prolongation of murine cardiac allograft survival and maintained generation of regulatory CD4+CD25+ cells

    Directory of Open Access Journals (Sweden)

    Uchiyama Masateru

    2012-03-01

    Full Text Available Abstract Background Interactions between the immune response and brain functions such as olfactory, auditory, and visual sensations are likely. This study investigated the effect of sounds on alloimmune responses in a murine model of cardiac allograft transplantation. Methods Naïve CBA mice (H2k underwent transplantation of a C57BL/6 (B6, H2b heart and were exposed to one of three types of music--opera (La Traviata, classical (Mozart, and New Age (Enya--or one of six different single sound frequencies, for 7 days. Additionally, we prepared two groups of CBA recipients with tympanic membrane perforation exposed to opera for 7 days and CBA recipients exposed to opera for 7 days before transplantation (pre-treatment. An adoptive transfer study was performed to determine whether regulatory cells were generated in allograft recipients. Immunohistochemical, cell-proliferation, cytokine, and flow cytometry assessments were also performed. Results CBA recipients of a B6 cardiac graft that were exposed to opera music and Mozart had significantly prolonged allograft survival (median survival times [MSTs], 26.5 and 20 days, respectively, whereas those exposed to a single sound frequency (100, 500, 1000, 5000, 10,000, or 20,000 Hz or Enya did not (MSTs, 7.5, 8, 9, 8, 7.5, 8.5 and 11 days, respectively. Untreated, CBA mice with tympanic membrane perforations and CBA recipients exposed to opera for 7 days before transplantation (pre-treatment rejected B6 cardiac grafts acutely (MSTs, 7, 8 and 8 days, respectively. Adoptive transfer of whole splenocytes, CD4+ cells, or CD4+CD25+ cells from opera-exposed primary allograft recipients resulted in significantly prolonged allograft survival in naive secondary recipients (MSTs, 36, 68, and > 100 days, respectively. Proliferation of splenocytes, interleukin (IL-2 and interferon (IFN-γ production was suppressed in opera-exposed mice, and production of IL-4 and IL-10 from opera-exposed transplant recipients increased

  8. Dysregulation of CD4+CD25+CD127lowFOXP3+ regulatory T cells in HIV-infected pregnant women

    DEFF Research Database (Denmark)

    Kolte, Lilian; Gaardbo, Julie C; Karlsson, Ingrid;

    2010-01-01

    Pregnancy represents a major challenge to immunologic tolerance. How the fetal "semiallograft" evades maternal immune attack is unknown. Pregnancy success may involve alteration of both central (thymic) and peripheral tolerance mechanisms. HIV infection is characterized by CD4(+) T-cell depletion......, chronic immune activation, and altered lymphocyte subsets. We studied immunologic consequences of pregnancy in 20 HIV-infected women receiving highly active antiretroviral therapy (HAART), and for comparison in 16 HIV-negative women. Lymphocyte subsets, thymic output, and cytokine profiles were measured...... prospectively during pregnancy and postpartum. A significant expansion of CD4(+)CD25(+)CD127(low)FoxP3(+) regulatory T cells indicating alteration of peripheral tolerance was seen during second trimester, but only in HIV-negative women. HIV-infected women had lower CD4 counts, lower thymic output and Th-2...

  9. FoxP3(+)CD4(+)CD25(+) T cells with regulatory properties can be cultured from colonic mucosa of patients with Crohn's disease

    DEFF Research Database (Denmark)

    Rømer, Johanne Lade

    2005-01-01

    /winged helix transcription factor FoxP3 is a master gene for T(reg) function and defects in the FoxP3 gene lead to a clinical picture similar to inflammatory bowel disease (IBD). Murine colitis can be cured by adoptive transfer of T(regs) and ex vivo-generated gut-specific T(regs) represent an attractive...... option for therapy in CD. Thus, defective T(regs) could contribute to the development of CD. We cultured biopsies of colonic mucosa in the presence of high concentrations of interleukin (IL)-2 and IL-4 to overcome the anergic nature of naturally occurring CD4(+)CD25(+) T(regs) in the mucosa. We...

  10. CD4+ CD25+ FoxP3+ regulatory T cells suppress cytotoxicity of CD8+ effector T cells: implications for their capacity to limit inflammatory central nervous system damage at the parenchymal level

    Directory of Open Access Journals (Sweden)

    Göbel Kerstin

    2012-02-01

    Full Text Available Abstract Background CD4+ CD25+ forkhead box P3 (FoxP3+ regulatory T cells (T reg cells are known to suppress adaptive immune responses, key control tolerance and autoimmunity. Methods We challenged the role of CD4+ T reg cells in suppressing established CD8+ T effector cell responses by using the OT-I/II system in vitro and an OT-I-mediated, oligodendrocyte directed ex vivo model (ODC-OVA model. Results CD4+ T reg cells dampened cytotoxicity of an ongoing CD8+ T effector cell attack in vitro and within intact central nervous system tissue ex vivo. However, their suppressive effect was limited by the strength of the antigen signal delivered to the CD8+ T effector cells and the ratio of regulatory to effector T cells. CD8+ T effector cell suppression required T cell receptor-mediated activation together with costimulation of CD4+ T reg cells, but following activation, suppression did not require restimulation and was antigen non-specific. Conclusions Our results suggest that CD4+ T reg cells are capable of suppressing CD8+ T effector cell responses at the parenchymal site, that is, limiting parenchymal damage in autoimmune central nervous system inflammation.

  11. Controlled expression of functional miR-122 with a ligand inducible expression system

    Directory of Open Access Journals (Sweden)

    Tzertzinis George

    2010-10-01

    Full Text Available Abstract Background To study the biological function of miRNAs, and to achieve sustained or conditional gene silencing with siRNAs, systems that allow controlled expression of these small RNAs are desirable. Methods for cell delivery of siRNAs include transient transfection of synthetic siRNAs and expression of siRNAs in the form of short hairpins using constitutive RNA polymerase III promoters. Systems employing constitutive RNA polymerase II promoters have been used to express miRNAs. However, for many experimental systems these methods do not offer sufficient control over expression. Results We present an inducible mammalian expression system that allows for the conditional expression of short hairpin RNAs that are processed in vivo to generate miRNAs or siRNAs. Using modified nuclear receptors in a two hybrid format and a synthetic ligand, the Rheoswitch system allows rapid and reversible induction of mRNA expression. We evaluated the system's properties using miR-122 as a model miRNA. A short hairpin encoding miR-122 cloned into the expression vector was correctly processed to yield mature miRNA upon induction with ligand and the amount of miRNA produced was commensurate with the concentration of ligand. miR-122 produced in this way was capable of silencing both endogenous target genes and appropriately designed reporter genes. Stable cell lines were obtained, resulting in heritable, consistent and reversible expression of miR-122, a significant advantage over transient transfection. Based on these results, obtained with a microRNA we adapted the method to produce a desired siRNA by designing short hairpins that can be accurately and efficiently processed. Conclusion We established an Inducible expression system with a miR-122 backbone that can be used for functional studies of miRNAs and their targets, in heterologous cells that do not normally express the miRNA. Additionally we demonstrate the feasibility of using the miR-122 backbone to

  12. Expression of PD-1 Molecule on Regulatory T Lymphocytes in Patients with Insulin-Dependent Diabetes Mellitus

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    Valentina Perri

    2015-09-01

    Full Text Available Type 1 diabetes is caused by autoreactive T cells that destroy pancreatic beta cells. Animal models suggested that a CD4+CD25+ population has a regulatory function capable of preventing activation and effector functions of autoreactive T cells. However, the role of CD4+CD25high T cells in autoimmunity and their molecular mechanisms remain the subject of investigation. We therefore evaluated T regulatory cell frequencies and their PD-1 expression in the peripheral blood of long-standing diabetics under basal conditions and after CD3/CD28 stimulation. Under basal conditions, the percentages of T regulatory cells were significantly higher while that of T effector cells were significantly lower in patients than in controls. The ratio of regulatory to effector T cells was higher in patients than that in controls, suggesting that T regulatory cells were functional in patients. Percentages of total PD-1+, PD-1low and PD-1high expressing T regulatory cells did not change in patients and in controls. After stimulation, a defect in T regulatory cell proliferation was observed in diabetics and the percentages of total PD-1+, PD-1low and PD-1high expressing cells were lower in patients. Our data suggest a defective activation of T regulatory cells in long-standing diabetics due to a lower expression of PD-1 on their surface.

  13. Study of Four-Week Aerobic Endurance Exercise on the Induction of CD4+CD25+Treg in Mouse Spleen%4周有氧耐力运动对小鼠脾脏CD4+CD25+调节性T细胞诱导表达的实验研究

    Institute of Scientific and Technical Information of China (English)

    王杰龙; 陈军; 吴明方; 张光波; 陈礼文; 赵建萍

    2012-01-01

    目的:通过对4周负重游泳运动前、后小鼠脾脏CD4- CD25+ Treg表达水平变化及相关细胞因子IL-2、IL-10和TGF-β1水平变化的分析,探讨和研究有氧耐力运动对小鼠脾脏CD4+ CD25+ Treg表达及细胞因子IL-2、IL-10和TGF-β的影响.方法:KM种雄性小鼠100只随机分为对照组(50只)和运动组(50只).运动各组分别按运动方案进行45 min的负重(5%体重)游泳运动每天1次,每周6次.分别于实验前、每周末次运动后24 h,处死小鼠,摘取脾脏和胸腺、采集血样.计算脾指数和胸腺指数,流式细胞仪检测小鼠脾脏CD4+CD25+ Treg的表达,ELISA法测定血清IL-2、IL-10和TGF-β含量.结果:在4周负重游泳运动过程中,运动组小鼠脾指数在第1周末增加并在随后2、3、4周表现出下降的趋势,胸腺指数表现为逐周减少.运动组小鼠脾脏CD4+ CD25+ Treg的表达率表现为逐周增高,运动3周组、4周组高于运动0周组和对照组(P<0.05).运动2周组和3周组血清IL-2水平升高,高于对照组(P<0.05);运动3周组和4周组血清IL-10和TGF-β1水平高于对照组(P<0.05).结论:4周有氧耐力游泳运动可延缓小鼠脾脏生长,加快胸腺萎缩,进而影响免疫器官的发育和功能.4周有氧耐力游泳运动可诱导小鼠T细胞活化,脾脏CD4+ CD25+ Treg表达水平提高,血清IL-10和TGF-β1水平升高,诱导免疫偏移.

  14. Role of glucagon-like peptide-1 analogue liraglutide played in the proliferation of CD4~+ CD25~- T cells in normal people and type 1 diabetic patients in vitro

    Institute of Scientific and Technical Information of China (English)

    胡瑛

    2013-01-01

    Objective To study the role of glucagon-like peptide-1 (GLP-1) analogue liraglutide played in the proliferation of CD4+CD25-T cells in normal people and newly-onset type 1 diabetic patients,and to evaluate the possible immune regulatory role of liraglutide in the

  15. CD4+CD25+CD127 regulatory cells play multiple roles in maintaining HIV-1 p24 production in patients on long-term treatment: HIV-1 p24-producing cells and suppression of anti-HIV immunity

    Directory of Open Access Journals (Sweden)

    Yan-Mei Jiao

    2015-08-01

    Conclusions: CD4+CD25+CD127 regulatory cells play multiple roles in maintaining HIV-1 p24 production in long-term ART patients. Treg cells may be a target for eliminating the latent HIV reservoir after effective long-term ART.

  16. Interplay of T Helper 17 Cells with CD4+CD25high FOXP3+ Tregs in Regulation of Allergic Asthma in Pediatric Patients

    OpenAIRE

    Amit Agarwal; Meenu Singh; Chatterjee, B. P.; Anil Chauhan; Anuradha Chakraborti

    2014-01-01

    Background. There is evidence that Tregs are important to prevent allergic diseases like asthma but limited literature exists on role of TH17 cells in allergic diseases. Methods. Fifty children with asthma and respiratory allergy (study group) and twenty healthy children (control group) were recruited in this study. Total IgE levels and pulmonary function tests were assessed. The expression of Tregs and cytokines was determined by flow cytometry. Results. The average level of total IgE in stu...

  17. Ex vivo generation of human alloantigen-specific regulatory T cells from CD4(posCD25(high T cells for immunotherapy.

    Directory of Open Access Journals (Sweden)

    Jorieke H Peters

    Full Text Available BACKGROUND: Regulatory T cell (Treg based immunotherapy is a potential treatment for several immune disorders. By now, this approach proved successful in preclinical animal transplantation and auto-immunity models. In these models the success of Treg based immunotherapy crucially depends on the antigen-specificity of the infused Treg population. For the human setting, information is lacking on how to generate Treg with direct antigen-specificity ex vivo to be used for immunotherapy. METHODOLOGY/PRINCIPAL FINDINGS: Here, we demonstrate that in as little as two stimulation cycles with HLA mismatched allogeneic stimulator cells and T cell growth factors a very high degree of alloantigen-specificity was reached in magnetic bead isolated human CD4(posCD25(high Treg. Efficient increases in cell numbers were obtained. Primary allogeneic stimulation appeared a prerequisite in the generation of alloantigen-specific Treg, while secondary allogeneic or polyclonal stimulation with anti-CD3 plus anti-CD28 monoclonal antibodies enriched alloantigen-specificity and cell yield to a similar extent. CONCLUSIONS/SIGNIFICANCE: The ex vivo expansion protocol that we describe will very likely increase the success of clinical Treg-based immunotherapy, and will help to induce tolerance to selected antigens, while minimizing general immune suppression. This approach is of particular interest for recipients of HLA mismatched transplants.

  18. CD4+CD25+调节性T细胞及其相关蛋白Foxp3在小儿支原体肺炎患者中的表达

    Institute of Scientific and Technical Information of China (English)

    袁浩; 周毅峰; 曹友德; 陈雪初; 马玲飞

    2013-01-01

    目的 探讨不同滴度时小儿肺炎支原体(MP)感染患儿外周血CD4+CD25+Treg细胞和CD4+Foxp3+Treg细胞变化及其临床意义.方法 采用流式细胞分析法分别检测MP感染患儿76例,其中MP抗体IgM阳性滴度1:80 26例,1:160 17例,1:320 15例,1:640 18例和24例健康体检者(对照组)外周血CD4+CD25+Treg细胞,CD4+Foxp3+Treg细胞的表达水平.结果 MP感染患儿1:80组CD4+CD25+Treg、CD4+Foxp3+Treg细胞均明显高于1:640组和对照组(P0.05),CD4+CD25+Treg细胞及CD4+Foxp3+Treg细胞的变化与滴度的变化均呈负相关(r=-0.402、-0.376,P<0.01),CD4+Foxp3+Treg细胞的变化与CD4+CD25+Treg细胞的变化呈正相关(r=0.627,P<0.01).结论 CD4+CD25+ Treg细胞和CD4+Foxp3+Treg细胞在小儿MP感染低滴度组发病初期显著升高,但在高滴度组升高并不明显,且两种细胞的变化与滴度的变化呈负相关,Foxp3蛋白可能是CD4+CD25+Treg特异性的标志.

  19. Expressive capabilities of the dialogue language in automated control systems

    Energy Technology Data Exchange (ETDEWEB)

    Lyubarskiy, Yu.Ya.

    1982-09-01

    Provisions for dialogue communication between operating personnel and a computer are of utmost importance in contemporary industrial automated control systems and in automated systems of dispatcher control. The most advanced dialogue systems are the question-answer systems which enable one to communicate with the computer in a language that is close to the natural professional language of the user. This article describes a method for construction of economical question-answer systems which could be realized with the help of minicomputers, and examination of methods for providing such QAs with the expressive capabilities possessed by a natural language. These capabilities include the ambiguity of meanings of words of the input language (polysemy), some elliptic constructions (surpression), and tropeic forms (different forms of metonymy and metaphors). 9 references.

  20. Accumulation of CCR4⁺CTLA-4 FOXP3⁺CD25(hi regulatory T cells in colon adenocarcinomas correlate to reduced activation of conventional T cells.

    Directory of Open Access Journals (Sweden)

    Helena Svensson

    Full Text Available BACKGROUND: Colorectal cancer usually gives rise to a specific anti-tumor immune response, but for unknown reasons the resulting immunity is not able to clear the tumor. Recruitment of activated effector lymphocytes to the tumor is important for efficient anti-tumor responses, while the presence of regulatory T cells (Treg down-modulate tumor-specific immunity. We therefore aimed to determine homing mechanisms and activation stage of Treg and effector T cell infiltrating colon tumors compared to cells from the unaffected mucosa in patients suffering from colon adenocarcinoma. METHODOLOGY/PRINCIPAL FINDINGS: Lymphocytes were isolated from unaffected and tumor mucosa from patients with colon adenocarcinoma, and flow cytometry, immunohistochemistry, and quantitative PCR was used to investigate the homing mechanisms and activation stage of infiltrating Treg and conventional lymphocytes. We detected significantly higher frequencies of CD25(highFOXP3⁺CD127(low putative Treg in tumors than unaffected mucosa, which had a complete demethylation in the FOXP3 promotor. Tumor-associated Treg had a high expression of CTLA-4, and some appeared to be antigen experienced effector/memory cells based on their expression of αEβ7 (CD103. There were also significantly fewer activated T cells and more CTLA-4⁺ conventional T cells susceptible to immune regulation in the tumor-associated mucosa. In contrast, CD8⁺granzyme B⁺ putative cytotoxic cells were efficiently recruited to the tumors. The frequencies of cells expressing α4β7 and the Th1 associated chemokine receptor CXCR3 were significantly decreased among CD4⁺ T cells in the tumor, while frequencies of CD4⁺CCR4⁺ lymphocytes were significantly increased. CONCLUSIONS/SIGNIFICANCE: This study shows that CCR4⁺CTLA4(hi Treg accumulate in colon tumors, while the frequencies of activated conventional Th1 type T cells are decreased. The altered lymphocyte composition in colon tumors will probably

  1. Expression of IL-2R on Peripheral Blood Lymphocytes of Patients with Colorectal Cancer and Its Clinical Significance

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective: To study expression of membrane receptors ofinterleukin-2 (CD25) on the peripheral blood lymphocytes (PBL) of patients with colorectal cancer and its clinical significance. Methods: CD25 percentages (CD25%) in PBL of 105 colorectal cancer patients before operation and 100 normal individuals were examined by flow cytometer, and the results were clinically and pathologically analyzed. Results: The mean of CD25% in PBL of the normal individuals was 17.24± 5.33, it was significantly lower (p<0.01) than that of the colon cancer patients (21.29± 7.95) or rectal cancer patients (21.62± 6.11). In contrast to the normal individuals, the means of CD25% in PBL in ulcer type (20.53± 6.50) or protruded type (21.56± 6.16) colorectal cancer patients were notably elevated (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was observed between the normal individuals and patients with less than 4 cm mass (22.10± 5.43) or 4cm- 8cm mass (20.90± 6.96). The significant difference (P<0.05) of means of CD25% in PBL was also observed between the normal individuals and patients with greater than 8 cm mass (21.56± 5.41). The mean of CD25% in PBL in patients with well differentiation colorectal cancer was 22.20± 5.50, it was significantly higher than that in normal individuals (P<0.05). The means of CD25% in PBL in patients with middle or poor differentiation colorectal cancer were 21.30± 6.89 and 22.15± 5.71 respectively, they were obviously higher than that in normal individuals (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was present between the colorectal cancer patients without metastatic lymph nodes (22.06± 6.90) and normal individuals. The significant difference (P<0.05) of means of CD25% in PBL was present between the colorectal cancer patients with metastatic lymph nodes (20.73± 6.40) and normal individuals. The means of CD25% in PBL in colorectal cancer patients in various clinic stages were significantly higher

  2. Rapamycin ameliorates experimental autoimmune uveoretinitis by inhibiting Th1/Th2/Th17 cells and upregulating CD4+CD25+ Foxp3 regulatory T cells

    Institute of Scientific and Technical Information of China (English)

    Li-Fei; Yuan; Guang-Da; Li; Xin-Jun; Ren; Hong; Nian; Xiao-Rong; Li; Xiao-Min; Zhang

    2015-01-01

    · AIM: To determine the effects of rapamycin on experimental autoimmune uveoretinitis(EAU) and investigate of role of rapamycin on T cell subsets in the disease.·METHODS: EAU was induced in rats using peptides1169 to 1191 of the interphotoreceptor binding protein(IRBP). Rapamycin(0.2 mg/kg/d) was administrated by intraperitoneal injection for a consecutive 7d after immunization. Th1/Th2/Th17 cytokines, TGF-β1, and IL-6produced by lymphocyteswere measured by ELISA, while Th17 cells and CD4 +CD25 + regulatory T cells(Tregs)from rat spleen were detected by flow cytometry.·RESULTS: Intraperitoneal treatment immediately after immunization dramatically ameliorated the clinical course of EAU. Clinical responses were associated with reduced retinal inflammatory cell infiltration and tissue destruction. Rapamycin induced suppression of Th1/Th2/Th17 cytokines, including IFN-γ, IL-2, IL-17, IL-4, and IL-10 release from T lymphocytes of EAU rats, in vitro.Rapamycin also significantly increased TGF-β1production but had no effect on IL-6 productionof T lymphocytes from EAU rats in vitro. Furthermore,rapamycin decreased the ratio of Th17 cells/CD4 +T cells and upregulated Tregs in EAU, as detected by flow cytometry.·CONCLUSION: Rapamycin effectively interferes with T cell mediated autoimmune uveitis by inhibiting antigen-specific T cell functions and enhancing Tregs in EAU.Rapamycin is a promising new alternative as an adjunct corticosteroid-sparing agent for treating uveitis.

  3. Adalimumab ameliorates OVA-induced airway inflammation in mice: Role of CD4(+) CD25(+) FOXP3(+) regulatory T-cells.

    Science.gov (United States)

    Elsakkar, Mohamed G; Sharaki, Olla A; Abdallah, Dina M; Mostafa, Dalia K; Shekondali, Fadia T

    2016-09-01

    Asthma is a chronic inflammatory heterogeneous disorder initiated by a dysregulated immune response which drives disease development in susceptible individuals. Though T helper 2 (TH2) biased responses are usually linked to eosinophilic asthma, other Th cell subsets induce neutrophilic airway inflammation which provokes the most severe asthmatic phenotypes. A growing evidence highlights the role of T regulatory (Treg) cells in damping abnormal Th responses and thus inhibiting allergy and asthma. Therefore, strategies to induce or augment Treg cells hold promise for treatment and prevention of allergic airway inflammation. Recently, the link between Tumor necrosis factor-α (TNF-α) and Treg has been uncovered, and TNF-α antagonists are increasingly used in many autoimmune diseases. Yet, their benefits in allergic airway inflammation is not clarified. We investigated the effect of Adalimumab, a TNF-α antagonist, on Ovalbumin (OVA)-induced allergic airway inflammation in CD1 mice and explored its impact on Treg cells. Our results showed that Adalimumab treatment attenuated the OVA-induced increase in serum IgE, TH2 and TH1 derived inflammatory cytokines (IL-4 and IFN-γ, respectively) in bronchoalveolar lavage (BAL) fluid, suppressed recruitment of inflammatory cells in BAL fluid and lung, and inhibited BAL fluid neutrophilia. It also ameliorated goblet cell metaplasia and bronchial fibrosis. Splenocytes flow cytometry revealed increased percentage of CD4(+) CD25(+) FOXP3(+) Treg cells by Adalimumab that was associated with increase in their suppressive activity as shown by elevated BAL fluid IL-10. We conclude that the beneficial effects of Adalimumab in this CD1 neutrophilic model of allergic airway inflammation are attributed to augmentation of Treg cell number and activity. PMID:27262379

  4. Effects of active bufadienolide compounds on human cancer cells and CD4+CD25+Foxp3+ regulatory T cells in mitogen-activated human peripheral blood mononuclear cells.

    Science.gov (United States)

    Yuan, Bo; He, Jing; Kisoh, Keishi; Hayashi, Hideki; Tanaka, Sachiko; Si, Nan; Zhao, Hai-Yu; Hirano, Toshihiko; Bian, Baolin; Takagi, Norio

    2016-09-01

    The growth inhibitory effects of bufadienolide compounds were investigated in two intractable cancer cells, a human glioblastoma cell line U-87 and a pancreatic cancer cell line SW1990. Among four bufadienolide compounds, a dose-dependent cytotoxicity was observed in these cancer cells after treatment with gamabufotalin and arenobufagin. The IC50 values of the two compounds were 3-5 times higher in normal peripheral blood mononuclear cells (PBMCs) than these values for both cancer cell lines. However, similar phenomena were not observed for two other bufadienolide compounds, telocinobufagin and bufalin. These results thus suggest that gamabufotalin and arenobufagin possess selective cytotoxic activity against tumor cells rather than normal cells. Moreover, a clear dose-dependent lactate dehydrogenase (LDH) release, a well-known hallmark of necrosis, was observed in both cancer cells treated with gamabufotalin, suggesting that gamabufotalin-mediated cell death is predominantly associated with a necrosis-like phenotype. Of most importance, treatment with as little as 8 ng/ml of gamabufotalin, even an almost non-toxic concentration to PBMCs, efficiently downregulated the percentages of CD4+CD25+Foxp3+ regulator T (Treg) cells in mitogen-activated PBMCs. Given that Treg cells play a critical role in tumor immunotolerance by suppressing antitumor immunity, these results suggest that gamabufotalin may serve as a promising candidate, as an adjuvant therapeutic agent by manipulating Treg cells to enhance the efficacy of conventional anticancer drugs and lessen their side-effects. These findings provide insights into the clinical application of gamabufotalin for cancer patients with glioblastoma/pancreatic cancer based on its cytocidal effect against tumor cells as well as its depletion of Treg cells. PMID:27431260

  5. An autonomous molecular computer for logical control of gene expression

    Science.gov (United States)

    Benenson, Yaakov; Gil, Binyamin; Ben-Dor, Uri; Adar, Rivka; Shapiro, Ehud

    2004-05-01

    Early biomolecular computer research focused on laboratory-scale, human-operated computers for complex computational problems. Recently, simple molecular-scale autonomous programmable computers were demonstrated allowing both input and output information to be in molecular form. Such computers, using biological molecules as input data and biologically active molecules as outputs, could produce a system for `logical' control of biological processes. Here we describe an autonomous biomolecular computer that, at least in vitro, logically analyses the levels of messenger RNA species, and in response produces a molecule capable of affecting levels of gene expression. The computer operates at a concentration of close to a trillion computers per microlitre and consists of three programmable modules: a computation module, that is, a stochastic molecular automaton; an input module, by which specific mRNA levels or point mutations regulate software molecule concentrations, and hence automaton transition probabilities; and an output module, capable of controlled release of a short single-stranded DNA molecule. This approach might be applied in vivo to biochemical sensing, genetic engineering and even medical diagnosis and treatment. As a proof of principle we programmed the computer to identify and analyse mRNA of disease-related genes associated with models of small-cell lung cancer and prostate cancer, and to produce a single-stranded DNA molecule modelled after an anticancer drug.

  6. Autoregulatory systems controlling translation factor expression: Thermostat-like control of translational accuracy

    OpenAIRE

    Betney, Russell; de Silva, Eric; Krishnan, Jawahar; Stansfield, Ian

    2010-01-01

    In both prokaryotes and eukaryotes, the expression of a large number of genes is controlled by negative feedback, in some cases operating at the level of translation of the mRNA transcript. Of particular interest are those cases where the proteins concerned have cell-wide function in recognizing a particular codon or RNA sequence. Examples include the bacterial translation termination release factor RF2, initiation factor IF3, and eukaryote poly(A) binding protein. The regulatory loops that c...

  7. Multiple controls affect arsenite oxidase gene expression in Herminiimonas arsenicoxydans

    Directory of Open Access Journals (Sweden)

    Coppée Jean-Yves

    2010-02-01

    Full Text Available Abstract Background Both the speciation and toxicity of arsenic are affected by bacterial transformations, i.e. oxidation, reduction or methylation. These transformations have a major impact on environmental contamination and more particularly on arsenic contamination of drinking water. Herminiimonas arsenicoxydans has been isolated from an arsenic- contaminated environment and has developed various mechanisms for coping with arsenic, including the oxidation of As(III to As(V as a detoxification mechanism. Results In the present study, a differential transcriptome analysis was used to identify genes, including arsenite oxidase encoding genes, involved in the response of H. arsenicoxydans to As(III. To get insight into the molecular mechanisms of this enzyme activity, a Tn5 transposon mutagenesis was performed. Transposon insertions resulting in a lack of arsenite oxidase activity disrupted aoxR and aoxS genes, showing that the aox operon transcription is regulated by the AoxRS two-component system. Remarkably, transposon insertions were also identified in rpoN coding for the alternative N sigma factor (σ54 of RNA polymerase and in dnaJ coding for the Hsp70 co-chaperone. Western blotting with anti-AoxB antibodies and quantitative RT-PCR experiments allowed us to demonstrate that the rpoN and dnaJ gene products are involved in the control of arsenite oxidase gene expression. Finally, the transcriptional start site of the aoxAB operon was determined using rapid amplification of cDNA ends (RACE and a putative -12/-24 σ54-dependent promoter motif was identified upstream of aoxAB coding sequences. Conclusion These results reveal the existence of novel molecular regulatory processes governing arsenite oxidase expression in H. arsenicoxydans. These data are summarized in a model that functionally integrates arsenite oxidation in the adaptive response to As(III in this microorganism.

  8. Real-time feedback control of gene expression

    OpenAIRE

    Uhlendorf, Jannis

    2013-01-01

    Gene expression is fundamental for the functioning of cellular processes and is tightly regulated. Inducible promoters allow one to perturb gene expression by changing the expression level of a protein from its physiological level. This is a common tool to decipher the functioning of biological processes: the expression level of a gene is changed and one observes how the perturbed cell behaves differently from an unperturbed cell. A shortcoming of inducible promoters is the difficulty to appl...

  9. Expressing emotions through vibration for perception and control

    OpenAIRE

    ur Réhman, Shafiq

    2010-01-01

    This thesis addresses a challenging problem: “how to let the visually impaired ‘see’ others emotions”. We, human beings, are heavily dependent on facial expressions to express ourselves. A smile shows that the person you are talking to is pleased, amused, relieved etc. People use emotional information from facial expressions to switch between conversation topics and to determine attitudes of individuals. Missing emotional information from facial expressions and head gestures makes the visuall...

  10. 哮喘小鼠CD4+CD25+调节性T细胞数量及功能的改变%The change of the CD4+CD25+ regulatory T cells and its function in a mouse model of asthma

    Institute of Scientific and Technical Information of China (English)

    吴奎; 孙鲲; 毕玉田; 夏俊波; 王长征

    2005-01-01

    目的观察哮喘小鼠脾脏CD4+CD25+ 调节性T细胞(简称Treg细胞)数量及功能的改变.方法以屋尘螨提取液复制小鼠哮喘模型后,分离小鼠脾脏CD4+ T细胞,采用流式细胞仪检测Treg细胞占CD4+ T细胞的比例;分离Treg细胞,与屋尘螨提取液共同培养后测定上清液中白细胞介素-4(IL-4)及IL-10的含量;将Treg细胞与CD4+ T细胞共同培养,观察其对CD4+ T细胞增殖及IL-4、IL-5、γ-干扰素(IFN-γ)分泌的影响.结果哮喘小鼠脾脏Treg细胞占CD4+ T细胞比例明显下降;与正常组相比,哮喘小鼠脾脏Treg细胞IL-4、IL-10的分泌无明显差异;Treg细胞可显著抑制哮喘小鼠CD4+ T细胞的增殖,并降低其IL-4的分泌,对IFN-γ的合成分泌无明显作用;使用anti-CD25 mAb后,哮喘和正常小鼠Treg细胞的作用均被显著抑制.结论哮喘小鼠Treg细胞功能与正常对照组小鼠无明显区别,但数量显著下降.哮喘发病过程中Th2型反应占优势,与Treg细胞数量减少,对Th2型反应的抑制作用降低有一定关系.

  11. Plasmid vector with temperature-controlled gene expression

    International Nuclear Information System (INIS)

    In plasmid pBR327, a fragment 169 b.p. long including promotor p3 of the bla gene has been deleted. The deletional derivative so obtained (pSP2) has been used to construct a recombinant plasmid bearing a fragment of phage λ DNA with the p/sub R/ promotor and the gene of the temperature-sensitive repressor cI. It has been shown that the plasmid vector so constructed (pCE119) with promotor cR performs repressor-cI-controlled transcription of the bla gene, as a result of which induction for an hour at 420C leads to an almost 100-fold increase in the amount of product of the bla gene as compared with that at 320C. The possibility of the use of plasmid cPE119 for the expression of other genes has been demonstrated for the case of the semisynthetic β-galactosidase gene of E. coli. In this case, on induction of the cells with recombinant plasmid pCEZ12 for 3 hours at 420C, a 300-fold increase in the amount of active β-galactosidase, as compared with that at 320C, was observed. It is important to point out that under these conditions (at 420C), at least 99% of the cells containing the plasmid retain the phenotype lacZ+, which indicates the stability of the proposed vector system

  12. Efficient control of gene expression by a tetracycline-dependent transactivator in single Dictyostelium discoideum cells

    NARCIS (Netherlands)

    Blaauw, M; Linskens, MHK; van Haastert, PJM

    2000-01-01

    We established a tetracycline-regulated gene expression system that tightly controls expression of genes in Dictyostelium discoideum. The control elements are contained in two plasmid vectors, one being an integrated plasmid encoding a chimeric tetracycline-controlled transcriptional activator prote

  13. CD44 expression positively correlates with Foxp3 expression and suppressive function of CD4+ Treg cells

    Directory of Open Access Journals (Sweden)

    König Rolf

    2009-10-01

    Full Text Available Abstract Background CD4+CD25+ regulatory T (Treg cells develop in the thymus and can suppress T cell proliferation, modulated by Foxp3 and cytokines; however, the relevance of CD44 in Treg cell development is less clear. To address this issue, we analyzed Foxp3 expression in CD44+ Treg cells by using multiple parameters, measured the levels of the immunoregulatory cytokine interleukin (IL-10 in various thymocyte subsets, and determined the suppressor activity in different splenic Treg cell populations. Results Within mouse thymocytes, we detected Treg cells with two novel phenotypes, namely the CD4+CD8-CD25+CD44+ and CD4+CD8-CD25+CD44- staining features. Additional multi-parameter analyses at the single-cell and molecular levels suggested to us that CD44 expression positively correlated with Foxp3 expression in thymocytes, the production of IL-10, and Treg activity in splenic CD4+CD25+ T cells. This suppressive effect of Treg cells on T cell proliferation could be blocked by using anti-IL-10 neutralizing antibodies. In addition, CD4+CD25+CD44+ Treg cells expressed higher levels of IL-10 and were more potent in suppressing effector T cell proliferation than were CD4+CD25+CD44- cells. Conclusion This study indicates the presence of two novel phenotypes of Treg cells in the thymus, the functional relevance of CD44 in defining Treg cell subsets, and the role of both IL-10 and Foxp3 in modulating the function of Treg cells. Reviewers This article was reviewed by Dr. M. Lenardo, Dr. L. Klein & G. Wirnsberger (nominated by Dr. JC Zungia-Pfluker, and Dr. E.M. Shevach.

  14. The effect of preference expression modality on self-control

    NARCIS (Netherlands)

    Klesse, A.K.; Levav, J.; Goukens, C.

    2015-01-01

    The marketplace affords consumers various modalities to express their preferences (e.g., by pressing a button on a vending machine or making an oral request at a restaurant). In this article, we compare speaking to manual preference expression modalities (button-pressing, writing, and taking) and st

  15. RNA Binding Proteins that Control Human Papillomavirus Gene Expression.

    OpenAIRE

    Naoko Kajitani; Stefan Schwartz

    2015-01-01

    The human papillomavirus (HPV) life cycle is strictly linked to the differentiation program of the infected mucosal epithelial cell. In the basal and lower levels of the epithelium, early genes coding for pro-mitotic proteins and viral replication factors are expressed, while terminal cell differentiation is required for activation of late gene expression and production of viral particles at the very top of the epithelium. Such productive infections are normally cleared within 18–24 months. I...

  16. HLA-DR expression on regulatory T cells is closely associated with the global immune activation in HIV-1 infected subjects na(i)ve to antiretroviral therapy

    Institute of Scientific and Technical Information of China (English)

    XIAO Jian; LI Ming-yuan; WANG Ying; QIAN Ke-lei; CAO Qing-hua; QIU Chen-li; QIU Cao; XUE Yi-le; ZHANG Xiao-yan; ZHONG Ping; XU Jian-qing

    2011-01-01

    Background The frequencies of regulatory T cells (Tregs) increased over the HIV infection but its counts actually decreased. We proposed that the decrease of Treg counts may cause the reduction of inhibitory effect and thereby account for the over-activation of Tregs during HIV infection. However, it remains unknown whether Tregs are also over-activated and thereafter the activation induced death may lead to the decrease of Tregs. Methods Tregs were defined as CD4+CD25+CD127lo/-T cells. Eighty-one HIV-1 infected patients were enrolled in our study, and twenty-two HIV-1 seronegative donors were recruited as the control. The levels of HLA-DR on Tregs were determined by FACSAria flow cytometer. ResultsCompared to HIV-1 seronegative donors, the levels of HLA-DR on CD4+CD25+CD127lo/- Tregs were significantly increased in HIV-1 infected patients, and its increase was positively associated with viral loads (r=0.3163,P=0.004) and negatively with CD4 T-cell counts (r=-0.4153, P<0.0001). In addition, significant associations between HLA-DR expression on CD4+CD25+CD127lo/-Tregs and the percentages of HLA-DR, CD38, Ki67 expressing CD4+ and CD8+ T cells were also identified. Conclusion HLA-DR on Tregs is a good marker for viral replication and disease progression. The over-activation of Tregs might result in the decrease of Tregs.

  17. ECAG 2008 Workshop: Facial and Bodily Expressions for Control and Adaptation of Games

    OpenAIRE

    Nijholt, Anton; Poppe, Ronald

    2008-01-01

    In this workshop of the 8th IEEE International Conference on Automatic Face and Gesture Recognition (FG 2008), the emphasis is on research on facial and bodily expressions for the control and adaptation of games. We distinguish between two forms of expressions, depending on whether the user has the initiative and consciously uses his or her movements and expressions to control the interface, or whether the application takes the initiative to adapt itself to the affective state of the user as ...

  18. IDO expressing fibroblasts promote the expansion of antigen specific regulatory T cells.

    Science.gov (United States)

    Curran, Terry-Ann; Jalili, Reza Baradar; Farrokhi, Ali; Ghahary, Aziz

    2014-01-01

    Regulatory CD4(+)CD25(+)Foxp3(+) T cells (Tregs) can be induced and expanded by dendritic cells (DCs) in the presence of the enzyme indoleamine 2,3-dioxygenase (IDO). Here we report that a possible alternative to DCs are IDO expressing dermal fibroblasts (DFs), which are easier to isolate and sustain in culture compared to DCs. When mouse splenocytes were co-cultured with IDO expressing DFs, a significant increase in frequency and the number of Tregs was found compared to those of control group (13.16%±1.8 vs. 5.53%±1.2, pa subset of Tregs which can be used to generate antigen-specific immune tolerance. PMID:23891282

  19. Human Adipose-Derived Stromal/Stem Cells Induce Functional CD4+CD25+FoxP3+CD127− Regulatory T Cells Under Low Oxygen Culture Conditions

    OpenAIRE

    Frazier, Trivia P.; McLachlan, James B.; Gimble, Jeffrey M.; Tucker, Hugh A; Brian G Rowan

    2014-01-01

    Human adipose tissue stromal/stem cells (ASCs) are known to induce proliferation of resting T cells under ambient (21%) O2 conditions; however, ASCs exist physiologically under lower oxygen (5% O2) conditions in adipose tissue. The effects of low oxygen levels on ASC immunomodulation of T cells are unknown. In this study, we show that ASCs stimulated proliferation of naive CD4+ T cells and the percentage of CD25+ T cells was significantly increased under both low and ambient O2. Forkhead box ...

  20. Increase of circulating CD4+CD25highFoxp3+ regulatory T cells in patients with metastatic renal cell carcinoma during treatment with dendritic cell vaccination and low-dose interleukin-2

    DEFF Research Database (Denmark)

    Berntsen, Annika; Brimnes, Marie Klinge; thor Straten, Per;

    2010-01-01

    Regulatory T cells (Treg) play an important role in the maintenance of immune tolerance and may be one of the obstacles of successful tumor immunotherapy. In this study, we analyzed the impact of administration of dendritic cell (DC) vaccination in combination with low-dose interleukin (IL)-2 in...... patients with metastatic renal cell carcinoma on the frequency of CD4+CD25highFoxp3+ Treg cells in peripheral blood. We found that the treatment increased the frequency of Treg cells more than 7-fold compared with pretreatment levels (P...

  1. Gene Expression Control by Glucocorticoid Receptors during Innate Immune Responses

    Directory of Open Access Journals (Sweden)

    André M. Xavier

    2016-04-01

    Full Text Available Glucocorticoids (GCs are potent anti-inflammatory compounds that have been extensively used in clinical practice for several decades. GCs effects on inflammation are generally mediated through GC receptors (GRs. Signal transduction through these nuclear receptors leads to dramatic changes in gene expression programs in different cell types, typically due to GR binding to DNA or to transcription modulators. During the last decade the view of GCs as exclusive anti-inflammatory molecules has been challenged. GR negative interference in pro-inflammatory gene expression was a landmark in terms of molecular mechanisms that suppress immune activity. In fact, GR can induce varied inhibitory molecules, including a negative regulator of Toll-like receptors (TLRs pathway, or subject key transcription factors, such as NF-B and AP-1, to a repressor mechanism. In contrast, the expression of some acute-phase proteins (APPs and other players of innate immunity generally requires GR signaling. Consequently, GRs must operate context-dependent inhibitory, permissive or stimulatory effects on host defense signaling triggered by pathogens or tissue damage. This review aims to disclose how contradictory or comparable effects on inflammatory gene expression can depend on pharmacological approach (including selective glucocorticoid receptor modulators; SEGRMs, cell culture, animal treatment or transgenic strategies used as models. Although the current view of GR-signaling integrated many advances in the field, some answers to important questions remain elusive.

  2. Gene Expression Control by Glucocorticoid Receptors during Innate Immune Responses

    Science.gov (United States)

    Xavier, Andre Machado; Anunciato, Aparecida Kataryna Olimpio; Rosenstock, Tatiana Rosado; Glezer, Isaias

    2016-01-01

    Glucocorticoids (GCs) are potent anti-inflammatory compounds that have been extensively used in clinical practice for several decades. GC’s effects on inflammation are generally mediated through GC receptors (GRs). Signal transduction through these nuclear receptors leads to dramatic changes in gene expression programs in different cell types, typically due to GR binding to DNA or to transcription modulators. During the last decade, the view of GCs as exclusive anti-inflammatory molecules has been challenged. GR negative interference in pro-inflammatory gene expression was a landmark in terms of molecular mechanisms that suppress immune activity. In fact, GR can induce varied inhibitory molecules, including a negative regulator of Toll-like receptors pathway, or subject key transcription factors, such as NF-κB and AP-1, to a repressor mechanism. In contrast, the expression of some acute-phase proteins and other players of innate immunity generally requires GR signaling. Consequently, GRs must operate context-dependent inhibitory, permissive, or stimulatory effects on host defense signaling triggered by pathogens or tissue damage. This review aims to disclose how contradictory or comparable effects on inflammatory gene expression can depend on pharmacological approach (including selective GC receptor modulators; SEGRMs), cell culture, animal treatment, or transgenic strategies used as models. Although the current view of GR-signaling integrated many advances in the field, some answers to important questions remain elusive. PMID:27148162

  3. The mRNA expression of pro- and anti-inflammatory cytokines in T regulatory cells in children with type 1 diabetes.

    Directory of Open Access Journals (Sweden)

    Maria Górska

    2010-06-01

    Full Text Available Type 1 diabetes mellitus (T1DM is caused by the autoimmune-mediated destruction of insulin-producing beta cells in the pancreas. T regulatory cells (Tregs represent an active mechanism of suppressing autoreactive T cells that escape central tolerance. The aim of our study was to test the hypothesis that T regulatory cells express pro- and anti-inflammatory cytokines, elements of cytotoxicity and OX40/4-1BB molecules. The examined group consisted of 50 children with T1DM. Fifty two healthy individuals (control group were enrolled into the study. A flow cytometric analysis of T-cell subpopulations was performed using the following markers: anti-CD3, anti-CD4, anti-CD25, anti-CD127, anti-CD134 and anti-CD137. Concurrently with the flow cytometric assessment of Tregs we separated CD4+CD25+CD127dim/- cells for further mRNA analysis. mRNA levels for transcription factor FoxP3, pro- and anti-inflammatory cytokines (interferon gamma, interleukin-2, interleukin-4, interleukin-10, transforming growth factor beta1 and tumor necrosis factor alpha, activatory molecules (OX40, 4-1BB and elements of cytotoxicity (granzyme B, perforin 1 were determined by real-time PCR technique. We found no alterations in the frequency of CD4+CD25highCD127low cells between diabetic and control children. Treg cells expressed mRNA for pro- and anti-inflammatory cytokines. Lower OX40 and higher 4-1BB mRNA but not protein levels in Treg cells in diabetic patients compared to the healthy children were noted. Our observations confirm the presence of mRNA for pro- and anti-inflammatory cytokines in CD4+CD25+CD127dim/- cells in the peripheral blood of children with T1DM. Further studies with the goal of developing new strategies to potentiate Treg function in autoimmune diseases are warranted.

  4. Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

    Directory of Open Access Journals (Sweden)

    D. Mesquita Júnior

    2014-08-01

    Full Text Available Regulatory T (TREG cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE. TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25+/highCD127Ø/lowFoxP3+, and effector T cells were defined as CD25+CD127+FoxP3Ø. The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4+TREG and CD28+TREG cells and an increased frequency of CD40L+TREG cells. There was a decrease in the TREG/effector-T ratio for GITR+, HLA-DR+, OX40+, and CD45RO+ cells, and an increased ratio of TREG/effector-T CD40L+ cells in patients with SLE. In addition, CD40L+TREG cell frequency correlated with the SLE disease activity index (P=0.0163. In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease.

  5. Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

    Energy Technology Data Exchange (ETDEWEB)

    Mesquita Júnior, D. [Disciplina de Reumatologia, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Cruvinel, W.M. [Disciplina de Reumatologia, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Departamento de Biomedicina, Universidade Católica de Goiás, Goiânia, GO (Brazil); Araujo, J.A.P. [Disciplina de Reumatologia, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Salmazi, K.C.; Kallas, E.G. [Disciplina de Imunologia Clínica e Alergia, Departamento de Clínica Médica, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP (Brazil); Andrade, L.E.C. [Disciplina de Reumatologia, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP (Brazil)

    2014-08-22

    Regulatory T (TREG) cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE). TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25{sup +/high}CD127{sup Ø/low}FoxP3{sup +}, and effector T cells were defined as CD25{sup +}CD127{sup +}FoxP3{sup Ø}. The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4{sup +}TREG and CD28{sup +}TREG cells and an increased frequency of CD40L{sup +}TREG cells. There was a decrease in the TREG/effector-T ratio for GITR{sup +}, HLA-DR{sup +}, OX40{sup +}, and CD45RO{sup +} cells, and an increased ratio of TREG/effector-T CD40L{sup +} cells in patients with SLE. In addition, CD40L{sup +}TREG cell frequency correlated with the SLE disease activity index (P=0.0163). In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease.

  6. Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

    International Nuclear Information System (INIS)

    Regulatory T (TREG) cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE). TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25+/highCD127Ø/lowFoxP3+, and effector T cells were defined as CD25+CD127+FoxP3Ø. The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4+TREG and CD28+TREG cells and an increased frequency of CD40L+TREG cells. There was a decrease in the TREG/effector-T ratio for GITR+, HLA-DR+, OX40+, and CD45RO+ cells, and an increased ratio of TREG/effector-T CD40L+ cells in patients with SLE. In addition, CD40L+TREG cell frequency correlated with the SLE disease activity index (P=0.0163). In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease

  7. Roles of GITR/GITRL signal system expression at bloodstream lymphocytes in children with asthma%哮喘患儿血淋巴细胞糖皮质激素诱导的肿瘤坏死因子受体及其配体信号系统的表达及意义

    Institute of Scientific and Technical Information of China (English)

    方丽; 李蓉蓉; 骆方军; 赵丽燕; 陈培英; 俞培锋

    2015-01-01

    目的:观察 GITR 及其配体(GITRL)在哮喘患儿外周血淋巴细胞的表达,探讨哮喘的部分炎症机制。方法50例儿童重度哮喘患儿(哮喘组)分别在急性期和缓解期取静脉血,流式细胞术检测CD +4 CD25+Treg 细胞 GITR 和 CD +4 T 细胞 GITRL 的平均荧光强度,与32例健康儿童作对照,进行统计分析。结果哮喘组急性期 CD +4 CD25+Treg 细胞 GITR 的表达水平为(24.2±8.2)MFI,显著低于对照组的(28.5±6.0)MFI(t =2.5,P <0.05)。哮喘组急性期与对照组 CD +4 T 细胞 GITRL 的表达水平分别为(5.2±3.6)MFI与(4.6±1.0)MFI,差异无统计学意义(t =1.1,P >0.05)。哮喘组 CD +4 CD25+Treg 细胞 GITR 的表达水平在缓解期[(29.5±8.3)MFI]显著高于急性期(t =-9.9,P <0.01);CD +4 T 细胞 GITRL 的表达水平在哮喘组急性期、缓解期(5.7±3.6)MFI 之间差异无统计学意义(t =-1.6,P >0.05)。CD +4 CD25+Treg GITR 与 CD +4 T GITRL 的表达之间无显著相关性(n =132,r =0.04,P >0.05)。结论哮喘急性期患儿 CD +4 CD25+Treg 细胞GITR 表达下降,在缓解期得到回升,而 CD +4 T 细胞 GITRL 的表达水平没有变化,GITR/GITRL 信号系统可能参与了哮喘的炎症过程。%Objective To investigate the expression of blood CD +4 CD +25 Treg GITR,CD +4 T cell GITRL in children with asthma,and the role of them in asthmatic inflammation.Methods 50 cases of severe asthma were selected,and were controlled with thirty two healthy children.The venous blood was collected both in the period of acute episode and clinic remission.The mean fluorescence intensity of CD +4 CD +25 Treg GITR and CD +4 T cell GITRL was detected by flow cytometry.Results The expression of CD +4 CD +25 Treg GITR in the asthma acute period group was (24.2 ±8.2)MFI,which was significantly lower than

  8. Different circadian oscillators control Ca2+ fluxes and Lhcb gene expression

    OpenAIRE

    Sai, Jiqing; Johnson, Carl Hirschie

    1999-01-01

    Circadian biological clocks control many biological events, but the pathways by which these events are controlled are largely unknown. Based on a model suggesting that cytosolic-free calcium levels control the expression of the Lhcb gene in plants, we tested whether the circadian oscillation of free calcium is responsible for driving the rhythm of Lhcb expression. We found that these rhythms free-run with different periods in tobacco seedlings in constant conditions. Moreover, robust oscillat...

  9. Topics and Signs: Defensive Control of Emotional Expression.

    Science.gov (United States)

    Horowitz, Mardi J.; And Others

    1993-01-01

    Single-case study examined frank disclosure of important topics in brief exploratory psychotherapy, including topics closely related to recent, unintegrated stressor life event. Quantitative measures of emotion and control variables showed heightened levels of both emotionality and defensive control during discourse on topic of stressor event.…

  10. A regulatory network controls nephrocan expression and midgut patterning

    OpenAIRE

    Hou, Juan; Wei, Wei; Saund, Ranajeet S.; Xiang, Ping; Cunningham, Thomas J.; Yi, Yuyin; Alder, Olivia; Lu, Daphne Y. D.; Savory, Joanne G. A.; Krentz, Nicole A. J.; Montpetit, Rachel; Cullum, Rebecca; Hofs, Nicole; Lohnes, David; Humphries, R. Keith

    2014-01-01

    Although many regulatory networks involved in defining definitive endoderm have been identified, the mechanisms through which these networks interact to pattern the endoderm are less well understood. To explore the mechanisms involved in midgut patterning, we dissected the transcriptional regulatory elements of nephrocan (Nepn), the earliest known midgut specific gene in mice. We observed that Nepn expression is dramatically reduced in Sox17−/− and Raldh2−/− embryos compared with wild-type em...

  11. A modified consumer inkjet for spatiotemporal control of gene expression.

    Directory of Open Access Journals (Sweden)

    Daniel J Cohen

    Full Text Available This paper presents a low-cost inkjet dosing system capable of continuous, two-dimensional spatiotemporal regulation of gene expression via delivery of diffusible regulators to a custom-mounted gel culture of E. coli. A consumer-grade, inkjet printer was adapted for chemical printing; E. coli cultures were grown on 750 microm thick agar embedded in micro-wells machined into commercial compact discs. Spatio-temporal regulation of the lac operon was demonstrated via the printing of patterns of lactose and glucose directly into the cultures; X-Gal blue patterns were used for visual feedback. We demonstrate how the bistable nature of the lac operon's feedback, when perturbed by patterning lactose (inducer and glucose (inhibitor, can lead to coordination of cell expression patterns across a field in ways that mimic motifs seen in developmental biology. Examples of this include sharp boundaries and the generation of traveling waves of mRNA expression. To our knowledge, this is the first demonstration of reaction-diffusion effects in the well-studied lac operon. A finite element reaction-diffusion model of the lac operon is also presented which predicts pattern formation with good fidelity.

  12. Differences in B7 and CD28 family gene expression in the peripheral blood between newly diagnosed young-onset and adult-onset type 1 diabetes patients.

    Science.gov (United States)

    Pruul, K; Kisand, K; Alnek, K; Metsküla, K; Reimand, K; Heilman, K; Peet, A; Varik, K; Peetsalu, M; Einberg, Ü; Tillmann, V; Uibo, R

    2015-09-01

    Type-1 diabetes (T1D) is a heterogeneous autoimmune disease, and there are pathogenetic differences between young- and adult-onset T1D patients. We hypothesized that the expressions of genes involved in costimulatory immune system pathways in peripheral blood are differently regulated in young- and adult-onset T1D. Study group I consisted of 80 children, adolescents, and young adults (age range 1.4-21.4 y; 31 controls and 49 T1D patients). Study group II consisted of 48 adults (age range 22.0-78.4 y; 30 controls and 18 T1D patients). The mRNA expression levels of CD86, CD28, CD25, CD226, CD40, BTLA, GITR, PDCD1, FoxP3, TGF-β, ICOS, sCTLA4, flCTLA4, and CD80 were measured in peripheral blood. Genetic polymorphisms (HLA haplotypes; rs231806, rs231775, and rs3087243 in CTLA4; rs763361 in CD226; and rs706778 in CD25) and T1D-associated autoantibodies were analyzed. In group I, there was significantly lower expression of CD226 in T1D patients than in the controls. In group II, there were significantly higher expression levels of CD86 and TGF-β in T1D patients than in the controls. In the T1D patients in group I, the upregulated CD80 expression correlated with the expression of both CTLA4 splice variants (sCTLA4 and flCTLA4). In contrast, in group II, upregulated CD86 correlated with TGF-β and CD25. In group I, the inhibitory CD80-CTLA4 pathway was activated, whereas, in group II, the activation CD86-CD28 pathway and TGF-β production were activated. These results emphasize the differences between young-onset and adult-onset T1D in the regulation of costimulatory pathways. These differences should be considered when developing novel treatments for T1D. PMID:25980680

  13. Pacientes com lúpus eritematoso sistêmico e síndrome antifosfolípide secundária possuem números reduzidos de células B CD4+ CD25+ Foxp3+ células Treg e células B CD3- CD19+ circulantes

    Directory of Open Access Journals (Sweden)

    Ester Rosári Raphaelli Dal Ben

    2014-06-01

    Full Text Available Introdução: A depleção de células T CD4+ CD25+ Foxp3+ regulatórias (células Treg foi descrita em pacientes com lúpus eritematoso sistêmico (LES e, recentemente, na síndrome antifosfolípide (SAF primária; até o momento, o tópico não tinha sido estudado em pacientes com LES e com SAF secundária (LES/SAFS. Objetivo: Quantificar linfócitos totais, células Treg, células T CD3+ CD19- e células B CD3- CD19+ em pacientes com LES/SAF e em controles saudáveis. Métodos: Subtipos celulares foram imunofenotipados utilizando anticorpos monoclonais específicos (antiCD3CY5, antiCD4FITC, antiCD25, antiFoxp3, antiCD19PE e citometria de fluxo. Resultados: Participaram do estudo 25 pacientes com LES/SAF (média de idade 43,5 anos, 96% mulheres, 96% da raça branca, duração média da doença 9,87 anos, SLEDAI médio 10±5,77 e 25 controles compatibilizados para idade e gênero. Foi constatado que os números de células Treg e de células B CD3- CD19+ estavam significativamente mais baixos em pacientes com LES/SAF, em comparação com controles (todos p0,05. Conclusões: Nesse estudo preliminar, pacientes com LES e com SAF secundária demonstraram depleção de células Treg e de células B CD3 CD19+; a redução numérica dos dois subtipos teve correlação com aumento de SLEDAI. A depleção de células Treg pode contribuir para a lesão autoimune observada em pacientes com LES/SAFS. O número reduzido de células B CD3 CD19+ observado nesses pacientes está a merecer estudos objetivando um aprofundamento em sua elucidação.

  14. Dual transcriptional-translational cascade permits cellular level tuneable expression control.

    Science.gov (United States)

    Morra, Rosa; Shankar, Jayendra; Robinson, Christopher J; Halliwell, Samantha; Butler, Lisa; Upton, Mathew; Hay, Sam; Micklefield, Jason; Dixon, Neil

    2016-02-18

    The ability to induce gene expression in a small molecule dependent manner has led to many applications in target discovery, functional elucidation and bio-production. To date these applications have relied on a limited set of protein-based control mechanisms operating at the level of transcription initiation. The discovery, design and reengineering of riboswitches offer an alternative means by which to control gene expression. Here we report the development and characterization of a novel tunable recombinant expression system, termed RiboTite, which operates at both the transcriptional and translational level. Using standard inducible promoters and orthogonal riboswitches, a multi-layered modular genetic control circuit was developed to control the expression of both bacteriophage T7 RNA polymerase and recombinant gene(s) of interest. The system was benchmarked against a number of commonly used E. coli expression systems, and shows tight basal control, precise analogue tunability of gene expression at the cellular level, dose-dependent regulation of protein production rates over extended growth periods and enhanced cell viability. This novel system expands the number of E. coli expression systems for use in recombinant protein production and represents a major performance enhancement over and above the most widely used expression systems. PMID:26405200

  15. Expression of transferred thymidine kinase genes is controlled by methylation.

    OpenAIRE

    Christy, B.; Scangos, G

    1982-01-01

    Plasmid pTKx-1, containing the herpes simplex virus gene for thymidine kinase (TK) inserted into the BamHI site of plasmid pBR322, was introduced into Ltk- cells by calcium phosphate precipitation in the absence of carrier DNA. Line 101 is a TK+ derivative of Ltk- that contains multiple copies of pTKx-1 in a multimeric structure. A derivative of 101 that retained but no longer expressed the herpes simplex TK genes (termed 101BU1) and derivatives of line 101BU1 that reexpressed the genes (term...

  16. A Modified Consumer Inkjet for Spatiotemporal Control of Gene Expression

    OpenAIRE

    Cohen, Daniel J.; Morfino, Roberto C.; Maharbiz, Michel M.

    2009-01-01

    This paper presents a low-cost inkjet dosing system capable of continuous, two-dimensional spatiotemporal regulation of gene expression via delivery of diffusible regulators to a custom-mounted gel culture of E. coli. A consumer-grade, inkjet printer was adapted for chemical printing; E. coli cultures were grown on 750 µm thick agar embedded in micro-wells machined into commercial compact discs. Spatio-temporal regulation of the lac operon was demonstrated via the printing of patterns of lact...

  17. act Operon Control of Developmental Gene Expression in Myxococcus xanthus

    OpenAIRE

    Gronewold, Thomas M. A.; Kaiser, Dale

    2002-01-01

    Cell-bound C-signal guides the building of a fruiting body and triggers the differentiation of myxospores. Earlier work has shown that transcription of the csgA gene, which encodes the C-signal, is directed by four genes of the act operon. To see how expression of the genes encoding components of the aggregation and sporulation processes depends on C-signaling, mutants with loss-of-function mutations in each of the act genes were investigated. These mutations were found to have no effect on g...

  18. VSG gene expression site control in insect form Trypanosoma brucei.

    OpenAIRE

    Rudenko, G; Blundell, P A; Taylor, M. C.; Kieft, R.; Borst, P

    1994-01-01

    When the African trypanosome Trypanosoma brucei is taken up from mammals by a tse-tse fly, it replaces its variant surface glycoprotein (VSG) coat by a procyclin coat. Transcription of VSG genes stops in the fly, but transcription of sequences derived from the promoter area of the VSG expression site(s) remains high. Whether this is due to continuing high activity of one promoter or to low activity of many promoters was unclear. We have used the small differences between the sequences of diff...

  19. Agonist-Driven Development of CD4+CD25+Foxp3+Regulatory T Cells Requires a Second Signal Mediated by Stat6

    DEFF Research Database (Denmark)

    Sanchez-Guajardo, Vanesa Maria; Tanshot, C.; O'Malley, J.T.;

    2007-01-01

    The factors that induce Foxp3 expression and regulatory T (Treg) cell development remain unknown. In this study, we investigated the role of STAT4 and STAT6 in agonist-driven generation of Ag-specific Foxp3-expressing Treg cells. Our findings indicate that fully efficient induction of Foxp3...... expression and development of Ag-specific Treg cells requires the synergistic action of two signals: a TCR-mediated signal and a second signal mediated by STAT6. Indeed, by comparing the development of wild-type and STAT4- and STAT6-deficient hemagglutinin-specific T cells in the presence of hemagglutinin Ag...... a role for the STAT6 pathway in Treg cell development and maintenance....

  20. Control of cyclooxygenase-2 expression and tumorigenesis by endogenous 5-methoxytryptophan

    OpenAIRE

    Cheng, Huei-Hsuan; Kuo, Cheng-Chin; Yan, Jiann-Long; Chen, Hua-Ling; Lin, Wei-Chung; Wang, Kai-Hsuan; Tsai, Kelvin K.-C.; Guvén, Hayrettin; Flaberg, Emilie; Szekely, Laszlo; Klein, George; Wu, Kenneth K.

    2012-01-01

    Cyclooxygenase-2 (COX-2) expression is induced by mitogenic and proinflammatory factors. Its overexpression plays a causal role in inflammation and tumorigenesis. COX-2 expression is tightly regulated, but the mechanisms are largely unclear. Here we show the control of COX-2 expression by an endogenous tryptophan metabolite, 5-methoxytryptophan (5-MTP). By using comparative metabolomic analysis and enzyme-immunoassay, our results reveal that normal fibroblasts produce and release 5-MTP into t...

  1. Genetic control of neuronal activity in mice conditionally expressing TRPV1

    OpenAIRE

    Arenkiel, Benjamin R.; Klein, Marguerita E; Davison, Ian G.; Katz, Lawrence C.; Ehlers, Michael D.

    2008-01-01

    Here we describe a knock-in mouse model for Cre-loxP–based conditional expression of TRPV1 in central nervous system neurons. Expression of Cre recombinase using biolistics, lentivirus or genetic intercrosses triggered heterologous expression of TRPV1 in a cell-specific manner. Application of the TRPV1 ligand capsaicin induced strong inward currents, triggered action potentials and activated stereotyped behaviors, allowing cell type–specific chemical genetic control of neuronal activity in vi...

  2. Genetic analysis of the phase variation control of expression of type 1 fimbriae in Escherichia coli.

    OpenAIRE

    Freitag, C S; Abraham, J. M.; Clements, J R; Eisenstein, B. I.

    1985-01-01

    Expression of type 1 fimbriae in Escherichia coli exhibits phase variation, whereby individual cells can alternate between states of organelle expression (Fim+) and nonexpression (Fim-). Strains with a fimD-lac operon fusion, in which lac, rather than fimD, expression is under the control of the fimD promoter, undergo Lac+ in equilibrium Lac- phase variation, instead. After positioning a lambda prophage adjacent to the operon fusion, we were able to isolate specialized lambda phage carrying b...

  3. [Express-method of assessing the status of hypertension control].

    Science.gov (United States)

    Glazunov, I S; Konstantinov, E N; Molchanov, V A; Gundarov, I A

    1986-01-01

    The authors describe a method for evaluating arterial hypertension control among the unorganized population visiting polyclinics and organized population taken care of at the medical centers. The method was tried in the city of Chelyabinsk as applicable to a randomized population sample of a medical territorial district and to a randomized sample (8%) of the workers from 9 shops of the steel plant. The method lies in the study of the population health status (arterial hypertension prevalence and knowledge, treatment coverage and efficacy) and of the documentation available at the treatment and prophylactic institutions concerning arterial hypertension control (coverage, registration, scope of examinations, and so forth). It is shown that the method is unsophisticated and feasible for examination of the population and documentation requires 5 days. It is suggested that the method should be used in the study of the situation concerning arterial hypertension control as well as in the study of the problems in the control of other non-infectious diseases. PMID:3824187

  4. AlgU controls expression of virulence genes in Pseudomonas syringae pv. tomato DC3000

    Science.gov (United States)

    Plant pathogenic bacteria are able to integrate information about their environment and adjust gene expression to provide adaptive functions. AlgU, an ECF sigma factor encoded by Pseudomonas syringae, controls expression of genes for alginate biosynthesis and is active while the bacteria are associa...

  5. The amygdalo-motor pathway and the control of facial expressions

    Directory of Open Access Journals (Sweden)

    Katalin M Gothard

    2014-03-01

    Full Text Available Facial expressions reflect decisions about the perceived meaning of social stimuli emitted by others and the expected socio-emotional outcome of the reciprocating expression. The decision to produce a facial expression emerges from the joint activity of a network of structures that include the amygdala and multiple, interconnected cortical and subcortical motor areas. Reciprocal transformations between sensory and motor signals give rise to distinct brain states that promote, or impede the production of facial expressions. The muscles of the upper and lower face are controlled by anatomically distinct motor areas and thus require distinct patterns of motor commands. Concomitantly multiple areas, including the amygdala, monitor the ongoing overt behavior (the expression of self and the covert, autonomic responses that accompany emotional expressions. Interoceptive signals and visceral states, therefore, should be incorporated into the formalisms of decision making in order account for decisions that govern the receiving-emitting cycle of facial expressions.

  6. Itch expression by Treg cells controls Th2 inflammatory responses

    OpenAIRE

    Jin, Hyung-Seung; Park, Yoon; Elly, Chris; Liu, Yun-Cai

    2013-01-01

    Regulatory T (Treg) cells maintain immune homeostasis by limiting autoimmune and inflammatory responses. Treg differentiation, maintenance, and function are controlled by the transcription factor Foxp3. However, the exact molecular mechanisms underlying Treg cell regulation remain elusive. Here, we show that Treg cell–specific ablation of the E3 ubiquitin ligase Itch in mice caused massive multiorgan lymphocyte infiltration and skin lesions, chronic T cell activation, and the development of s...

  7. Gene Expression of Leptin and Long Leptin Receptor Isoform in Endometriosis: A Case-Control Study

    OpenAIRE

    Andrea Prestes Nácul; Sheila Bunecker Lecke; Maria Isabel Edelweiss; Débora Martinho Morsch; Poli Mara Spritzer

    2013-01-01

    In this study, leptin/BMI ratio in serum and peritoneal fluid and gene expression of leptin and long form leptin receptor (OB-RL) were assessed in eutopic and ectopic endometria of women with endometriosis and controls. Increased serum leptin/BMI ratio was found in endometriosis patients. Leptin and OB-RL gene expression was significantly higher in ectopic versus eutopic endometrium of patients and controls. A positive, significant correlation was observed between leptin and OB-RL transcripts...

  8. Prediction of Children’s Empathy-Related Responding From Their Effortful Control and Parents’ Expressivity

    OpenAIRE

    Valiente, Carlos; Eisenberg, Nancy; Fabes, Richard A.; Shepard, Stephanie A.; Cumberland, Amanda; Losoya, Sandra H.

    2004-01-01

    In this study, the linear and interactive relations of children’s effortful control and parents’ emotional expressivity to children’s empathy-related responses were examined. Participants were 214 children, 4.5 to 8 years old. Children’s effortful control was negatively related to their personal distress and was positively related to their sympathy. Parents’ positive expressivity was marginally negatively related to children’s personal distress and was marginally positively related to childre...

  9. A Novel Control Algorithm Expressions Set for not Negligible Resistive Parameters PM Brushless AC Motors

    OpenAIRE

    Rizzo, Renato; Andrea DEL PIZZO; Ivan SPINA

    2012-01-01

    This paper deals with Permanent Magnet Brushless Motors. In particular is proposed a new set of control algorithm expressions that is realized taking into account resistive parameters of the motor, differently from simplified models of this type of motors where these parameters are usually neglected. The control is set up and an analysis of the performance is reported in the paper, where the validation of the new expressions is done with reference to a motor prototype particularly compact bec...

  10. Mind-controlled transgene expression by a wireless-powered optogenetic designer cell implant

    OpenAIRE

    Marc Folcher; Sabine Oesterle; Katharina Zwicky; Thushara Thekkottil; Julie Heymoz; Muriel Hohmann; Matthias Christen; Marie Daoud El-Baba; Peter Buchmann; Martin Fussenegger

    2014-01-01

    Synthetic devices for traceless remote control of gene expression may provide new treatment opportunities in future gene- and cell-based therapies. Here we report the design of a synthetic mind-controlled gene switch that enables human brain activities and mental states to wirelessly programme the transgene expression in human cells. An electroencephalography (EEG)-based brain–computer interface (BCI) processing mental state-specific brain waves programs an inductively linked wireless-powered...

  11. Dynamic features of gene expression control by small regulatory RNAs.

    Science.gov (United States)

    Mitarai, Namiko; Benjamin, Julie-Anna M; Krishna, Sandeep; Semsey, Szabolcs; Csiszovszki, Zsolt; Massé, Eric; Sneppen, Kim

    2009-06-30

    Small regulatory RNAs (sRNAs) in eukaryotes and bacteria play an important role in the regulation of gene expression either by binding to regulatory proteins or directly to target mRNAs. Two of the best-characterized bacterial sRNAs, Spot42 and RyhB, form a complementary pair with the ribosome binding region of their target mRNAs, thereby inhibiting translation or promoting mRNA degradation. To investigate the steady-state and dynamic potential of such sRNAs, we examine the 2 key parameters characterizing sRNA regulation: the capacity to overexpress the sRNA relative to its target mRNA and the speed at which the target mRNA is irreversibly inactivated. We demonstrate different methods to determine these 2 key parameters, for Spot42 and RyhB, which combine biochemical and genetic experiments with computational analysis. We have developed a mathematical model that describes the functional properties of sRNAs with various characteristic parameters. We observed that Spot42 and RyhB function in distinctive parameter regimes, which result in divergent mechanisms. PMID:19541626

  12. Controlled expression of enhanced green fluorescent protein and hepatitis B virus precore protein in mammalian cells

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A novel tetracycline regulation expression system was used to regulate the expression of enhanced green fluorescent protein (EGFP) and hepatitis B virus precore protein in the mammalian cell lines with lipofectAMINE. Flow cytometry assays showed that application of the system resulted in about 18-fold induction of EGFP expression in CHO cell lines and 5-fold induction in SSMC-7721 cells and about 2-fold in the HEK293 cells. Furthermore, the effective use of this system for the controlled expression of HBV precore protein gene in hepatocellular carcinoma cells was tested.

  13. Quality Control Usage in High-Density Microarrays Reveals Differential Gene Expression Profiles in Ovarian Cancer.

    Science.gov (United States)

    Villegas-Ruiz, Vanessa; Moreno, Jose; Jacome-Lopez, Karina; Zentella-Dehesa, Alejandro; Juarez-Mendez, Sergio

    2016-01-01

    There are several existing reports of microarray chip use for assessment of altered gene expression in different diseases. In fact, there have been over 1.5 million assays of this kind performed over the last twenty years, which have influenced clinical and translational research studies. The most commonly used DNA microarray platforms are Affymetrix GeneChip and Quality Control Software along with their GeneChip Probe Arrays. These chips are created using several quality controls to confirm the success of each assay, but their actual impact on gene expression profiles had not been previously analyzed until the appearance of several bioinformatics tools for this purpose. We here performed a data mining analysis, in this case specifically focused on ovarian cancer, as well as healthy ovarian tissue and ovarian cell lines, in order to confirm quality control results and associated variation in gene expression profiles. The microarray data used in our research were downloaded from ArrayExpress and Gene Expression Omnibus (GEO) and analyzed with Expression Console Software using RMA, MAS5 and Plier algorithms. The gene expression profiles were obtained using Partek Genomics Suite v6.6 and data were visualized using principal component analysis, heat map, and Venn diagrams. Microarray quality control analysis showed that roughly 40% of the microarray files were false negative, demonstrating over- and under-estimation of expressed genes. Additionally, we confirmed the results performing second analysis using independent samples. About 70% of the significant expressed genes were correlated in both analyses. These results demonstrate the importance of appropriate microarray processing to obtain a reliable gene expression profile. PMID:27268623

  14. Marek's Disease Viral Interleukin-8 Promotes Lymphoma Formation through Targeted Recruitment of B Cells and CD4+ CD25+ T Cells

    OpenAIRE

    Engel, Annemarie T.; Selvaraj, Ramesh K.; Kamil, Jeremy P.; Osterrieder, Nikolaus; Kaufer, Benedikt B

    2012-01-01

    Marek's disease virus (MDV) is a cell-associated and highly oncogenic alphaherpesvirus that infects chickens. During lytic and latent MDV infection, a CXC chemokine termed viral interleukin-8 (vIL-8) is expressed. Deletion of the entire vIL-8 open reading frame (ORF) was shown to severely impair disease progression and tumor development; however, it was unclear whether this phenotype was due to loss of secreted vIL-8 or of splice variants that fuse exons II and III of vIL-8 to certain upstrea...

  15. Flt3-L Increases CD4+CD25+Foxp3+ICOS+ Cells in the Lungs of Cockroach-Sensitized and -Challenged Mice

    OpenAIRE

    McGee, Halvor S; Edwan, Jehad H.; Devendra K Agrawal

    2009-01-01

    We previously reported in an ovalbumin-induced model of allergic asthma that Fms-like tyrosine kinase 3 ligand (Flt3-L) reversed airway hyperresponsiveness (AHR) and airway inflammation, and increased the number of regulatory CD11chighCD8αhighCD11blow dendritic cells in the lung. In this study, we investigated the effect of Flt3-L in a clinically relevant aeroallergen-induced asthma on the phenotypic expression of lung T cells. Balb/c mice were sensitized and challenged with cockroach antigen...

  16. 维生素A和布地奈德对哮喘大鼠外周血T细胞亚群和调节性T细胞的影响%The effects of vitamin A and budesonide on serum T-lymphocyte subsets and CD4 + CD25 + regulatory T cells in rat asthma model

    Institute of Scientific and Technical Information of China (English)

    何金根; 潘家华; 楼皖玲; 刘辉

    2011-01-01

    目的:探讨维生素A(Vit A)和布地奈德对哮喘大鼠外周血T细胞亚群和CD4+ CD25+ 调节性T细胞(CD4+ CD25+regulatory T cell,CD4+ CD25+ Treg)的影响,为其在哮喘防治中的应用提供实验依据.方法:将42只SD大鼠用卵蛋白建立哮喘模型,随机分成布地奈德治疗组、Vit A治疗组和哮喘对照组,每组各14只,各组干预1周,于第4周和第5周取外周血50μl,流式细胞术检测T细胞亚群及Treg细胞变化.结果:激发后第4周,Vit A组CD3+T细胞、CD4+ CD25+ Treg细胞显著高于布地奈德组(P<0.05),CD3+ CD8+ T细胞均低于布地奈德组(P<0.05);布地奈德组CD4+ CD25+ Treg细胞显著低于哮喘组(P<0.01);激发后第5周,布地奈德组CD4+ CD25+ Treg细胞低于哮喘组(P<0.05).结论:雾化吸入Vit A可减轻哮喘大鼠炎症反应,这一作用可能与哮喘大鼠外周血CD3+ T细胞和CIM+ CD25+ Treg细胞升高、CD8+ T细胞降低有关.%Objective:To investigate the effects of vitamin A and budesonide on peripheral blood T-lymphocyte subsets and CD4 +CD25 + Treg cells in rat asthma model, which provide the experimental basis for vitamin A in the prevention and treatment of bronchial asthma. Methods :Forty-two adult female SD rats were divided into three groups, the budesonide group, vitamin A group and asthma group. There were 14 rats in each group. The peripheral blood was conducted to determinate T-lymphocyte subsets and CD4 + CD25 + Treg by flow cytometry. Results: After the 4th of stimulation, CD3 + T cell and CD4 + CD25 + Treg cells in vitamin A group were significantly higher than those in budesonide group. CD3 + CD8 + T cell in vitamin A group was significantly lower than that in asthma group ( P < 0.05 ). CD4 + CD25 + Treg cells of budesonide group was significantly lower than that in asthma group ( P < 0.01 ). After the 5th of stimulation, CD4 + CD25 + Treg cells in budesonide group was significantly lower than that in asthma group ( P < 0. 05 ).Conclusions

  17. Prediction of Children's Empathy-Related Responding from Their Effortful Control and Parents' Expressivity

    Science.gov (United States)

    Valiente, Carlos; Eisenberg, Nancy; Fabes, Richard A.; Shepard, Stephanie A.; Cumberland, Amanda; Losoya, Sandra H.

    2004-01-01

    In this study, the linear and interactive relations of children's effortful control and parents' emotional expressivity to children's empathy-related responses were examined. Participants were 214 children, 4.5 to 8 years old. Children's effortful control was negatively related to their personal distress and was positively related to their…

  18. Early pregnancy peripheral blood gene expression and risk of preterm delivery: a nested case control study

    Directory of Open Access Journals (Sweden)

    Muhie Seid Y

    2009-12-01

    Full Text Available Abstract Background Preterm delivery (PTD is a significant public health problem associated with greater risk of mortality and morbidity in infants and mothers. Pathophysiologic processes that may lead to PTD start early in pregnancy. We investigated early pregnancy peripheral blood global gene expression and PTD risk. Methods As part of a prospective study, ribonucleic acid was extracted from blood samples (collected at 16 weeks gestational age from 14 women who had PTD (cases and 16 women who delivered at term (controls. Gene expressions were measured using the GeneChip® Human Genome U133 Plus 2.0 Array. Student's T-test and fold change analysis were used to identify differentially expressed genes. We used hierarchical clustering and principle components analysis to characterize signature gene expression patterns among cases and controls. Pathway and promoter sequence analyses were used to investigate functions and functional relationships as well as regulatory regions of differentially expressed genes. Results A total of 209 genes, including potential candidate genes (e.g. PTGDS, prostaglandin D2 synthase 21 kDa, were differentially expressed. A set of these genes achieved accurate pre-diagnostic separation of cases and controls. These genes participate in functions related to immune system and inflammation, organ development, metabolism (lipid, carbohydrate and amino acid and cell signaling. Binding sites of putative transcription factors such as EGR1 (early growth response 1, TFAP2A (transcription factor AP2A, Sp1 (specificity protein 1 and Sp3 (specificity protein 3 were over represented in promoter regions of differentially expressed genes. Real-time PCR confirmed microarray expression measurements of selected genes. Conclusions PTD is associated with maternal early pregnancy peripheral blood gene expression changes. Maternal early pregnancy peripheral blood gene expression patterns may be useful for better understanding of PTD

  19. Effortful control, positive emotional expression, and behavior problems in children born preterm.

    Science.gov (United States)

    Burnson, Cynthia; Poehlmann, Julie; Schwichtenberg, A J

    2013-12-01

    The present study focused on the role of high effortful control in the expression of positive emotion and development of behavior problems in children born preterm (mean gestational age = 31.4 weeks). Using data from a prospective longitudinal study, the present study assessed effortful control and behavior problems at 24 and 36 months and positive emotional expression at 24 months in a sample of 173 children born preterm. Less positive emotional expression was associated with higher effortful control for boys but not girls. Higher effortful control was associated with fewer total behavior problems, but this relation was attenuated when socioeconomic assets were included in the model. More socioeconomic assets were associated with fewer behavior problems for both boys and girls and higher effortful control for girls. Socioeconomic assets appear to be an important factor in the development of effortful control and behavior problems in children born preterm regardless of gender, whereas positive emotional expression was important for boys. Future intervention research should examine fostering adaptive levels of effortful control in high-risk populations as a means to facilitate resilience processes. PMID:23810984

  20. Stressor Controllability and Fos Expression in Stress Regulatory Regions in Mice

    OpenAIRE

    Liu, X.; Tang, X.; Sanford, LD

    2009-01-01

    Controllability is an important determinant of the effects of stress on behavior. We trained mice with escapable (ES) and inescapable (IS) shock and examined behavioral freezing and Fos expression in brain regions involved in stress to determine whether stressor controllability produced differential activation of these regions. Mice (C57BL/6J) were trained to escape footshock by moving to a safe chamber in a shuttlebox. This terminated shock for both ES mice (n=5) and yoked-control mice recei...

  1. Does CD4+CD25+foxp3+ cell (Treg) and IL-10 profile determine susceptibility to immune reconstitution inflammatory syndrome (IRIS) in HIV disease?

    OpenAIRE

    Saravanan Shanmugam; Lloyd Charmaine AC; Balakrishnan Pachamuthu; Sekar Ramalingam; Murugavel Kailapuri G; Velu Vijayakumar; Vignesh Ramachandran; Shankar Esaki; Solomon Suniti; Kumarasamy Nagalingeswaran

    2008-01-01

    Abstract HIV-specific T-lymphocyte responses that underlie IRIS are incomplete and largely remain hypothetical. Of the several mechanisms presented by the host to control host immunological damage, Treg cells are believed to play a critical role. Using the available experimental evidence, it is proposed that enormous synthesis of conventional FoxP3- Th cells (responsive) often renders subjects inherently vulnerable to IRIS, whereas that of natural FoxP3+ Treg cell synthesis predominate among ...

  2. IL2RA/CD25 Gene Polymorphisms: Uneven Association with Multiple Sclerosis (MS) and Type 1 Diabetes (T1D)

    Science.gov (United States)

    Alcina, Antonio; Fedetz, María; Ndagire, Dorothy; Fernández, Oscar; Leyva, Laura; Guerrero, Miguel; Abad-Grau, María M.; Arnal, Carmen; Delgado, Concepción; Lucas, Miguel; Izquierdo, Guillermo; Matesanz, Fuencisla

    2009-01-01

    Background IL-2 receptor (IL2R) alpha is the specific component of the high affinity IL2R system involved in the immune response and in the control of autoimmunity. Methods and Results Here we perform a replication and fine mapping of the IL2RA gene region analyzing 3 SNPs previously associated with multiple sclerosis (MS) and 5 SNPs associated with type 1 diabetes (T1D) in a collection of 798 MS patients and 927 matched Caucasian controls from the south of Spain. We observed association with MS in 6 of 8 SNPs. The rs1570538, at the 3′- UTR extreme of the gene, previously reported to have a weak association with MS, is replicated here (P = 0.032). The most associated T1D SNP (rs41295061) was not associated with MS in the present study. However, the rs35285258, belonging to another independent group of SNPs associated with T1D, showed the maximal association in this study but different risk allele. We replicated the association of only one (rs2104286) of the two IL2RA SNPs identified in the recently performed genome-wide association study of MS. Conclusions These findings confirm and extend the association of this gene with MS and reveal a genetic heterogeneity of the associated polymorphisms and risk alleles between MS and T1D suggesting different immunopathological roles of IL2RA in these two diseases. PMID:19125193

  3. Gene expression analysis in prostate cancer: the importance of the endogenous control.

    LENUS (Irish Health Repository)

    Vajda, Alice

    2013-03-01

    Aberrant gene expression is a hallmark of cancer. Quantitative reverse-transcription PCR (qRT-PCR) is the gold-standard for quantifying gene expression, and commonly employs a house-keeping gene (HKG) as an endogenous control to normalize results; the choice of which is critical for accurate data interpretation. Many factors, including sample type, pathological state, and oxygen levels influence gene expression including putative HKGs. The aim of this study was to determine the suitability of commonly used HKGs for qRT-PCR in prostate cancer.

  4. Photocaged Arabinose: A Novel Optogenetic Switch for Rapid and Gradual Control of Microbial Gene Expression.

    Science.gov (United States)

    Binder, Dennis; Bier, Claus; Grünberger, Alexander; Drobietz, Dagmar; Hage-Hülsmann, Jennifer; Wandrey, Georg; Büchs, Jochen; Kohlheyer, Dietrich; Loeschcke, Anita; Wiechert, Wolfgang; Jaeger, Karl-Erich; Pietruszka, Jörg; Drepper, Thomas

    2016-02-15

    Controlling cellular functions by light allows simple triggering of biological processes in a non-invasive fashion with high spatiotemporal resolution. In this context, light-regulated gene expression has enormous potential for achieving optogenetic control over almost any cellular process. Here, we report on two novel one-step cleavable photocaged arabinose compounds, which were applied as light-sensitive inducers of transcription in bacteria. Exposure of caged arabinose to UV-A light resulted in rapid activation of protein production, as demonstrated for GFP and the complete violacein biosynthetic pathway. Moreover, single-cell analysis revealed that intrinsic heterogeneity of arabinose-mediated induction of gene expression was overcome when using photocaged arabinose. We have thus established a novel phototrigger for synthetic bio(techno)logy applications that enables precise and homogeneous control of bacterial target gene expression. PMID:26677142

  5. Neural Correlates of Conflict Control on Facial Expressions with a Flanker Paradigm

    DEFF Research Database (Denmark)

    Liu, T.; Xiao, T; Shi, Jiannong

    2013-01-01

    results manifested that participants exhibited faster response speed in identifying neutral target face when it was flanked by neutral distractors than by happy distractors. Electrophysiological results showed that happy target expression induced larger N2 amplitude when flanked by sad distractors than by...... happy distractors and scramble blocks during the conflict monitoring processing. During the attentional control processing, happy target expression induced faster P3 response when it was flanked by happy distractors than by sad distractors, and sad target expression evoked larger P3 amplitude when it...... was flanked by happy distractors comparing with sad distractors. Taken together, the current findings of temporal dynamic of brain activity during cognitive control on affective conflicts shed light on the essential relationship between cognitive control and affective information processing....

  6. Thioperamide induces CD4+ CD25+ Foxp3+ regulatory T lymphocytes in the lung mucosa of allergic mice through its action on dendritic cells

    Directory of Open Access Journals (Sweden)

    Geffner J

    2011-09-01

    Full Text Available Maria Marta Amaral1*, Carolina A Alvarez1*, Cecilia Langellotti2, Carolina Jancic1, Gabriela Salamone1, Jorge Geffner1, Mónica Vermeulen1,1Institute of Hematologic Research, National Academy of Medicine, Buenos Aires, Argentina; 2Institute of Virology, CICVyA, INTA, Castelar, Argentina *Authors contributed equally to this workBackground: Histamine is an important mediator in the development of allergic reactions. The biological effects of histamine are mediated through four histaminergic receptors. In recent years, an important role has been assigned to the proinflammatory functions of histamine regarding the H4 receptor. Previously, we have demonstrated that injection of immature dendritic cells treated with histamine into allergic mice promotes an increase in CD8+ Tc2 lymphocytes, which are involved in the worsening of allergy symptoms during the chronic phase of the disease. The aim of this study was to evaluate the role of the H3/H4 receptor antagonist, thioperamide, in allergy.Methods: Ovalbumin-allergized mice and nonallergized mice were injected with phosphate-buffered saline, dendritic cells, or thioperamide-treated dendritic cells. After treatment, the lungs of the mice were obtained and analyzed for changes in the populations of dendritic cells and T lymphocytes, as well as the expression of H and H4 receptors in mononuclear lung cells.Results: We found an increase in regulatory T cells in the lungs of allergic mice intratracheally injected with dendritic cells which had their H3/H4 receptors blocked with thioperamide. We also found an increase in the production of interleukin-10 by dendritic cells of the lung. Finally, we observed a decrease in serum levels of specific anti-IgE and a reduction of eosinophils in bronchoalveolar lavage from allergic mice.Conclusion: Thioperamide induces a significant improvement in symptoms of allergic reaction perhaps via induction of regulatory T lymphocytes. These findings may become relevant in the

  7. A Novel Control Algorithm Expressions Set for not Negligible Resistive Parameters PM Brushless AC Motors

    Directory of Open Access Journals (Sweden)

    Renato RIZZO

    2012-08-01

    Full Text Available This paper deals with Permanent Magnet Brushless Motors. In particular is proposed a new set of control algorithm expressions that is realized taking into account resistive parameters of the motor, differently from simplified models of this type of motors where these parameters are usually neglected. The control is set up and an analysis of the performance is reported in the paper, where the validation of the new expressions is done with reference to a motor prototype particularly compact because is foreseen for application on tram propulsion drives. The results are evidenced in the last part of the paper.

  8. Triple-controlled oncolytic adenovirus expressing melittin to exert inhibitory efficacy on hepatocellular carcinoma

    OpenAIRE

    Qian, Chun-Yu; Wang, Kai-Li; Fang, Fan-Fu; Gu, Wei; Huang, Feng; Wang, Fu-Zhe; Li, Bai; Wang, Li-Na

    2015-01-01

    Hepatocellular carcinoma (HCC) is a highly malignant disease, and its outcome of routine therapies is poor. Comprehensive treatment including gene therapy is an important way to improve patients’ prognosis and survival. In this study, we successfully constructed a triple-controlled cancer-selective oncolytic adenovirus, QG511-HA-Melittin, carrying melittin gene, in which the hybrid promoter, hypoxia-response element (HRE)-AFP promoter, was used to control viral E1a expression targeting AFP-po...

  9. Controlling transgene expression in subcutaneous implants using a skin lotion containing the apple metabolite phloretin

    OpenAIRE

    Gitzinger, Marc; Kemmer, Christian; El-Baba, Marie Daoud; Weber, Wilfried; Fussenegger, Martin

    2009-01-01

    Adjustable control of therapeutic transgenes in engineered cell implants after transdermal and topical delivery of nontoxic trigger molecules would increase convenience, patient compliance, and elimination of hepatic first-pass effect in future therapies. Pseudomonas putida DOT-T1E has evolved the flavonoid-triggered TtgR operon, which controls expression of a multisubstrate-specific efflux pump (TtgABC) to resist plant-derived defense metabolites in its rhizosphere habitat. Taking advantage ...

  10. Perceiving emotions: Cueing social categorization processes and attentional control through facial expressions.

    Science.gov (United States)

    Cañadas, Elena; Lupiáñez, Juan; Kawakami, Kerry; Niedenthal, Paula M; Rodríguez-Bailón, Rosa

    2016-09-01

    Individuals spontaneously categorise other people on the basis of their gender, ethnicity and age. But what about the emotions they express? In two studies we tested the hypothesis that facial expressions are similar to other social categories in that they can function as contextual cues to control attention. In Experiment 1 we associated expressions of anger and happiness with specific proportions of congruent/incongruent flanker trials. We also created consistent and inconsistent category members within each of these two general contexts. The results demonstrated that participants exhibited a larger congruency effect when presented with faces in the emotional group associated with a high proportion of congruent trials. Notably, this effect transferred to inconsistent members of the group. In Experiment 2 we replicated the effects with faces depicting true and false smiles. Together these findings provide consistent evidence that individuals spontaneously utilise emotions to categorise others and that such categories determine the allocation of attentional control. PMID:26197208

  11. Coordinating gene expression and axon assembly to control axon growth: potential role of GSK3 signaling

    Directory of Open Access Journals (Sweden)

    Fengquan Zhou

    2012-02-01

    Full Text Available Axon growth requires coordinated regulation of gene expression in the neuronal soma, anterograde transport of synthesized raw materials along the axon, and assembly of cytoskeleton and membranes in the nerve growth cone. Glycogen synthase kinase 3 (GSK3 signaling has recently been shown to play key roles in regulation of axonal transport and cytoskeletal assembly during axon growth. GSK3 signaling is also known to regulate gene expression via controlling the functions of many transcription factors, suggesting that GSK3 may be an important regulator of gene transcription supporting axon growth. Here we will review signaling pathways that control local axon assembly at the growth cone and gene expression in the soma during developmental or regenerative axon growth and discuss the potential involvement of GSK3 signaling in these processes, with a particular focus on how GSK3 signaling modulates the function of axon growth-associated transcription factors.

  12. Expression

    Directory of Open Access Journals (Sweden)

    Wang-Xia Wang

    2014-02-01

    Full Text Available The miR-15/107 family comprises a group of 10 paralogous microRNAs (miRNAs, sharing a 5′ AGCAGC sequence. These miRNAs have overlapping targets. In order to characterize the expression of miR-15/107 family miRNAs, we employed customized TaqMan Low-Density micro-fluid PCR-array to investigate the expression of miR-15/107 family members, and other selected miRNAs, in 11 human tissues obtained at autopsy including the cerebral cortex, frontal cortex, primary visual cortex, thalamus, heart, lung, liver, kidney, spleen, stomach and skeletal muscle. miR-103, miR-195 and miR-497 were expressed at similar levels across various tissues, whereas miR-107 is enriched in brain samples. We also examined the expression patterns of evolutionarily conserved miR-15/107 miRNAs in three distinct primary rat brain cell preparations (enriched for cortical neurons, astrocytes and microglia, respectively. In primary cultures of rat brain cells, several members of the miR-15/107 family are enriched in neurons compared to other cell types in the central nervous system (CNS. In addition to mature miRNAs, we also examined the expression of precursors (pri-miRNAs. Our data suggested a generally poor correlation between the expression of mature miRNAs and their precursors. In summary, we provide a detailed study of the tissue and cell type-specific expression profile of this highly expressed and phylogenetically conserved family of miRNA genes.

  13. Mind-controlled transgene expression by a wireless-powered optogenetic designer cell implant.

    Science.gov (United States)

    Folcher, Marc; Oesterle, Sabine; Zwicky, Katharina; Thekkottil, Thushara; Heymoz, Julie; Hohmann, Muriel; Christen, Matthias; Daoud El-Baba, Marie; Buchmann, Peter; Fussenegger, Martin

    2014-01-01

    Synthetic devices for traceless remote control of gene expression may provide new treatment opportunities in future gene- and cell-based therapies. Here we report the design of a synthetic mind-controlled gene switch that enables human brain activities and mental states to wirelessly programme the transgene expression in human cells. An electroencephalography (EEG)-based brain-computer interface (BCI) processing mental state-specific brain waves programs an inductively linked wireless-powered optogenetic implant containing designer cells engineered for near-infrared (NIR) light-adjustable expression of the human glycoprotein SEAP (secreted alkaline phosphatase). The synthetic optogenetic signalling pathway interfacing the BCI with target gene expression consists of an engineered NIR light-activated bacterial diguanylate cyclase (DGCL) producing the orthogonal second messenger cyclic diguanosine monophosphate (c-di-GMP), which triggers the stimulator of interferon genes (STING)-dependent induction of synthetic interferon-β promoters. Humans generating different mental states (biofeedback control, concentration, meditation) can differentially control SEAP production of the designer cells in culture and of subcutaneous wireless-powered optogenetic implants in mice. PMID:25386727

  14. Host plant preference of spider mites on Bt-expressing and control potatoes

    Czech Academy of Sciences Publication Activity Database

    Zemek, Rostislav

    Coventry : University of Warwick, 2008. s. 61-61. [Annual Meeting of the Society for Invertebrate Pathology /41./ and International Conference on Bacillus thuringiensis Incorporating COST 862 Action: Bacterial Toxins for Insect Control /9./. 03.08.2008-07.08.2008, Coventry] Institutional research plan: CEZ:AV0Z50070508 Keywords : Bt-expressing Subject RIV: EH - Ecology, Behaviour

  15. Islet amyloid polypeptide and insulin expression are controlled differently in primary and transformed islet cells

    DEFF Research Database (Denmark)

    Madsen, O D; Michelsen, Bo Thomas; Westermark, P;

    1991-01-01

    the tissue specificity of expressions of IAPP and insulin are controlled differently, and that coexpression of IAPP with hormones different from insulin may be a marker for pluripotent transformed rat islet cell clones, which are able to activate insulin gene transcription during passage in vivo....

  16. Combinatorial control of temporal gene expression in the Drosophila wing by enhancers and core promoters

    OpenAIRE

    O’Keefe, David D.; Thomas, Sean R; Bolin, Kelsey; Griggs, Ellen; Edgar, Bruce A.; Buttitta, Laura A

    2012-01-01

    Abstract Background The transformation of a developing epithelium into an adult structure is a complex process, which often involves coordinated changes in cell proliferation, metabolism, adhesion, and shape. To identify genetic mechanisms that control epithelial differentiation, we analyzed the temporal patterns of gene expression during metamorphosis of the Drosophila wing. Results We found...

  17. Sources of variation in baseline gene expression levels from toxicogenomics study control animals across multiple laboratories

    Directory of Open Access Journals (Sweden)

    Lee Janice S

    2008-06-01

    Full Text Available Abstract Background The use of gene expression profiling in both clinical and laboratory settings would be enhanced by better characterization of variance due to individual, environmental, and technical factors. Meta-analysis of microarray data from untreated or vehicle-treated animals within the control arm of toxicogenomics studies could yield useful information on baseline fluctuations in gene expression, although control animal data has not been available on a scale and in a form best served for data-mining. Results A dataset of control animal microarray expression data was assembled by a working group of the Health and Environmental Sciences Institute's Technical Committee on the Application of Genomics in Mechanism Based Risk Assessment in order to provide a public resource for assessments of variability in baseline gene expression. Data from over 500 Affymetrix microarrays from control rat liver and kidney were collected from 16 different institutions. Thirty-five biological and technical factors were obtained for each animal, describing a wide range of study characteristics, and a subset were evaluated in detail for their contribution to total variability using multivariate statistical and graphical techniques. Conclusion The study factors that emerged as key sources of variability included gender, organ section, strain, and fasting state. These and other study factors were identified as key descriptors that should be included in the minimal information about a toxicogenomics study needed for interpretation of results by an independent source. Genes that are the most and least variable, gender-selective, or altered by fasting were also identified and functionally categorized. Better characterization of gene expression variability in control animals will aid in the design of toxicogenomics studies and in the interpretation of their results.

  18. Development of gene microarray in screening differently expressed genes in keloid and normal-control skin

    Institute of Scientific and Technical Information of China (English)

    陈伟; 付小兵; 葛世丽; 孙晓庆; 周岗; 赵志力; 盛志勇

    2004-01-01

    Background Keloid is an intricate lesion that is probably regulated by many genes. In this study, the authors used the technique of complementary DNA (cDNA) microarray to analyse abnormal gene expression in keloids and normal control skins. Methods The polymerase chain reaction (PCR) products of 8400 genes were spotted in an array on chemical-material-coated-glass plates. The DNAs were fixed on the glass plates. The total RNAs were isolated from freshly excised human keloid and normal control skins, and the mRNAs were then purified. The mRNA from both keloid and normal control skins were reversely transcribed to cDNAs, with the incorporation of fluorescent dUTP, for preparing the hybridisation probes. The mixed probes were then hybridised to the cDNA microarray. After thorough washing, the cDNA microarray was scanned for differing fluorescent signals from two types of tissues. Gene expression of tissue growth factor-β1 (TGF-β1) and of c-myc was detected with both RT-PCR and Northern blot hybridisation to confirm the effectiveness of cDNA microarray. Results Among the 8400 human genes, 402 were detected with different expression levels between keloid and normal control skins. Two hundred and fifty genes, including TGF-β1 and c-myc, were up-regulated and 152 genes were down-regulated. Higher expressions of TGF-β1 and c-myc in keloid were also revealed using RT-PCR and Northern blot methods. Conclusion cDNA microarray analysis provides a powerful tool for investigating differential gene expression in keloid and normal control skins. Keloid is a complicated lesion with many genes involved.

  19. GABAA receptor subunit gene expression in human prefrontal cortex: comparison of schizophrenics and controls

    Science.gov (United States)

    Akbarian, S.; Huntsman, M. M.; Kim, J. J.; Tafazzoli, A.; Potkin, S. G.; Bunney, W. E. Jr; Jones, E. G.; Bloom, F. E. (Principal Investigator)

    1995-01-01

    The prefrontal cortex of schizophrenics is hypoactive and displays changes related to inhibitory, GABAergic neurons, and GABAergic synapses. These changes include decreased levels of glutamic acid decarboxylase (GAD), the enzyme for GABA synthesis, upregulation of muscimol binding, and downregulation of benzodiazepine binding to GABAA receptors. Studies in the visual cortex of nonhuman primates have demonstrated that gene expression for GAD and for several GABAA receptor subunit polypeptides is under control of neuronal activity, raising the possibility that similar mechanisms in the hypoactive prefrontal cortex of schizophrenics may explain the abnormalities in GAD and in GABAA receptor regulation. In the present study, which is the first of its type on human cerebral cortex, levels of mRNAs for six GABAA receptor subunits (alpha 1, alpha 2, alpha 5, beta 1, beta 2, gamma 2) and their laminar expression patterns were analyzed in the prefrontal cortex of schizophrenics and matched controls, using in situ hybridization histochemistry and densitometry. Three types of laminar expression pattern were observed: mRNAs for the alpha 1, beta 2, and gamma 2 subunits, which are the predominant receptor subunits expressed in the mature cortex, were expressed at comparatively high levels by cells of all six cortical layers, but most intensely by cells in lower layer III and layer IV. mRNAs for the alpha 2, alpha 5, and beta 1 subunits were expressed at lower levels; alpha 2 and beta 1 were expressed predominantly by cells in layers II, III, and IV; alpha 5 was expressed predominantly in layers IV, V, and VI. There were no significant changes in overall mRNA levels for any of the receptor subunits in the prefrontal cortex of schizophrenics, and the laminar expression pattern of all six receptor subunit mRNAs did not differ between schizophrenics and controls. Because gene expression for GABAA receptor subunits is not consistently altered in the prefrontal cortex of

  20. Automated optogenetic feedback control for precise and robust regulation of gene expression and cell growth.

    Science.gov (United States)

    Milias-Argeitis, Andreas; Rullan, Marc; Aoki, Stephanie K; Buchmann, Peter; Khammash, Mustafa

    2016-01-01

    Dynamic control of gene expression can have far-reaching implications for biotechnological applications and biological discovery. Thanks to the advantages of light, optogenetics has emerged as an ideal technology for this task. Current state-of-the-art methods for optical expression control fail to combine precision with repeatability and cannot withstand changing operating culture conditions. Here, we present a novel fully automatic experimental platform for the robust and precise long-term optogenetic regulation of protein production in liquid Escherichia coli cultures. Using a computer-controlled light-responsive two-component system, we accurately track prescribed dynamic green fluorescent protein expression profiles through the application of feedback control, and show that the system adapts to global perturbations such as nutrient and temperature changes. We demonstrate the efficacy and potential utility of our approach by placing a key metabolic enzyme under optogenetic control, thus enabling dynamic regulation of the culture growth rate with potential applications in bacterial physiology studies and biotechnology. PMID:27562138

  1. An aminoglycoside sensing riboswitch controls the expression of aminoglycoside resistance acetyltransferase and adenyltransferases.

    Science.gov (United States)

    Chen, Dongrong; Murchie, Alastair I H

    2014-10-01

    The emergence of antibiotic resistance in human pathogens is an increasing threat to public health. The fundamental mechanisms that control the high levels of expression of antibiotic resistance genes are not yet completely understood. The aminoglycosides are one of the earliest classes of antibiotics that were introduced in the 1940s. In the clinic aminoglycoside resistance is conferred most commonly through enzymatic modification of the drug although resistance through enzymatic modification of the target rRNA through methylation or the overexpression of efflux pumps is also appearing. An aminoglycoside sensing riboswitch has been identified that controls expression of the aminoglycoside resistance genes that encode the aminoglycoside acetyltransferase (AAC) and aminoglycoside nucleotidyltransferase (ANT) (adenyltransferase (AAD)) enzymes. AAC and ANT cause resistance to aminoglycoside antibiotics through modification of the drugs. Expression of the AAC and ANT resistance genes is regulated by aminoglycoside binding to the 5' leader RNA of the aac/aad genes. The aminoglycoside sensing RNA is also associated with the integron cassette system that captures antibiotic resistance genes. Specific aminoglycoside binding to the leader RNA induces a structural transition in the leader RNA, and consequently induction of resistance protein expression. Reporter gene expression, direct measurements of drug RNA binding, chemical probing and UV cross-linking combined with mutational analysis demonstrated that the leader RNA functioned as an aminoglycoside sensing riboswitch in which drug binding to the leader RNA leads to the induction of aminoglycoside antibiotic resistance. This article is part of a Special Issue entitled: Riboswitches. PMID:24631585

  2. Significance of serum CD25/serum ferritin in the diagnosis of lymphoma-associated hemophagocytic syndrome%可溶性白细胞介素2受体/血清铁蛋白对诊断淋巴瘤相关性噬血细胞综合征的意义

    Institute of Scientific and Technical Information of China (English)

    吴道香; 王昭; 王晓琳; 林榕榕; 刘暖; 张嘉

    2012-01-01

    目的 探讨可溶性白细胞介素2受体( sCD25)/血清铁蛋白在诊断淋巴瘤相关性噬血细胞综合征( LAHS)中的意义,为进一步提高LAHS诊断率,及时诊断和治疗LAHS提供临床依据.方法 收集2008年10月至2011年6月首都医科大学附属北京友谊医院经诊的确诊噬血细胞综合征患者70例,包括31例LAHS和39例非LAHS.收集所有患者血清,采用酶联免疫吸附法(ELISA)检测其sCD25水平,并于当日检测血清铁蛋白水平,然后计算sCD25和血清铁蛋白的比值,并比较LAHS和非LAHS之间血清sCD25、血清铁蛋白以及sCD25/血清铁蛋白有无差异.结果 LAHS和非LAHS患者sCD25质量浓度分别为(15760.52±7851.74)、(12727.41±11285.28) pg/ml(t=-1.78,P=0.075,血清铁蛋白中位质量浓度分别为1750.00、2947.00 ng/ml(Z=-1.490,P=0.136),sCD25/血清铁蛋白中位比值分别为8.57×10-3、2.84×10-3,差异具有统计学意义(Z=-2.106,P=0.035).结论 sCD25/血清铁蛋白在LAHS中明显升高,对诊断LAHS具有重要的提示意义,有可能成为诊断LAHS的重要指标.%Objective To investigate the significance of serum CD25 / serum ferritin in the diagnosis of lymphoma-associated hemophagocytic syndrome (LAHS),so as to provide the clinical basis for improving its recognition and giving effective therapy. Methods The serums were collected from 70 patients with hemophagocytic syndrom in Beijing Friendship Hospital, Capital Medical University during the period from October 2008 to June 2011, including 31 LAHS cases and 39 other disease-associated HPS cases. The serum CD25 level in HPS patients was measured with enzyme-linked immunosorbent assay (ELISA), and the serum ferritin was measured on the same day. Then the serum CD25/ferritin ratio was calculated and the differences of the serum CD25, serum ferritin and serum CD25/ferritin ratio between two groups were compared. Results The variance of serum CD25 [(15760.52±7851.74) pg/ml vs (12727.41±11285.28) pg/ml,t=-1.78,P

  3. Thymic retention of CD4+CD25+FoxP3+ T regulatory cells is associated with their peripheral deficiency and thrombocytopenia in a murine model of immune thrombocytopenia.

    Science.gov (United States)

    Aslam, Rukhsana; Hu, Yu; Gebremeskel, Simon; Segel, George B; Speck, Edwin R; Guo, Li; Kim, Michael; Ni, Heyu; Freedman, John; Semple, John W

    2012-09-01

    Immune thrombocytopenia (ITP) is a bleeding disorder in which antibodies and/or T cells lead to enhanced peripheral platelet destruction and reduced bone marrow platelet production. Several reports have observed that ITP is associated with a peripheral deficiency of tolerance-inducing CD4+CD25+FoxP3+ T regulatory cells (Tregs). Using a murine model of ITP, we analyzed Tregs in the spleen and thymus. CD61 knockout mice were immunized against wild-type (CD61+) platelets, and their splenocytes were transferred into severe combined immunodeficient (SCID) mice. Compared with SCID mice receiving naive splenocytes, within 2 weeks after transfer, the ITP SCID mice became thrombocytopenic (< 200 × 10(9) platelets/L) and had increased serum anti-CD61 antibodies. The quantity of thymic Tregs by 2 weeks after transfer was significantly elevated, whereas Tregs in the spleens were significantly reduced. Treatment of the ITP mice with 2 g/kg intravenous immunoglobulin raised the platelet counts, reduced antibody production, and normalized the thymic and splenic Treg populations. Compared with thymocytes from ITP mice treated with intravenous immunoglobulin, thymocytes from untreated ITP mice delayed the onset of ITP when administered before engraftment with immune splenocytes. These results suggest that ITP in mice is associated with a peripheral Treg deficiency because of thymic retention and therapy normalizes the Tregs. PMID:22760780

  4. Uptake of donor lymphocytes treated with 8-methoxypsoralen and ultraviolet A light by recipient dendritic cells induces CD4{sup +}CD25{sup +}Foxp3{sup +} regulatory T cells and down-regulates cardiac allograft rejection

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, De-Hua [Organ Transplant Center, Chinese PLA 309th Hospital, No. 17A Hei-Shan-Hu Road, Beijing 100091 (China); Dou, Li-Ping [Department of Hematology, Chinese PLA General Hospital, No. 28 Fu-Xing Road, Beijing 100853 (China); Wei, Yu-Xiang; Du, Guo-Sheng; Zou, Yi-Ping; Song, Ji-Yong; Zhu, Zhi-Dong; Cai, Ming; Qian, Ye-Yong [Organ Transplant Center, Chinese PLA 309th Hospital, No. 17A Hei-Shan-Hu Road, Beijing 100091 (China); Shi, Bing-Yi, E-mail: shibingyi@medmail.com.cn [Organ Transplant Center, Chinese PLA 309th Hospital, No. 17A Hei-Shan-Hu Road, Beijing 100091 (China)

    2010-05-14

    Extracorporeal photopheresis (ECP) is an effective immunomodulatory therapy and has been demonstrated to be beneficial for graft-vs-host disease and solid-organ allograft rejection. ECP involves reinfusion of a patient's autologous peripheral blood leukocytes treated ex vivo with 8-methoxypsoralen and UVA light radiation (PUVA). Previous studies focused only on ECP treatment of recipient immune cells. Our study is the first to extend the target of ECP treatment to donor immune cells. The results of in vitro co-culture experiments demonstrate uptake of donor PUVA-treated splenic lymphocytes (PUVA-SPs) by recipient immature dendritic cells (DCs). Phagocytosis of donor PUVA-SPs does not stimulate phenotype maturation of recipient DCs. In the same co-culture system, donor PUVA-SPs enhanced production of interleukin-10 and interferon-{gamma} by recipient DCs and impaired the subsequent capability of recipient DCs to stimulate recipient naive T cells. Phagocytosis of donor PUVA-SP (PUVA-SP DCs) by recipient DCs shifted T-cell responses in favor of T helper 2 cells. Infusion of PUVA-SP DCs inhibited cardiac allograft rejection in an antigen-specific manner and induced CD4{sup +}CD25{sup high}Foxp3{sup +} regulatory T cells. In conclusion, PUVA-SP DCs simultaneously deliver the donor antigen and the regulatory signal to the transplant recipient, and thus can be used to develop a novel DC vaccine for negative immune regulation and immune tolerance induction.

  5. Main Regulatory Factors for Differentiation,Development and Function of Naturally Occurred CD4+CD25+Regulatory T Cells——Review%天然调节性T细胞的调节因素研究进展

    Institute of Scientific and Technical Information of China (English)

    吴敏; 谢彦晖

    2008-01-01

    天然CD4+CD25+调节T细胞来源于胸腺,通过直接接触机制抑制效应细胞的增殖,调节自身免疫和移植免疫.本文主要综述了影响天然调节T细胞分化、发育和功能的主要因素和可能机制.Foxp3是Treg的标志,检测其表达可以作为判定Treg的方法.IL-2主要通过IL-2Rα及STAT5途径促进Treg的增殖活化.膜型和分泌型TGF-β具有不同功能,膜型TGF-β1可能主要介导Treg的抑制功能,而分泌型TGF-β可能主要促进Treg的增殖.树突状细胞由于作用途径不同,对Treg既有正调节,也有负调节.CTLA-4途径可能通过作用于Treg自身、DC或效应细胞直接或间接地调节Treg的功能.

  6. Uptake of donor lymphocytes treated with 8-methoxypsoralen and ultraviolet A light by recipient dendritic cells induces CD4+CD25+Foxp3+ regulatory T cells and down-regulates cardiac allograft rejection

    International Nuclear Information System (INIS)

    Extracorporeal photopheresis (ECP) is an effective immunomodulatory therapy and has been demonstrated to be beneficial for graft-vs-host disease and solid-organ allograft rejection. ECP involves reinfusion of a patient's autologous peripheral blood leukocytes treated ex vivo with 8-methoxypsoralen and UVA light radiation (PUVA). Previous studies focused only on ECP treatment of recipient immune cells. Our study is the first to extend the target of ECP treatment to donor immune cells. The results of in vitro co-culture experiments demonstrate uptake of donor PUVA-treated splenic lymphocytes (PUVA-SPs) by recipient immature dendritic cells (DCs). Phagocytosis of donor PUVA-SPs does not stimulate phenotype maturation of recipient DCs. In the same co-culture system, donor PUVA-SPs enhanced production of interleukin-10 and interferon-γ by recipient DCs and impaired the subsequent capability of recipient DCs to stimulate recipient naive T cells. Phagocytosis of donor PUVA-SP (PUVA-SP DCs) by recipient DCs shifted T-cell responses in favor of T helper 2 cells. Infusion of PUVA-SP DCs inhibited cardiac allograft rejection in an antigen-specific manner and induced CD4+CD25highFoxp3+ regulatory T cells. In conclusion, PUVA-SP DCs simultaneously deliver the donor antigen and the regulatory signal to the transplant recipient, and thus can be used to develop a novel DC vaccine for negative immune regulation and immune tolerance induction.

  7. Translational control is a major contributor to hypoxia induced gene expression

    International Nuclear Information System (INIS)

    Background and purpose: Hypoxia is a common feature of solid tumors that is associated with an aggressive phenotype, resistance to therapy and poor prognosis. Major contributors to these adverse effects are the transcriptional program activated by the HIF family of transcription factors as well as the translational response mediated by PERK-dependent phosphorylation of eIF2α and inhibition of mTORC1 activity. In this study we determined the relative contribution of both transcriptional and translational responses to changes in hypoxia induced gene expression. Material and methods: Total and efficiently translated (polysomal) mRNA was isolated from DU145 prostate carcinoma cells that were exposed for up to 24 h of hypoxia (2). Changes in transcription and translation were assessed using affymetrix microarray technology. Results: Our data reveal an unexpectedly large contribution of translation control on both induced and repressed gene expression at all hypoxic time points, particularly during acute hypoxia (2-4 h). Gene ontology analysis revealed that gene classes like transcription and signal transduction are stimulated by translational control whereas expression of genes involved in cell growth and protein metabolism are repressed during hypoxic conditions by translational control. Conclusions: Our data indicate that translation influences gene expression during hypoxia on a scale comparable to that of transcription.

  8. Stable Expression of Lentiviral Antigens by Quality-Controlled Recombinant Mycobacterium bovis BCG Vectors.

    Science.gov (United States)

    Hart, Bryan E; Asrican, Rose; Lim, So-Yon; Sixsmith, Jaimie D; Lukose, Regy; Souther, Sommer J R; Rayasam, Swati D G; Saelens, Joseph W; Chen, Ching-Ju; Seay, Sarah A; Berney-Meyer, Linda; Magtanong, Leslie; Vermeul, Kim; Pajanirassa, Priyadharshini; Jimenez, Amanda E; Ng, Tony W; Tobin, David M; Porcelli, Steven A; Larsen, Michelle H; Schmitz, Joern E; Haynes, Barton F; Jacobs, William R; Lee, Sunhee; Frothingham, Richard

    2015-07-01

    The well-established safety profile of the tuberculosis vaccine strain, Mycobacterium bovis bacille Calmette-Guérin (BCG), makes it an attractive vehicle for heterologous expression of antigens from clinically relevant pathogens. However, successful generation of recombinant BCG strains possessing consistent insert expression has encountered challenges in stability. Here, we describe a method for the development of large recombinant BCG accession lots which stably express the lentiviral antigens, human immunodeficiency virus (HIV) gp120 and simian immunodeficiency virus (SIV) Gag, using selectable leucine auxotrophic complementation. Successful establishment of vaccine stability stems from stringent quality control criteria which not only screen for highly stable complemented BCG ΔleuCD transformants but also thoroughly characterize postproduction quality. These parameters include consistent production of correctly sized antigen, retention of sequence-pure plasmid DNA, freeze-thaw recovery, enumeration of CFU, and assessment of cellular aggregates. Importantly, these quality assurance procedures were indicative of overall vaccine stability, were predictive for successful antigen expression in subsequent passaging both in vitro and in vivo, and correlated with induction of immune responses in murine models. This study has yielded a quality-controlled BCG ΔleuCD vaccine expressing HIV gp120 that retained stable full-length expression after 10(24)-fold amplification in vitro and following 60 days of growth in mice. A second vaccine lot expressed full-length SIV Gag for >10(68)-fold amplification in vitro and induced potent antigen-specific T cell populations in vaccinated mice. Production of large, well-defined recombinant BCG ΔleuCD lots can allow confidence that vaccine materials for immunogenicity and protection studies are not negatively affected by instability or differences between freshly grown production batches. PMID:25924766

  9. A SINGLE TETRACYCLINE-REGULATED VECTOR DEVISED FOR CONTROLLED INSULIN GENE EXPRESSION

    Institute of Scientific and Technical Information of China (English)

    Xue-yang Zhang; Ben-li Su; Hong Li; Ran Bai; Zhao-hui Xu; Chang-chen Li

    2004-01-01

    Objective To construct a single plasmid vector mediating doxycycline-inducible recombined human insulin gene expression in myotube cell line.Methods An expression cassette of rtTAnls driven by promoter of human cytomegalovirus and a furin-cuttable recom bined human insulin expression cassette driven by a reverse poly-tetO DNA motif were cloned into a single plasmid vector (prTR-tetO-mINS). The prTR-tetO-mINS and pLNCX were co-transfected into a myotube cell line (C2C12) and pLNCX vector were used as a control. After selection with G418, the transfected cells were induced with doxycycline at concentrations of 0, 2, and 10 μg/mL. RT-PCR was used to determine expression levels of recombinant insulin mRNA at the 5th day.Insulin production in cell cultures medium (at different incubation time) and cell extracts (at the 7th day) were analyzed with human pro/insulin RIA kits.Results Immune reactive insulin (IRI) level in cell medium was found increased at 24 hours of doxycycline incubation,and still increased at the 5th day. After withdrawn of doxycycline, IRI decreased sharply and was at baseline three days later. IRI and human insulin mRNA levels were positively related to different levels of doxycycline. A 25-fold increase in IRI was found against background expression at the 7th day.Conclusion Human insulin expression can be successfully regulated by doxycycline and the background was very low.This single ret-on insulin expression system may provide a new approach to a controlled insulin gene therapy in skeletal muscle.

  10. MiRNA-mediated control of HLA-G expression and function.

    Directory of Open Access Journals (Sweden)

    Irit Manaster

    Full Text Available HLA-G is a non-classical HLA class-Ib molecule expressed mainly by the extravillous cytotrophoblasts (EVT of the placenta. The expression of HLA-G on these fetal cells protects the EVT cells from immune rejection and is therefore important for a healthy pregnancy. The mechanisms controlling HLA-G expression are largely unknown. Here we demonstrate that miR-148a and miR-152 down-regulate HLA-G expression by binding its 3'UTR and that this down-regulation of HLA-G affects LILRB1 recognition and consequently, abolishes the LILRB1-mediated inhibition of NK cell killing. We further demonstrate that the C/G polymorphism at position +3142 of HLA-G 3'UTR has no effect on the miRNA targeting of HLA-G. We show that in the placenta both miR-148a and miR-152 miRNAs are expressed at relatively low levels, compared to other healthy tissues, and that the mRNA levels of HLA-G are particularly high and we therefore suggest that this might enable the tissue specific expression of HLA-G.

  11. Assessment of CcpA-mediated catabolite control of gene expression in Bacillus cereus ATCC 14579

    Directory of Open Access Journals (Sweden)

    Buist Girbe

    2008-04-01

    Full Text Available Abstract Background The catabolite control protein CcpA is a transcriptional regulator conserved in many Gram-positives, controlling the efficiency of glucose metabolism. Here we studied the role of Bacillus cereus ATCC 14579 CcpA in regulation of metabolic pathways and expression of enterotoxin genes by comparative transcriptome analysis of the wild-type and a ccpA-deletion strain. Results Comparative analysis revealed the growth performance and glucose consumption rates to be lower in the B. cereus ATCC 14579 ccpA deletion strain than in the wild-type. In exponentially grown cells, the expression of glycolytic genes, including a non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase that mediates conversion of D-glyceraldehyde 3-phosphate to 3-phospho-D-glycerate in one single step, was down-regulated and expression of gluconeogenic genes and genes encoding the citric acid cycle was up-regulated in the B. cereus ccpA deletion strain. Furthermore, putative CRE-sites, that act as binding sites for CcpA, were identified to be present for these genes. These results indicate CcpA to be involved in the regulation of glucose metabolism, thereby optimizing the efficiency of glucose catabolism. Other genes of which the expression was affected by ccpA deletion and for which putative CRE-sites could be identified, included genes with an annotated function in the catabolism of ribose, histidine and possibly fucose/arabinose and aspartate. Notably, expression of the operons encoding non-hemolytic enterotoxin (Nhe and hemolytic enterotoxin (Hbl was affected by ccpA deletion, and putative CRE-sites were identified, which suggests catabolite repression of the enterotoxin operons to be CcpA-dependent. Conclusion The catabolite control protein CcpA in B. cereus ATCC 14579 is involved in optimizing the catabolism of glucose with concomitant repression of gluconeogenesis and alternative metabolic pathways. Furthermore, the results point to metabolic control

  12. Expression of tropodithietic acid biosynthesis is controlled by a novel autoinducer.

    Science.gov (United States)

    Geng, Haifeng; Belas, Robert

    2010-09-01

    The interactions between marine prokaryotic and eukaryotic microorganisms are crucial to many biological and biogeochemical processes in the oceans. Often the interactions are mutualistic, as in the symbiosis between phytoplankton, e.g., the dinoflagellate Pfiesteria piscicida and Silicibacter sp. TM1040, a member of the Roseobacter taxonomic lineage. It is hypothesized that an important component of this symbiosis is bacterial production of tropodithietic acid (TDA), a biologically active tropolone compound whose synthesis requires the expression of tdaABCDEF (tdaA-F), as well as six additional genes (cysI, malY, paaIJK, and tdaH). The factors controlling tda gene expression are not known, although growth in laboratory standing liquid cultures drastically increases TDA levels. In this report, we measured the transcription of tda genes to gain a greater understanding of the factors controlling their expression. While the expression of tdaAB was constitutive, tdaCDE and tdaF mRNA increased significantly (3.7- and 17.4-fold, respectively) when cells were grown in standing liquid broth compared to their levels with shaking liquid culturing. No transcription of tdaC was detected when a tdaCp::lacZ transcriptional fusion was placed in 11 of the 12 Tda(-) mutant backgrounds, with cysI being the sole exception. The expression of tdaC could be restored to 9 of the remaining 11 Tda(-) mutants-tdaA and tdaH failed to respond-by placing wild-type (Tda(+)) strains in close proximity or by supplying exogenous TDA to the mutant, suggesting that TDA induces tda gene expression. These results indicate that TDA acts as an autoinducer of its own synthesis and suggest that roseobacters may use TDA as a quorum signal. PMID:20601479

  13. Expression of Tropodithietic Acid Biosynthesis Is Controlled by a Novel Autoinducer▿ †

    Science.gov (United States)

    Geng, Haifeng; Belas, Robert

    2010-01-01

    The interactions between marine prokaryotic and eukaryotic microorganisms are crucial to many biological and biogeochemical processes in the oceans. Often the interactions are mutualistic, as in the symbiosis between phytoplankton, e.g., the dinoflagellate Pfiesteria piscicida and Silicibacter sp. TM1040, a member of the Roseobacter taxonomic lineage. It is hypothesized that an important component of this symbiosis is bacterial production of tropodithietic acid (TDA), a biologically active tropolone compound whose synthesis requires the expression of tdaABCDEF (tdaA-F), as well as six additional genes (cysI, malY, paaIJK, and tdaH). The factors controlling tda gene expression are not known, although growth in laboratory standing liquid cultures drastically increases TDA levels. In this report, we measured the transcription of tda genes to gain a greater understanding of the factors controlling their expression. While the expression of tdaAB was constitutive, tdaCDE and tdaF mRNA increased significantly (3.7- and 17.4-fold, respectively) when cells were grown in standing liquid broth compared to their levels with shaking liquid culturing. No transcription of tdaC was detected when a tdaCp::lacZ transcriptional fusion was placed in 11 of the 12 Tda− mutant backgrounds, with cysI being the sole exception. The expression of tdaC could be restored to 9 of the remaining 11 Tda− mutants—tdaA and tdaH failed to respond—by placing wild-type (Tda+) strains in close proximity or by supplying exogenous TDA to the mutant, suggesting that TDA induces tda gene expression. These results indicate that TDA acts as an autoinducer of its own synthesis and suggest that roseobacters may use TDA as a quorum signal. PMID:20601479

  14. Expression of the Arf tumor suppressor gene is controlled by Tgfβ2 during development

    Science.gov (United States)

    Freeman-Anderson, Natalie E.; Zheng, Yanbin; McCalla-Martin, Amy C.; Treanor, Louise M.; Zhao, Yi D.; Garfin, Phillip M.; He, Tong-Chuan; Mary, Michelle N.; Thornton, J. Derek; Anderson, Colleen; Gibbons, Melissa; Saab, Raya; Baumer, Shannon H.; Cunningham, John M.; Skapek, Stephen X.

    2009-01-01

    Summary The Arf tumor suppressor (also known as Cdkn2a) acts as an oncogene sensor induced by `abnormal' mitogenic signals in incipient cancer cells. It also plays a crucial role in embryonic development: newborn mice lacking Arf are blind due to a pathological process resembling severe persistent hyperplastic primary vitreous (PHPV), a human eye disease. The cell-intrinsic mechanism implied in the oncogene sensor model seems unlikely to explain Arf regulation during embryo development. Instead, transforming growth factor β2 (Tgfβ2) might control Arf expression, as we show that mice lacking Tgfβ2 have primary vitreous hyperplasia similar to Arf-/- mice. Consistent with a potential linear pathway, Tgfβ2 induces Arf transcription and p19Arf expression in cultured mouse embryo fibroblasts (MEFs); and Tgfβ2-dependent cell cycle arrest in MEFs is maintained in an Arf-dependent manner. Using a new model in which Arf expression can be tracked by β-galactosidase activity in ArflacZ/+ mice, we show that Tgfβ2 is required for Arf transcription in the developing vitreous as well as in the cornea and the umbilical arteries, two previously unrecognized sites of Arf expression. Chemical and genetic strategies show that Arf promoter induction depends on Tgfβ receptor activation of Smad proteins; the induction correlates with Smad2 phosphorylation in MEFs and Arf-expressing cells in vivo. Chromatin immunoprecipitation shows that Smads bind to genomic DNA proximal to Arf exon 1β. In summary, Tgfβ2 and p19Arf act in a linear pathway during embryonic development. We present the first evidence that p19Arf expression can be coupled to extracellular cues in normal cells and suggest a new mechanism for Arf control in tumor cells. PMID:19465598

  15. Synergistic and Dose-Controlled Regulation of Cellulase Gene Expression in Penicillium oxalicum.

    Science.gov (United States)

    Li, Zhonghai; Yao, Guangshan; Wu, Ruimei; Gao, Liwei; Kan, Qinbiao; Liu, Meng; Yang, Piao; Liu, Guodong; Qin, Yuqi; Song, Xin; Zhong, Yaohua; Fang, Xu; Qu, Yinbo

    2015-09-01

    Filamentous fungus Penicillium oxalicum produces diverse lignocellulolytic enzymes, which are regulated by the combinations of many transcription factors. Here, a single-gene disruptant library for 470 transcription factors was constructed and systematically screened for cellulase production. Twenty transcription factors (including ClrB, CreA, XlnR, Ace1, AmyR, and 15 unknown proteins) were identified to play putative roles in the activation or repression of cellulase synthesis. Most of these regulators have not been characterized in any fungi before. We identified the ClrB, CreA, XlnR, and AmyR transcription factors as critical dose-dependent regulators of cellulase expression, the core regulons of which were identified by analyzing several transcriptomes and/or secretomes. Synergistic and additive modes of combinatorial control of each cellulase gene by these regulatory factors were achieved, and cellulase expression was fine-tuned in a proper and controlled manner. With one of these targets, the expression of the major intracellular β-glucosidase Bgl2 was found to be dependent on ClrB. The Bgl2-deficient background resulted in a substantial gene activation by ClrB and proved to be closely correlated with the relief of repression mediated by CreA and AmyR during cellulase induction. Our results also signify that probing the synergistic and dose-controlled regulation mechanisms of cellulolytic regulators and using it for reconstruction of expression regulation network (RERN) may be a promising strategy for cellulolytic fungi to develop enzyme hyper-producers. Based on our data, ClrB was identified as focal point for the synergistic activation regulation of cellulase expression by integrating cellulolytic regulators and their target genes, which refined our understanding of transcriptional-regulatory network as a "seesaw model" in which the coordinated regulation of cellulolytic genes is established by counteracting activators and repressors. PMID:26360497

  16. Enhanced Chemokine Receptor Expression on Leukocytes of Patients with Alzheimer's Disease.

    Directory of Open Access Journals (Sweden)

    David Goldeck

    Full Text Available Although primarily a neurological complaint, systemic inflammation is present in Alzheimer's Disease, with higher than normal levels of proinflammatory cytokines and chemokines in the periphery as well as the brain. A gradient of these factors may enhance recruitment of activated immune cells into the brain via chemotaxis. Here, we investigated the phenotypes of circulating immune cells in AD patients with multi-colour flow cytometry to determine whether their expression of chemokine receptors is consistent with this hypothesis. In this study, we confirmed our previously reported data on the shift of early- to late-differentiated CD4+ T-cells in AD patients. The percentage of cells expressing CD25, a marker of acute T-cell activation, was higher in patients than in age-matched controls, and percentages of CCR6+ cells were elevated. This chemokine receptor is primarily expressed on pro-inflammatory memory cells and Th17 cells. The proportion of cells expressing CCR4 (expressed on Th2 cells and CCR5 (Th1 cells and dendritic cells was also greater in patients, and was more pronounced on CD4+ than CD8+ T-cells. These findings allow a more detailed insight into the systemic immune status of patients with Alzheimer's disease and suggest possible novel targets for immune therapy.

  17. Indoleamine 2,3-dioxygenase expression in patients with allergic rhinitis: a case-control study

    Directory of Open Access Journals (Sweden)

    Luukkainen Annika

    2011-12-01

    Full Text Available Abstract Background Indoleamine 2,3-dioxygenase (IDO is a tryptophan catalyzing enzyme. It has been suggested that it has a role in lower airway allergic inflammations, but its role in allergic rhinitis has not been investigated. Objective Our aim was to evaluate the expression of IDO in the nasal mucosa of allergic rhinitis patients allergic to birch pollen during peak exposure to birch pollen allergen and compare it to non-atopic patients. Methods IDO expression was immunohistochemically evaluated from nasal specimens obtained in- and off-season from otherwise healthy non-smoking volunteers both allergic to birch pollen (having mild or moderate allergic rhinoconjunctivitis and non-allergic controls. Results: The IDO expression levels were low in healthy controls and remained low also in patients allergic to birch pollen. There were no differences in the expression of IDO in- and off-season in either healthy or allergic subjects. Conclusions There is a controversy in the role of IDO in upper and lower airways during allergic airway disease. It seems that IDO is associated to allergic inflammations of the lower airways, but does not have a local role in the nasal cavity at least in mild or moderate forms of allergic rhinitis.

  18. CLONING AND EXPRESSING TRYPSIN MODULATING OOSTATIC FACTOR IN Chlorella desiccata TO CONTROL MOSQUITO LARVAE.

    Science.gov (United States)

    Borovsky, Dov; Sterner, Andeas; Powell, Charles A

    2016-01-01

    The insect peptide hormone trypsin modulating oostatic factor (TMOF), a decapeptide that is synthesized by the mosquito ovary and controls the translation of the gut's trypsin mRNA was cloned and expressed in the marine alga Chlorella desiccata. To express Aedes aegypti TMOF gene (tmfA) in C. desiccata cells, two plasmids (pYES2/TMOF and pYDB4-tmfA) were engineered with pKYLX71 DNA (5 Kb) carrying the cauliflower mosaic virus (CaMV) promoter 35S(2) and the kanamycin resistant gene (neo), as well as, a 8 Kb nitrate reductase gene (nit) from Chlorella vulgaris. Transforming C. desiccata with pYES2/TMOF and pYDB4-tmfA show that the engineered algal cells express TMOF (20 ± 4 μg ± SEM and 17 ± 3 μg ± SEM, respectively in 3 × 10(8) cells) and feeding the cells to mosquito larvae kill 75 and 60% of Ae. aegypti larvae in 4 days, respectively. Southern and Northern blots analyses show that tmfA integrated into the genome of C. desiccata by homologous recombination using the yeast 2 μ circle of replication and the nit in pYES2/TMOF and pYDB4-tmfA, respectively, and the transformed algal cells express tmfA transcript. Using these algal cells it will be possible in the future to control mosquito larvae in the marsh. PMID:26440910

  19. RAE-1 is expressed in the adult subventricular zone and controls cell proliferation of neurospheres

    DEFF Research Database (Denmark)

    Popa, Natalia; Cédile, Oriane; Pollet-Villard, Xavier;

    2011-01-01

    Improving and controlling the capacity of endogenous or grafted adult neural stem cells to repair the nervous system relies on a better knowledge of interactions between immune cells and neural stem cells. Class I major histocompatibility complex (MHC) family members comprise numerous proteins pl...... reveal an unexpected role of RAE-1 in regulating adult SVZ neurogenesis by supporting stem/progenitor cells proliferation.......Improving and controlling the capacity of endogenous or grafted adult neural stem cells to repair the nervous system relies on a better knowledge of interactions between immune cells and neural stem cells. Class I major histocompatibility complex (MHC) family members comprise numerous proteins...... of two MHC class I-related members by neural stem/progenitor cells: retinoic acid early induced transcript (RAE)-1 and CD1d. The expression of RAE-1 but not CD1d disappears when differentiation of neurosphere cells is induced. Interestingly, RAE-1 transcripts are expressed in the brain during...

  20. Transgenic mice expressing yellow fluorescent protein under control of the human tyrosine hydroxylase promoter.

    Science.gov (United States)

    Choi, Eun Yang; Yang, Jae Won; Park, Myung Sun; Sun, Woong; Kim, Hyun; Kim, Seung U; Lee, Myung Ae

    2012-10-01

    Pathogenesis of Parkinson's disease and related catecholaminergic neurological disorders is closely associated with changes in the levels of tyrosine hydroxylase (TH). Therefore, investigation of the regulation of the TH gene system should assist in understanding the pathomechanisms involved in these neurological disorders. To identify regulatory domains that direct human TH expression in the central nervous system (CNS), we generated two transgenic mouse lines in which enhanced yellow fluorescent protein (EYFP) is expressed under the control of either 3.2-kb (hTHP-EYFP construct) human TH promoter or 3.2-kb promoter with 2-kb 3'-flanking regions (hTHP-ex3-EYFP construct) of the TH gene. In the adult transgenic mouse brain, the hTHP-EYFP construct directs neuron-specific EYFP expression in various CNS areas, such as olfactory bulb, striatum, interpeduncular nucleus, cerebral cortex, hippocampus, and particularly dentate gyrus. Although these EYFP-positive cells were identified as mature neurons, few EYFP-positive cells were TH-positive neurons. On the other hand, we could detect the EYFP mRNA expression in a subset of neurons in the olfactory bulb, midbrain, and cerebellum, in which expression of endogenous TH is enriched, with hTHP-ex3-EYFP transgenic mice. These results indicate that the 3.2-kb sequence upstream of the TH gene is not sufficient for proper expression and that the 2-kb sequence from the translation start site to exon 3 is necessary for expression of EYFP in a subset of catecholaminergic neurons. PMID:22714400

  1. Comprehensive translational control of tyrosine kinase expression by upstream open reading frames

    OpenAIRE

    Wethmar, K; J. Schulz; Muro, E.M.; Talyan, S.; Andrade-Navarro, M A; Leutz, A

    2016-01-01

    Post-transcriptional control has emerged as a major regulatory event in gene expression and often occurs at the level of translation initiation. Although overexpression or constitutive activation of tyrosine kinases (TKs) through gene amplification, translocation or mutation are well-characterized oncogenic events, current knowledge about translational mechanisms of TK activation is scarce. Here, we report the presence of translational cis-regulatory upstream open reading frames (uORFs) in th...

  2. Genetic control of antibody responses induced by recombinant Mycobacterium bovis BCG expressing a foreign antigen.

    OpenAIRE

    Lagranderie, M; Lo-Man, R; Dériaud, E; Gicquel, B; Gheorghiu, M; Leclerc, C

    1997-01-01

    Recombinant Mycobacterium bovis BCG expressing foreign antigens represents a promising candidate for the development of future vaccines and was shown in several experimental models to induce protective immunity against bacterial or parasitic infections. Innate resistance to BCG infection is under genetic control and could modify the immune responses induced against an antigen delivered by such engineered microorganisms. To investigate this question, we analyzed the immune responses of various...

  3. Quality control in microarray assessment of gene expression in human airway epithelium

    OpenAIRE

    Attiyeh Marc A; Harvey Ben-Gary; Wang Wei; Hackett Neil R; O'Connor Timothy P; Raman Tina; Dang David T; Teater Matthew; Crystal Ronald G

    2009-01-01

    Abstract Background Microarray technology provides a powerful tool for defining gene expression profiles of airway epithelium that lend insight into the pathogenesis of human airway disorders. The focus of this study was to establish rigorous quality control parameters to ensure that microarray assessment of the airway epithelium is not confounded by experimental artifact. Samples (total n = 223) of trachea, large and small airway epithelium were collected by fiberoptic bronchoscopy of 144 in...

  4. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression

    OpenAIRE

    Qi, Lei S.; Larson, Matthew H.; Gilbert, Luke A.; Doudna, Jennifer A.; Weissman, Jonathan S.; Arkin, Adam P; Lim, Wendell A.

    2013-01-01

    Targeted gene regulation on a genome-wide scale is a powerful strategy for interrogating, perturbing, and engineering cellular systems. Here, we develop a method for controlling gene expression based on Cas9, an RNA-guided DNA endonuclease from a type II CRISPR system. We show that a catalytically dead Cas9 lacking endonuclease activity, when coexpressed with a guide RNA, generates a DNA recognition complex that can specifically interfere with transcriptional elongation, RNA polymerase bindin...

  5. Inertial Sensor-Based Touch and Shake Metaphor for Expressive Control of 3D Virtual Avatars

    OpenAIRE

    Shashidhar Patil; Harinadha Reddy Chintalapalli; Dubeom Kim; Youngho Chai

    2015-01-01

    In this paper, we present an inertial sensor-based touch and shake metaphor for expressive control of a 3D virtual avatar in a virtual environment. An intuitive six degrees-of-freedom wireless inertial motion sensor is used as a gesture and motion control input device with a sensor fusion algorithm. The algorithm enables user hand motions to be tracked in 3D space via magnetic, angular rate, and gravity sensors. A quaternion-based complementary filter is implemented to reduce noise and drift...

  6. Epigenetic control of effector gene expression in the plant pathogenic fungus Leptosphaeria maculans.

    Directory of Open Access Journals (Sweden)

    Jessica L Soyer

    2014-03-01

    Full Text Available Plant pathogens secrete an arsenal of small secreted proteins (SSPs acting as effectors that modulate host immunity to facilitate infection. SSP-encoding genes are often located in particular genomic environments and show waves of concerted expression at diverse stages of plant infection. To date, little is known about the regulation of their expression. The genome of the Ascomycete Leptosphaeria maculans comprises alternating gene-rich GC-isochores and gene-poor AT-isochores. The AT-isochores harbor mosaics of transposable elements, encompassing one-third of the genome, and are enriched in putative effector genes that present similar expression patterns, namely no expression or low-level expression during axenic cultures compared to strong induction of expression during primary infection of oilseed rape (Brassica napus. Here, we investigated the involvement of one specific histone modification, histone H3 lysine 9 methylation (H3K9me3, in epigenetic regulation of concerted effector gene expression in L. maculans. For this purpose, we silenced the expression of two key players in heterochromatin assembly and maintenance, HP1 and DIM-5 by RNAi. By using HP1-GFP as a heterochromatin marker, we observed that almost no chromatin condensation is visible in strains in which LmDIM5 was silenced by RNAi. By whole genome oligoarrays we observed overexpression of 369 or 390 genes, respectively, in the silenced-LmHP1 and -LmDIM5 transformants during growth in axenic culture, clearly favouring expression of SSP-encoding genes within AT-isochores. The ectopic integration of four effector genes in GC-isochores led to their overexpression during growth in axenic culture. These data strongly suggest that epigenetic control, mediated by HP1 and DIM-5, represses the expression of at least part of the effector genes located in AT-isochores during growth in axenic culture. Our hypothesis is that changes of lifestyle and a switch toward pathogenesis lift chromatin

  7. Epigenetic control of effector gene expression in the plant pathogenic fungus Leptosphaeria maculans.

    Science.gov (United States)

    Soyer, Jessica L; El Ghalid, Mennat; Glaser, Nicolas; Ollivier, Bénédicte; Linglin, Juliette; Grandaubert, Jonathan; Balesdent, Marie-Hélène; Connolly, Lanelle R; Freitag, Michael; Rouxel, Thierry; Fudal, Isabelle

    2014-03-01

    Plant pathogens secrete an arsenal of small secreted proteins (SSPs) acting as effectors that modulate host immunity to facilitate infection. SSP-encoding genes are often located in particular genomic environments and show waves of concerted expression at diverse stages of plant infection. To date, little is known about the regulation of their expression. The genome of the Ascomycete Leptosphaeria maculans comprises alternating gene-rich GC-isochores and gene-poor AT-isochores. The AT-isochores harbor mosaics of transposable elements, encompassing one-third of the genome, and are enriched in putative effector genes that present similar expression patterns, namely no expression or low-level expression during axenic cultures compared to strong induction of expression during primary infection of oilseed rape (Brassica napus). Here, we investigated the involvement of one specific histone modification, histone H3 lysine 9 methylation (H3K9me3), in epigenetic regulation of concerted effector gene expression in L. maculans. For this purpose, we silenced the expression of two key players in heterochromatin assembly and maintenance, HP1 and DIM-5 by RNAi. By using HP1-GFP as a heterochromatin marker, we observed that almost no chromatin condensation is visible in strains in which LmDIM5 was silenced by RNAi. By whole genome oligoarrays we observed overexpression of 369 or 390 genes, respectively, in the silenced-LmHP1 and -LmDIM5 transformants during growth in axenic culture, clearly favouring expression of SSP-encoding genes within AT-isochores. The ectopic integration of four effector genes in GC-isochores led to their overexpression during growth in axenic culture. These data strongly suggest that epigenetic control, mediated by HP1 and DIM-5, represses the expression of at least part of the effector genes located in AT-isochores during growth in axenic culture. Our hypothesis is that changes of lifestyle and a switch toward pathogenesis lift chromatin

  8. Cultural differences in the links between parental control and children's emotional expressivity.

    Science.gov (United States)

    Louie, Jennifer Y; Oh, Brian J; Lau, Anna S

    2013-10-01

    Research suggests that parental control may be motivated by various socialization goals and contributes to children's adjustment in diverse ways depending on cultural context. The present study examined whether parental psychological control was differentially related to children's emotional expressivity in a sample of 127 Korean, Asian American (AA), and European American (EA) preschoolers. Results indicated that Korean and AA parents endorsed more parental control (emotion suppression, shaming) than EA parents. Similarly, Korean and AA children displayed less observable sadness and exuberance during emotion-eliciting tasks than EA children. Furthermore, moderation analyses revealed that for EA families, parental control was positively correlated with child anger and exuberance; however, the associations were not significant for AA and Korean families. PMID:23834255

  9. PPP1, a plant-specific regulator of transcription controls Arabidopsis development and PIN expression.

    Science.gov (United States)

    Benjamins, René; Barbez, Elke; Ortbauer, Martina; Terpstra, Inez; Lucyshyn, Doris; Moulinier-Anzola, Jeanette; Khan, Muhammad Asaf; Leitner, Johannes; Malenica, Nenad; Butt, Haroon; Korbei, Barbara; Scheres, Ben; Kleine-Vehn, Jürgen; Luschnig, Christian

    2016-01-01

    Directional transport of auxin is essential for plant development, with PIN auxin transport proteins representing an integral part of the machinery that controls hormone distribution. However, unlike the rapidly emerging framework of molecular determinants regulating PIN protein abundance and subcellular localization, insights into mechanisms controlling PIN transcription are still limited. Here we describe PIN2 PROMOTER BINDING PROTEIN 1 (PPP1), an evolutionary conserved plant-specific DNA binding protein that acts on transcription of PIN genes. Consistent with PPP1 DNA-binding activity, PPP1 reporter proteins are nuclear localized and analysis of PPP1 null alleles and knockdown lines indicated a function as a positive regulator of PIN expression. Furthermore, we show that ppp1 pleiotropic mutant phenotypes are partially reverted by PIN overexpression, and results are presented that underline a role of PPP1-PIN promoter interaction in PIN expression control. Collectively, our findings identify an elementary, thus far unknown, plant-specific DNA-binding protein required for post-embryonic plant development, in general, and correct expression of PIN genes, in particular. PMID:27553690

  10. Prognostic Impact of Erythropoietin Expression and Erythropoietin Receptor Expression on Locoregional Control and Survival of Patients Irradiated for Stage II/III Non-Small-Cell Lung Cancer

    International Nuclear Information System (INIS)

    Purpose: Prognostic factors can guide the physician in selecting the optimal treatment for an individual patient. This study investigates the prognostic value of erythropoietin (EPO) and EPO receptor (EPO-R) expression of tumor cells for locoregional control and survival in non-small-cell lung cancer (NSCLC) patients. Methods and Materials: Fourteen factors were investigated in 62 patients irradiated for stage II/III NSCLC, as follows: age, gender, Karnofsky performance score (KPS), histology, grading, TNM/American Joint Committee on Cancer (AJCC) stage, surgery, chemotherapy, pack years (average number of packages of cigarettes smoked per day multiplied by the number of years smoked), smoking during radiotherapy, hemoglobin levels during radiotherapy, EPO expression, and EPO-R expression. Additionally, patients with tumors expressing both EPO and EPO-R were compared to those expressing either EPO or EPO-R and to those expressing neither EPO nor EPO-R. Results: On univariate analysis, improved locoregional control was associated with AJCC stage II cancer (p 70 (p = 0.08), an N stage of 0 to 1 (p = 0.07), and no EPO-R expression (p = 0.10). On multivariate analysis, AJCC stage II and no EPO expression remained significant. No smoking during radiotherapy was almost significant. On univariate analysis, improved survival was associated with N stage 0 to 1 (p = 0.009), surgery (p = 0.039), hemoglobin levels of ≥12 g/d (p = 0.016), and no EPO expression (p = 0.001). On multivariate analysis, N stage 0 to 1 and no EPO expression maintained significance. Hemoglobin levels of ≥12 g/d were almost significant. On subgroup analyses, patients with tumors expressing both EPO and EPO-R had worse outcomes than those expressing either EPO or EPO-R and those expressing neither EPO nor RPO-R. Conclusions: EPO expression of tumor cells was an independent prognostic factor for locoregional control and survival in patients irradiated for NSCLC. EPO-R expression showed a trend

  11. Effect of medical castration on CD4+ CD25+ T cells, CD8+ T cell IFN-gamma expression, and NK cells: a physiological role for testosterone and/or its metabolites.

    Science.gov (United States)

    Page, Stephanie T; Plymate, Stephen R; Bremner, William J; Matsumoto, Alvin M; Hess, David L; Lin, Daniel W; Amory, John K; Nelson, Peter S; Wu, Jennifer D

    2006-05-01

    The higher prevalence of autoimmune disease among women compared with men suggests that steroids impact immune regulation. To investigate how sex steroids modulate cellular immune function, we conducted a randomized trial in 12 healthy men aged 35-55 yr treated for 28 days with placebo, a GnRH antagonist, acyline to induce medical castration, or acyline plus daily testosterone (T) gel to replace serum T, followed by a 28-day recovery period. Serum hormones were measured weekly and peripheral blood lymphocytes (PBLs) were collected biweekly for analyses of thymus-derived lymphocyte (T cell) subtypes and natural killer (NK) cells. Compared with the other groups and to baseline throughout the drug exposure period, men receiving acyline alone had significant reductions in serum T (near or below castrate levels), dihydrotestosterone, and estradiol (P physiological balance of autoimmunity and protective immunity by preserving the number of regulatory T cells and the activation of CD8+ T cells. In addition, sex steroids suppress NK cell proliferation. This study supports a complex physiological role for T and/or its metabolites in immune regulation. PMID:16352669

  12. The expression of CD25, CD11b, SWC1, SWC7, MHC-II, and family of CD45 molecules can be used to characterize different stages of gamma delta T lymphocytes in pigs

    Czech Academy of Sciences Publication Activity Database

    Štěpánová, Kateřina; Šinkora, Marek

    2012-01-01

    Roč. 36, č. 4 (2012), s. 728-740. ISSN 0145-305X R&D Projects: GA ČR GA524/07/0087 Institutional research plan: CEZ:AV0Z50200510 Keywords : Porcine immune system * Cell surface molecules * Lymphocyte subpopulations Subject RIV: EC - Immunology Impact factor: 3.238, year: 2012

  13. A high proportion of bone marrow T cells with regulatory phenotype (CD4+CD25hiFoxP3+) in Ewing sarcoma patients is associated with metastatic disease.

    Science.gov (United States)

    Brinkrolf, Peter; Landmeier, Silke; Altvater, Bianca; Chen, Christiane; Pscherer, Sibylle; Rosemann, Annegret; Ranft, Andreas; Dirksen, Uta; Juergens, Heribert; Rossig, Claudia

    2009-08-15

    Immunosuppressive CD4+CD25(hi)FoxP3+ T cells (T(reg) cells) have been found at increased densities within the tumor microenvironment in many malignancies and interfere with protective antitumor immune responses. Osseous Ewing sarcomas (ESs) are thought to derive from a bone marrow (BM) mesenchymal cell of origin, and microscopic marrow involvement defines a subpopulation of patients at a high risk of relapse. We hypothesized that BM-resident T cells may contribute to a permissive milieu for immune escape of ESs. Using 6-color-flow cytometry, we investigated the pattern of immune cell subset distribution including NK cells, gammadelta T cells, central and effector memory CD8+ and CD4+ T cells as well as T cells with regulatory phenotype (T(reg) cells) in BM obtained at diagnosis from 45 primary or relapsed ES patients treated within standardized protocols. Although patients at relapse had an inverted CD4:CD8 T-cell ratio, neither CD8+ effector/memory T-cell subsets nor T(reg) cells significantly differed from patients at diagnosis. No significant associations of innate and effector/memory T-cell subpopulations with known risk factors were found, including age, gender, tumor site, primary metastases and histological tumor response. By contrast, T(reg) cells were found at significantly higher frequencies in patients with primary metastatic disease compared with localized ESs (5.0 vs. 3.3%, p = 0.01). Thus, increased BM T(reg) cells in patients with metastasized ES may reflect an immune escape mechanism that contributes to the development of metastatic disease. Immunotherapeutic strategies will have to adequately consider the regulatory milieu within areas of Ewing tumor-immune interactions. PMID:19480009

  14. Alteration in frequency and function of CD4⁺CD25⁺FOXP3⁺ regulatory T cells in patients with immune thrombocytopenic purpura.

    Directory of Open Access Journals (Sweden)

    Nargess Arandi

    2014-04-01

    Full Text Available Immune thrombocytopenic purpura (ITP is an autoimmune bleeding disorder characterized by production of auto-antibodies against platelet antigens. It is obvious that regulatory T cells (Tregs have a major role in controlling immune homeostasis and preventing autoimmunity.To investigate the frequency and functions of Tregs, twenty ITP patients and twenty age- and sex-matched healthy controls were recruited. The peripheral blood mononuclear cells were isolated and the proportion of Tregs was defined by flow cytometry method. The expression of immune-regulatory markers, cytotoxic T-lymphocyte associated antigen-4 (CTLA-4 and glucocorticoid induced tumor necrosis factor receptor (GITR were also assessed by quantitative Real-time PCR TaqMan method. For evaluation of Treg function, Tregs were enriched and their ability to inhibit proliferation of T cells was measured and levels of immune-regulatory cytokines IL-10 and TGF-β were also measured.Results showed that the frequency of Tregs and the mean fluorescence intensity of FOXP3 protein significantly decreased in ITP patients compared to those in healthy controls. In addition, there was a significant reduction in relative expression of both CTLA-4 and GITR mRNA in ITP patients (P=0.02 and P=0.006, respectively. The suppressive function of Tregs also diminished in ITP patients compared to that in controls. Both IL-10 and TGF-β cytokines were produced in lower amounts in ITP patients than controls.It could be concluded that alteration in Treg frequency and functional characteristics might be responsible for loss of self-tolerance and subsequently destructive immune responses observed in ITP patients.

  15. Validation of endogenous control reference genes for normalizing gene expression studies in endometrial carcinoma.

    Science.gov (United States)

    Ayakannu, Thangesweran; Taylor, Anthony H; Willets, Jonathon M; Brown, Laurence; Lambert, David G; McDonald, John; Davies, Quentin; Moss, Esther L; Konje, Justin C

    2015-09-01

    Real-time quantitative RT-PCR (qRT-PCR) is a powerful technique used for the relative quantification of target genes, using reference (housekeeping) genes for normalization to ensure the generation of accurate and robust data. A systematic examination of the suitability of endogenous reference genes for gene expression studies in endometrial cancer tissues is absent. The aims of this study were therefore to identify and evaluate from the thirty-two possible reference genes from a TaqMan(®) array panel their suitability as an internal control gene. The mathematical software packages geNorm qBasePLUS identified Pumilio homolog 1 (Drosophila) (PUM1), ubiquitin C (UBC), phosphoglycerate kinase (PGK1), mitochondrial ribosomal protein L19 (MRPL19) and peptidylpropyl isomerase A (cyclophilin A) (PPIA) as the best reference gene combination, whilst NormFinder identified MRPL19 as the best single reference gene, with importin 8 (IPO8) and PPIA being the best combination of two reference genes. BestKeeper ranked MRPL19 as the most stably expressed gene. In addition, the study was validated by examining the relative expression of a test gene, which encodes the cannabinoid receptor 1 (CB1). A significant difference in CB1 mRNA expression between malignant and normal endometrium using MRPL19, PPIA, and IP08 in combination was observed. The use of MRPL19, IPO8 and PPIA was identified as the best reference gene combination for the normalization of gene expression levels in endometrial carcinoma. This study demonstrates that the arbitrary selection of endogenous control reference genes for normalization in qRT-PCR studies of endometrial carcinoma, without validation, risks the production of inaccurate data and should therefore be discouraged. PMID:26124453

  16. Evaluation of external RNA controls for the standardisation of gene expression biomarker measurements

    Directory of Open Access Journals (Sweden)

    Elaswarapu Ramnath

    2010-11-01

    Full Text Available Abstract Background Gene expression profiling is an important approach for detecting diagnostic and prognostic biomarkers, and predicting drug safety. The development of a wide range of technologies and platforms for measuring mRNA expression makes the evaluation and standardization of transcriptomic data problematic due to differences in protocols, data processing and analysis methods. Thus, universal RNA standards, such as those developed by the External RNA Controls Consortium (ERCC, are proposed to aid validation of research findings from diverse platforms such as microarrays and RT-qPCR, and play a role in quality control (QC processes as transcriptomic profiling becomes more commonplace in the clinical setting. Results Panels of ERCC RNA standards were constructed in order to test the utility of these reference materials (RMs for performance characterization of two selected gene expression platforms, and for discrimination of biomarker profiles between groups. The linear range, limits of detection and reproducibility of microarray and RT-qPCR measurements were evaluated using panels of RNA standards. Transcripts of low abundance (≤ 10 copies/ng total RNA showed more than double the technical variability compared to higher copy number transcripts on both platforms. Microarray profiling of two simulated 'normal' and 'disease' panels, each consisting of eight different RNA standards, yielded robust discrimination between the panels and between standards with varying fold change ratios, showing no systematic effects due to different labelling and hybridization runs. Also, comparison of microarray and RT-qPCR data for fold changes showed agreement for the two platforms. Conclusions ERCC RNA standards provide a generic means of evaluating different aspects of platform performance, and can provide information on the technical variation associated with quantification of biomarkers expressed at different levels of physiological abundance

  17. An animal model allowing controlled receptor expression for molecular ultrasound imaging.

    Science.gov (United States)

    Saini, Reshu; Sorace, Anna G; Warram, Jason M; Mahoney, Marshall J; Zinn, Kurt R; Hoyt, Kenneth

    2013-01-01

    Reported in this study is an animal model system for evaluating targeted ultrasound (US) contrast agents binding using adenoviral (Ad) vectors to modulate cellular receptor expression. An Ad vector encoding an extracellular hemagglutinin (HA) epitope tag and a green fluorescent protein (GFP) reporter was used to regulate receptor expression. A low and high receptor density (in breast cancer tumor bearing mice) was achieved by varying the Ad dose with a low plaque forming unit (PFU) on day 1 and high PFU on day 2 of experimentation. Targeted US contrast agents, or microbubbles (MB), were created by conjugating either biotinylated anti-HA or IgG isotype control antibodies to the MB surface with biotin-streptavidin linkage. Targeted and control MBs were administered on both days of experimentation and contrast-enhanced US (CEUS) was performed on each mouse using MB flash destruction technique. Signal intensities from MBs retained within tumor vasculature were analyzed through a custom Matlab program. Results showed intratumoral enhancement attributable to targeted MB accumulation was significantly increased from the low Ad vector dosing and the high Ad vector dosing (p = 0.001). Control MBs showed no significant differences between day 1 and day 2 imaging (p = 0.96). Additionally, targeted MBs showed a 10.5-fold increase in intratumoral image intensity on day 1 and an 18.8-fold increase in image intensity on day 2 compared with their control MB counterparts. PMID:23122640

  18. Reducing the expression of implicit stereotypes: reflexive control through implementation intentions.

    Science.gov (United States)

    Mendoza, Saaid A; Gollwitzer, Peter M; Amodio, David M

    2010-04-01

    The authors tested the effectiveness of implementation intentions as a strategy for limiting the behavioral expression of implicit stereotypes. Implementation intentions are if-then plans that link an intended response to an anticipated situational cue, thereby enabling a reflexive form of control. The authors examined whether two different types of implementation intentions could improve response accuracy on the Shooter Task, a reaction time measure of implicit stereotyping. In Study 1, participants used a distraction-inhibiting implementation intention designed to engage control over the perception of goal-irrelevant stimuli (e.g., race). In Study 2, participants used a response-facilitating implementation intention designed to promote goal-directed action. Across studies, implementation intentions improved accuracy, thereby limiting the behavioral expression of implicit stereotypes. Furthermore, process dissociation analyses indicated that the distraction-inhibiting implementation intention increased controlled processing while reducing automatic stereotype activation, whereas the response-facilitating implementation intention increased only controlled processing. Implications for goal strategy approaches to reducing prejudice are discussed. PMID:20363905

  19. Thyroid hormone and adrenergic signaling interact to control pineal expression of the dopamine receptor D4 gene (Drd4)

    DEFF Research Database (Denmark)

    Kim, Jong-So; Bailey, Michael J; Weller, Joan L;

    2009-01-01

    Dopamine plays diverse and important roles in vertebrate biology, impacting behavior and physiology through actions mediated by specific G-protein-coupled receptors, one of which is the dopamine receptor D4 (Drd4). Here we present studies on the >100-fold daily rhythm in rat pineal Drd4 expression....... Our studies indicate that Drd4 is the dominant dopamine receptor gene expressed in the pineal gland. The gene is expressed in pinealocytes at levels which are approximately 100-fold greater than in other tissues, except the retina, in which transcript levels are similar. Pineal Drd4 expression is...... circadian in nature and under photoneural control. Whereas most rhythmically expressed genes in the pineal are controlled by adrenergic/cAMP signaling, Drd4 expression also requires thyroid hormone. This advance raises the questions of whether Drd4 expression is regulated by this mechanism in other systems...

  20. EXPRESS

    International Nuclear Information System (INIS)

    This paper presents EXPRESS, an expert system developed for the automation of reliability studies. The first part consists in the description of the method for static thermohydraulic systems. In this step, the authors define the knowledge representation based on the two inference engines - ALOUETTE and LCR developed by EDF. They explain all the process to construct a fault tree from a topological and functional description of the system. Numerous examples are exhibited in illustration of the method. This is followed by the lessons derived from the studies performed on some safety systems of the PALUEL nuclear plant. The development of the same approach for electric power systems is described, insisting on the difference resulting from the sequential nature of these systems. Finally, they show the main advantages identified during the studies

  1. Distinct Tlr4-expressing cell compartments control neutrophilic and eosinophilic airway inflammation.

    Science.gov (United States)

    McAlees, J W; Whitehead, G S; Harley, I T W; Cappelletti, M; Rewerts, C L; Holdcroft, A M; Divanovic, S; Wills-Karp, M; Finkelman, F D; Karp, C L; Cook, D N

    2015-07-01

    Allergic asthma is a chronic, inflammatory lung disease. Some forms of allergic asthma are characterized by T helper type 2 (Th2)-driven eosinophilia, whereas others are distinguished by Th17-driven neutrophilia. Stimulation of Toll-like receptor 4 (TLR4) on hematopoietic and airway epithelial cells (AECs) contributes to the inflammatory response to lipopolysaccharide (LPS) and allergens, but the specific contribution of TLR4 in these cell compartments to airway inflammatory responses remains poorly understood. We used novel, conditionally mutant Tlr4(fl/fl) mice to define the relative contributions of AEC and hematopoietic cell Tlr4 expression to LPS- and allergen-induced airway inflammation. We found that Tlr4 expression by hematopoietic cells is critical for neutrophilic airway inflammation following LPS exposure and for Th17-driven neutrophilic responses to the house dust mite (HDM) lysates and ovalbumin (OVA). Conversely, Tlr4 expression by AECs was found to be important for robust eosinophilic airway inflammation following sensitization and challenge with these same allergens. Thus, Tlr4 expression by hematopoietic and airway epithelial cells controls distinct arms of the immune response to inhaled allergens. PMID:25465099

  2. Cryptic Transcription and Early Termination in the Control of Gene Expression

    Directory of Open Access Journals (Sweden)

    Jessie Colin

    2011-01-01

    Full Text Available Recent studies on yeast transcriptome have revealed the presence of a large set of RNA polymerase II transcripts mapping to intergenic and antisense regions or overlapping canonical genes. Most of these ncRNAs (ncRNAs are subject to termination by the Nrd1-dependent pathway and rapid degradation by the nuclear exosome and have been dubbed cryptic unstable transcripts (CUTs. CUTs are often considered as by-products of transcriptional noise, but in an increasing number of cases they play a central role in the control of gene expression. Regulatory mechanisms involving expression of a CUT are diverse and include attenuation, transcriptional interference, and alternative transcription start site choice. This review focuses on the impact of cryptic transcription on gene expression, describes the role of the Nrd1-complex as the main actor in preventing nonfunctional and potentially harmful transcription, and details a few systems where expression of a CUT has an essential regulatory function. We also summarize the most recent studies concerning other types of ncRNAs and their possible role in regulation.

  3. THE EXPRESSION AND CLINICAL VALUE OF APOPTOSIS CONTROL GENE Bcl-2 AND Bax IN BREAST CANCER

    Institute of Scientific and Technical Information of China (English)

    ZHENG Jun; YAO Zhen-xiang; ZHANG Jing

    1999-01-01

    Objective: To study the expression and clinical value of apoptosis control gene bcl-2 and bax in breast cancer.Methods: Protein bax and bcl-2 in 41 breast cancers obtained from operations in our hospital in 1996 were detected using ABC immunohistochemical stain assay and compared with 10 cases with normal breast tissues.Results: The positive rate of bax in normal breast tissue was 90% and in breast cancer was 59%, with a significant statistical difference between them (P<0.05), but there was no statistical difference in bcl-2 protein expression. Among the 41 breast cancer, the group with lymph node metastasis (21 cases) had obviously low bax expression (43%) and high bcl-2 expression (76%), showing significant difference to the group without lymph node metastasis (P<0.05).Conclusion: The antiapoptosis function of bcl-2 was stronger than bax in breast cancer. Protein bax and bcl-2 assay may be useful in understanding the biological behaviors of breast cancer.

  4. ABA-mediated ROS in mitochondria regulate root meristem activity by controlling PLETHORA expression in Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Li Yang

    2014-12-01

    Full Text Available Although research has determined that reactive oxygen species (ROS function as signaling molecules in plant development, the molecular mechanism by which ROS regulate plant growth is not well known. An aba overly sensitive mutant, abo8-1, which is defective in a pentatricopeptide repeat (PPR protein responsible for the splicing of NAD4 intron 3 in mitochondrial complex I, accumulates more ROS in root tips than the wild type, and the ROS accumulation is further enhanced by ABA treatment. The ABO8 mutation reduces root meristem activity, which can be enhanced by ABA treatment and reversibly recovered by addition of certain concentrations of the reducing agent GSH. As indicated by low ProDR5:GUS expression, auxin accumulation/signaling was reduced in abo8-1. We also found that ABA inhibits the expression of PLETHORA1 (PLT1 and PLT2, and that root growth is more sensitive to ABA in the plt1 and plt2 mutants than in the wild type. The expression of PLT1 and PLT2 is significantly reduced in the abo8-1 mutant. Overexpression of PLT2 in an inducible system can largely rescue root apical meristem (RAM-defective phenotype of abo8-1 with and without ABA treatment. These results suggest that ABA-promoted ROS in the mitochondria of root tips are important retrograde signals that regulate root meristem activity by controlling auxin accumulation/signaling and PLT expression in Arabidopsis.

  5. Expression of Arabidopsis hexokinase in citrus guard cells controls stomatal aperture and reduces transpiration

    Directory of Open Access Journals (Sweden)

    Nitsan eLugassi

    2015-12-01

    Full Text Available Hexokinase (HXK is a sugar-phosphorylating enzyme involved in sugar-sensing. It has recently been shown that HXK in guard cells mediates stomatal closure and coordinates photosynthesis with transpiration in the annual species tomato and Arabidopsis. To examine the role of HXK in the control of the stomatal movement of perennial plants, we generated citrus plants that express Arabidopsis HXK1 (AtHXK1 under KST1, a guard cell-specific promoter. The expression of KST1 in the guard cells of citrus plants has been verified using GFP as a reporter gene. The expression of AtHXK1 in the guard cells of citrus reduced stomatal conductance and transpiration with no negative effect on the rate of photosynthesis, leading to increased water-use efficiency. The effects of light intensity and humidity on stomatal behavior were examined in rooted leaves of the citrus plants. The optimal intensity of photosynthetically active radiation and lower humidity enhanced stomatal closure of AtHXK1-expressing leaves, supporting the role of sugar in the regulation of citrus stomata. These results suggest that HXK coordinates photosynthesis and transpiration and stimulates stomatal closure not only in annual species, but also in perennial species.

  6. Nucleolar dominance and maternal control of 45S rDNA expression.

    Science.gov (United States)

    Michalak, Katarzyna; Maciak, Sebastian; Kim, Young Bun; Santopietro, Graciela; Oh, Jung Hun; Kang, Lin; Garner, Harold R; Michalak, Pawel

    2015-12-01

    Using a system of interspecies hybrids, trihybrids, and recombinants with varying proportions of genomes from three distinct Xenopus species, we provide evidence for de novo epigenetic silencing of paternal 45 S ribosomal ribonucleic acid (rRNA) genes and their species-dependent expression dominance that escapes transcriptional inactivation after homologous recombination. The same pattern of imprinting is maintained in the offspring from mothers being genetic males (ZZ) sex-reversed to females, indicating that maternal control of ribosomal deoxyribonucleic acid (rDNA) expression is not sex-chromosome linked. Nucleolar dominance (nucleolus underdevelopment) in Xenopus hybrids appears to be associated with a major non-Mendelian reduction in the number of 45 S rDNA gene copies rather than a specific pattern of their expression. The loss of rRNA gene copies in F1 hybrids was non-random with respect to the parental species, with the transcriptionally dominant variant preferentially removed from hybrid zygotes. This dramatic disruption in the structure and function of 45 S rDNA impacts transcriptome patterns of small nucleolar RNAs and messenger RNAs, with genes from the ribosome and oxidative stress pathways being among the most affected. Unorthodoxies of rDNA inheritance and expression may be interpreted as hallmarks of genetic conflicts between parental genomes, as well as defensive epigenetic mechanisms employed to restore genome integrity. PMID:26645200

  7. Control of radiation sensitivity of mammalian cells. Regulation of expression of DNA repair genes

    International Nuclear Information System (INIS)

    This review describes authors' investigations concerning regulation of expression of DNA repair genes for the purpose of control of radiosensitivity of mammalian cells for cancer radiotherapy. One of their experiments concerns the enhancement of sensitivity to radiation and anti-tumor agents by suppressing the expression of mammalian Rad51 gene which playing a central role in recombination repair against DNA double-strand break, by RNA interference (RNAi). Described are the mode of action of RNAi, mechanism of suppression of Rad51 gene expression by it, enhancing effect in radiosensitivity, stable suppression and enhancement by hairpin RNA and its possible usefulness in cancer therapy. The other concerns the histone H2AX gene, which delivering the repair signal post phosphorylation in chromatin against the double-strand break. Experimental results of suppression of the histone H2AX gene by tet-off system, enhancement of radiosensitivity by the suppression and functional recovery by the gene transfer are described, and the radiosensitivity can be thus artificially controlled by tetracycline in authors' F9 2AX (tet/tet) cells. (N.I.)

  8. Electrotransfer parameters as a tool for controlled and targeted gene expression in skin.

    Science.gov (United States)

    Kos, Spela; Blagus, Tanja; Cemazar, Maja; Lampreht Tratar, Ursa; Stimac, Monika; Prosen, Lara; Dolinsek, Tanja; Kamensek, Urska; Kranjc, Simona; Steinstraesser, Lars; Vandermeulen, Gaëlle; Préat, Véronique; Sersa, Gregor

    2016-01-01

    Skin is an attractive target for gene electrotransfer. It consists of different cell types that can be transfected, leading to various responses to gene electrotransfer. We demonstrate that these responses could be controlled by selecting the appropriate electrotransfer parameters. Specifically, the application of low or high electric pulses, applied by multi-electrode array, provided the possibility to control the depth of the transfection in the skin, the duration and the level of gene expression, as well as the local or systemic distribution of the transgene. The influence of electric pulse type was first studied using a plasmid encoding a reporter gene (DsRed). Then, plasmids encoding therapeutic genes (IL-12, shRNA against endoglin, shRNA against melanoma cell adhesion molecule) were used, and their effects on wound healing and cutaneous B16F10 melanoma tumors were investigated. The high-voltage pulses resulted in gene expression that was restricted to superficial skin layers and induced a local response. In contrast, the low-voltage electric pulses promoted transfection into the deeper skin layers, resulting in prolonged gene expression and higher transgene production, possibly with systemic distribution. Therefore, in the translation into the clinics, it will be of the utmost importance to adjust the electrotransfer parameters for different therapeutic approaches and specific mode of action of the therapeutic gene. PMID:27574782

  9. Inositol phosphate pathway controls transcription of telomeric expression sites in trypanosomes.

    Science.gov (United States)

    Cestari, Igor; Stuart, Ken

    2015-05-26

    African trypanosomes evade clearance by host antibodies by periodically changing their variant surface glycoprotein (VSG) coat. They transcribe only one VSG gene at a time from 1 of about 20 telomeric expression sites (ESs). They undergo antigenic variation by switching transcription between telomeric ESs or by recombination of the VSG gene expressed. We show that the inositol phosphate (IP) pathway controls transcription of telomeric ESs and VSG antigenic switching in Trypanosoma brucei. Conditional knockdown of phosphatidylinositol 5-kinase (TbPIP5K) or phosphatidylinositol 5-phosphatase (TbPIP5Pase) or overexpression of phospholipase C (TbPLC) derepresses numerous silent ESs in T. brucei bloodstream forms. The derepression is specific to telomeric ESs, and it coincides with an increase in the number of colocalizing telomeric and RNA polymerase I foci in the nucleus. Monoallelic VSG transcription resumes after reexpression of TbPIP5K; however, most of the resultant cells switched the VSG gene expressed. TbPIP5K, TbPLC, their substrates, and products localize to the plasma membrane, whereas TbPIP5Pase localizes to the nucleus proximal to telomeres. TbPIP5Pase associates with repressor/activator protein 1 (TbRAP1), and their telomeric silencing function is altered by TbPIP5K knockdown. These results show that specific steps in the IP pathway control ES transcription and antigenic switching in T. brucei by epigenetic regulation of telomere silencing. PMID:25964327

  10. Expression liver-directed genes by employing synthetic transcriptional control units

    Institute of Scientific and Technical Information of China (English)

    Marie-Luise Lemken; Wolfgang A. Wybranietz; Ulrike Schmidt; Florian Graepler; Sorin Armeanu; Michael Bitzer; Ulrich M. Lauer

    2005-01-01

    AIM: To generate and characterize the synthetic transcriptional control units for transcriptional targeting of the liver,thereby compensating for the lack of specificity of currently available gene therapeutic vector systems.METHODS: Synthetic transcriptional control unit constructs were generated and analyzed for transcriptional activities in different cell types by FACS quantification, semi-quantitative RT-PCR, and Western blotting. RESULTS: A new bifunctionally-enhanced green fluorescent protein (EGFP)/neor fusion gene cassette was generated,and could flexibly be used both for transcript quantification and for selection of stable cell clones. Then, numerous synthetic transcriptional control units consisting of a minimal promoter linked to "naturally" derived composite enhancer elements from liver-specific expressed genes or binding sites of liver-specific transcription factors were inserted upstream of this reporter cassette. Following liposome-mediated transfection, EGFP reporter protein quantification by FACS analysis identified constructs encoding multimerized composite elements of the apolipoprotein B100 (ApoB) promoter or the ornithin transcarbamoylase (OTC) enhancer to exhibit maximum transcriptional activities in liver originating cell lines, but only background levels in non-liver originating cell lines. In contrast, constructs encoding only singular binding sites of liver-specific transcription factors, namely hepatocyte nuclear factor (HNF)1, HNF3, HNF4, HNF5, or CAAT/enhancer binding protein (C/EBP) only achieved background levels of EGFP expression. Finally, both semi-quantitative RT-PCR and Western blotting analysis of Hep3B cells demonstrated maximum transcriptional activities for a multimeric 4xApoB cassette construct, which fully complied with the data obtained by initial FACS analysis.CONCLUSION: Synthetic transcriptional control unit constructs not only exhibit a superb degree of structural compactness, but also provide new means for liver

  11. Computational design of a Zn2+ receptor that controls bacterial gene expression

    Science.gov (United States)

    Dwyer, M. A.; Looger, L. L.; Hellinga, H. W.

    2003-09-01

    The control of cellular physiology and gene expression in response to extracellular signals is a basic property of living systems. We have constructed a synthetic bacterial signal transduction pathway in which gene expression is controlled by extracellular Zn2+. In this system a computationally designed Zn2+-binding periplasmic receptor senses the extracellular solute and triggers a two-component signal transduction pathway via a chimeric transmembrane protein, resulting in transcriptional up-regulation of a -galactosidase reporter gene. The Zn2+-binding site in the designed receptor is based on a four-coordinate, tetrahedral primary coordination sphere consisting of histidines and glutamates. In addition, mutations were introduced in a secondary coordination sphere to satisfy the residual hydrogen-bonding potential of the histidines coordinated to the metal. The importance of the secondary shell interactions is demonstrated by their effect on metal affinity and selectivity, as well as protein stability. Three designed protein sequences, comprising two distinct metal-binding positions, were all shown to bind Zn2+ and to function in the cell-based assay, indicating the generality of the design methodology. These experiments demonstrate that biological systems can be manipulated with computationally designed proteins that have drastically altered ligand-binding specificities, thereby extending the repertoire of genetic control by extracellular signals.

  12. Inertial Sensor-Based Touch and Shake Metaphor for Expressive Control of 3D Virtual Avatars.

    Science.gov (United States)

    Patil, Shashidhar; Chintalapalli, Harinadha Reddy; Kim, Dubeom; Chai, Youngho

    2015-01-01

    In this paper, we present an inertial sensor-based touch and shake metaphor for expressive control of a 3D virtual avatar in a virtual environment. An intuitive six degrees-of-freedom wireless inertial motion sensor is used as a gesture and motion control input device with a sensor fusion algorithm. The algorithm enables user hand motions to be tracked in 3D space via magnetic, angular rate, and gravity sensors. A quaternion-based complementary filter is implemented to reduce noise and drift. An algorithm based on dynamic time-warping is developed for efficient recognition of dynamic hand gestures with real-time automatic hand gesture segmentation. Our approach enables the recognition of gestures and estimates gesture variations for continuous interaction. We demonstrate the gesture expressivity using an interactive flexible gesture mapping interface for authoring and controlling a 3D virtual avatar and its motion by tracking user dynamic hand gestures. This synthesizes stylistic variations in a 3D virtual avatar, producing motions that are not present in the motion database using hand gesture sequences from a single inertial motion sensor. PMID:26094629

  13. Inertial Sensor-Based Touch and Shake Metaphor for Expressive Control of 3D Virtual Avatars

    Directory of Open Access Journals (Sweden)

    Shashidhar Patil

    2015-06-01

    Full Text Available In this paper, we present an inertial sensor-based touch and shake metaphor for expressive control of a 3D virtual avatar in a virtual environment. An intuitive six degrees-of-freedom wireless inertial motion sensor is used as a gesture and motion control input device with a sensor fusion algorithm. The algorithm enables user hand motions to be tracked in 3D space via magnetic, angular rate, and gravity sensors. A quaternion-based complementary filter is implemented to reduce noise and drift. An algorithm based on dynamic time-warping is developed for efficient recognition of dynamic hand gestures with real-time automatic hand gesture segmentation. Our approach enables the recognition of gestures and estimates gesture variations for continuous interaction. We demonstrate the gesture expressivity using an interactive flexible gesture mapping interface for authoring and controlling a 3D virtual avatar and its motion by tracking user dynamic hand gestures. This synthesizes stylistic variations in a 3D virtual avatar, producing motions that are not present in the motion database using hand gesture sequences from a single inertial motion sensor.

  14. Control of myosin heavy chain expression: interaction of hypothyroidism and hindlimb suspension.

    Science.gov (United States)

    Diffee, G M; Haddad, F; Herrick, R E; Baldwin, K M

    1991-12-01

    The aim of this study was to contrast competing influences, hypothyroidism and hindlimb suspension, on myosin heavy chain (MHC) expression studied at the protein level and mRNA level. Female Sprague-Dawley rats were assigned to either normal control (NC), normal suspended (NS), or hypothyroid (thyroidectomized) control (TC) and suspended (TS) groups. NS and TS animals were suspended for 14 days following which myofibrils and total RNA were purified from the hindlimb muscles. In the soleus and vastus intermedius (VI), there was an increase in type I MHC and a decrease in type IIa MHC in both the TC and TS groups and a decrease in type I and increase in type IIa MHC in the NS group. At the mRNA level, similar shifts were observed with the exception that 1) the increased type IIa MHC seen in the soleus and VI of the NS animals was not accompanied by an increase in IIa mRNA and 2) type IIb mRNA was increased in the NS soleus without concomitant changes in IIb protein levels. These data suggest the following: 1) a hypothyroid state predominates over mechanical unweighting factors in the control of MHC distribution in slow muscles; and 2) translational or posttranslational factors may be important in the regulation of type IIa and IIb MHC expression during hindlimb suspension. PMID:1767813

  15. RNA expression patterns in serum microvesicles from patients with glioblastoma multiforme and controls

    Directory of Open Access Journals (Sweden)

    Noerholm Mikkel

    2012-01-01

    Full Text Available Abstract Background RNA from exosomes and other microvesicles contain transcripts of tumour origin. In this study we sought to identify biomarkers of glioblastoma multiforme in microvesicle RNA from serum of affected patients. Methods Microvesicle RNA from serum from patients with de-novo primary glioblastoma multiforme (N = 9 and normal controls (N = 7 were analyzed by microarray analysis. Samples were collected according to protocols approved by the Institutional Review Board. Differential expressions were validated by qRT-PCR in a separate set of samples (N = 10 in both groups. Results Expression profiles of microvesicle RNA correctly separated individuals in two groups by unsupervised clustering. The most significant differences pertained to down-regulated genes (121 genes > 2-fold down in the glioblastoma multiforme patient microvesicle RNA, validated by qRT-PCR on several genes. Overall, yields of microvesicle RNA from patients was higher than from normal controls, but the additional RNA was primarily of size Conclusions Serum microvesicle RNA from patients with glioblastoma multiforme has significantly down-regulated levels of RNAs coding for ribosome production, compared to normal healthy controls, but a large overabundance of RNA of unknown origin with size

  16. RNA expression patterns in serum microvesicles from patients with glioblastoma multiforme and controls

    International Nuclear Information System (INIS)

    RNA from exosomes and other microvesicles contain transcripts of tumour origin. In this study we sought to identify biomarkers of glioblastoma multiforme in microvesicle RNA from serum of affected patients. Microvesicle RNA from serum from patients with de-novo primary glioblastoma multiforme (N = 9) and normal controls (N = 7) were analyzed by microarray analysis. Samples were collected according to protocols approved by the Institutional Review Board. Differential expressions were validated by qRT-PCR in a separate set of samples (N = 10 in both groups). Expression profiles of microvesicle RNA correctly separated individuals in two groups by unsupervised clustering. The most significant differences pertained to down-regulated genes (121 genes > 2-fold down) in the glioblastoma multiforme patient microvesicle RNA, validated by qRT-PCR on several genes. Overall, yields of microvesicle RNA from patients was higher than from normal controls, but the additional RNA was primarily of size < 500 nt. Gene ontology of the down-regulated genes indicated these are coding for ribosomal proteins and genes related to ribosome production. Serum microvesicle RNA from patients with glioblastoma multiforme has significantly down-regulated levels of RNAs coding for ribosome production, compared to normal healthy controls, but a large overabundance of RNA of unknown origin with size < 500 nt

  17. Single muscle fiber gene expression in human skeletal muscle: validation of internal control with exercise

    International Nuclear Information System (INIS)

    Reverse transcription and real-time PCR have become the method of choice for the detection of low-abundance mRNA transcripts obtained from small human muscle biopsy samples. GAPDH, β-actin, β-2M, and 18S rRNA are widely employed as endogenous control genes, with the assumption that their expression is unregulated and constant for given experimental conditions. The aim of this study was to determine if mRNA transcripts could be performed on isolated human single muscle fibers and to determine reliable housekeeping genes (HKGs) using quantitative gene expression protocols at rest and in response to an acute exercise bout. Muscle biopsies were obtained from the gastrocnemius of three adult males before, immediately after, and 4 h following 30 min of treadmill running at 70% of VO2max. A total of 40 single fibers (MHC I and IIa) were examined for GAPDH, β-actin, β-2M, and 18S rRNA using quantitative RT-PCR and SYBR Green detection. All analyzed single fiber segments showed ribosomal RNA (28S/18S). No degradation or additional bands below ribosomal were detected (rRNA ratio 1.5-1.8). Also, no high or low-molecular weight genomic DNA contamination was observed. For each housekeeping gene the duplicate average SD was ±0.13 with a CV of 0.58%. Stable expression of GAPDH was observed at all time points for each fiber type (MHC I and IIa). Inconsistent expression of β-actin, β-2M, and 18S rRNA was observed during the post-exercise time points for each fiber type. These data indicate that successful extraction of high quality RNA from human single muscle fibers along with quantification of mRNA of selected genes can be performed. Furthermore, exercise does influence the expression of certain HKGs with GAPDH being the most stable

  18. Validation of endogenous control genes for gene expression studies on human ocular surface epithelium.

    Directory of Open Access Journals (Sweden)

    Bina Kulkarni

    Full Text Available PURPOSE: To evaluate a panel of ten known endogenous control genes (ECG with quantitative reverse transcription PCR (qPCR, for identification of stably expressed endogenous control genes in the ocular surface (OS epithelial regions including cornea, limbus, limbal epithelial crypt and conjunctiva to normalise the quantitative reverse transcription PCR data of genes of interest expressed in above-mentioned regions. METHOD: The lasermicrodissected (LMD OS epithelial regions of cryosectioned corneoscleral buttons from the cadaver eyes were processed for RNA extraction and cDNA synthesis to detect genes of interest with qPCR. Gene expression of 10 known ECG--glyceraldehyde-3-phosphate dehydrogenase (GAPDH, beta actin (ACTB, peptidylprolyl isomerase (PPIA, TATA-box binding protein (TBP1, hypoxanthine guanine phosphoribosyl transferase (HPRT1, beta glucuronidase (GUSB, Eucaryotic 18S ribosomal RNA (18S, phosphoglycerate kinase (PGK1, beta-2-microglobulin (B2M, ribosomal protein, large, P0 (RPLP0--was measured in the OS epithelial regions by qPCR method and the data collected was further analysed using geNorm software. RESULTS: The expression stability of ecgs in the os epithelial regions in increasing order as determined with genorm software is as follows: ACTB<18Sexpressed in individual OS epithelial regions: HPRT1-TBP in cornea, GUSB-PPIA in limbus, B2M-PPIA and RPLP0-TBP in LEC and conjunctiva respectively. However, across the entire ocular surface including all the regions mentioned above, PPIA-RPLP0 pair was shown to be most stable. CONCLUSION: This study has identified stably expressed ECGs on the OS epithelial regions for effective qPCR results in genes of interest. The results from this study are broadly applicable to quantitative reverse transcription PCR studies on human OS epithelium and provide evidence for the use

  19. Evaluation of real-time PCR endogenous control genes for analysis of gene expression in bovine endometrium

    Directory of Open Access Journals (Sweden)

    Mitchell Murray D

    2009-11-01

    Full Text Available Abstract Background Quantitative real-time PCR gene expression results are generally normalised using endogenous control genes. These reference genes should be expressed at a constant level across all sample groups in a study, and should not be influenced by study treatments or conditions. There has been no systematic investigation of endogenous control genes for bovine endometrium to date. The suitability of both commonly used and novel endogenous control genes was evaluated in this study, with the latter being selected from stably expressed transcripts identified through microarray analysis of bovine endometrium. Fifteen candidate endogenous control genes were assessed across different tissue subtypes in pregnant and cycling Holstein-Friesian dairy cows from two divergent genetic backgrounds. Results The expression profiles of five commonly used endogenous control genes (GAPDH, PPIA, RPS9, RPS15A, and UXT and 10 experimentally derived candidate endogenous control genes (SUZ12, C2ORF29, ZNF131, ACTR1A, HDAC1, SLC30A6, CNOT7, DNAJC17, BBS2, and RANBP10 were analysed across 44 samples to determine the most stably expressed gene. Gene stability was assessed using the statistical algorithms GeNorm and Normfinder. All genes presented with low overall variability (0.87 to 1.48% CV of Cq. However, when used to normalise a differentially expressed gene (oxytocin receptor - OXTR in the samples, the reported relative gene expression levels were significantly affected by the control gene chosen. Based on the results of this analysis, SUZ12 is proposed as the most appropriate control gene for use in bovine endometrium during early pregnancy or the oestrus cycle. Conclusion This study establishes the suitability of novel endogenous control genes for comparing expression levels in endometrial tissues of pregnant and cycling bovines, and demonstrates the utility of microarray analysis as a method for identifying endogenous control gene candidates.

  20. Optimization of the Lactococcus lactis nisin-controlled gene expression system NICE for industrial applications

    Directory of Open Access Journals (Sweden)

    Mond James

    2005-05-01

    Full Text Available Abstract Background The nisin-controlled gene expression system NICE of Lactococcus lactis is one of the most widely used expression systems in Gram-positive bacteria. Despite its widespread use, no optimization of the culture conditions and nisin induction has been carried out to obtain maximum yields. As a model system induced production of lysostaphin, an antibacterial protein (mainly against Staphylococcus aureus produced by S. simulans biovar. Staphylolyticus, was used. Three main areas need optimization for maximum yields: cell density, nisin-controlled induction and protein production, and parameters specific for the target-protein. Results In a series of pH-controlled fermentations the following parameters were optimized: pH of the culture, use of NaOH or NH4OH as neutralizing agent, the addition of zinc and phosphate, the fermentation temperature, the time point of induction (cell density of the culture, the amount of nisin added for induction and the amount of three basic medium components, i.e. yeast extract, peptone and lactose. For each culture growth and lysostaphin production was followed. Lysostaphin production yields depended on all parameters that were varied. In the course of the optimization a three-fold increase in lysostaphin yield was achieved from 100 mg/l to 300 mg/l. Conclusion Protein production with the NICE gene expression system in L. lactis strongly depends on the medium composition, the fermentation parameters and the amount of nisin added for induction. Careful optimization of key parameters lead to a significant increase in the yield of the target protein.

  1. Genetic control of eosinophilia in mice: gene(s) expressed in bone marrow-derived cells control high responsiveness

    International Nuclear Information System (INIS)

    A heterogeneity in the capacity of strains of mice to mount eosinophilia is described. BALB/c and C3H are eosinophil high responder strains (EO-HR) and CBA and A/J are eosinophil low responder strains (EO-LR), judged by the response of blood eosinophils to Ascaris suum, and the response of blood, bone marrow, and spleen eosinophils to keyhole limpet hemocyanin given 2 days after 150 mg/kg cyclophosphamide. Some of the gene(s) for high responsiveness appear to be dominant because (EO-HR x EO-LR)F1 mice were intermediate to high responders. This gene is expressed in bone marrow-derived cells because radiation chimeras of the type EO-HR→F1 were high responders and EO-LR→F1 were low responders. This description of a genetic control of eosinophilia in mice may be useful in understanding the role of this cell in parasite immunity and allergy

  2. Control of gene expression in Helicobacter pylori using the Tet repressor

    OpenAIRE

    McClain, Mark S.; Duncan, Stacy S.; Gaddy, Jennifer A.; Cover, Timothy L.

    2013-01-01

    The lack of a versatile system to control gene expression in Helicobacter pylori has hampered efforts to study H. pylori physiology and pathogenesis. To overcome these limitations, we evaluated the utility of an inducible system based on the well-characterized Tet repressor (TetR) and Tet operator (tetO). As validation of this system, we introduced three copies of tetO into the promoter region upstream of the cagUT operon (encoding two virulence factors required for function of the H. pylori ...

  3. The role of stromal mast cells in the modification of CD4 CD25 Foxp3 regulatory T cells, Th17 lymphocytes and cytotoxic lymphocytes Tc1 in the development and progression of tumor

    OpenAIRE

    Katarzyna Starska; Ewa Brzezińska-Błaszczyk

    2010-01-01

    Despite the lack of direct evidence that immune surveillance cells protect against tumor development, indirect clinical observations and experimental studies indicate activity in the immune response against cancer cells of various origin. Little is known about the effects of the stromal tumor mast cell (MC) in the activity of immune cells, i.e. CD4[sup] [/sup]CD25[sup] [/sup]Foxp3[sup] [/sup] regulatory T cells, Th17 lymphocytes, cytotoxic lymphocytes Tc1 and their mutual modulatory function ...

  4. Adaptive Representations for Improving Evolvability, Parameter Control, and Parallelization of Gene Expression Programming

    Directory of Open Access Journals (Sweden)

    Nigel P. A. Browne

    2010-01-01

    Full Text Available Gene Expression Programming (GEP is a genetic algorithm that evolves linear chromosomes encoding nonlinear (tree-like structures. In the original GEP algorithm, the genome size is problem specific and is determined through trial and error. In this work, a method for adaptive control of the genome size is presented. The approach introduces mutation, transposition, and recombination operators that enable a population of heterogeneously structured chromosomes, something the original GEP algorithm does not support. This permits crossbreeding between normally incompatible individuals, speciation within a population, increases the evolvability of the representations, and enhances parallel GEP. To test our approach, an assortment of problems were used, including symbolic regression, classification, and parameter optimization. Our experimental results show that our approach provides a solution for the problem of self-adaptive control of the genome size of GEP's representation.

  5. Tissue-Expressed B7-H1 Critically Controls Intestinal Inflammation

    Directory of Open Access Journals (Sweden)

    Lisa Scandiuzzi

    2014-02-01

    Full Text Available B7-H1 (PD-L1 on immune cells plays an important role in T cell coinhibition by binding its receptor PD-1. Here, we show that both human and mouse intestinal epithelium express B7-H1 and that B7-H1-deficient mice are highly susceptible to dextran sodium sulfate (DSS- or trinitrobenzenesulfonic acid (TNBS-induced gut injury. B7-H1 deficiency during intestinal inflammation leads to high mortality and morbidity, which are associated with severe pathological manifestations in the colon, including loss of epithelial integrity and overgrowth of commensal bacteria. Results from bone marrow chimeric and knockout mice show that B7-H1 expressed on intestinal parenchyma, but not on hematopoietic cells, controls intestinal inflammation in an adaptive immunity-independent fashion. Finally, we demonstrate that B7-H1 dampened intestinal inflammation by inhibiting tumor necrosis factor α (TNF-α production and by stimulating interleukin 22 secretion from CD11c+CD11b+ lamina propria cells. Thus, our data uncover a mechanism through which intestinal tissue-expressed B7-H1 functions as an essential ligand for innate immune cells to prevent gut inflammation.

  6. Determination of internal controls for quantitative gene expression of Isochrysis zhangjiangensis at nitrogen stress condition

    Science.gov (United States)

    Wu, Shuang; Zhou, Jiannan; Cao, Xupeng; Xue, Song

    2016-02-01

    Isochrysis zhangjiangensis is a potential marine microalga for biodiesel production, which accumulates lipid under nitrogen limitation conditions, but the mechanism on molecular level is veiled. Quantitative real-time polymerase chain reaction (qPCR) provides the possibility to investigate the gene expression levels, and a valid reference for data normalization is an essential prerequisite for firing up the analysis. In this study, five housekeeping genes, actin (ACT), α-tubulin (TUA), ß-tubulin (TUB), ubiquitin (UBI), 18S rRNA (18S) and one target gene, diacylglycerol acyltransferase (DGAT), were used for determining the reference. By analyzing the stabilities based on calculation of the stability index and on operating the two types of software, geNorm and bestkeeper, it showed that the reference genes widely used in higher plant and microalgae, such as UBI, TUA and 18S, were not the most stable ones in nitrogen-stressed I. zhangjiangensis, and thus are not suitable for exploring the mRNA expression levels under these experimental conditions. Our results show that ACT together with TUB is the most feasible internal control for investigating gene expression under nitrogen-stressed conditions. Our findings will contribute not only to future qPCR studies of I. zhangjiangensis, but also to verification of comparative transcriptomics studies of the microalgae under similar conditions.

  7. Identification and characterization of enhancers controlling the inflammatory gene expression program in macrophages.

    Science.gov (United States)

    Ghisletti, Serena; Barozzi, Iros; Mietton, Flore; Polletti, Sara; De Santa, Francesca; Venturini, Elisa; Gregory, Lorna; Lonie, Lorne; Chew, Adeline; Wei, Chia-Lin; Ragoussis, Jiannis; Natoli, Gioacchino

    2010-03-26

    Enhancers determine tissue-specific gene expression programs. Enhancers are marked by high histone H3 lysine 4 mono-methylation (H3K4me1) and by the acetyl-transferase p300, which has allowed genome-wide enhancer identification. However, the regulatory principles by which subsets of enhancers become active in specific developmental and/or environmental contexts are unknown. We exploited inducible p300 binding to chromatin to identify, and then mechanistically dissect, enhancers controlling endotoxin-stimulated gene expression in macrophages. In these enhancers, binding sites for the lineage-restricted and constitutive Ets protein PU.1 coexisted with those for ubiquitous stress-inducible transcription factors such as NF-kappaB, IRF, and AP-1. PU.1 was required for maintaining H3K4me1 at macrophage-specific enhancers. Reciprocally, ectopic expression of PU.1 reactivated these enhancers in fibroblasts. Thus, the combinatorial assembly of tissue- and signal-specific transcription factors determines the activity of a distinct group of enhancers. We suggest that this may represent a general paradigm in tissue-restricted and stimulus-responsive gene regulation. PMID:20206554

  8. MALT1 Protease Activity Controls the Expression of Inflammatory Genes in Keratinocytes upon Zymosan Stimulation.

    Science.gov (United States)

    Schmitt, Anja; Grondona, Paula; Maier, Tabea; Brändle, Marc; Schönfeld, Caroline; Jäger, Günter; Kosnopfel, Corinna; Eberle, Franziska C; Schittek, Birgit; Schulze-Osthoff, Klaus; Yazdi, Amir S; Hailfinger, Stephan

    2016-04-01

    The protease activity of the paracaspase mucosa-associated lymphoid tissue lymphoma translocation gene 1 (MALT1) plays an important role in antigen receptor-mediated lymphocyte activation by controlling the activity of the transcription factor nuclear factor-κB and is thus essential for the expression of inflammatory target genes. MALT1 is not only present in cells of the hematopoietic lineage, but is ubiquitously expressed. Here we report that stimulation with zymosan or Staphylococcus aureus induced MALT1 protease activity in human primary keratinocytes. Inhibition of the Src family of kinases or novel protein kinase C isoforms as well as silencing of CARMA2 or BCL10 interfered with activation of MALT1 protease. Silencing or inhibition of MALT1 protease strongly decreased the expression of important inflammatory genes such as TNFα, IL-17C, CXCL8 and HBD-2. MALT1-inhibited cells were unable to mount an antimicrobial response upon zymosan stimulation or phorbolester/ionomycin treatment, demonstrating a central role of MALT1 protease activity in keratinocyte immunity and suggesting MALT1 as a potential target in inflammatory skin diseases. PMID:26767426

  9. Fgf15 regulates thalamic development by controlling the expression of proneural genes.

    Science.gov (United States)

    Martinez-Ferre, Almudena; Lloret-Quesada, Cosme; Prakash, Nilima; Wurst, Wolfgang; Rubenstein, John L R; Martinez, Salvador

    2016-07-01

    The establishment of the brain structural complexity requires a precisely orchestrated interplay between extrinsic and intrinsic signals modulating cellular mechanisms to guide neuronal differentiation. However, little is known about the nature of these signals in the diencephalon, a complex brain region that processes and relays sensory and motor information to and from the cerebral cortex and subcortical structures. Morphogenetic signals from brain organizers regulate histogenetic processes such as cellular proliferation, migration, and differentiation. Sonic hedgehog (Shh) in the key signal of the ZLI, identified as the diencephalic organizer. Fgf15, the mouse gene orthologous of human, chick, and zebrafish Fgf19, is induced by Shh signal and expressed in the diencephalic alar plate progenitors during histogenetic developmental stages. This work investigates the role of Fgf15 signal in diencephalic development. In the absence of Fgf15, the complementary expression pattern of proneural genes: Ascl1 and Nng2, is disrupted and the GABAergic thalamic cells do not differentiate; in addition dorsal thalamic progenitors failed to exit from the mitotic cycle and to differentiate into neurons. Therefore, our findings indicate that Fgf15 is the Shh downstream signal to control thalamic regionalization, neurogenesis, and neuronal differentiation by regulating the expression and mutual segregation of neurogenic and proneural regulatory genes. PMID:26311466

  10. Epigenetic control of virulence gene expression in Pseudomonas aeruginosa by a LysR-type transcription regulator.

    Directory of Open Access Journals (Sweden)

    Keith H Turner

    2009-12-01

    Full Text Available Phenotypic variation within an isogenic bacterial population is thought to ensure the survival of a subset of cells in adverse conditions. The opportunistic pathogen Pseudomonas aeruginosa variably expresses several phenotypes, including antibiotic resistance, biofilm formation, and the production of CupA fimbriae. Here we describe a previously unidentified bistable switch in P. aeruginosa. This switch controls the expression of a diverse set of genes, including aprA, which encodes the secreted virulence factor alkaline protease. We present evidence that bistable expression of PA2432, herein named bexR (bistable expression regulator, which encodes a LysR-type transcription regulator, controls this switch. In particular, using DNA microarrays, quantitative RT-PCR analysis, chromatin immunoprecipitation, and reporter gene fusions, we identify genes directly under the control of BexR and show that these genes are bistably expressed. Furthermore, we show that bexR is itself bistably expressed and positively autoregulated. Finally, using single-cell analyses of a GFP reporter fusion, we present evidence that positive autoregulation of bexR is necessary for bistable expression of the BexR regulon. Our findings suggest that a positive feedback loop involving a LysR-type transcription regulator serves as the basis for an epigenetic switch that controls virulence gene expression in P. aeruginosa.

  11. Rodent Aanat: Intronic E-box sequences control tissue specificity but not rhythmic expression in the pineal gland

    OpenAIRE

    2007-01-01

    Rodent Aanat: Intronic E-box sequences control tissue specificity but not rhythmic expression in the pineal gland UNITED KINGDOM (Humphries, Ann) UNITED KINGDOM Received: 2006-12-30 Revised: 2007-02-07 Accepted: 2007-02-07

  12. Role of long purine stretches in controlling the expression of genes associated with neurological disorders.

    Science.gov (United States)

    Singh, Himanshu Narayan; Rajeswari, Moganty R

    2015-11-10

    Purine repeat sequences present in the human genome are known to act as hotspots for mutations leading to chromosomal imbalances. It is established that large purine repeats (PRs) form stable DNA triplex structure which can inhibit gene expression. Friedreich's ataxia (FRDA), the autosomal neurodegenerative disorder is the only human disease known so far, where a large purine (GAA) repeat in the FXN gene is known to inhibit the expression of frataxin protein. We explored the hidden purine repeats (PRn with n ≥ 200) if any, in the human genome to find out how they are associated with neurological disorders. The results showed 28 PRs, which are mostly restricted to the intronic regions. Interestingly, the transcriptome expression analysis of PR-carrying genes (PR-genes) revealed that most of them are down-regulated in neurological disorders (autism, Alzheimer's disease, schizophrenia, epilepsy, mental retardation, Parkinson's disease, brain tumor) as compared to that in healthy controls. The altered gene expression in brain disorders can be interpreted in terms of a possible expansion of purine repeats leading to formation of very stable DNA-triplex and/or alleviation of the repair enzymes and/or other unknown cellular factors. Interactome analysis identified four PR-genes in signaling pathways whose dysregulation is correlated directly with pathogenesis: GRK5 and KLK6 in Alzheimer's disease; FGF14 in craniosynostosis, mental retardation and FLT1 in neuroferritinopathy. By virtue of being mutational hotspots and their ability to form DNA-triplex, purine repeats in genome disturb the genome integrity and interfere with the transcriptional regulation. However, validation of the disease linkage of PR-genes can be validated using knock-out techniques. PMID:26149656

  13. E2F Transcription Factors Control the Roller Coaster Ride of Cell Cycle Gene Expression.

    Science.gov (United States)

    Thurlings, Ingrid; de Bruin, Alain

    2016-01-01

    Initially, the E2F transcription factor was discovered as a factor able to bind the adenovirus E2 promoter and activate viral genes. Afterwards it was shown that E2F also binds to promoters of nonviral genes such as C-MYC and DHFR, which were already known at that time to be important for cell growth and DNA metabolism, respectively. These findings provided the first clues that the E2F transcription factor might be an important regulator of the cell cycle. Since this initial discovery in 1987, several additional E2F family members have been identified, and more than 100 targets genes have been shown to be directly regulated by E2Fs, the majority of these are important for controlling the cell cycle. The progression of a cell through the cell cycle is accompanied with the increased expression of a specific set of genes during one phase of the cell cycle and the decrease of the same set of genes during a later phase of the cell cycle. This roller coaster ride, or oscillation, of gene expression is essential for the proper progression through the cell cycle to allow accurate DNA replication and cell division. The E2F transcription factors have been shown to be critical for the temporal expression of the oscillating cell cycle genes. This review will focus on how the oscillation of E2Fs and their targets is regulated by transcriptional, post-transcriptional and post-translational mechanism in mammals, yeast, flies, and worms. Furthermore, we will discuss the functional impact of E2Fs on the cell cycle progression and outline the consequences when E2F expression is disturbed. PMID:26254918

  14. Persistence, Localization, and External Control of Transgene Expression After Single Injection of Adeno-Associated Virus into Injured Joints

    OpenAIRE

    Lee, Hannah H.; O'Malley, Michael J.; Friel, Nicole A.; Payne, Karin A.; Qiao, Chunping; Xiao, Xiao(Institute for Strings, Cosmology and Astroparticle Physics (ISCAP) and Physics Department, Columbia University, 538 West 120th Street, New York, NY, 10027 U.S.A.); Chu, Constance R.

    2013-01-01

    A single intra-articular injection of adeno-associated virus (AAV) results in stable and controllable transgene expression in normal rat knees. Because undamaged joints are unlikely to require treatment, the study of AAV delivery in joint injury models is crucial to potential therapeutic applications. This study tests the hypotheses that persistent and controllable AAV-transgene expression are (1) highly localized to the cartilage when AAV is injected postinjury and (2) localized to the intra...

  15. Neurophysiology of spontaneous facial expressions: I. Motor control of the upper and lower face is behaviorally independent in adults.

    Science.gov (United States)

    Ross, Elliott D; Gupta, Smita S; Adnan, Asif M; Holden, Thomas L; Havlicek, Joseph; Radhakrishnan, Sridhar

    2016-03-01

    Facial expressions are described traditionally as monolithic entities. However, humans have the capacity to produce facial blends, in which the upper and lower face simultaneously display different emotional expressions. This, in turn, has led to the Component Theory of facial expressions. Recent neuroanatomical studies in monkeys have demonstrated that there are separate cortical motor areas for controlling the upper and lower face that, presumably, also occur in humans. The lower face is represented on the posterior ventrolateral surface of the frontal lobes in the primary motor and premotor cortices and the upper face is represented on the medial surface of the posterior frontal lobes in the supplementary motor and anterior cingulate cortices. Our laboratory has been engaged in a series of studies exploring the perception and production of facial blends. Using high-speed videography, we began measuring the temporal aspects of facial expressions to develop a more complete understanding of the neurophysiology underlying facial expressions and facial blends. The goal of the research presented here was to determine if spontaneous facial expressions in adults are predominantly monolithic or exhibit independent motor control of the upper and lower face. We found that spontaneous facial expressions are very complex and that the motor control of the upper and lower face is overwhelmingly independent, thus robustly supporting the Component Theory of facial expressions. Seemingly monolithic expressions, be they full facial or facial blends, are most likely the result of a timing coincident rather than a synchronous coordination between the ventrolateral and medial cortical motor areas responsible for controlling the lower and upper face, respectively. In addition, we found evidence that the right and left face may also exhibit independent motor control, thus supporting the concept that spontaneous facial expressions are organized predominantly across the horizontal facial

  16. Epigenetic control of group V phospholipase A2 expression in human malignant cells.

    Science.gov (United States)

    Menschikowski, Mario; Hagelgans, Albert; Nacke, Brit; Jandeck, Carsten; Mareninova, Olga A; Asatryan, Liana; Siegert, Gabriele

    2016-06-01

    Secreted phospholipases A2 (sPLA2) are suggested to play an important role in inflammation and tumorigenesis. Different mechanisms of epigenetic regulation are involved in the control of group IIA, III and X sPLA2s expression in cancer cells, but group V sPLA2 (GV-PLA2) in this respect has not been studied. Here, we demonstrate the role of epigenetic mechanisms in regulation of GV-PLA2 expression in different cell lines originating from leukaemia and solid cancers. In blood leukocytes from leukaemic patients, levels of GV-PLA2 transcripts were significantly lower in comparison to those from healthy individuals. Similarly, in DU-145 and PC-3 prostate and CAL-51 and MCF-7 mammary cancer cell lines, levels of GV-PLA2 transcripts were significantly lower in relation to those found in normal epithelial cells of prostate or mammary. By sequencing and methylation-specific high-resolution melting (MS-HRM) analyses of bisulphite-modified DNA, distinct CpG sites in the GV-PLA2 promoter region were identified that were differentially methylated in cancer cells in comparison to normal epithelial and endothelial cells. Spearman rank order analysis revealed a significant negative correlation between the methylation degree and the cellular expression of GV-PLA2 (r = -0.697; p = 0.01). The effects of demethylating agent (5-aza-2'-deoxycytidine) and histone deacetylase inhibitor (trichostatin A) on GV-PLA2 transcription in the analysed cells confirmed the importance of DNA methylation and histone modification in the regulation of the GV-PLA2 gene expression in leukaemic, prostate and mammary cancer cell lines. The exposure of tumour cells to human recombinant GV-PLA2 resulted in a reduced colony forming activity of MCF-7, HepG2 and PC-3 cells, but not of DU-145 cells suggesting a cell-type-dependent effect of GV-PLA2 on cell growth. In conclusion, our results suggest that epigenetic mechanisms such as DNA methylation and histone modification play an important role in

  17. Comparison of gene expression and genome-wide DNA methylation profiling between phenotypically normal cloned pigs and conventionally bred controls

    DEFF Research Database (Denmark)

    Fei, Gao; Luo, Yonglun; Li, Shengting;

    2011-01-01

    -specific digital karyotyping (MMSDK) and Solexa sequencing technology. Typical tissue-specific differences with respect to both gene expression and DNA methylation were observed in muscle and liver from cloned as well as control pigs. Gene expression profiles were highly similar between cloned pigs and controls......Animal breeding via Somatic Cell Nuclear Transfer (SCNT) has enormous potential in agriculture and biomedicine. However, concerns about whether SCNT animals are as healthy or epigenetically normal as conventionally bred ones are raised as the efficiency of cloning by SCNT is much lower than natural...... breeding or In-vitro fertilization (IVF). Thus, we have conducted a genome-wide gene expression and DNA methylation profiling between phenotypically normal cloned pigs and control pigs in two tissues (muscle and liver), using Affymetrix Porcine expression array as well as modified methylation...

  18. MK3 controls Polycomb target gene expression via negative feedback on ERK

    Directory of Open Access Journals (Sweden)

    Prickaerts Peggy

    2012-08-01

    Full Text Available Abstract Background Gene-environment interactions are mediated by epigenetic mechanisms. Polycomb Group proteins constitute part of an epigenetic cellular transcriptional memory system that is subject to dynamic modulation during differentiation. Molecular insight in processes that control dynamic chromatin association and dissociation of Polycomb repressive complexes during and beyond development is limited. We recently showed that MK3 interacts with Polycomb repressive complex 1 (PRC1. The functional relevance of this interaction, however, remained poorly understood. MK3 is activated downstream of mitogen- and stress-activated protein kinases (M/SAPKs, all of which fulfill crucial roles during development. We here use activation of the immediate-early response gene ATF3, a bona fide PRC1 target gene, as a model to study how MK3 and its effector kinases MAPK/ERK and SAPK/P38 are involved in regulation of PRC1-dependent ATF3 transcription. Results Our current data show that mitogenic signaling through ERK, P38 and MK3 regulates ATF3 expression by PRC1/chromatin dissociation and epigenetic modulation. Mitogenic stimulation results in transient P38-dependent H3S28 phosphorylation and ERK-driven PRC1/chromatin dissociation at PRC1 targets. H3S28 phosphorylation by itself appears not sufficient to induce PRC1/chromatin dissociation, nor ATF3 transcription, as inhibition of MEK/ERK signaling blocks BMI1/chromatin dissociation and ATF3 expression, despite induced H3S28 phosphorylation. In addition, we establish that concomitant loss of local H3K27me3 promoter marking is not required for ATF3 activation. We identify pERK as a novel signaling-induced binding partner of PRC1, and provide evidence that MK3 controls ATF3 expression in cultured cells via negative regulatory feedback on M/SAPKs. Dramatically increased ectopic wing vein formation in the absence of Drosophila MK in a Drosophila ERK gain-of-function wing vein patterning model, supports the

  19. Positive control of lac operon expression in vitro by guanosine 5'-diphosphate 3'-diphosphate.

    Science.gov (United States)

    Primakoff, P; Artz, S W

    1979-04-01

    Maximal expression of the Escherichia coli lactose operon in a coupled in vitro transcription-translation system from a Salmonella typhimurium relA mutant was strongly dependent upon addition of guanosine 5'-diphosphate 3'-diphosphate (ppGpp). Without added ppGpp, at saturating 3',5'-cyclic AMP (cAMP) concentrations, synthesis of beta-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) was reproducibly only 5-7% of that which can be obtained with 0.5-0.8 mM ppGpp. Experiments in which transcription was uncoupled from translation indicated that this 14- to 20-fold stimulation by ppGpp occurred at the level of transcription. When coupled beta-galactosidase synthesis was primed with a template containing a well-characterized mutant lac promoter (lacP(r)L8UV5), the dependence on ppGpp was greatly reduced. This result provides an important experimental control previously unavailable for verifying the significance of ppGpp effects on gene regulation in vitro; it indicates that activation of lacP(+) expression by ppGpp is specifically an effect of increased transcription initiations. Furthermore, the large ppGpp stimulation of lacP(+) DNA enabled the level of expression of this template to approach that of lacP(r)L8UV5 DNA, an observation expected from results in vivo but not obtained with other transcription-translation systems in vitro. The importance of these results is considered with respect to previous ideas on the physiological role of ppGpp as a supercontrol molecule in bacterial regulation. PMID:109832

  20. Genetic control of eosinophilia in mice: gene(s) expressed in bone marrow-derived cells control high responsiveness

    Energy Technology Data Exchange (ETDEWEB)

    Vadas, M.A.

    1982-02-01

    A heterogeneity in the capacity of strains of mice to mount eosinophilia is described. BALB/c and C3H are eosinophil high responder strains (EO-HR) and CBA and A/J are eosinophil low responder strains (EO-LR), judged by the response of blood eosinophils to Ascaris suum, and the response of blood, bone marrow, and spleen eosinophils to keyhole limpet hemocyanin given 2 days after 150 mg/kg cyclophosphamide. Some of the gene(s) for high responsiveness appear to be dominant because (EO-HR x EO-LR)F/sub 1/ mice were intermediate to high responders. This gene is expressed in bone marrow-derived cells because radiation chimeras of the type EO-HR..-->..F/sub 1/ were high responders and EO-LR..-->..F/sub 1/ were low responders. This description of a genetic control of eosinophilia in mice may be useful in understanding the role of this cell in parasite immunity and allergy.

  1. Programmable control of bacterial gene expression with the combined CRISPR and antisense RNA system

    Science.gov (United States)

    Lee, Young Je; Hoynes-O'Connor, Allison; Leong, Matthew C.; Moon, Tae Seok

    2016-01-01

    A central goal of synthetic biology is to implement diverse cellular functions by predictably controlling gene expression. Though research has focused more on protein regulators than RNA regulators, recent advances in our understanding of RNA folding and functions have motivated the use of RNA regulators. RNA regulators provide an advantage because they are easier to design and engineer than protein regulators, potentially have a lower burden on the cell and are highly orthogonal. Here, we combine the CRISPR system from Streptococcus pyogenes and synthetic antisense RNAs (asRNAs) in Escherichia coli strains to repress or derepress a target gene in a programmable manner. Specifically, we demonstrate for the first time that the gene target repressed by the CRISPR system can be derepressed by expressing an asRNA that sequesters a small guide RNA (sgRNA). Furthermore, we demonstrate that tunable levels of derepression can be achieved (up to 95%) by designing asRNAs that target different regions of a sgRNA and by altering the hybridization free energy of the sgRNA–asRNA complex. This new system, which we call the combined CRISPR and asRNA system, can be used to reversibly repress or derepress multiple target genes simultaneously, allowing for rational reprogramming of cellular functions. PMID:26837577

  2. Heat-inducible translationally controlled tumor protein of Trichinella pseudospiralis: cloning and regulation of gene expression.

    Science.gov (United States)

    Mak, C H; Poon, M W; Lun, H M; Kwok, P Y; Ko, R C

    2007-04-01

    To elucidate the mechanism of inducing translationally controlled tumor protein (TCTP) in stress adaptation of adenophorean nematodes, the complete coding sequence of TCTP of the infective-stage larvae of Trichinella pseudospiralis was characterized. Two cDNA clones with different 3' untranslated region were identified. Tp-TCTP contained an open reading frame of 534 bp encoding 177 residues. The gene with five introns was expressed as histidine-tagged fusion protein having a molecular mass of 17.5 kDa. Quantitative reverse transcriptase polymerase chain reaction (RT-PCR) analysis showed that TCTP RNA was not accumulated when the infective-stage larvae were heat-shocked for 1 h at 45 or 60 degrees C. Using enzyme-linked immunosorbent assay and antiserum against the fusion protein, the expression of TCTP was found to be up-regulated at the translational level. The data suggest that translational regulation of TCTP may play an important role in the early heat-stress adaptation of the trichinellid. Cluster analysis demonstrated that the TCTP sequence of T. pseudospiralis is closely related to that of T. spiralis, but is diverged from the secernentean species. PMID:17149606

  3. PTTG expression in different experimental and human prolactinomas in relation to dopaminergic control of lactotropes

    Directory of Open Access Journals (Sweden)

    Bronstein Marcello D

    2007-01-01

    Full Text Available Abstract Background Pituitary tumor transforming gene (pttg is a novel oncogene that is expressed at higher level in most of the tumors analyzed to date compared to normal tissues. Nevertheless, its expression in prolactinomas and its relation with the pituitary dopamine receptor 2 (D2R are not well defined. We sought to determine the pituitary level of pttg in three different experimental models of prolactinomas with altered dopaminergic control of the pituitary: the dopaminergic D2R knockout female mouse, the estrogen-treated rat, and the senescent female rat. These three models shared the characteristics of increased pituitary weight, hyperprolactinemia, lactotrope hyperplasia and reduced or absent dopaminergic action at the pituitary level. We also studied samples from human macroprolactinomas, which were characterized as responsive or resistant to dopamine agonist therapy. Results When compared to female wild-type mice, pituitaries from female D2R knockout mice had decreased PTTG concentration, while no difference in pttg mRNA level was found. In senescent rats no difference in pituitary PTTG protein expression was found when compared to young rats. But, in young female rats treated with a synthetic estrogen (Diethylstylbestrol, 20 mg PTTG protein expression was enhanced (P = 0.029. Therefore, in the three experimental models of prolactinomas, pituitary size was increased and there was hyperprolactinemia, but PTTG levels followed different patterns. Patients with macroprolactinomas were divided in those in which dopaminergic therapy normalized or failed to normalize prolactin levels (responsive and resistant, respectively. When pituitary pttg mRNA level was analyzed in these macroprolactinomas, no differences were found. We next analyzed estrogen action at the pituitary by measuring pituitary estrogen receptor α levels. The D2R knockout female mice have low estrogen levels and in accordance, pituitary estrogen receptors were increased (P

  4. Flexible control of plant architecture and yield via switchable expression of Arabidopsis gai.

    Science.gov (United States)

    Ait-ali, Tahar; Rands, Carley; Harberd, Nicholas P

    2003-09-01

    The growth of plants is repressed by DELLA proteins, nuclear regulators whose activities are opposed by the growth-promoting phytohormone gibberellin (GA). Mutations affecting DELLA protein function were previously used by plant breeders to create the high-yielding semidwarf wheat varieties of the green revolution. gai is an Arabidopsis mutant DELLA protein-encoding orthologue of the wheat semidwarfing genes. Here we describe the development of a transgene that confers ethanol-inducible gai expression. Transient induction of gai causes transient growth repression: growth prior to and after treatment is unaffected. Appropriate ethanol treatments result in dwarf plants that produce the same numbers of seeds as untreated controls. This new technology represents a substantial advance in the applicability of genes encoding mutant DELLA proteins to agricultural and horticultural improvement, enhancing the flexibity with which these genes can be used for the sustainable achievement of increased crop plant yields. PMID:17166132

  5. Control of gene expression and mitochondrial biogenesis in the muscular adaption to endurance exercise

    DEFF Research Database (Denmark)

    Joseph, A. M.; Pilegaard, H.; Leick, L.;

    2006-01-01

    adaptations is an increase in mitochondrial content, which confers a greater resistance to muscle fatigue. This essay reviews current knowledge on the regulation of exercise-induced mitochondrial biogenesis at the molecular level. The major steps involved include, (i) transcriptional regulation of nuclear......-encoded genes encoding mitochondrial proteins by the coactivator peroxisome-proliferatoractivated receptor coactivator-1, (ii) control of mitochondrial DNA gene 1To whom correspondence should be addressed (email dhood@yorku.ca). 13 © 2006 The Biochemical Society Ch-02_essbiochem_hood.indd Page 13 11/13/06 10......:27:15 PM elhi /Volumes/ju108/POIN001/essbiochem_indd%0/Chapter 2 © 2006 The Biochemical Society 14 Essays in Biochemistry volume 42 2006 expression by the transcription factor Tfam, (iii) mitochondrial fi ssion and fusion mechanisms, and (iv) import of nuclear-derived gene products into the mitochondrion...

  6. Persistence, localization, and external control of transgene expression after single injection of adeno-associated virus into injured joints.

    Science.gov (United States)

    Lee, Hannah H; O'Malley, Michael J; Friel, Nicole A; Payne, Karin A; Qiao, Chunping; Xiao, Xiao; Chu, Constance R

    2013-04-01

    A single intra-articular injection of adeno-associated virus (AAV) results in stable and controllable transgene expression in normal rat knees. Because undamaged joints are unlikely to require treatment, the study of AAV delivery in joint injury models is crucial to potential therapeutic applications. This study tests the hypotheses that persistent and controllable AAV-transgene expression are (1) highly localized to the cartilage when AAV is injected postinjury and (2) localized to the intra-articular soft tissues when AAV is injected preinjury. Two AAV injection time points, postinjury and preinjury, were investigated in osteochondral defect and anterior cruciate ligament transection models of joint injury. Rats injected with AAV tetracycline response element (TRE)-luciferase received oral doxycycline for 7 days. Luciferase expression was evaluated longitudinally for 6 months. Transgene expression was persistent and controllable with oral doxycycline for 6 months in all groups. However, the location of transgene expression was different: postinjury AAV-injected knees had luciferase expression highly localized to the cartilage, while preinjury AAV-injected knees had more widespread signal from intra-articular soft tissues. The differential transgene localization between preinjury and postinjury injection can be used to optimize treatment strategies. Highly localized postinjury injection appears advantageous for treatments targeting repair cells. The more generalized and controllable reservoir of transgene expression following AAV injection before anterior cruciate ligament transection (ACLT) suggests an intriguing concept for prophylactic delivery of joint protective factors to individuals at high risk for early osteoarthritis (OA). Successful external control of intra-articular transgene expression provides an added margin of safety for these potential clinical applications. PMID:23496155

  7. Establishment of an artificial β-cell line expressing insulin under the control of doxycycline

    Institute of Scientific and Technical Information of China (English)

    Xin-Yu Qin; Kun-Tang Shen; Xin Zhang; Zhi-Hong Cheng; Xiang-Ru Xu; Ze-Guang Han

    2002-01-01

    AIM: Artificial β-cell lines may offer an abundant source of calls for the treatment of type Ⅰ diabetes, but insulin secretion in β-cells is tightly regulwted in physiological conditions The Tet-On system is a "gene switch" system,which can induce gene expression by administration of tetracycline (Tet) derivatives such as doxcycline (D ox).Using this system, we established 293 cells to an artificial cell line secreting insulin in response to stimulation by Dox.METHODS: The mutated proinsulin cDNA was obtained fromplasmid pcDNA3.1/C-mlNS by the polymerase chain reaction(PCR), and was inserted downstream from the promoter onthe expression vector pTRE2, to construct a recombinedexpression vector pTRE2mlNS. The promoter on pTRE2consists of the tetracycline-response element and the CMVminimal promoter and is thus activated by the reversetetracycline-controlled transactivator (rtTA) when Dox isadministrated. pTRE2mlNS and plasmid pTK-Hyg encodinghygromycin were co-transfected in the tet293 cells, whichexpress rtTA stably. Following hygromycin screening, thesurvived cells expressing insulin were selected andenriched. Dox was used to control the expression of insulinin these cells. At the levels of mRNA and protein, theregulating effect of Dox in culture medium on the expressionof proinsulin gene was estimated respectively with Northernblot, RT-PCR, and radioimmunoassay.RESULTS: From the 28 hygromycin-resistant cell strains, weselected one cell strain (tet293/Ins6) secreting insulin notonly automatically, but in response to stimulation by Dox.The amount on insulin secretion was dependent on the Doxdose (0,10,100,200,400,800 and 1000 μg@ L-1 ), the level ofinsulin secreted by the cells treated with Dox ( 1000μg. L-1 )wes 241.0 pU@d1 @cell-1 , which was 25-fold that of 9.7 pU@d1@ cell-1 without Dox treatment. Northern blot analyses andRT-PCR further confinned that the transcription of insulingene had already been up-regulated after exposing tet293/Ins6 cells to Dox for

  8. Quality control parameters on a large dataset of regionally dissected human control brains for whole genome expression studies

    OpenAIRE

    Trabzuni, Daniah; Ryten, Mina; Walker, Robert; Smith, Colin; Imran, Sabaena; Ramasamy, Adaikalavan; Weale, Michael E; Hardy, John

    2011-01-01

    We are building an open-access database of regional human brain expression designed to allow the genome-wide assessment of genetic variability on expression. Array and RNA sequencing technologies make assessment of genome-wide expression possible. Human brain tissue is a challenging source for this work because it can only be obtained several and variable hours post-mortem and after varying agonal states. These variables alter RNA integrity in a complex manner. In this report, we assess the e...

  9. Expansion of peripheral and intratumoral regulatory T-cells in hepatocellular carcinoma: A case-control study

    Directory of Open Access Journals (Sweden)

    Sapna Thakur

    2011-01-01

    Full Text Available Background: Hepatocellular carcinoma (HCC is notorious for poor prognosis with limited therapeutic options. A better understanding of the role of regulatory T-cells (Tregs in HCC is important for design of immunotherapy based clinical protocol. The objective of the present study was to evaluate the presence of Tregs in tumor microenvironment in patients with HCC compared to chronic hepatitis (CH. Materials and Methods: The frequency of CD4 + CD25 + Treg cells was evaluated from peripheral blood (PB of 28 patients of HCC and 30 controls including CH cases and healthy donors using flowcytometry. Intratumoral Treg were also analyzed in tissue samples from 17 HCC cases and 15 CH cases. In addition the expression of FOXP3 and CTLA-4 was also studied by RT-PCR. Results: Frequency of CD4 + CD25 + cells in the PBMCs of HCC cases was significantly higher than in HC (10.8 ± 7.64 vs 3.05 ± 1.30, P < 0.005 and CH patients (2.88 ± 1.92, P < 0.005. Also Treg population was significantly higher in HCC tumor microenvironment compared to CH biopsies (15.8 ± 5.32 vs 5.51 ± 3.40, P < 0.05. Expression of FOXP3 and CTLA-4 was also significantly higher in HCC patients ( P < 0.05 compared to CH group. Conclusions: We provide evidence of an increased population of Treg not only in the PB but also in tumor microenvironment of HCC patients, suggesting association of enhanced Treg activity with poor immune responses to tumor antigens. These findings may in future play a significant role in designing immunotherapeutic approaches in HCC.

  10. CRISPR-based genome editing and expression