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Sample records for anti-human cd25 antibody

  1. 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.

  2. Effects of in vivo injection of anti-chicken CD25 monoclonal antibody on regulatory T cell depletion and CD4+CD25- T cell properties in chickens.

    Science.gov (United States)

    Shanmugasundaram, Revathi; Selvaraj, Ramesh K

    2012-03-01

    Regulatory T cells (Tregs) are defined as CD4(+)CD25(+) cells in chickens. This study examined the effects of an anti-chicken CD25 monoclonal antibody injection (0.5 mg/bird) on in vivo depletion of Tregs and the properties of CD4(+)CD25(-) cells in Treg-depleted birds. The CD4(+)CD25(+) cell percentage in the blood was lower at 8 d post injection than at 0 d. Anti-CD25-mediated CD4(+)CD25(+) cell depletion in blood was maximum at 12 d post injection. The anti-CD25 antibody injection depleted CD4(+)CD25(+) cells in the spleen and cecal tonsils, but not in the thymus, at 12 d post antibody injection. CD4(+)CD25(-) cells from the spleen and cecal tonsils of birds injected with the anti-chicken CD25 antibody had higher proliferation and higher IL-2 and IFNγ mRNA amounts than the controls at 12 d post injection. At 20 d post injection, CD4(+)CD25(+) cell percentages in the blood, spleen and thymus were comparable to that of the 0 d post injection. It could be concluded that anti-chicken CD25 injection temporarily depleted Treg population and increased and IL-2 and IFNγ mRNA amounts in CD4(+)CD25(-) cells at 12d post injection.

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

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    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.

  4. Recombinant anti-human melanoma antibodies are versatile molecules.

    Science.gov (United States)

    Neri, D; Natali, P G; Petrul, H; Soldani, P; Nicotra, M R; Vola, R; Rivella, A; Creighton, A M; Neri, P; Mariani, M

    1996-08-01

    The low cost, high versatility, and reliable production of bacterially produced recombinant antibody fragments speeds up the development of tumor-targeting agents. High-quality recombinant anti-melanoma antibodies are much sought after in the scientific community. We cloned the murine antibody 225.28S, currently used in radioimmunoimaging of human melanoma lesions, in single-chain Fv configuration (scFv) for soluble expression in bacteria. The recombinant antibody fragment conserved the binding specificity of the parental antibody. In order to arm the scFv(225.28S) with biologically useful effector functions, we developed vectors for soluble expression of scFv(225.28S) in bacteria that allow both covalent and noncovalent chemical antibody modification at positions that do not interfere with antigen binding. An expression vector was developed that appends a cysteine residue at the C-terminal extremity of the recombinant antibody, thus allowing reaction with thiol-specific reagents, including 99mTc labeling, at a position that does not interfere with antigen binding. The scFv(225.28S) was also successfully expressed with a casein kinase II substrate tag that enables efficient and stable 32P labeling. For noncovalent antibody modification, we developed an expression vector that appends the human calmodulin gene at the C-terminal extremity of scFv(225.28S). The calmodulin domain is poorly immunogenic and can be targeted with chemically modified high-affinity calmodulin ligands. The recombinant anti-human melanoma antibodies described in this article should prove useful "building blocks" for the development of anti-melanoma diagnostic and therapeutic strategies.

  5. 抗人CD25嵌合抗体基因的构建及其瞬时表达研究%Study on construction and transient expression of human-mouse chimeric antibody gene against human CD25

    Institute of Scientific and Technical Information of China (English)

    胡迪超; 张爱华; 潘勇兵; 詹珊珊; 杨晓明

    2011-01-01

    目的:构建抗人CD25嵌合抗体基因并在哺乳动物细胞中进行瞬时表达和初步鉴定.方法:采用RLM-RACE法克隆WuTac抗体可变区和信号肽序列,并利用基因拼接法构建嵌合抗体基因.用脂质体法瞬时转染三种哺乳动物细胞,并使用ELISA、FCM、WB、Dot blot和免疫荧光法进行检测.结果:成功克隆WuTac抗体可变区和信号肽序列,并构建了抗人CD25嵌合抗体表达质粒.瞬时转染结果表明所表达的嵌合抗体保留了亲本抗体WuTac的抗原结合力.结论:成功构建了抗人CD25嵌合抗体基因,为其进一步研究打下基础.%Objective:To construct chimeric antibody gene against human an CD25 angigen,and prelin inarily identify the expressed prod-ucts produced from transiently transfected mammalian cells in order to facilitate the further study of stable expression.Methods:The RLM-RACE was employed to clone variable region genes and leader sequences,and the Overlap PVR method was used to construct the chimeric anti-body gene.After transiently transfected in three mammalian cells with liposome method, the expressed products were determined by ELISA,FCM,W B,Dotblot and immunofluorescence assay.Results:The variable region genes and leader sequences were successfully amplified,and the eukayotic expression plasmids were constructed.The results from transient transfection indicate the expressed products retain the antigen binding capacity of parental antibody WuTac.Conclusion:The successfully constructed chimeric antibody gene against human CD25 lays sound foun-dation for further study.

  6. 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.

  7. An anti-human ICAM-1 antibody inhibits rhinovirus-induced exacerbations of lung inflammation.

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    Stephanie Traub

    Full Text Available Human rhinoviruses (HRV cause the majority of common colds and acute exacerbations of asthma and chronic obstructive pulmonary disease (COPD. Effective therapies are urgently needed, but no licensed treatments or vaccines currently exist. Of the 100 identified serotypes, ∼90% bind domain 1 of human intercellular adhesion molecule-1 (ICAM-1 as their cellular receptor, making this an attractive target for development of therapies; however, ICAM-1 domain 1 is also required for host defence and regulation of cell trafficking, principally via its major ligand LFA-1. Using a mouse anti-human ICAM-1 antibody (14C11 that specifically binds domain 1 of human ICAM-1, we show that 14C11 administered topically or systemically prevented entry of two major groups of rhinoviruses, HRV16 and HRV14, and reduced cellular inflammation, pro-inflammatory cytokine induction and virus load in vivo. 14C11 also reduced cellular inflammation and Th2 cytokine/chemokine production in a model of major group HRV-induced asthma exacerbation. Interestingly, 14C11 did not prevent cell adhesion via human ICAM-1/LFA-1 interactions in vitro, suggesting the epitope targeted by 14C11 was specific for viral entry. Thus a human ICAM-1 domain-1-specific antibody can prevent major group HRV entry and induction of airway inflammation in vivo.

  8. Humanized versus murine anti-human epidermal growth factor receptor monoclonal antibodies for immunoscintigraphic studies

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    Morales, Alejo A. Morales; Duconge, Jorge; Alvarez-Ruiz, Daniel; Becquer-Viart, Maria de Los Angeles; Nunez-Gandolff, Gilda; Fernandez, Eduardo; Caballero-Torres, Idania; Iznaga-Escobar, Normando

    2000-02-01

    The anti-human epidermal growth factor receptor (EGF-R) humanized antibody h-R3 (IgG{sub 1}), which binds to an extracellular domain of EGF-R, was used to evaluate the biodistribution on nude mice xenografted with A431 epidermoid carcinoma cell line. Results are compared with its murine version ior egf/r3 monoclonal antibody (mAb). Twenty-one athymic female 4NMRI nu/nu mice were injected intravenously with 10 {mu}g/100 {mu}Ci of {sup 99m}Tc-labeled mAbs. The mAb ior C5 that recognizes an antigen expressed preferentially on the surface of malignant and cytoplasm of normal colorectal cells was used as negative control. Immunoreactivity of {sup 99m}Tc-labeled mAbs was measured by enzyme linked immunosorbent assay on A431 cell line and the immunoreactive fractions determined by Lindmo method. Among all organs significant accumulation was found in tumor (6.14{+-}2.50 %ID/g, 5.06{+-}2.61 %ID/g for murine and humanized mAbs, respectively) 4 h after injection. The immunoreactive fractions were found to be 0.88 and 0.81 for murine and humanized mAb, respectively. Thus, we expect better results using the humanized mAb h-R3 for diagnostic immunoscintigraphy.

  9. Combinative application CD4+ CD25+ regulatory T cells test and irregular antibody screening in the diagnosis of invalid clinical transfusion%联合检测CD4+ CD25+调节性T细胞和血清不规则抗体在红细胞输注无效诊断的应用研究

    Institute of Scientific and Technical Information of China (English)

    李忱炜

    2016-01-01

    Objective To investigate whether the T cells (CD4+ CD25+) levels and the irregular antibodies screening cloud improve the diagnosis of invalid red blood cells transfusion.Methods Thirty-one patients with red blood cell invalid transfusion in the People's Hospital of Yubei District of Chongqing were selected.Flow cytometry was used to detect the changes of CD4+CD25+ regulatory T cells.Standard cells Ⅰ,Ⅱ,Ⅲ were used to screen on irregular antibodies in red blood cells (RBC).Results The rate of invalid RBC transfusion was 7.52% (31/412) in Yubei District.The incidence rate of medical diseases was 77.42% (24/31),much higher than surgical disease (22.58% (7/31,P =0.002),and the major departments were oncology department,hematological department and infectious department.The CD4+ CD25+ regulatory T cells were decrease from (22.18±1.58) % to (16.57±1.77) %(P=0.023).Conclusion Combin CD4+CD25+ regulatory T cells test and irregular antibody screening can help prevent and reduce the invalid transfusionof red blood cells.%目的 通过联合检测红细胞输注前后调节性T细胞(CD4+ CD25+)的变化和不规则抗体存在情况,探讨其是否有利于提高患者红细胞输注无效的检出率.方法 选取渝北区人民医院红细胞输注无效患者31例,采用流式细胞术检测输血前后CD4+ CD25+调节性T细胞变化,采用标准筛查细胞Ⅰ、Ⅱ、Ⅲ检测血清低效价、低亲和力不规则抗体.结果 渝北区红细胞输注无效发生率为7.52%(31/412),内科系统患者红细胞输注无效构成比为77.42%(24/31),远大于外科系统患者的22.58%(7/31) (P=0.002),红细胞输注无效发生以肿瘤科、血液内科、感染科为主;红细胞输注无效患者CD4+CD25+调节性T细胞表达由(22.18±1.58)%下降至(16.57±1.77)% (P=0.023).结论 联合检测CD4+ CD25+T细胞的变化和不规则抗体表达,有利于提高红细胞输注无效的检出率,预防和减少红细胞输注无效.

  10. Cetuximab in combination with anti-human IgG antibodies efficiently down-regulates the EGF receptor by macropinocytosis

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    Berger, Christian [Department of Pathology, Oslo University Hospital, Rikshospitalet, Post box 4950 Nydalen, 0424 Oslo (Norway); Madshus, Inger Helene [Institute of Pathology, University of Oslo, Rikshospitalet, 0027 Oslo (Norway); Department of Pathology, Oslo University Hospital, Rikshospitalet, Post box 4950 Nydalen, 0424 Oslo (Norway); Stang, Espen, E-mail: espsta@rr-research.no [Department of Pathology, Oslo University Hospital, Rikshospitalet, Post box 4950 Nydalen, 0424 Oslo (Norway)

    2012-12-10

    The monoclonal antibody C225 (Cetuximab) blocks binding of ligand to the epidermal growth factor receptor (EGFR). In addition, it is known that incubation with C225 induces endocytosis of the EGFR. This endocytosis has previously been shown to be increased when C225 is combined with an additional monoclonal anti-EGFR antibody. However, the effects of antibody combinations on EGFR activation, endocytosis, trafficking and degradation have been unclear. By binding a secondary antibody to the C225-EGFR complex, we here demonstrate that a combination of antibodies can efficiently internalize and degrade the EGFR. Although the combination of antibodies activated the EGFR kinase and induced ubiquitination of the EGFR, the kinase activity was not required for internalization of the EGFR. In contrast to EGF-induced EGFR down-regulation, the antibody combination efficiently degraded the EGFR without initiating downstream proliferative signaling. The antibody-induced internalization of EGFR was found not to depend on clathrin and/or dynamin, but depended on actin polymerization, suggesting induction of macropinocytosis. Macropinocytosis may cause internalization of large membrane areas, and this could explain the highly efficient internalization of the EGFR induced by combination of antibodies. -- Highlight: Black-Right-Pointing-Pointer Cetuximab induced endocytosis of EGFR increases upon combination with anti-human IgG. Black-Right-Pointing-Pointer Antibody combination causes internalization of EGFR by macropinocytosis. Black-Right-Pointing-Pointer Antibody-induced internalization of EGFR is independent of EGFR kinase activity. Black-Right-Pointing-Pointer Antibody combination may have a zipper effect and cross-link EGFRs on neighboring cells.

  11. A Novel Anti-Human Syndecan-1(CD138) Monoclonal Antibody 4B3: Characterization and Application

    Institute of Scientific and Technical Information of China (English)

    Wanping Sun; Fengming Wang; Fang Xie; Guoqing Wang; Jin Sun; Gehua Yu; Yuhua Qiu; Xueguang Zhang

    2007-01-01

    Syndecan-1 (CD138), a member of integral membrane heparin sulfate proteoglycans, is an essential matrix receptor for maintaining the normal morphological phenotypes. In this study, we generated a specific mouse anti-human syndecan-1 monoclonal antibody (mAb) 4B3 and identified it by competition assay with the available syndecan-1 mAb (BB4). Stained by 4B3, the expression of syndecan-1 was detected on tumor cell lines, such as 8226,U266, XG-1, XG-2, Daudi and Jurkat. The expression was also found on neuron stem cells. It was established that 4B3 mAb could inhibit XG-1 and XG-2 proliferation. The data not only determined that 4B3 mAb was a functional anti-human syndecan-1 mAb, but also indicated that syndecan-1 might be a valuable surface antigen and play an important role in regulation of tumor pathology and differentiation of neural stem cells. This novel antibody 4B3 may be value of study of tumor proliferation/survival mechanism and contributes to diagnosis and treatment of diverse diseases.

  12. Flow cytometric immunophenotyping of feline bone marrow cells and haematopoietic progenitor cells using anti-human antibodies.

    Science.gov (United States)

    Araghi, Atefeh; Nassiri, Seyed Mahdi; Atyabi, Nahid; Rahbarghazi, Reza; Mohammadi, Elham

    2014-04-01

    There is a paucity of species-specific antibodies available for feline haematopoietic conditions. The purpose of this study was to broaden the panel of antibodies available for use in the immunophenotypic characterisation of feline haematopoietic cells by testing clones of anti-human monoclonal antibodies (mAbs) on normal, neoplastic and cultured feline haematopoietic progenitors to determine cross-reactivity to feline counterparts. In this study, 24 clones of anti-human mAbs were tested on normal or neoplastic feline bone marrow and peripheral blood cells. Six of these mAbs, including anti-cluster of differentiation (CD)61, anti-CD18, anti-CD14, anti-CD235a, anti-CD41 and anti-CD29, cross-reacted with normal feline bone marrow cells, whereas anti-CD33 and anti-CD117 cross-reacted with the blast cells in the bone marrow of two cats with myelodysplastic syndrome, and anti-CD71, anti-235a, anti-41 and anti-42 cross-reacted with immature erythroid cells in a cat with erythroleukaemia. In a feline immunodeficiency virus-positive cat, bone marrow cells were labelled with anti-CD33, anti-14 and anti-45. Anti-CD18, anti-CD14, anti-CD41 and anti-CD61 also reacted with the peripheral blood cells of the healthy cats. The feline haematopoietic progenitors formed colonies in the methylcellulose-based semisolid medium with significant enrichment of colony-forming unit-granulocyte, monocyte and burst-forming unit-erythroid. A panel of six anti-feline mAbs (anti-CD21-like, anti-T lymphocytes, anti-CD172a, anti-granulocyte, anti-CD45-like and anti-CD18) and eight anti-human antibodies (anti-CD71, anti-CD33, anti-CD235a, anti-CD41, anti-CD61, anti-CD117, anti-CD38 and anti-CD34) were used for the immunophenotypic characterisation of the feline bone marrow progenitors. CD45, CD33, CD235a and CD18 were expressed by the feline haematopoietic progenitor cells, with the highest expression level for CD45.

  13. Reactivity of eleven anti-human leucocyte monoclonal antibodies with lymphocytes from several domestic animals

    DEFF Research Database (Denmark)

    Aasted, Bent; Blixenkrone-Møller, Merete; Larsen, Else Bang;

    1988-01-01

    Nine commercially available monoclonal antibodies and two monoclonal antibodies from The American Type Culture Collection, raised against various human leucocyte surface antigens, were tested on lymphocytes from cow, sheep, goat, swine, horse, cat, dog, mink, and rabbit as well as man. Four antib...

  14. Generation of mouse anti-human urate anion exchanger antibody by genetic immunization and its identification

    Institute of Scientific and Technical Information of China (English)

    XU Guo-shuang; WU Di; CHEN Xiang-mei; SHI Suo-zhu; HONG Quan; ZHANG Ping; LU Yang

    2005-01-01

    Background Human urate anion exchanger (hURAT1) as a major urate transporter expressed on renal tubular epithelial cells regulates blood urate level by reabsorbing uric acid. Antibody is an important tool to study hURAT1. This study aimed, by genetic immunization, to produce mouse anti-hURAT1 polyclonal antibody with high throughput and high specificity and to detect the location of hURAT1 in human kidney.Methods Human renal total RNA was isolated and the entire cDNA of hURAT1 was amplified by RT-PCR. The sequence of intracellular high antigenicity fragment (A280 to R349) was chosen by prediction software of protein antigenicity, and its cDNA was amplified from cDNA of hURAT1, and then cloned into pBQAP-TT vector to construct recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization. Mice were inoculated with this recombinant plasmid and two other adjuvant plasmids, pCMVi-GMCSF and pCMVi-Flt3L, which helped to enhance the antibody’s generation. After four weeks, the mice were sacrificed to obtain the anti-hURAT1 antibody from serum. The antibody was identified by western blot analysis and immunohistochemistry. At the same time, rabbit anti-hURAT1 antibody was produced by protein immunization. The specificity and efficiency between the rabbit and mouse anti-hURAT1 antibody were compared by western blot analysis and immunohistochemistry.Results The entire cDNA of hURAT1 and cDNA of its intracellular high immunogenic fragment were amplified successfully. Recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization was confirmed by restriction digestion and sequencing. Both the mouse anti-hURAT1 antibody and rabbit anti-hURAT1 antibody recognized 58kD hURAT1 and 64kD glycosylated hURAT1 protein bands in western blot. Immunohistochemically, hURAT1 was located at the brush border membrane of renal proximal tubular cells. In addition, the throughput and specificity of the mouse anti-hURAT1 antibody were higher than those of the rabbit anti-hURAT1 antibody

  15. Effect of variations in peptide sequence on anti-human milk fat globule membrane antibody reactions.

    Science.gov (United States)

    Xing, P X; Reynolds, K; Pietersz, G A; McKenzie, I F

    1991-02-01

    Monoclonal anti-mucine antibodies BC1, BC2 and BC3 produced using human milk fat globule membrane react with a synthetic peptide p1-24 (PDTRPAPGSTAPPAHGVTSAPDTR) representing the repeating amino acid sequence of the mucin core protein. The minimum epitope recognized by these three monoclonal antibodies (mAb) in p1-24 was contained in the five amino acids APDTR. To analyse the variation of position of the epitope, various modifications of the APDTR sequence were made by synthesizing peptides and testing by direct binding and inhibition enzyme-linked immunosorbent assays. Firstly, peptides p13-32 and C-p13-32, in which the epitope APDTR was placed in the middle instead of the C-terminal as in p1-24, were examined. These peptides had a greater reaction with mAb BC1, BC2 and BC3 compared with the reaction with p1-24. Secondly, A-p1-24 and TSA-p1-24 were made wherein two APDTR epitopes were present--these peptides were shown to bind two IgG antibody molecules. Finally, the contribution of each amino acid in the APDTR epitope was studied using the pepscan polyethylene rods, making all 20 of the amino acid substitutions in each position for SAPDTR (the minimum epitope APDTR with an adjacent amino acid S). In the 120 peptides examined there were some 'permissible' substitutions in A, D and T but not in P or R for BC1 and BC2; there were more 'permissible' substitutions for BC3; different substitution patterns were found with each antibody and some substitutions gave an increased reaction compared with the native peptide SAPDTR. The studies are of value in analysing the reaction of antibodies with epitopes expressed in breast cancer and in determining the antigenicity of synthetic peptides.

  16. Production of the Polyclonal Anti-human Metallothionein 2A Antibody with Recombinant Protein Technology

    Institute of Scientific and Technical Information of China (English)

    Faiz M.M.T.MARIKAR; Qi-Ming SUN; Zi-Chun HUA

    2006-01-01

    Metallothionein 2A (MT2A) is a small stress response protein that can be induced by exposure to toxic metals. It is highly expressed in breast cancer cells. In this study, the eDNA encoding the human MT2A protein was expressed as glutathione S-transferase (GST) fusion protein in Escherichia coli.Recombinant MT2A proteins were loaded onto 12% sodium dodecylsulfate-polyacrylamide gel and separated by electrophoresis, the recombinant protein was visualized by Coomassie blue staining and the 33 kDa recombinant GST-MT2A fusion protein band was cut out from the gel. The gel slice was minced and used to generate polyclonal antisera. Immunization of rabbit against MT2A protein allowed the production of high titer polyclonal antiserum. This new polyclonal antibody recognized recombinant MT2A protein in Western blot analysis. This low-cost antibody will be useful for detection in various immuno-assays.

  17. Cloning and Sequence Analysis of Light Variable Region Gene of Anti-human Retinoblastoma Monoclonal Antibody

    Institute of Scientific and Technical Information of China (English)

    Xiufeng Zhong; Yongping Li; Shuqi Huang; Bo Ning; Chunyan Zhang; Jianliang Zheng; Guanguang Feng

    2002-01-01

    Purpose: To clone the variable region gene of light chain of monoclonal antibody against human retinoblastoma and to analyze the characterization of its nucleotide sequence as well as amino acid sequence.Methods: Total RNA was extracted from 3C6 hybridoma cells secreting specific monoclonal antibody(McAb)against human retinoblastoma(RB), then transcripted reversely into cDNA with olig-dT primers.The variable region of the light chain (VL) gene fragments was amplified using polymeerase chain reaction(PCR) and further cloned into pGEM(R) -T Easy vector. Then, 3C6 VL cDNA was sequenced by Sanger's method.Homologous analysis was done by NCBI BLAST.Results: The complete nucleotide sequence of 3C6 VL cDNA consisted of 321 bp encoding 107 amino acid residues, containing four workframe regions(FRs)and three complementarity-determining regions (CDRs) as well as the typical structure of two cys residues. The sequence is most homological to a member of the Vk9 gene family, and its chain utilizes the Jkl gene segment.Conclusion: The light chain variable region gene of the McAb against human RB was amplified successfully , which belongs to the Vk9 gene family and utilizes Vk-Jk1 gene rearrangement. This study lays a good basis for constructing a recombinant antibody and for making a new targeted therapeutic agents against retinoblastoma.

  18. Preparation and functional studies of hydroxyethyl chitosan nanoparticles loaded with anti-human death receptor 5 single-chain antibody

    Directory of Open Access Journals (Sweden)

    Yang J

    2014-05-01

    Full Text Available Jingjing Yang,1,3,* Xiaoping Huang,1,3,* Fanghong Luo,1 Xiaofeng Cheng,3 Lianna Cheng,3 Bin Liu,4 Lihong Chen,2 Ruyi Hu,1,3 Chunyan Shi,1,3 Guohong Zhuang,1,3 Ping Yin2 1Anti-Cancer Research Center, Medical College, Xiamen University, Fujian, People's Republic of China, 2The Department of Pathology, Zhongshan Hospital, Xiamen University, Xiamen, People's Republic of China, 3Organ transplantation institution, Xiamen University, Xiamen, People's Republic of China, 4Jilin Vocational College of Industry and Technology, Jilin, People's Republic of China  *These authors contributed equally to this work Objective: To prepare hydroxyethyl chitosan nanoparticles loaded with anti-human death receptor 5 single-chain antibody, and study their characteristics, functions, and mechanisms of action. Materials and methods: The anti-human death receptor 5 single-chain antibody was constructed and expressed. Protein-loaded hydroxyethyl chitosan nanoparticles were prepared, and their size, morphology, particle-size distribution and surface zeta potential were measured by scanning electron microscopy and laser particle-size analysis. Mouse H22 hepatocellular carcinoma cells were cultured, and growth inhibition was examined using the CellTiter-Blue cell-viability assay. Flow cytometry and Hoechst 33342 were employed to measure cell apoptosis. Kunming mice with H22 tumor models were treated with protein-loaded hydroxyethyl chitosan nanoparticles, and their body weight and tumor size were measured, while hematoxylin and eosin staining was used to detect antitumor effects in vivo and side effects from tumors. Results: The protein-loaded hydroxyethyl chitosan nanoparticles had good stability; the zeta potential was -24.2±0.205, and the dispersion index was 0.203. The inhibition of the protein-loaded hydroxyethyl chitosan nanoparticles on H22 growth was both time- and dose-dependent. Increased expressions of active caspase 8, active caspase 3, and BAX were detected

  19. Induction of Murine Mucosal CCR5-Reactive Antibodies as an Anti-Human Immunodeficiency Virus Strategy

    Science.gov (United States)

    Barassi, C.; Soprana, E.; Pastori, C.; Longhi, R.; Buratti, E.; Lillo, F.; Marenzi, C.; Lazzarin, A.; Siccardi, A. G.; Lopalco, L.

    2005-01-01

    The genital mucosa is the main site of initial human immunodeficiency virus type 1 (HIV-1) contact with its host. In spite of repeated sexual exposure, some individuals remain seronegative, and a small fraction of them produce immunoglobulin G (IgG) and IgA autoantibodies directed against CCR5, which is probably the cause of the CCR5-minus phenotype observed in the peripheral blood mononuclear cells of these subjects. These antibodies recognize the 89-to-102 extracellular loop of CCR5 in its native conformation. The aim of this study was to induce infection-preventing mucosal anti-CCR5 autoantibodies in individuals at high risk of HIV infection. Thus, we generated chimeric immunogens containing the relevant CCR5 peptide in the context of the capsid protein of Flock House virus, a presentation system in which it is possible to engineer conformationally constrained peptide in a highly immunogenic form. Administered in mice via the systemic or mucosal route, the immunogens elicited anti-CCR5 IgG and IgA (in sera and vaginal fluids). Analogous to exposed seronegative individuals, mice producing anti-CCR5 autoantibodies express significantly reduced levels of CCR5 on the surfaces of CD4+ cells from peripheral blood and vaginal washes. In vitro studies have shown that murine IgG and IgA (i) specifically bind human and mouse CD4+ lymphocytes and the CCR5-transfected U87 cell line, (ii) down-regulate CCR5 expression of CD4+ cells from both humans and untreated mice, (iii) inhibit Mip-1β chemotaxis of CD4+ CCR5+ lymphocytes, and (iv) neutralize HIV R5 strains. These data suggest that immune strategies aimed at generating anti-CCR5 antibodies at the level of the genital mucosa might be feasible and represent a strategy to induce mucosal HIV-protective immunity. PMID:15890924

  20. Construction and Analysis of Three-dimensional Graphic Model of Single-chain Fv Derived from an Anti-human Placental Acidic Isoferritin Monoclonal Antibody by Computer

    Institute of Scientific and Technical Information of China (English)

    ZHOU Chun; SHEN Guanxin; ZHU Huifen; YANG Jing; ZHANG Yue; FENG Jiannan; SHEN Beifen

    2000-01-01

    A three-dimensional (3D) graphic model of a single-chain Fv (scFv) which was derived from an anti-human placental acidic isoferritin (PAF) monoclonal antibody (Mab) was constructed by a homologous protein-predicting computer algorithm on Silicon graphic computer station.The structure, surface static electricity and hydrophobicity of scFv were investigated. Computer graphic modelling indicated that all regions of scFv including the linker, variable regions of the heavy (VH) and light (VL) chains were suitable. The VH region and the VL region were involved in composing the "hydrophobic pocket". The linker was drifted away VH and VL regions. The complementarity determining regions (CDRs) of VH and VL regions surrounded the "hydrophobic pocket". This study provides a theory basis for improving antibody affinity, investigating antibody structure and analyzing the functions of VH and VL regions in antibody activity.

  1. Production and characterization of murine monoclonal anti-human DNase II antibodies, and their use for immunoaffinity purification of DNase II from human liver and urine.

    Science.gov (United States)

    Nakajima, Tamiko; Yasuda, Toshihiro; Takeshita, Haruo; Mori, Shinjiro; Mogi, Kouichi; Kaneko, Yasushi; Nakazato, Emiko; Kishi, Koichiro

    2002-04-15

    Four murine monoclonal anti-human deoxyribonuclease II (DNase II) antibodies were obtained from BALB/c mice immunized with human DNase II purified from human liver. Both single radial enzyme diffusion (SRED) and DNA-cast polyacrylamide gel electrophoresis (DNA-cast PAGE) were very useful for obtaining the DNase II-specific antibodies. All of the antibodies showed specific inhibition of human DNase II enzyme activity and specific immunostaining of the 32-kDa enzyme band, which is one of the three non-identical subunits of human DNase II molecule separated by sodium dodecyl sulfate (SDS)-PAGE followed by blotting on a transfer membrane. A formyl-cellulofine resin conjugated with each antibody specifically adsorbed and efficiently desorbed the active DNase II enzyme. Insertion of the immunoaffinity step in our purification procedure made the purification of human DNase II easier, faster and more effective than the conventional procedure.

  2. Study of chronic hemolytic anaemia patients in Rio de Janeiro: prevalence of anti-human parvovirus B19 IgG antibodies and the developement aplastic crises

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    SANT'ANNA Anadayr L.M.

    2002-01-01

    Full Text Available The prevalence of anti-human parvovirus B19 IgG antibodies was determined in sera from 165 chronic hemolytic anemia patients, receiving medical care at Instituto Estadual de Hematologia (IEHE, Rio de Janeiro, during the year of 1994. This sample represents around 10% of the chronic hemolytic anemia patients attending at IEHE. Most of these patients (140 have sickle cell disease. Anti-B19 IgG antibodies were detected in 32.1% of patients. No statistically significant difference (p > 0.05 was seen between IgG antibody prevalence in male (27.8% and female (35.5% patients. Anti-B19 IgG antibodies were more frequent in older (37.6% than younger (28.2% than 20 years old patients, although this difference had no statistical significance (p > 0.05. Anti-B19 IgG antibody prevalence showed that 67.9% of patients enrolled in the study were susceptible to B19 acute infection. With the aim to detect acute B19 infection, patients follow up continued until February 1996. During this period four patients presented transient aplastic crisis due to human parvovirus B19 as confirmed by the detection of specific IgM antibodies. All four patients were younger than 20 years old, and 3 were younger than 10 years old. Three of them were sickle cell disease patients. Three of the four acute B19 infection occurred during 1994 springtime.

  3. Study of chronic hemolytic anaemia patients in Rio de Janeiro: prevalence of anti-human parvovirus B19 IgG antibodies and the development aplastic crises.

    Science.gov (United States)

    Sant'Anna, Anadayr L M; Garcia, Rita de Cássia N Cubel; Marzoche, Mônica; da Rocha, Heloisa Helena A Gallo; Paula, Maria Tereza M; Lobo, Clarisse C; Nascimento, Jussara P

    2002-01-01

    The prevalence of anti-human parvovirus B19 IgG antibodies was determined in sera from 165 chronic hemolytic anemia patients, receiving medical care at Instituto Estadual de Hematologia (IEHE), Rio de Janeiro, during the year of 1994. This sample represents around 10% of the chronic hemolytic anemia patients attending at IEHE. Most of these patients (140) have sickle cell disease. Anti-B19 IgG antibodies were detected in 32.1% of patients. No statistically significant difference (p > 0.05) was seen between IgG antibody prevalence in male (27.8%) and female (35.5%) patients. Anti-B19 IgG antibodies were more frequent in older (37.6%) than younger (28.2%) than 20 years old patients, although this difference had no statistical significance (p > 0.05). Anti-B19 IgG antibody prevalence showed that 67.9% of patients enrolled in the study were susceptible to B19 acute infection. With the aim to detect acute B19 infection, patients follow up continued until February 1996. During this period four patients presented transient aplastic crisis due to human parvovirus B19 as confirmed by the detection of specific IgM antibodies. All four patients were younger than 20 years old, and 3 were younger than 10 years old. Three of them were sickle cell disease patients. Three of the four acute B19 infection occurred during 1994 springtime.

  4. Anti-human Vascular Endothelial Growth Factor (VEGF) Antibody Selection for Immunohistochemical Staining of Proliferating Blood Vessels

    NARCIS (Netherlands)

    C.M. van der Loos; L.B. Meijer-Jorna; M.E.C. Broekmans; H.P.H.M. Ploegmakers; P. Teeling; O.J. de Boer; A.C. van der Wal

    2010-01-01

    Nine commercially available VEGF antibodies are investigated for their ability to immunostain vascular malformations (VM) with or without immature capillary proliferation. First, all antibodies were optimized for their performance in immunohistochemistry with placenta and colon adenocarcinoma as pos

  5. Duodenal enteroglucagonoma revealed by differential comparison of serum and tissue glucagon reactivity with Siemens' Double Glucagon Antibody and DakoCytomation's Polyclonal Rabbit Anti-Human Glucagon: a case report

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    Abouljoud Marwan S

    2010-06-01

    Full Text Available Abstract Introduction This case report demonstrates that the differential immunohistochemical reactivities of Siemens' Double Antibody Glucagon compared to DakoCytomation's Polyclonal Rabbit Anti-Human Glucagon allow for pathologic distinction of enteral versus pancreatic glucagonoma. Case presentation A 64-year-old Caucasian man was diagnosed with a duodenal enteroglucagonoma following presentation with obstructive jaundice. He had a low serum glucagon level using Siemens' Double Antibody Glucagon, a clinical syndrome consistent with glucagon hypersecretion. A periampullary mass biopsy proved to be a neuroendocrine tumor, with positive immunohistochemical reactivity to DakoCytomation's Polyclonal Rabbit Anti-Human Glucagon. Conclusions Differential comparison of the immunohistochemical reactivities of Siemens' Double Antibody Glucagon and DakoCytomation's Polyclonal Rabbit Anti-Human Glucagon discerns enteroglucagon from pancreatic glucagon.

  6. The effectiveness of an anti-human IL-6 receptor monoclonal antibody combined with chemotherapy to target colon cancer stem-like cells.

    Science.gov (United States)

    Ying, Jin; Tsujii, Masahiko; Kondo, Jumpei; Hayashi, Yoshito; Kato, Motohiko; Akasaka, Tomofumi; Inoue, Takuta; Shiraishi, Eri; Inoue, Tahahiro; Hiyama, Satoshi; Tsujii, Yoshiki; Maekawa, Akira; Kawai, Shoichiro; Fujinaga, Tetsuji; Araki, Maekawa; Shinzaki, Shinichiro; Watabe, Kenji; Nishida, Tsutomu; Iijima, Hideki; Takehara, Tetsuo

    2015-04-01

    Recent studies have demonstrated that cancer stem cells (CSCs) can initiate and sustain tumor growth and exhibit resistance to clinical cytotoxic therapies. Therefore, CSCs represent the main target of anticancer therapy. Interleukin-6 (IL-6) promotes cellular proliferation and drug resistance in colorectal cancer, and its serum levels correlate with patient survival. Therefore, IL-6 and its downstream signaling molecule the signal transducer and activator of transcription-3 (STAT3) represent potential molecular targets. In the present study, we investigated the effects of IL-6 and its downstream signaling components on stem cell biology, particularly the chemoresistance of CSCs, to explore potential molecular targets for cancer therapy. The colon cancer cell line WiDr was cultured in serum-free, non-adherent, and three-dimensional spheroid-forming conditions to enrich the stem cell-like population. Spheroid-forming cells slowly proliferated and expressed high levels of Oct-4, Klf4, Bmi-1, Lgr5, IL-6, and Notch 3 compared with adherent cells. Treatment with an anti-human IL-6 receptor monoclonal antibody reduced spheroid formation, stem cell-related gene expression, and 5-fluorouracil (5-FU) resistance. In addition, IL-6 treatment enhanced the levels of p-STAT3 (Tyr705), the expression of Oct-4, Klf4, Lgr5, and Notch 3, and chemoresistance to 5-FU. siRNA targeting Notch 3 suppressed spheroid formation, Oct-4 and Lgr5 expression, and 5-FU chemoresistance, whereas STAT3 inhibition enhanced Oct-4, Klf4, Lgr5, and Notch 3 expression and 5-FU chemoresistance along with reduced spheroid growth. Taken together, these results indicate that IL-6 functions in dichotomous pathways involving Notch 3 induction and STAT3 activation. The former pathway is involved in cancer stem-like cell biology and enhanced chemoresistance, and the latter pathway leads to accelerated proliferation and reduced chemoresistance. Thus, an anti-human IL-6 receptor monoclonal antibody or Notch 3

  7. Pharmacokinetics, biodistribution and dosimetry of 99mTc-labeled anti-human epidermal growth factor receptor humanized monoclonal antibody R3 in rats.

    Science.gov (United States)

    Iznaga Escobar, N; Morales, A M; Ducongé, J; Torres, I C; Fernández, E; Gómez, J A

    1998-01-01

    The pharmacokinetics, biodistribution and dosimetry of 99mTc-labeled anti-human epidermal growth factor receptor (anti-hEGF-r) humanized monoclonal antibody (MAb) R3 was investigated following intravenous injection in normal Wistar rats. Serum disappearance curves were best fit by a two-compartment model having a mean distribution half-life (t 1/2alpha) of 0.250 h and a mean elimination (t 1/2beta) of 13.89 h. Among the various organs, a little accumulation of the radiolabeled antibody was found only in kidneys. Biodistribution and dosimetry studies in humans were performed by extrapolation of the animal data to humans. Absorbed dose to normal organs and the remainder of the whole body were estimated using the medical internal radiation dose formula, and dose contributions from radioactivity in transit through the gastrointestinal tract were estimated using a compartment model. Extrapolated values of radiation absorbed dose to normal organs in rads per millicurie administered were whole body, 0.0085; lower large intestine wall, 0.0898; small intestine, 0.0530; upper large intestine wall, 0.0731; and kidneys, 0.0455. The effective dose equivalent predicted was 0.0162 rem/mCi and the effective dose was found to be 0.015 rem/mCi. On the basis of the pharmacokinetics, biodistribution and internal radiation dosimetry information obtained in this study, a diagnostic phase I clinical trial with 99mTc-labeled humanized MAb R3 conjugate in patients should be supported.

  8. Anti-human tissue factor antibody ameliorated intestinal ischemia reperfusion-induced acute lung injury in human tissue factor knock-in mice.

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    Xiaolin He

    Full Text Available BACKGROUND: Interaction between the coagulation and inflammation systems plays an important role in the development of acute respiratory distress syndrome (ARDS. Anti-coagulation is an attractive option for ARDS treatment, and this has promoted development of new antibodies. However, preclinical trials for these antibodies are often limited by the high cost and availability of non-human primates. In the present study, we developed a novel alternative method to test the role of a humanized anti-tissue factor mAb in acute lung injury with transgenic mice. METHODOLOGY/PRINCIPAL FINDINGS: Human tissue factor knock-in (hTF-KI transgenic mice and a novel humanized anti-human tissue factor mAb (anti-hTF mAb, CNTO859 were developed. The hTF-KI mice showed a normal and functional expression of hTF. The anti-hTF mAb specifically blocked the pro-coagulation activity of brain extracts from the hTF-KI mice and human, but not from wild type mice. An extrapulmonary ARDS model was used by intestinal ischemia-reperfusion. Significant lung tissue damage in hTF-KI mice was observed after 2 h reperfusion. Administration of CNTO859 (5 mg/kg, i.v. attenuated the severity of lung tissue injury, decreased the total cell counts and protein concentration in bronchoalveolar lavage fluid, and reduced Evans blue leakage. In addition, the treatment significantly reduced alveolar fibrin deposition, and decreased tissue factor and plasminogen activator inhibitor-1 activity in the serum. This treatment also down-regulated cytokine expression and reduced cell death in the lung. CONCLUSIONS: This novel anti-hTF antibody showed beneficial effects on intestinal ischemia-reperfusion induced acute lung injury, which merits further investigation for clinical usage. In addition, the use of knock-in transgenic mice to test the efficacy of antibodies against human-specific proteins is a novel strategy for preclinical studies.

  9. Isolation and identification of CD4+CD25+ regulatory T cells in rat

    Institute of Scientific and Technical Information of China (English)

    Ling Lü; Feng Zhang; Liyong Pu; Chao Jiang

    2006-01-01

    Objective: To establish a stable and high efficient method for collection of CD4+CD25+ regulatory T cells from rats in vitro. Methods: CD4+CD25+ regulatory T cells were isolated from the rat splenic cells through two steps by magic cell sorting (MACS) system. The first step was negative selection of CD4+ T cells by cocktail antibodies and anti-IgG magic microbeads, and the second step was positive selection of CD25+ T cells by anti-CD25 PE and anti-PE magic microbeads. The purity and viability of separated cells were measured by flow cytometry (FACS) and Trypan blue staining. The suppressive ability of seperated cells on the proliferation of CD4+CD25- T cells was assessed by cell proliferation assay. Results: The purity of negatively enriched CD4+ T cells was 79%-87% (83.6% ± 2.5%) , and the purity of positively enriched CD4+CD25+ T cells was 86%-93% ( 90.2 ± 1.8%) with the viability of 92%-95% (92.8% ± 3.4%). The enriched cells significantly suppressed the proliferation of CD4+CD25- T cells in mixed lymphocyte culture (P < 0.05). Conclusion: An effective method can be established for enrichment of CD4+CD25+ regulatory T cells in two steps by MACS, with satisfied cell purity, viability and function.

  10. ARG098, a novel anti-human Fas antibody, suppresses synovial hyperplasia and prevents cartilage destruction in a severe combined immunodeficient-HuRAg mouse model

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    Matsubara Tsukasa

    2010-09-01

    Full Text Available Abstract Background The anti-human Fas/APO-1/CD95 (Fas mouse/human chimeric monoclonal IgM antibody ARG098 (ARG098 targets the human Fas molecule. The cytotoxic effects of ARG098 on cells isolated from RA patients, on normal cells in vitro, and on RA synovial tissue and cartilage in vivo using implanted rheumatoid tissues in an SCID mouse model (SCID-HuRAg were investigated to examine the potential of ARG098 as a therapy for RA. Methods ARG098 binding to each cell was analyzed by cytometry. The effects of ARG098 on several cells were assessed by a cell viability assay in vitro. Effects on the RA synovium, lymphocytes, and cartilage were assessed in vivo using the SCID-HuRAg mouse model. Results ARG098 bound to cell surface Fas molecules, and induced apoptosis in Fas-expressing RA synoviocytes and infiltrating lymphocytes in the RA synovium in a dose-dependent manner. However, ARG098 did not affect the cell viability of peripheral blood mononuclear cells of RA patients or normal chondrocytes. ARG098 also induced apoptosis in RA synoviocytes and infiltrating lymphocytes in the RA synovium in vivo. The destruction of cartilage due to synovial invasion was inhibited by ARG098 injection in the modified SCID-HuRAg mouse model. Conclusions ARG098 treatment suppressed RA synovial hyperplasia through the induction of apoptosis and prevented cartilage destruction in vivo. These results suggest that ARG098 might become a new therapy for RA.

  11. HAHA--nothing to laugh about. Measuring the immunogenicity (human anti-human antibody response) induced by humanized monoclonal antibodies applying ELISA and SPR technology.

    Science.gov (United States)

    Nechansky, Andreas

    2010-01-05

    Immunogenicity induced by passively applied proteins is a serious issue because it is directly related to the patient's safety. The out-come of an immune reaction to a therapeutic protein can range from transient appearance of antibodies without any clinical significance to severe life threatening conditions. Within this article, enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) methodology to measure immunogenicity are compared and the pros and cons are discussed.

  12. Expansion of CD25+ Innate Lymphoid Cells Reduces Atherosclerosis

    Science.gov (United States)

    Engelbertsen, Daniel; Foks, Amanda C.; Alberts-Grill, Noah; Kuperwaser, Felicia; Chen, Tao; Lederer, James A.; Jarolim, Petr; Grabie, Nir; Lichtman, Andrew H.

    2015-01-01

    Objective Innate lymphoid cells (ILCs) are a newly discovered subset of immune cells that promote tissue homeostasis and protect against pathogens. ILCs produce cytokines also produced by T lymphocytes that have been shown to affect atherosclerosis, but the influence of ILCs on atherosclerosis has not been explored. Approach and Results We demonstrate that CD25+ ILCs that produce type 2 cytokines (ILC2s) are present in the aorta of atherosclerotic immunodeficient ldlr−/−rag1−/− mice. To investigate the role of ILCs in atherosclerosis, ldlr−/−rag1−/− mice were concurrently fed an atherogenic diet and treated with either ILC-depleting anti-CD90.2 antibodies or with IL-2/anti-IL-2 complexes that expand CD25+ ILCs. Lesion development was not affected by anti-CD90.2 treatment, but was reduced in IL-2/anti-IL-2 -treated mice. These IL-2 treated mice had reduced VLDL cholesterol and increased triglycerides compared to controls and reduced apolipoprotein B100 gene expression in the liver. IL-2/anti-IL-2 treatment caused expansion of ILC2s in aorta and other tissues, elevated levels of IL-5, systemic eosinophila and hepatic eosinophilic inflammation. Blockade of IL-5 reversed the IL-2-complex-induced eosinophilia but did not change lesion size. Conclusions This study demonstrates that expansion of CD25-expressing ILCs by IL-2/anti-IL-2 complexes leads to a reduction in VLDL cholesterol and atherosclerosis. Global depletion of ILCs by anti-CD90.2 did not significantly affect lesion size indicating that different ILC subsets may have divergent effects on atherosclerosis. PMID:26494229

  13. Influence of CD4+CD25+ T cells on Plasmodium berghei NK65 infection in BALB/c mice.

    Science.gov (United States)

    Long, Ton That Ai; Nakazawa, Shusuke; Onizuka, Shozaburo; Huaman, Maria Cecilia; Kanbara, Hiroji

    2003-02-01

    CD4(+) T cells co-expressing CD25 (CD4(+)CD25(+) T cells) have been identified as immunoregulatory suppressors modulating autoimmune response. Beside that, autoimmune response was supposed to be associated with malaria infection. Based on these data, we hypothesised that CD4(+)CD25(+) T cells may influence protective immunity to malaria parasites, while suppressing autoimmune response arising throughout the course of malarial infection. To test this possibility, we evaluated the kinetics of CD4(+)CD25(+) T cells during malaria infection and investigated the influence of CD25 depletion by anti-mouse CD25 monoclonal antibody (PC61) on the infection, using a mouse model of premunition to Plasmodium berghei NK65 malaria. The results showed that, during exacerbation of P. berghei NK65 infection, the proportion of CD4(+)CD25(+) T cells among CD4(+) T cells decreased, although that of CD4(+) T cells increased. CD25 depletion clearly delayed the growth of parasitaemia during parasite challenge, particularly in immunised mice. These findings demonstrated that CD4(+)CD25(+) T cells are able to influence protective immunity underlying premunition to P. berghei NK65 parasites.

  14. Efficient induction of CD25- iTreg by co-immunization requires strongly antigenic epitopes for T cells

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    Li Jinyao

    2011-05-01

    Full Text Available Abstract Background We previously showed that co-immunization with a protein antigen and a DNA vaccine coding for the same antigen induces CD40low IL-10high tolerogenic DCs, which in turn stimulates the expansion of antigen-specific CD4+CD25-Foxp3+ regulatory T cells (CD25- iTreg. However, it was unclear how to choose the antigen sequence to maximize tolerogenic antigen presentation and, consequently, CD25- iTreg induction. Results In the present study, we demonstrated the requirement of highly antigenic epitopes for CD25- iTreg induction. Firstly, we showed that the induction of CD25- iTreg by tolerogenic DC can be blocked by anti-MHC-II antibody. Next, both the number and the suppressive activity of CD25- iTreg correlated positively with the overt antigenicity of an epitope to activate T cells. Finally, in a mouse model of dermatitis, highly antigenic epitopes derived from a flea allergen not only induced more CD25- iTreg, but also more effectively prevented allergenic reaction to the allergen than did weakly antigenic epitopes. Conclusions Our data thus indicate that efficient induction of CD25- iTreg requires highly antigenic peptide epitopes. This finding suggests that highly antigenic epitopes should be used for efficient induction of CD25- iTreg for clinical applications such as flea allergic dermatitis.

  15. CD4+CD25high Regulatory Cells in Peripheral Blood of NSCLC Patients

    Institute of Scientific and Technical Information of China (English)

    LIU Li; YAO Junxia; DING Qian; HUANG Shiang

    2006-01-01

    The proportion and changes of CD4+CD25high regulatory T cells (Trs) in peripheral blood of non-small cell lung cancer (NSCLC) patients were analyzed and their clinical significance explored. The peripheral blood was collected from 61 patients with NSCLC and 15 healthy controls. By using monoclonal antibodies, the blood samples were evaluated with the flow cytometry for lymphocyte subsets (CD3+, CD4+ and CD8+) and CD4+CD25high Tr cells. The results showed that the proportion of CD4+CD25high Tr cells in NSCLC group was significantly higher than in control group [(4.36±2.07) % vs (2.04±1.03) %, P<0.01]. The proportion of CD4+CD25 high Tr cells in late stage was higher than that in early stage [stages Ⅰ + Ⅱ (2.26±0.6) %; stage Ⅲ (3.28±1.38) %; stage Ⅳ(6.06±4.08) %] (P<0.05). Kaplan-Meier survival analysis revealed that the prognosis of the patients who had higher proportion of CD4+CD25high Tr cells in peripheral blood was worse (P=0.0026). In conclusion, the relative increase in CD4+CD25high Tr cells in peripheral blood may be related to immunosuppression and tumor progression in patients with NSCLC. This finding suggests that CD4+CD25+high Tr cells in peripheral blood of NSCLC may be positive for prognosis analysis. The use of depletion of the CD4+CD25high Tr cell therapy to treat NSCLC patients may be an effective strategy.

  16. Depletion of CD25+ cells during acute toxoplasmosis does not significantly increase mortality in Swiss OF1 mice

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    Haroon Akbar

    2012-03-01

    Full Text Available The interleukin (IL-2R alpha chain (CD25 is expressed on regulatory T cells (Treg, which constitute more than 85% of the CD25+ T cell population in a naïve mouse. CD25 is also expressed on effector T cells in mice suffering from an acute infection by the obligate intracellular protozoan parasite, Toxoplasma gondii. Lethal toxoplasmosis is accompanied by a significant loss of Treg in mice naturally susceptible to toxoplasmosis. The present study was done to explore the role of Treg cells using an anti-CD25 antibody-mediated depletion in mice naturally resistant to toxoplasmosis. Although a significant decrease in the percentage of Treg cells was observed following anti-CD25 monoclonal antibody injections, the depletion of CD25+ cells during acute toxoplasmosis did not significantly increase the mortality of Swiss OF1 mice and no significant difference was observed in the brain parasitic load between the mice in the depleted-infected and isotype-infected groups. We found no significant difference between the titres of total IgG in the sera of the mice from the two groups in the chronic phase. However, CD25+ cells depletion was followed by significantly higher levels of IL-12 in the serum of depleted mice than in that of mice injected with the isotype control antibody.

  17. Large Scale Generation and Characterization of Anti-Human IgA Monoclonal Antibody in Ascitic Fluid of Balb/c Mice

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    Fatemeh Ezzatifar

    2015-03-01

    Full Text Available Purpose: Monoclonal antibodies are potentially powerful tools used in biomedical research, diagnosis, and treatment of infectious diseases and cancers. The monoclonal antibody against Human IgA can be used as a diagnostic application to detect infectious diseases. The aim of this study was to improve an appropriate protocol for large-scale production of mAbs against IgA. Methods: For large-scale production of the monoclonal antibody, hybridoma cells that produce monoclonal antibodies against Human IgA were injected intraperitoneally into Balb/c mice that were previously primed with 0.5 ml Pristane. After ten days, ascitic fluid was harvested from the peritoneum of each mouse. The ELISA method was carried out for evaluation of the titration of produced mAbs. The ascitic fluid was investigated in terms of class and subclass by a mouse mAb isotyping kit. MAb was purified from the ascitic fluid by ion exchange chromatography. The purity of the monoclonal antibody was confirmed by SDS-PAGE, and the purified monoclonal antibody was conjugated with HRP. Results: Monoclonal antibodies with high specificity and sensitivity against Human IgA were prepared by hybridoma technology. The subclass of antibody was IgG1 and its light chain was the kappa type. Conclusion: This conjugated monoclonal antibody could have applications in designing ELISA kits in order to diagnose different infectious diseases such as toxoplasmosis and H. Pylori.

  18. Large Scale Production and Characterization of Anti-Human IgG Monoclonal Antibody in Peritoneum of Balb/c MICE

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    B. Baradaran

    2005-01-01

    Full Text Available Monoclonal antibodies are key reagents that are used in biomedical researches, diagnosis of immunodeficiency diseases such as IgG subclasses deficiency and treatment of diseases like infections and cancers .For large scale production of monoclonal antibody, hybridoma cells that produce monoclonal antibody against human IgG were injected into the peritoneum of the Balb/c mice which have previously been primed with 0.5 ml Pristane. After ten days, approximately 3 ml ascitic fluid was harvested from the peritoneum of each mouse. Ascitic fluid was assayed for the titer of monoclonal antibody in reaction with human IgG and its cross reactivity in reaction with IgM & IgA. The titer of mAb was 100,000 and didn't show cross reactivity with IgM & IgA. Immunobloting was done for confirming the ELISA method. In immunobloting, only one sharp band in the heavy chain position of IgG was developed. The subclass of antibody was IgG1 and its light chain was kappa. Ascitic fluid was purified by ion exchange chromatography and the purified monoclonal antibody was conjugated with HRP. The conjugated monoclonal antibody could have application in diagnosis of infectious diseases like Toxoplasmosis, Rubella and IgG class of all other infectious diseases.

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

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    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.

  20. Effect of an anti-human Co-029/tspan8/Tspan8 mouse monoclonal antibody on tumour growth in a nude mouse model

    Directory of Open Access Journals (Sweden)

    Naouel eAilane

    2014-09-01

    Full Text Available New therapeutic agents are needed in digestive tract tumours. Co-029/tspan8 is a tetraspanin frequently expressed on human colorectal tumours, In this work, we report the effects of the monoclonal antibody Ts29.2, targeting Co-029/tspan8, on colorectal tumor cells in vitro and after implantation in nude mice. HT29, Isreco1 and SW480 colorectal tumor cell lines were used for this study. HT29 has a strong endogenous expression of Co-029/tspan8, whereas Isreco1 cells don’t express Co-029/tspan8 and SW480 has only a weak expression. Isreco1 and SW480 were transduced to express Co-029/tspan8 at the same level as HT29. In order to check the specificity of the effect of monoclonal antibody Ts29.2, low Co029/tspan8 expressing SW480 cells were injected simultaneously with transduced cells in the back, on the left and right sides of the mice. With an early treatment, Ts29.2 mAb inhibited growth of tumors expressing Co-029/tspan8 up to 70%, whereas a delayed treatment was less efficient. No effect of the antibody on cell proliferation or apoptosis induction was detected in vitro. No increase of activated caspase 3 labeling was observed in vivo and areas occupied by vessels were not significantly different between treated mice and controls. This suggests that the action of Ts29.2 is linked neither to cellular toxicity nor to the inhibition of the previously reported angiogenic properties of Co-029/tspan8. An inhibition of cell proliferation in vivo is demonstrated by a reduction of the mitotic index in HT29 tumors of Ts29.2 treated mice. The discrepancy between in vitro and in vivo data on cell proliferation suggests that the binding of Ts29.2 to tumour cells may modify their response to signals issued from the microenvironment. Given the restricted pattern of tissue expression of the tetraspanin Co-029/tspan8, these preliminary results put forth for consideration the antibody targeting of this tetraspanin in further investigations for therapeutic

  1. Mother-infant transfer of anti-human papillomavirus (HPV) antibodies following vaccination with the quadrivalent HPV (type 6/11/16/18) virus-like particle vaccine.

    Science.gov (United States)

    Matys, Katie; Mallary, Sara; Bautista, Oliver; Vuocolo, Scott; Manalastas, Ricardo; Pitisuttithum, Punee; Saah, Alfred

    2012-06-01

    The exploratory immunogenicity objective of this analysis was to characterize the titer of vaccine human papillomavirus (HPV)-type immunoglobulins in both peripartum maternal blood and the cord blood of infants born to women who received blinded therapy. Data were derived from a randomized, placebo-controlled, double-blind safety, immunogenicity, and efficacy study (protocol 019; NCT00090220). This study enrolled 3,819 women between the ages of 24 and 45 years from 38 international study sites between 18 June 2004 and 30 April 2005. Data in the current analysis are from subjects enrolled in Philippines and Thailand. For each of HPV types 6, 11, 16, and 18, maternal anti-HPV was found in cord blood samples. Furthermore, HPV titers in cord blood samples were highly positively correlated with maternal HPV titers. Additionally, there were instances when anti-HPV antibodies were no longer detectable in maternal serum samples and yet were detected in matched cord blood samples. These results demonstrate that quadrivalent HPV (qHPV) vaccine-induced antibodies cross the placenta and could potentially provide some benefit against vaccine-type HPV infection and related diseases such as recurrent respiratory papillomatosis.

  2. 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.

  3. 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.

  4. 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.

  5. Analysis of Epstein-Barr viral DNA load, EBV-LMP2 specific cytotoxic T-lymphocytes and levels of CD4+CD25+T cells in patients with nasopharyngeal carcinomas positive for IgA antibody to EBV viral capsid antigen

    Institute of Scientific and Technical Information of China (English)

    MO Wu-ning; TANG An-zhou; ZHOU Ling; HUANG Guang-wu; WANG Zhan; ZENG Yi

    2009-01-01

    Background Epstein-Barr virus (EBV) is a herpesvirus commonly associated with several malignant diseases including nasopharyngeal carcinoma (NPC), which is a common cancer in Southeastem Asia. Previous studies showed that plasma levels of EBV-DNA might be a sensitive and reliable biomarker for the diagnosis, staging and evaluating of therapy for NPC. There are a few analyses of the levels of EBV-latent membrane protein 2 (LMP2)-specific cytotoxic T-lymphocytes (CTLs) in patients with NPC. This study was conducted to investigate the levels of EBV-LMP2-specific CTLs, EBV-DNA load and the level of CD4+CD25+T cells in such patients.Methods From February 2006 to April 2006, 62 patients with NPC, 40 healthy virus carders positive for EBV viral capsid antigen (EBV-IgA-VCA) and 40 controls were enrolled in the study. We used a highly sensitive ELISPOT assay,real-time polymerase chain reaction (PCR) and flow cytometry to measure the EBV-LMP2-specific CTL response, the EBV DNA load and the level of CD4+CD25+T cells, respectively.Results The EBV-LMP2-specific CTL responses of the samples from the control, healthy virus carders and patients with NPC were significantly different from the LMP2 epitopes, with the control and healthy virus carder samples displaying a stronger response in three cases. There were significant differences in EBV DNA load in serum between NPC and the healthy groups; patients with NPC at stages Ⅲ or Ⅳ had significantly higher viral loads compared with those at stages Ⅰ or Ⅱ. A significantly higher percentage of CD4+CD25+ T lymphocytes were detected in the patients, compared with healthy virus carriers and healthy controls. Moreover, patients with advanced stages of NPC (Ⅲ and Ⅲ) had significantly higher percentages than the patients with early stages (Ⅰ and Ⅱ).Conclusions Patients with NPC are frequently unable to establish or maintain sufficient immunosurveillance to control proliferating B cells harboring EBV and to destroy the tumor

  6. 调节性CD4~+CD25~+T细胞的分离纯化及免疫功能研究%Isolation purification and function analysis of CD4~+CD25~+regulatory T cells

    Institute of Scientific and Technical Information of China (English)

    杨阳; 韩岩; 杨麦贵; 夏炜; 庄然; 郭树忠

    2009-01-01

    Objective To prepare CD4~+CD25~+regulatory T cells (Treg) and analyze their immune function in order to prevent and treat rejection of composite tissue allotransplantation (CTA) by inducing local immunotolence. Methods The CD4~+CD25~ +Treg (1×10~6) were isolated from the rat spleens,and sorted with magnetic bead by magnetic activated cell sorting (MACS) system. The activity of CD4~+CD25~+Treg was assayed by Trypan blue (2% ) staining,and their purity was analyzed by flow cytometry (FCM). The relationship between IL-2 (200 U/ml) and CD4~+CD25~+Treg reactivity to stimuli by CD3 (5 mg/L) antibody was observed. Results The mean activity and purity of CD4~+CD25~+Treg isolated from 8 male rats were (97.90±0.36)% and (96.05±0.41)% respectively. The lower reactivity to CD3 stimuli was found with cultivation inhibition ratio of 89%. IL-2 could reverse the inhibition. Conclusion The CD4~+CD25~+Treg with higher purity can be isolated quickly by MACS, and the cells are anergic immuno-suppression in vitro. This can fit the demand of investigation on animal CTA rejection.%目的 制备调节性CD~+ CD25~+T细胞(Treg)分析其免疫功能,诱导局部免疫耐受防治同种异体复合组织移植(CTA)排斥反应.方法 采用免疫磁珠法(MACS)从雄性大鼠脾脏细胞分离CD4~+CD25~+Treg(1×10~6),2%锥虫蓝染色检测活性、流式细胞术分析其纯度,在5 mg/L抗CD3的刺激下观察其反应性、增殖及其与200 U/ml细胞介素(IL)-2的关系.结果 从8只雄性大鼠脾脏分选出的CD4~+CD25~+Treg活性平均为(97.90±0.36)%及纯度为(96.05±0.41)%,CD3刺激呈低反应,按比例培养抑制率为89%,IL-2可使CD4~+CD25~-抑制逆转.结论 MACS能快速分选出较高纯度的CD4~+CD25~+Treg,并且活性良好在体外具有免疫无能及免疫抑制作用,能满足动物CTA排斥反应研究的需要.

  7. Influence of membrane CD25 stability on T lymphocyte activity: implications for immunoregulation.

    Directory of Open Access Journals (Sweden)

    Todd M Brusko

    Full Text Available BACKGROUND: CD25, a component of the IL-2 receptor, is important in T cell proliferation, activation induced cell death, as well as the actions of both regulatory (Treg and effector (Teff T cells. Recent genome wide association studies have implicated the CD25 locus as an important region for genetic susceptibility to a number of autoimmune disorders, with serum levels of soluble CD25 receptor (sCD25 serving as a potential phenotypic marker for this association. However, the functional impact of CD25 cleavage, as well as the influence of sCD25 on immunoregulatory activities, remain largely unknown and form the basis of this effort. METHODOLOGY/PRINCIPAL FINDINGS: The generation of sCD25 by Treg (CD4(+CD25(+ and Teff (CD4(+CD25(- cells was examined during in vitro suppression assays, efforts that demonstrated constitutive and stable surface CD25 expression on Treg throughout the period of in vitro assessment. In contrast, Teff cells increased CD25 expression during the process of in vitro suppression, with supernatant sCD25 levels correlating to the amount of cellular proliferation. Interestingly, under serum-free conditions, Tregs partially lost their characteristic anergic and suppressive properties. sCD25 supplementation at physiological concentrations to serum free in vitro suppression assays reduced Teff proliferation without specifically influencing suppression. Indeed, sCD25 production within these cultures correlated with cell death. CONCLUSIONS/SIGNIFICANCE: These results support the notion that sCD25 functions as both a surrogate marker of T cell activation as well as an indicator of subsequent cellular death. In addition, the role of CD25 in immunomodulation is likely dependent on the local inflammatory milieu, with molecules capable of modulating surface CD25 expression playing a key role in defining immune responsiveness.

  8. TLR5 signaling enhances the proliferation of human allogeneic CD40-activated B cell induced CD4hiCD25+ regulatory T cells.

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    Ping-Lung Chan

    Full Text Available Although diverse functions of different toll-like receptors (TLR on human natural regulatory T cells have been demonstrated recently, the role of TLR-related signals on human induced regulatory T cells remain elusive. Previously our group developed an ex vivo high-efficient system in generating human alloantigen-specific CD4(hiCD25(+ regulatory T cells from naïve CD4(+CD25(- T cells using allogeneic CD40-activated B cells as stimulators. In this study, we investigated the role of TLR5-related signals on the generation and function of these novel CD4(hiCD25(+ regulatory T cells. It was found that induced CD4(hiCD25(+ regulatory T cells expressed an up-regulated level of TLR5 compared to their precursors. The blockade of TLR5 using anti-TLR5 antibodies during the co-culture decreased CD4(hiCD25(+ regulatory T cells proliferation by induction of S phase arrest. The S phase arrest was associated with reduced ERK1/2 phosphorylation. However, TLR5 blockade did not decrease the CTLA-4, GITR and FOXP3 expressions, and the suppressive function of CD4(hiCD25(+ regulatory T cells. In conclusion, we discovered a novel function of TLR5-related signaling in enhancing the proliferation of CD4(hiCD25(+ regulatory T cells by promoting S phase progress but not involved in the suppressive function of human CD40-activated B cell-induced CD4(hiCD25(+ regulatory T cells, suggesting a novel role of TLR5-related signals in the generation of induced regulatory T cells.

  9. CD4+CD25+ regulatory T lymphocytes in tuberculous pleural effusion

    Institute of Scientific and Technical Information of China (English)

    QIN Xue-jun; SHI Huan-zhong; LIANG Qiu-li; HUANG Lu-ying; YANG Hai-bo

    2008-01-01

    Background Active suppression by CD4+CD25+ regulatory T lymphocytes plays an important role in the down-regulation of T cell responses to foreign and self-antigens.This study was conducted to analyze whether the CD4+CD25+ regulatory T cells exist and function normally in tuberculous pleural effusion.Methods The percentages of CD4+CD25+ T cells in pleural effusion and peripheral blood from patients with tuberculous pleurisy and peripheral blood from healthy control subjects were determined by flow cytometry.The expression of forkhead transcription factor Foxp3 was also examined.CD4+CD25+ and CD4+CD25- T cells from pleural effusion and blood were isolated,and were cultured to observe the effects Of CD4+CD25+ T cells on proliferation response of CD4+CD25- T cells in Vitro.Results There were increased numbers of CD4+CD25+ T cells in tuberculous pleural effusion compared with peripheral blood from both patients with tuberculous pleurisy and normal subjects,and these cells demonstrated a constitutive high-level expression of Foxp3.Moreover,CD4+CD25+ T cells mediated potent inhibition of proliferation response of CD4+CD25- T cells.Conclusion The increased CD4+CD25+ T cells in tuberculous pleu ral effusion express a high level of Foxp3 transcription factor,while potently suppressing the proliferation of CD4+CD25- T cells.

  10. The Influence of Propofol-Isoflurane Combined Anaesthesia on the T Cell Subpopulations and CD25 Expression%异丙酚-异氟醚复合麻醉对CD25表达及T细胞亚群影响的临床研究

    Institute of Scientific and Technical Information of China (English)

    刘菊英; 田玉科; 骆明恩; 张吉才

    2001-01-01

    The changes of T cell subpopulations (CD+4,CD+8 cell) and CD25+cells and CD25 expression in 40 patients undergoing elective cholecystectomy using either epidural or propofol and isoflurane combined anaesthesia (general anaesthesia) were investigated. Mononuclear cells and erythrocytes were simultaneously isolated before anaesthesia and during operation (60 min after anesthesia induction). Monoclonal antibodies to CD+3,CD+4,CD+ 8 T cells of the T cell subpopulations were identified. CD25 was assessed by immunohistochemistry method. In the patients receiving general anaesthesia,the percentages CD+3,CD+4 CD+8 of the cells were all increased,which were accompanied with a rise of CD25+ cell numbers (P0.05)。CD+3细胞变化两组间具有显著差异(P0.05)。但全麻组麻醉后CD25细胞明显增加(P<0.05)。认为两种麻醉方法对T细胞亚群及淋巴细胞CD25的影响不同。与硬膜外麻醉相比,异丙酚-异氟醚静吸复合全麻使T细胞免疫处于易激活状态。

  11. Characterization of Foxp3+CD4+CD25+ and IL-10-secreting CD4+CD25+ T cells during cure of colitis.

    Science.gov (United States)

    Uhlig, Holm H; Coombes, Janine; Mottet, Christian; Izcue, Ana; Thompson, Claire; Fanger, Andrea; Tannapfel, Andrea; Fontenot, Jason D; Ramsdell, Fred; Powrie, Fiona

    2006-11-01

    CD4+CD25+ regulatory T cells can prevent and resolve intestinal inflammation in the murine T cell transfer model of colitis. Using Foxp3 as a marker of regulatory T cell activity, we now provide a comprehensive analysis of the in vivo distribution of Foxp3+CD4+CD25+ cells in wild-type mice, and during cure of experimental colitis. In both cases, Foxp3+CD4+CD25+ cells were found to accumulate in the colon and secondary lymphoid organs. Importantly, Foxp3+ cells were present at increased density in colon samples from patients with ulcerative colitis or Crohn's disease, suggesting similarities in the behavior of murine and human regulatory cells under inflammatory conditions. Cure of murine colitis was dependent on the presence of IL-10, and IL-10-producing CD4+CD25+ T cells were enriched within the colon during cure of colitis and also under steady state conditions. Our data indicate that although CD4+CD25+ T cells expressing Foxp3 are present within both lymphoid organs and the colon, subsets of IL-10-producing CD4+CD25+ T cells are present mainly within the intestinal lamina propria suggesting compartmentalization of the regulatory T cell response at effector sites.

  12. 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

    . Upon enteroantigen activation, the CD4+ CD25- T cells secrete IL-4, IL-5, IL-10, granulocyte macrophage colony-stimulating factor, tumor necrosis factor-alpha and IFN-gamma. Clonotype mapping of the TCRBV regions 1-18 of enteroantigen-reactive CD4+ CD25- T cells by TCR clonotype mapping revealed...

  13. 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.

  14. A phase I study of CD25/regulatory T-cell-depleted donor lymphocyte infusion for relapse after allogeneic stem cell transplantation

    Science.gov (United States)

    Nikiforow, Sarah; Kim, Haesook T.; Daley, Heather; Reynolds, Carol; Jones, Kyle Thomas; Armand, Philippe; Ho, Vincent T.; Alyea, Edwin P.; Cutler, Corey S.; Ritz, Jerome; Antin, Joseph H.; Soiffer, Robert J.; Koreth, John

    2016-01-01

    Donor lymphocyte infusions are used to treat relapse after allogeneic hematopoietic stem cell transplantation, but responses are inadequate. In addition to effector cells, infusions contain CD25+ regulatory T cells (Treg) that may suppress graft-versus-tumor responses. We undertook a phase I study of donor lymphocyte infusions depleted of CD25+ T cells in patients with hematologic malignancies who had relapsed after transplantation. Twenty-one subjects received CD25/Treg-depleted infusions following removal of CD25+ cells using antibody-conjugated magnetic beads. Sixteen subjects received prior cytoreductive therapy. Four were in complete remission at the time of infusion. Two dose levels were administered: 1×107 (n=6) and 3×107 CD3+ cells/kg (n=15). A median 2.3 log-depletion of CD4+CD25+FOXP3+ Treg was achieved. Seven subjects (33%) developed clinically significant graft-versus-host disease by 1 year, including one patient who died. At dose level 1, five subjects had progressive disease and one had stable disease. At dose level 2, nine subjects (60%) achieved or maintained responses (8 complete responses, 1 partial response), including seven with active disease at the time of infusion. A shorter period between relapse and infusion was associated with response at dose level 2 (P=0.016). The 1-year survival rate was 53% among patients treated with dose level 2. Four of eight subjects with acute myeloid leukemia remained in remission at 1 year. When compared to unmodified donor lymphocyte infusions in 14 contemporaneous patients meeting study eligibility, CD25/Treg depletion was associated with a better response rate and improved event-free survival. Circulating naïve and central memory CD4+ T cells increased after CD25/Treg-depleted infusion, but no immunophenotypic signature for response was noted. CD25/Treg-depleted donor infusion appears feasible and capable of inducing graft-versus-tumor responses without excessive graft-versus-host disease. (Clinical

  15. The 1.7 A X-ray crystal structure of the porcine factor VIII C2 domain and binding analysis to anti-human C2 domain antibodies and phospholipid surfaces.

    Directory of Open Access Journals (Sweden)

    Caileen M Brison

    Full Text Available The factor VIII C2 domain is essential for binding to activated platelet surfaces as well as the cofactor activity of factor VIII in blood coagulation. Inhibitory antibodies against the C2 domain commonly develop following factor VIII replacement therapy for hemophilia A patients, or they may spontaneously arise in cases of acquired hemophilia. Porcine factor VIII is an effective therapeutic for hemophilia patients with inhibitor due to its low cross-reactivity; however, the molecular basis for this behavior is poorly understood. In this study, the X-ray crystal structure of the porcine factor VIII C2 domain was determined, and superposition of the human and porcine C2 domains demonstrates that most surface-exposed differences cluster on the face harboring the "non-classical" antibody epitopes. Furthermore, antibody-binding results illustrate that the "classical" 3E6 antibody can bind both the human and porcine C2 domains, although the inhibitory titer to human factor VIII is 41 Bethesda Units (BU/mg IgG versus 0.8 BU/mg IgG to porcine factor VIII, while the non-classical G99 antibody does not bind to the porcine C2 domain nor inhibit porcine factor VIII activity. Further structural analysis of differences between the electrostatic surface potentials suggest that the C2 domain binds to the negatively charged phospholipid surfaces of activated platelets primarily through the 3E6 epitope region. In contrast, the G99 face, which contains residue 2227, should be distal to the membrane surface. Phospholipid binding assays indicate that both porcine and human factor VIII C2 domains bind with comparable affinities, and the human K2227A and K2227E mutants bind to phospholipid surfaces with similar affinities as well. Lastly, the G99 IgG bound to PS-immobilized factor VIII C2 domain with an apparent dissociation constant of 15.5 nM, whereas 3E6 antibody binding to PS-bound C2 domain was not observed.

  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. T cells and T-cell subsets in mycosis fungoides and parapsoriasis. A study of 18 cases with anti-human T-cell monoclonal antibodies and histochemical techniques.

    Science.gov (United States)

    Buechner, S A; Winkelmann, R K; Banks, P M

    1984-07-01

    Skin lesions from 15 patients with mycosis fungoides (MF) and from three with parapsoriasis were studied immunohistochemically with monoclonal antibodies against T cells (Leu 1) and against T-cell subsets (Leu 2a, Leu 3a). Lymphoid cell reactivity was diverse among these sampled cases. In two cases of parapsoriasis and nine of MF, there was a predominance of helper/inducer (Leu-3a-reactive) cells over suppressor/cytotoxic (Leu-2a-reactive) cells. In one case of parapsoriasis and one (advanced tumor stage) of MF, there was suppressor/cytotoxic cell predominance. One case of MF showed strong reactivity for both T-cell subset markers. Four cases of MF (two plaque-stage and two tumor-stage) featured a predominant cell type in the dermis which was nonreactive for all three antibodies. The intraepidermal lymphoid cellularity was Leu-1-reactive in ten cases of MF and two of parapsoriasis. Among these 12 cases, the intraepidermal cellularity was Leu-2a-reactive in four and Leu-3a-reactive in three. The use of such studies of T-cell subsets on in situ cutaneous lymphoid infiltrates may demonstrate a correlation with cytomorphology, clinical stage, and disease prognosis.

  18. 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.

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

    Science.gov (United States)

    Jun, Cao; Qingshu, Li; Ke, Wei; Ping, Li; Jun, Dong; Jie, Luo; Su, Min

    2015-01-01

    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. PMID:26273136

  20. Daily subcutaneous injections of peptide induce CD4+ CD25+ T regulatory cells.

    Science.gov (United States)

    Dahlberg, P E; Schartner, J M; Timmel, A; Seroogy, C M

    2007-08-01

    Peptide immunotherapy is being explored to modulate varied disease states; however, the mechanism of action remains poorly understood. In this study, we investigated the ability of a subcutaneous peptide immunization schedule to induce of CD4(+) CD25(+) T regulatory cells. DO11.10 T cell receptor (TCR) transgenic mice on a Rag 2(-/-) background were injected subcutaneously with varied doses of purified ovalbumin (OVA(323-339)) peptide daily for 16 days. While these mice have no CD4(+) CD25(+) T regulatory cells, following this injection schedule up to 30% of the CD4(+) cells were found to express CD25. Real-time quantitative polymerase chain reaction (QPCR) analysis of the induced CD4(+) CD25(+) T cells revealed increased expression of forkhead box P3 (FoxP3), suggesting that these cells may have a regulatory function. Proliferation and suppression assays in vitro utilizing the induced CD4(+) CD25(+) T cells revealed a profound anergic phenotype in addition to potent suppressive capability. Importantly, co-injection of the induced CD4(+) CD25(+) T cells with 5,6-carboxy-succinimidyl-fluorescence-ester (CFSE)-labelled naive CD4(+) T cells (responder cells) into BALB/c recipient mice reduced proliferation and differentiation of the responder cells in response to challenge with OVA(323-339) peptide plus adjuvant. We conclude that repeated subcutaneous exposure to low-dose peptide leads to de novo induction of CD4(+) CD25(+) FoxP3(+) T regulatory cells with potent in vitro and in vivo suppressive capability, thereby suggesting that one mechanism of peptide immunotherapy appears to be induction of CD4(+) CD25(+) Foxp3(+) T regulatory cells.

  1. CD+4CD+25调节性T细胞与肿瘤的相关性研究%CD+4 CD+25 regulatory T cells and lung cancer

    Institute of Scientific and Technical Information of China (English)

    翟晋芳; 韩福才

    2009-01-01

    通过了解CD+4 CD+25调节性T细胞(CD+4 CD+25Treg)表面分子的特性和CD+4 CD+25Treg在外周血和组织中的表达,认识CD+4 CD+25 Treg在肿瘤免疫凋节中的作用,探索其作用的分子机制.%To know CD+4 CD+25 regulatory T cells' s function in tumor immunological regulation and to search for its functionary molecule mechanism by reviewing researches associated with the characteristics of CD+4 CD+25 regulatory T cells surface molecules and the expression of CD+4 CD+25 regulatory T cells in the peripheral blood and tissues.

  2. 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.

  3. 鼠抗人血小板CD36分子单克隆抗体的制备及活性分析%Preparation and Activity Analysis of Mouse Anti Human Platelet CD36 Monoclonal Antibody

    Institute of Scientific and Technical Information of China (English)

    陈麟凤; 张杰; 杨嘉慧; 罗圆圆; 庄远; 李卉; 冯倩; 汪德清

    2013-01-01

    amplified by RT-PCR, sequencing of the cDNA indicated the sequance was exactly the same to that in Genbank NM_001001547.2. The HEK293 cells with the plasmid were transfected, and SDS-PAGE confirmed that the transfect HEK293 cells expressed the human CD36 antigen extracellular protein fragments. Western-blot showed that the monoclonal antibody could recognize the recombinant CD36 with the sensitivity of 8 ng. It is concluded that the CD36 Gly30-Asn439 can be highly expressed by human embryonic kidney cells (HEK293), and the monoclonal antibody with biological activity has been obtained, which provide the basis for further study on platelet transfusion refractoriness.

  4. Evaluation of PLGA containing anti-CTLA4 inhibited endometriosis progression by regulating CD4+CD25+Treg cells in peritoneal fluid of mouse endometriosis model.

    Science.gov (United States)

    Liu, Qi; Ma, Pingchuan; Liu, Lanxia; Ma, Guilei; Ma, Jingjing; Liu, Xiaoxuan; Liu, Yijin; Lin, Wanjun; Zhu, Yingjun

    2017-01-01

    Our study investigated poly(lactic-co-glycolic acid) (PLGA) as protein delivery vehicles encapsulate CTLA-4-antibody (anti-CTLA-4) which is essential for CD4+CD25+Treg cells suppressive function exposing superior potential for inhibiting endometriosis progress in mouse model than single anti-CTLA-4. Anti-CTLA-4 loaded PLGA combined to ligands CTLA-4 in surface of CD4+CD25+Treg cells which distributed in peritoneal fluid of mouse endometriosis model. The particle size, zeta potential of the anti-CTLA-4 loaded nanoparticles was detected by dynamic light scattering. Morphology of nanoparticles was evaluated by transmission electron microscopy (TEM). Confocal laser scanning microscopy (CLSM) indicated distribution of anti-CTLA-4 with PLGA or without in peritoneal fluid. Cumulative anti-CTLA-4 release from nanoparticles was evaluated by Micro BCA assay. The percentage of CD4+CD25+Treg cells in peritoneal fluid was demonstrated by flow cytometer. In vitro experiment we co-culture ectopic endometrial cells (EEC) with isolated CD4+CD25+Treg cells in peritoneal fluid (PF), proliferation and invasion of ectopic endometrial cells (EEC) was measured by BrdU ELISA assay and Matrigel invasion assay. In comparison with anti-CTLA-4 without nanoparticles, the bioconjugates PLGA/anti-CTLA-4 were tolerated in peritoneal fluid with a controlled release of anti-CTLA-4 in 3, 7, 14days. Moreover, PLGA/anti-CTLA-4 had superior protective regulation ability to reduce level of CD4+CD25+Treg cells in peritoneal fluid. Most strikingly, in vitro experiment, PLGA/anti-CTLA-4 exhibited better ability in inhibiting proliferation and invasion of ectopic endometrial cells in co-culture system compared with anti-CTLA-4. Progressively, PLGA/anti-CTLA-4 had better suppressive activity to inhibited IL-10 and TGF-beta secreted by CD4+CD25+Treg cells which indicating that PLGA/anti-CTLA-4 suppressed cells proliferation and invasion through reduced IL-10 and TGF-beta production. Thus, PLGA/anti-CTLA-4 may

  5. Lenalidomide potentiates CD4(+)CD25(+)Treg-related suppression of lymphoma B-cell proliferation.

    Science.gov (United States)

    Grygorowicz, Monika Anna; Borycka, Ilona Sara; Nowak, Eliza; Paszkiewicz-Kozik, Ewa; Rymkiewicz, Grzegorz; Błachnio, Katarzyna; Biernacka, Marzena; Bujko, Mateusz; Walewski, Jan; Markowicz, Sergiusz

    2016-03-10

    We have previously found that ex vivo expanded human CD4(+)CD25(+)Treg cells suppress proliferation of lymphoma B-cell lines. Here we demonstrate that the immunomodulatory drug lenalidomide potentiates suppression of lymphoma B-cell proliferation by freshly isolated CD4(+)CD25(+)Tregs, as well as suppression by Tregs expanded polyclonally in the presence of rapamycin from CD4(+)CD25(+)T cells or CD4(+)CD25(+)CD127(lo)T cells. The regulation of lymphoma cell proliferation by Tregs pre-expanded with "third-party" allogeneic MoDCs in the presence of rapamycin was also potentiated by lenalidomide. Lenalidomide contributed to the suppression exerted by Tregs despite concomitant downregulation of Treg proliferation. Lenalidomide did not reduce the suppression of conventional T cells by expanded Tregs. The exposure of polyclonally expanded Tregs to lenalidomide did not significantly alter their phenotype. There was no uniform pattern of lenalidomide effect on Treg-mediated regulation of lymphoma B cells freshly isolated from patients. Freshly isolated lymphoma cells activated with multimeric CD40L and IL-4 to support their survival in vitro varied in their sensitivity to lenalidomide, and the regulatory effect of Tregs on such lymphoma cells ranged from suppression to help in individual patients. Lenalidomide potentiated or attenuated Treg effects on the survival of freshly isolated lymphoma cells. A combination of lenalidomide treatment with adoptive transfer of CD4(+)CD25(+)Tregs or CD4(+)CD25(+)CD127(lo)Tregs expanded ex vivo could be used to suppress proliferation of residual lymphoma in select patients with lymphoma responsive to the regulation by Tregs and sensitive to lenalidomide.

  6. 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.

  7. 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.

  8. Detection of CD4+CD25+ T cells in peripheral blood of patients with aplastic anemia%再生障碍性贫血患者外周血CD4+CD25+ T细胞的检测

    Institute of Scientific and Technical Information of China (English)

    王卫国; 马黎丽; 马芳; 李玉云

    2012-01-01

    Objective: To explore the relationship between the level of CD4 + CD25 + T cells and pathogenesis or prognosis of aplastic anemia by detecting CD4 + CD25 + T cells in peripheral blood. Methods: The proportion of CD4 + CD25 + T cells and CD4 + CD25high T cells in peripheral blood of four groups, including 20 cases of healthy individuals as normal controls ,30 cases of patients with first onset of aplastic anemia, 10 cases of patients accepted effective treatment and 10 cases of patients accepted invalid treatment, were determined by flow cytometry. Results: The rate of CD4 + CD25 + T cells in peripheral blood of four groups was not significantly different ( P > 0.05) ; Compared with group of normal controls, the rate of CD4+ CD25high T cells in patients with first onset of aplastic anemia decreased significantly ( P 0. 05 ). Conclusions: The decreasing of CD4 + CD25highT cells is one of the reasons causing breakdown of immune tolerance in patients with aplastic anemia,the rate of CD4 CD25 T cells may not reflect the prognosis of the patients.%目的:通过对外周血CD4+CD25+T细胞的检测,初步探讨CD4+CD25+T细胞水平与再生障碍性贫血(aplastic anemia,AA)发病和预后的关系.方法:流式细胞术分别检测20例正常对照、30例初发AA患者、10例治疗有效AA患者和10例治疗无效AA患者的CD4+CD25+T细胞和CD4+CD25highT细胞水平.结果:4组患者CD4+CD25+T细胞比例差异无统计学意义(P>0.05);初发AA和治疗无效组AA患者CD4+CD25highT细胞比例均低于正常对照组(P0.05).结论:CD4+CD25highT细胞比例下降是造成AA患者免疫耐受破坏的原因之一,CD4+CD25+T细胞比例可能并不能反映AA患者的预后状况.

  9. 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.

  10. Identification of lesional CD4+ CD25+ Foxp3+ regulatory T cells in Psoriasis.

    NARCIS (Netherlands)

    Bovenschen, H.J.; Vlijmen-Willems, I.M.J.J. van; Kerkhof, P.C.M. van de; Erp, P.E.J. van

    2006-01-01

    BACKGROUND: Depletion of CD4+ CD25+ Foxp3+ naturally occurring regulatory T cells (T(reg)) induces autoimmune phenomena. These cells have not yet been fully characterized in the skin of psoriatic patients. OBJECTIVES: To prove that the Zenon immunofluorescent labeling technique is suitable for the d

  11. IL-35, an anti-inflammatory cytokine which expands CD4+CD25+ Treg Cells.

    Science.gov (United States)

    Castellani, Maria Luisa; Anogeianaki, A; Felaco, P; Toniato, E; De Lutiis, M A; Shaik, B; Fulcheri, M; Vecchiet, J; Tetè, S; Salini, V; Theoharides, T C; Caraffa, A; Antinolfi, P; Frydas, I; Conti, P; Cuccurullo, C; Ciampoli, C; Cerulli, G; Kempuraj, D

    2010-01-01

    Interleukin 12 (IL 12) p35/p40 is a heterodimeric cytokine which plays a critical role in inflammation, immunity and tissue proliferation, and also plays a relevant function in T helper (Th) cell polarization and Th1 T-cell differentiation. IL-12 family members, IL-12p70, IL-23, IL-27 and IL-35, play an important role in influencing helper T-cell differentiation. EBV-induced gene 3 can be associated with the p35 subunit of IL-12 to form the EBI3/p35 heterodimer, also called IL-35. It has been shown that IL-35 has biological activity and able to expand CD4+CD25+ Treg cells, suppress the proliferation of CD4+CD25- effector cells and inhibit Th17 cell polarization. IL-35 has been shown to be constitutively expressed by regulatory T (Treg) cells CD4(+)CD25(+)Foxp3(+) and suggested to contribute to their suppressive activity. IL-35 is a crucial mediator which provokes CD4+CD25+ T cell proliferation and IL-10 generation, another well-known anti-inflammatory cytokine, along with TGFbeta cytokine. These studies suggest that IL-35, together with other successfully discovered cytokine inhibitors, represents a new potential therapeutic cytokine for chronic inflammation, autoimmunity and other immunological disorders.

  12. Antibody

    Science.gov (United States)

    An antibody is a protein produced by the body's immune system when it detects harmful substances, called antigens. Examples ... microorganisms (bacteria, fungi, parasites, and viruses) and chemicals. Antibodies may be produced when the immune system mistakenly ...

  13. 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

  14. 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.

  15. 一株识别肿瘤细胞上CD40突变体分子的单克隆抗体的研制及其生物学功能研究%Preparation and characterization of a mouse anti-human CD40 mutant monoclonal antibody

    Institute of Scientific and Technical Information of China (English)

    郑舒丹; 马泓冰; 高超; 汪家敏; 孙静; 罗先富; 张学光

    2009-01-01

    AIM: To prepare and characterize a mouse anti-human CD40 mutant monoclonal mAb. METHODS: Female BALB/c mice of 6-8 weeks old were immunized with CD40 mutant transfectant (L929-CD40mu) as immunogen. The spleen B cells of the mice were fused with Sp2/0 myeloma cells. The hybridoma cells were screened with CD40 mutant transfectant (L929-CD40mu) by FCM. Faststrip analysis was performed to identify Ig subclass of this mAb. The epitope recognized by this mAb was detected by Bio-5C11 competitive assay. Western blot technique was adopted to identify the mAb. The proliferation of tumor cells in vitro was analyzed by MTT assay and apoptosis of tumor cells in vitro was analyzed by PI-annexin V assay. RESULTS: One hybridoma cell line named 10C5 was obtained, which had the property of secreting anti-human CD40 mutant monoclonal antibody continuously and steadily. This mAb specifically recognized human CD40 mutant molecule and induced the apoptosis of tumor cells in vitro. CONCLUSION: One hybridoma cell line which can secret a mouse anti-human CD40 mutant mAb has been prepared successfully. This mAb can inhibit the growth of tumor cells expressing CD40 mutant and induce their apoptosis in vitro.%目的:以本科室发现肿瘤细胞上表达的CD40 787AA突变为基础,研制识别肿瘤细胞上CD40突变体分子的单克隆抗体(mAb),并对其生物学特性作初步分析.方法:以转人CD40突变体转基因细胞L929-CD40mu为免疫原,免疫6~8周龄的雌性BALB/c小鼠;采用B淋巴细胞融合技术,将免疫小鼠脾脏细胞与Sp2/0融合,以L929-CD40mu转基因细胞为抗体筛选阳性细胞,免疫荧光标记法对杂交瘤进行反复筛选和多次的克隆化培养;采用快速定性试纸法及竞争抑制结合试验分析该mAb的亚类及抗原识别位点;免疫印迹法对该mAb进行鉴定;采用MTT法分析mAb在体外对肿瘤细胞的抑制增殖效应以及PI-annexin V方法进行细胞凋亡测定.结果:获得1株稳定分泌鼠抗人CD40mu m

  16. CD4(+)CD25(+)Tregs express an increased LAG-3 and CTLA-4 in anterior chamber-associated immune deviation

    NARCIS (Netherlands)

    Zhu, X.F.; Yang, P.Z.; Zhou, H.Y.; Li, B.; Huang, X.K.; Meng, Q.L.; Wang, L.; Kijlstra, A.

    2007-01-01

    Background Regulatory CD4+CD25+ T cells have been proven to be essential for maintenance of peripheral tolerance and autoimmune diseases. ACAID is a model of immune privilege in the eye. Relatively little is known about the role and phenotype of these regulatory CD4+CD25+ T cells in ACAID. Methods I

  17. Effect of dendritic cells on the generation of CD4+CD25+Foxp3+ T cells.

    OpenAIRE

    Ivo Marguti

    2007-01-01

    As células dendríticas (DCs) são as principais células apresentadoras de antígeno do sistema imune. No entanto, trabalhos têm demonstrado seu envolvimento na manutenção da tolerância imunológica. As células T CD4+CD25+Foxp3+ possuem a capacidade de suprimir respostas imunes. Neste estudo avaliamos as alterações ocorridas na população de células T CD4+CD25+Foxp3+ após co-cultura de células de linfonodo com DCs. Nossos resultados demonstram que após a co-cultura há um aumento da população de cé...

  18. Evaluation of CD25-positive cells in relation to the subtypes and prognoses in various lymphoid tumours in dogs.

    Science.gov (United States)

    Mizutani, Noriyuki; Goto-Koshino, Yuko; Tsuboi, Masaya; Kagawa, Yumiko; Ohno, Koichi; Uchida, Kazuyuki; Tsujimoto, Hajime

    2016-05-01

    Interleukin-2 receptor alpha chain (CD25) expression has been reported in human lymphoid tumours and suggested to correlate with the prognosis. In this study, we detected CD25-positive cells in various types of lymphoid tumours in dogs. Immunohistochemical analyses of the tissues from diffuse large B-cell lymphoma (DLBCL) (n = 6), T-zone lymphoma (TZL) (n = 5), and follicular lymphoma (FL) (n = 2) revealed that cells strongly positive for CD25 were observed generally in accordance with lymphoma cell localization. CD25-positive cells were consistently detected in TZL and FL cases; however, the number of CD25-positive cells was variable among DLBCL cases. Furthermore, we evaluated the rate of CD25-positive cells by flow cytometric analysis in 29 dogs with lymphoid malignancies, including high-grade B-cell lymphoma (n = 17), TZL (n = 5), FL (n = 2), cutaneous lymphoma (n=2), and acute lymphoblastic leukaemia (ALL) (n = 3). CD25-positivity in the lymph node cells was significantly higher in dogs with high-grade B-cell lymphoma (mean ± SD, 49.6 ± 31.3%) or TZL (mean ± SD, 80.2 ± 10.0%) than that in healthy dogs (mean ± SD, 9.8 ± 2.8%). In prognostic analysis of 15 cases with high-grade B-cell lymphoma, the progression-free survival was significantly shorter in CD25-high group than that in CD25-low group. The results obtained in this study are useful for subtype differentiation and prognostic analysis of canine lymphomas and future development of molecular-targeted therapy directed at CD25.

  19. CD4+CD25+T细胞的扩增方法与临床应用%The proliferation and clinical application of CD4+ CD25+ regulatory T-cell

    Institute of Scientific and Technical Information of China (English)

    王慧

    2008-01-01

    CD4+CD25+T细胞是最重要的一类调节性T细胞(Tr).体内固有CD4+CD25+T细胞的自然扩增率极低,不能满足临床治疗的需要.通过采用FoxP3基因转染技术、阻断细胞活化信号、DC诱导、加入细胞因子等方法,对CD4+CD25+T细胞的数量和功能进行扩增,使其在器官移植、自身免疫性疾病和肿瘤免疫等领域具有广泛的临床应用前景.%CD4+ CD25+ T cells are the most important cells among the regulatory T-cells. It is known that thenatural proliferation of natural-occurring CD4+ CD25+ Tcell is very low, and is not satisfied for the clinical ther-apies. However, the quantity and function of CD4+ CD25+ T could be increased through FoxP3 gene transfec-tion, activation signal blockade, dendritil cell induction, and cytokine stimulation. Thus the clinical applicationof CD4+ CD25+ T in organ transplantation, autoimmune disease and tumor immunity areas will be possitble.

  20. 免疫磁珠两步法分离小鼠脾脏CD4+CD25+调节性T细胞%Concentration of Mouse Splenic CD4+CD25+ Regulatory T Cell by Magnetic Cell Sorting System

    Institute of Scientific and Technical Information of China (English)

    印永祥; 韩晓枫; 何阳

    2009-01-01

    体外分离CD4+CD25+调节性T细胞(CD4+CD25+Treg)并进行初步鉴定.用磁性细胞分离器(MiniMACS)分离CD4+CD25+Treg细胞,流式细胞术(flow cytometry,FCM)分析细胞纯度和Foxp3蛋白表达,体外检测细胞因子.MACS分离的CD4+CD25+Treg细胞纯度大于90%,细胞存活率大于93%,并且特异性表达Foxp3蛋白,体外能抑制CD4+CD25-Treg分泌IFN-γ,同时CD4+CD25+Treg能分泌抑制性细胞因子TGF-β1、IL-10.结果显示,通过MACS可分离高纯度、高活性的CD4+CD25+Treg细胞.

  1. Association of the IL2RA/CD25 Gene With Juvenile Idiopathic Arthritis

    Science.gov (United States)

    Hinks, Anne; Ke, Xiayi; Barton, Anne; Eyre, Steve; Bowes, John; Worthington, Jane; Thompson, Susan D; Langefeld, Carl D; Glass, David N; Thomson, Wendy

    2009-01-01

    Objective IL2RA/CD25, the gene for interleukin-2 receptor α, is emerging as a general susceptibility gene for autoimmune diseases because of its role in the development and function of regulatory T cells and the association of single-nucleotide polymorphisms (SNPs) within this gene with type 1 diabetes mellitus (DM), Graves' disease, rheumatoid arthritis (RA), and multiple sclerosis (MS). The aim of this study was to determine whether SNPs within the IL2RA/CD25 gene are associated with juvenile idiopathic arthritis (JIA). Methods Three SNPs within the IL2RA/CD25 gene, that previously showed evidence of an association with either RA, MS, or type 1 DM, were selected for genotyping in UK JIA cases (n = 654) and controls (n = 3,849). Data for 1 SNP (rs2104286) were also available from North American JIA cases (n = 747) and controls (n = 1,161). Association analyses were performed using Plink software. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. Results SNP rs2104286 within the IL2RA/CD25 gene was significantly associated with UK JIA cases (OR for the allele 0.76 [95% CI 0.66–0.88], P for trend = 0.0002). A second SNP (rs41295061) also showed modest evidence for association with JIA (OR 0.80 [95% CI 0.63–1.0], P = 0.05). Association with rs2104286 was convincingly replicated in the North American JIA cohort (OR 0.84 [95% CI 0.65–0.99], P for trend = 0.05). Meta-analysis of the 2 cohorts yielded highly significant evidence of association with JIA (OR 0.76 [95% CI 0.62–0.88], P = 4.9 × 10−5). Conclusion These results provide strong evidence that the IL2RA/CD25 gene represents a JIA susceptibility locus. Further investigation of the gene using both genetic and functional approaches is now required. PMID:19116909

  2. Analysis of human T-cell lymphotropic virus in CD25+ anaplastic large cell lymphoma in children.

    Science.gov (United States)

    Gualco, Gabriela; Chioato, Lucimara; Weiss, Lawrence M; Harrington, William J; Bacchi, Carlos E

    2009-07-01

    Anaplastic large cell lymphoma (ALCL) is recognized as 2 distinct diseases: anaplastic lymphoma kinase (ALK)+ ALCL and ALK- ALCL. ALK+ ALCL occurs in younger patients and has a better prognosis. Human T-cell lymphotropic virus (HTLV-1) is linked to the development of adult T-cell leukemia/lymphoma (ATLL), which frequently expresses CD25. CD25 is significantly expressed in childhood ALCL. In Brazil, HTLV-1 infection is endemic, and vertical transmission is responsible for spread to children. Of HTLV-1 carriers, 90% or more remain asymptomatic. Some cases of adult HTLV-1-related lymphomas have characteristics of ALCL but are considered CD30+ ATLL subtypes. No similar cases have been described in children. We analyzed 33 cases of pediatric ALCL, CD25+ and CD25-, for proviral HTLV-1 DNA. All cases corresponded to the common histologic ALCL type and were CD30+ in virtually all neoplastic cells. ALK expression was observed in 31 (94%) of 33 cases; CD25 was positive in 27 (82%), including 1 ALK- ALCL case. There was a strong positive correlation between ALK and CD25 expression. None of the cases showed proviral HTLV-1 DNA. ALCL in children has no relationship with HTLV-1; the frequent CD25 expression must be explained by a mechanism different from that in ATLL.

  3. [In vitro amplification of CD4(+) CD25(+) regulatory T cells and identification of amplified T cell immunosuppressive function].

    Science.gov (United States)

    Weng, Wen-Jun; Pan, Li; Fang, Jian-Pei; Xu, Lv-Hong

    2013-10-01

    This study was purposed to compare the effect of 3 different cell components for expanding CD4(+) CD25(+) Treg in vitro, and identify their immunosuppressive function. CD4(+) T cells, CD4(+) CD25(-)T cells and CD4(+) CD25(+)T cells were isolated from mouse splenocytes by MACS and then expanded in vitro. Phenotype of the T cell lines and expression of the FOXP3 was determined by flow cytometry. The inhibitory effect of expanded CD4(+) CD25(+) T cells on CD4(+) CD25(-)T cells was tested by MLR method. The results showed that the Treg cells from all the three groups were expanded significantly after culture for 2 weeks. In the CD4(+) T cells group, the proliferation rate was (77.8 ± 5.32) folds with a percentage of Treg cells increasing from (6.61 ± 1.00)% to (15.33 ± 1.31)%. The proliferation rate in the CD4(+) CD25(-) T cells group was (95.20 ± 7.67) folds, with the percentage of CD4(+) CD25(+) T cells raising from (0.37 ± 0.13)% to (9.84 ± 0.98)%. The proliferation rate in the CD4(+) CD25(+) T cells group was (41.20 ± 6.92) folds, the proportion of Treg cells decreased from (86.75 ± 1.25)% to (85.32 ± 1.62)%, and the expression of Foxp3 decreased from (76.92 ± 1.72)% to (75.33 ± 2.11)% during the culture, there were not significant differences in the cell purity and the expression of Foxp3, compared with pre-amplification. The inhibitory test showed that the expanded CD4(+) CD25(+) T cells could inhibit the proliferation of CD4(+) CD25(-) T cells in vitro in a cell dose-dependent manner. It is concluded that the amplification of CD4(+) CD25(+) Treg cells is successful in vitro, especially in the CD4(+) CD25(+) T cells group, the cell purity and Foxp3 gene is not obviously changes after amplification.

  4. 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

  5. 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

  6. Do CD4+CD25+ Immunoregulatory T Cells Hinder Tumor Immunotherapy?

    OpenAIRE

    Antony, Paul Andrew; Restifo, Nicholas P.

    2002-01-01

    After years of banishment from mainstream immunology, the notion that one subset of T cells can exert regulatory effects on other T lymphocytes is back in fashion. Recent work in knockout and transgenic mice has begun to bring molecular definition to our understanding of immunoregulatory CD4+CD25+ T cells (Treg/Th3/Tr1). The identification of the glucocorticoid-induced tumor necrosis factor receptor family-related gene (GITR, also known as TNFRSF18) expressed on T regulatory cells might affor...

  7. The origin of thymic CD4+CD25+ regulatory T cells and their co-stimulatory requirements are determined after elimination of recirculating peripheral CD4+ cells.

    Science.gov (United States)

    Zhan, Yifan; Bourges, Dorothee; Dromey, James A; Harrison, Leonard C; Lew, Andrew M

    2007-04-01

    Studies on the thymic ontogeny of naturally arising CD4(+)CD25(+) regulatory T cells (TR cells) are complicated by the contamination of recirculating cells from the periphery (both activated CD4(+) T and TR cells). We investigated TR cells in anti-CD4 antibody transgenic (Tg) (GK) mice that continuously deplete peripheral CD4 T cells but not thymocytes so that the generation of thymic TR cells and their developmental requirement can be accurately assessed. We show that in the thymuses of mice that lack peripheral CD4(+) cells, TR cells were present but were fewer in number compared with wild-type (WT) mice. Therefore, we show that peripheral TR cells do re-enter the thymus, comprising 20% of TR cells in the normal thymus. TR cells from both WT and GK mice expressed Foxp3 and GITR, and suppressed the proliferation of CD25(-)CD4(+) T cells. Furthermore, the co-stimulation requirements for TR generation were evaluated in mice with or without peripheral CD4 cells. Splenic TR cells in CD40L(-/-) mice and CTLA4Ig Tg mice were fewer compared with WT mice. Mice deficient in both co-stimulatory pathways had further reduction in splenic TR cells. Unlike the periphery, the reduction in thymic TR cells was only seen for CD40L(-/-) but not for CTLA4Ig Tg mice. Therefore, we found that the co-stimulation requirements for the thymic development of TR cells differed from those for peripheral homeostasis.

  8. Detection of CD4 + CD25 + Tree Cells and Foxp3 in Pathogenesis of Preeclampsia%子痫前期发病中CD4+CD25+Treg细胞与Foxp3的检测及意义

    Institute of Scientific and Technical Information of China (English)

    王莲莲; 曹霞; 魏军

    2011-01-01

    [目的]研究CD4+ CD25+调节性T细胞(CD4+ CD25+Treg细胞)和Foxp3基因表达与子痫前期发病的相关性.[方法]选择住院确诊的妊娠期高血压孕妇12例(A组)、轻度子痫前期(MPE)孕妇14例(B组)、重度子痫前期(SPE)孕妇15例(C组),30例正常妊娠晚期孕妇为对照组(D组),应用流式细胞术检测并比较四组外周血中CD4+ CD25+ Treg和 Foxp3的表达水平.[结果]A组、B组和C组孕妇外周血CD4+ CD25+ Treg 和Foxp3表达水平均显著低于对照组(P<0.05);C组外周血CD4+ CD25+ Treg和Foxp3表达水平显著低于A组和B组(P<0.05).[结论]子痫前期患者体内存在CD4+ CD25+ Treg细胞的异常,CD4+ CD25+Treg细胞的数量的减少和Foxp3表达的降低使CD4+ CD25+Treg细胞免疫抑制功能减弱,从而使母胎免疫耐受失衡,导致子痫前期的发生.%[Objective] To investigate the relationship between CD4+ CD25 + Treg cells, Foxp3 gene ex pression and the pathogenesis of preeclampsia. [Methods] Twelve pregnant women with gestational hyperten sion(group A) , 14 mild preeclampsia(MPE) pregnant women(group B) , 15 severe preeclampsia(SPE) preg nant women(group C) and 30 normal pregnant women(group D) were chosen in hospital. Flow cytometry was used to detect and compare CD4 + CD25 + Treg cells and expression level of Foxp3 in peripheral blood among 4 groups. [Results] The CD4 + CD25 + Treg cells and Foxp3 expression level in peripheral blood in group A, B and C were significantly lower than those in control group( P <0.05). The CD4 + CD25 + Treg cells and Foxp3 expression level in peripheral blood in group C were significantly lower than those in group ∧ and B( P <0.05). [Conclusion] Preeclampsia patients have abnormal CD4 + CD25 + Treg cells. The decrease of CD4 + CD25 +Treg cells and Foxp3 expression attenuates the immune suppression function of CD4 + CD25 + Treg cells, thus maternal-fetal immune tolerance becomes imbalance which results in the occurrence of preeclampsia.

  9. 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.

  10. 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.

  11. 宫颈疾病患者外周血CD4+CD25+C-D127-/low Treg细胞的检测及临床意义%Detection of the expression of CD4+CD25+CD127-/low Treg cells in peripheral blood in patients with cervix diseases and its clinical significance

    Institute of Scientific and Technical Information of China (English)

    袁冬兰; 钱华

    2012-01-01

    目的 检测宫颈癌、宫颈上皮内瘤变患者外周血CD4+CD25+CD127-/low Treg细胞的表达,分析其临床意义.方法 荧光抗体CD127-FITC、CD4-PECY5、CD25-PE标记T细胞,流式细胞术测定50例宫颈癌患者和49例宫颈上皮内瘤变患者中CD4+CD25+CD127-/low Treg细胞的表达.结果 与正常对照组[n=33,(8.07±2.18)%]比较,宫颈癌组[n=50,(11.15±1.97)%]、宫颈上皮内瘤变组[n=49,(10.61±2.62)%]CD4+CD25+CD127-low均显著升高,两者比较差异有统计学意义(t=6.018,P<0.001;t=4.943,P<0.001);与宫颈上皮内瘤变Ⅰ级组[n=16,(8.96±0.65)%]比较,宫颈上皮内瘤变Ⅱ~Ⅲ级组[n=33,(11.54±2.72)%]显著升高,两者比较有差异统计学意义(t=4.114,P<0.001);与宫颈癌Ⅰ期组[n=20,(10.34±1.79)%]比较,官颈癌Ⅱ期组[n=30,(11.68±1.97)%]显著升高,两者比较差异有统计学意义(t=2.424,P<0.05).结论 外周血CD4+CD25+CD127-/low Treg细胞的表达升高可能在宫颈癌和宫颈上皮内瘤变发病的发生、发展过程中起一定作用.%Objective To detect the expression of CD4+CD25+CD127-flow Treg cells in peripheral blood in patients with cervix cancer and cervical intraepithelial neoplasia (CIN), and to explore its clinical significance. Methods T cells were marked with fluorescence antibody CD127-FITC, CD4-PECY5, CD25-PE. The levels of CD4+CD25+CD127-flow Treg cells were detected by flow cytometry in 50 patients with cervix cancer (group A), 49 patients with CIN (group B), and 33 healthy individuals (the control group). Results Compared with the control group [(8.07±2.18)%], the levels of CD4+CD25+CD127-flow Treg cells in group A [(11.15±1.97)%] and group B [(10.61±2.62)%] were significantly higher (t=6.018,P<0.001;t=4.943 ,P<0.001). The level of CD4+CD25+CD127-flow Treg cells in patients with CIN of grade II ~ III [n=33, (11.54±2.72)%] was significantly higher than that in patients with CIN of grade I [n=16, (8.96±0.65)%], with t=4.114 and P<0

  12. Docosahexaenoic acid reduces suppressive and migratory functions of CD4CD25 regulatory T-cells

    Science.gov (United States)

    Yessoufou, Akadiri; Plé, Aude; Moutairou, Kabirou; Hichami, Aziz; Khan, Naim Akhtar

    2009-01-01

    Immunological tolerance is one of the fundamental aspects of the immune system. The CD4+CD25+ regulatory T (Treg) cells have emerged as key players in the development of tolerance to self and foreign antigens. However, little is known about the endogenous factors and mechanisms controlling their suppressive capacity on immune response. In this study, we observed that docosahexaenoic acid (DHA), an n-3 polyunsaturated fatty acid, diminished, in a dose-dependent manner, the capacity of Treg cells to inhibit the CD4+CD25− effector T-cell proliferation. DHA not only reduced the migration of Treg cells toward chemokines but also downregulated the mRNA expression of CCR-4 and CXCR-4 in Treg cells. DHA also curtailed ERK1/2 and Akt phosphorylation and downregulated the Smad7 levels in these cells. Contradictorily, DHA upregulated the mRNA expression of Foxp3, CTLA-4, TGF-β, and IL-10; nonetheless, this fatty acid increased the expression of p27KIP1 mRNA, known to be involved in Treg cell unresponsiveness. In Foxp3-immunoprepitated nuclear proteins, DHA upregulated histone desacetylase 7 levels that would again participate in the unresposnsiveness of these cells. Finally, a DHA-enriched diet also diminished, ex vivo, the suppressive capacity of Treg cells. Altogether, these results suggest that DHA, by diminishing Treg cell functions, may play a key role in health and disease. PMID:19561360

  13. 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".

  14. Glucosamine Modulates T Cell Differentiation through Down-regulating N-Linked Glycosylation of CD25.

    Science.gov (United States)

    Chien, Ming-Wei; Lin, Ming-Hong; Huang, Shing-Hwa; Fu, Shin-Huei; Hsu, Chao-Yuan; Yen, B Lin-Ju; Chen, Jiann-Torng; Chang, Deh-Ming; Sytwu, Huey-Kang

    2015-12-04

    Glucosamine has immunomodulatory effects on autoimmune diseases. However, the mechanism(s) through which glucosamine modulates different T cell subsets and diseases remain unclear. We demonstrate that glucosamine impedes Th1, Th2, and iTreg but promotes Th17 differentiation through down-regulating N-linked glycosylation of CD25 and subsequently inhibiting its downstream Stat5 signaling in a dose-dependent manner. The effect of glucosamine on T helper cell differentiation was similar to that induced by anti-IL-2 treatment, further supporting an IL-2 signaling-dependent modulation. Interestingly, excess glucose rescued this glucosamine-mediated regulation, suggesting a functional competition between glucose and glucosamine. High-dose glucosamine significantly decreased Glut1 N-glycosylation in Th1-polarized cells. This finding suggests that both down-regulated IL-2 signaling and Glut1-dependent glycolytic metabolism contribute to the inhibition of Th1 differentiation by glucosamine. Finally, glucosamine treatment inhibited Th1 cells in vivo, prolonged the survival of islet grafts in diabetic recipients, and exacerbated the severity of EAE. Taken together, our results indicate that glucosamine interferes with N-glycosylation of CD25, and thereby attenuates IL-2 downstream signaling. These effects suggest that glucosamine may be an important modulator of T cell differentiation and immune homeostasis.

  15. Long-term toxicity of fully humanized anti-human tumor necrosis factor-αmonoclonal antibody for injection in cynomolgus monkeys%全人源抗人肿瘤坏死因子α单克隆抗体注射液对食蟹猴的长期毒性试验

    Institute of Scientific and Technical Information of China (English)

    张囡; 王炯; 张雅婷; 宋刚; 詹珊珊; 潘勇兵

    2015-01-01

    OBJECTIVE To evaluate the long-term toxicity of fully human anti-human tumor necrosis factor-α monoclonal antibody(anti-hTNF-α FHMA)for injection in cynomolgus monkeys. METHODS Forty cynomolgus monkeys were randomly divided into 5 groups (4 males and 4 females in each group):negative control group,adalimumab 10 mg·kg-1 group,anti-hTNF-αFHMA 2,10 and 50 mg·kg-1 groups. Cynomolgus monkeys in each group were injected sc once a week for 5 consecutive times, followed by 4 weeks of recovery. During the test,general clinical observation,body mass,body temperature,electrocardiogram(ECG),hematology,coagulation function,blood biochemistry,urine, ophthalmology,immune index,and pathological changes in organs and tissues were observed. At the same time,plasma drug concentrations were detected and the toxicokinetics parameters were analyzed. RESULTS No significant toxicological changes related to drugs were observed in general clinical observation,body mass,body temperature,ECG,ophthalmic examination,blood cell counts,coagu⁃lation function,blood biochemistry,urine analysis,lymphocyte subsets,cytokines,serum immuno⁃globulin,serum complement. Neutralizing anti-drug antibody(ADA)could be detected in adalimumab group and anti-hTNF-αFHMA groups. Anti-hTNF-αFHMA showed linear dynamic characteristics in cyno⁃molgus monkeys. At the same dose(10 mg·kg-1),anti-hTNF-αFHMA had similar immunogenicity and kinetics characteristics to adalimumab. CONCLUSION The level of anti-hTNF-α FHMA at which no adverse effect was observed was 50 mg · kg-1,which is equivalent to 75 times clinical dosage of quasi (0.67 mg·kg-1),which suggests that anti-hTNF-αFHMA be safe in clinical use.%目的:评价全人源抗人肿瘤坏死因子α单克隆抗体(抗-hTNF-αFHMA)注射液对食蟹猴的长期毒性。方法将40只食蟹猴随机分成5组,分别为阴性对照组、阿达木单抗10 mg·kg-1对照组和抗-hTNF-αFHMA 2,10和50 mg·kg-1组,经皮下注射每周给药1

  16. Isolation, purification and function of mouse CD4 + CD25+ regulatory T lymphocytes in vitro%小鼠CD4+CD25+T调节细胞的分离、纯化及其功能检测

    Institute of Scientific and Technical Information of China (English)

    王百林; 刘增军; 翟淑萍; 陈义发; 秦欢; 刘秋玲

    2010-01-01

    Objective To establish a method for isolation and purification of CD4+ CD25+ regulatory T lymphocytes (Treg), and to identify partial functions of these cells. Methods Lymphocytes were isolated from the mouse spleens and then CD4+ CD25+ T cells were sorted by magnetic bead cell sorting (MACS) system. The purity of CD4+ CD25+ T cells were analyzed by flow cytometry (FCM). The activity of them was detected by trypan blue staining. Interleukin (IL)-2 and IL-10 levels in culture supernatant were determined by enzyme linked immunosorbent assay (ELISA). Results The purity of CD4+ CD25+ T cells sorted by MACS was 83%-96%. There was significant difference in the levels of IL-2 and IL-10 secreted by lymphocytes among CD4+ CD25+ Treg group [ (10.25±3.31), (40.32±8.05) ng/L], CD4+ CD25- T group [(58.21±13.05),(11.52±3.01)ng/L] and mixed culture group [ (39.54±13.82), (31.25±4.36)ng/L (P<0.05,P<0.01).Conclusion High purity of CD4+ CD25+ Treg with immune regulatory function can be isolated by MACS. The immunosuppression of CD4+ CD25+ Treg to CD4+ CD25- T was related to the regulation of IL-10 through IL-2.%目的 探讨小鼠CD4+CD25+T调节细胞(Treg)的分离培养、纯化及其部分功能检测.方法 采用免疫磁珠分离法(MACS)对分离小鼠的脾淋巴细胞进行分选CD4+CD25+Treg细胞,锥虫蓝细胞染色检测其活性,流式细胞仪检测分选所得活性细胞的纯度,酶联免疫吸附试验(ELISA)检测培养上清液中白细胞介素(IL)-2、IL-10水平的浓度.结果 MACS分离的CD4+CD25+Treg细胞的纯度达83%~96%.体外培养中Treg组、T组和混合组IL-2和IL-10的平均水平分别为:(10.25±2.31)、(40.32±8.05)ng/L;(5 8.21±13.05)、(11.52±3.01)ng/L;(39.54±12.82)、(31.25±4.36)ng/L,数据差异有统计学意义(P<0.05,P<0.01).结论 采用MACS系统两步法,可获得高纯度、具有免疫抑制功能的Treg细胞,该细胞对CD4+CD25-T细胞的免疫抑制作用可能是通过IL-10对IL-2的调节作用实现的.

  17. 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....

  18. 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....

  19. 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

    , 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......' results suggest that CCL20 is used by CCR6 regulatory T cells in the complex process of controlling colitis because transcripts for this chemokine were expressed to a higher level in protected animals. The chemokine pathways identified in the present study may be of importance for the development of new....../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...

  20. Dexamethasone Regulates Macrophage and Cd4+Cd25+ Cell Numbers in the Chicken Spleen

    Directory of Open Access Journals (Sweden)

    AS Calefi

    2016-03-01

    Full Text Available Abstract Dexamethasone (DEX is a corticoid hormone that is experimentally used to mimic the effects of increased levels of endogenous corticosterone observed during the stress response. Currently, stress is considered one of the major predisposing factors for diseases in the poultry industry. The aim of this study was to analyze the effects of DEX and/or of a 20-fold coccidial vaccine dose on leukocyte phenotypes in the spleen and cecal tonsils of chickens. Twenty specific-pathogen-free (SPF Leghorn chickens were divided into four groups: a non-treated group (NT, a DEX-treated group (Dex, a vaccinated group (V and a DEX-treated+vaccinated group (Dex+V. On experimental day (ED 42, each bird in the vaccinated groups received a anti-coccidial vaccine. DEX was injected in the birds of the Dex and Dex+V groups (0.9 mg/kg onED42 and ED45. The immunophenotyping was performed by flow cytometry analysis of splenocytes and cecal tonsils cells onED48. DEX treatment per se was unable to change CD4+CD8+, CD4+CD8+ and CD4-CD8+ populations with TCRgd or CD28 in the spleen, or macrophages and T lymphocytes in the cecal tonsils. V group birds presented higher numbers of splenic macrophages compared with those measured in the Dex+V group. The number of CD4+CD25+ cells in the spleen of birds of the V group was higher than those measured in the other experimental groups. Our data suggest that CD4+CD25+ cells and macrophages might be influenced by DEX treatment in spleen, but not in the cecal tonsils of chickens inoculated with Eimeria.

  1. Human CD4+ CD25+ Foxp3+ regulatory T cells do not constitutively express IL-35.

    Science.gov (United States)

    Bardel, Emilie; Larousserie, Frédérique; Charlot-Rabiega, Pascaline; Coulomb-L'Herminé, Aurore; Devergne, Odile

    2008-11-15

    EBV-induced gene 3 (EBI3) can associate with p28 to form the heterodimeric cytokine IL-27, or with the p35 subunit of IL-12 to form the EBI3/p35 heterodimer, recently named IL-35. In mice, IL-35 has been shown to be constitutively expressed by CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg cells) and suggested to contribute to their suppressive activity. In this study, we investigated whether human Treg cells express IL-35. Double-staining analysis of human thymuses showed that neither Foxp3(+) nor CD25(+) cells coexpressed EBI3. Similarly, Foxp3(+) cells present in human lymph nodes, tonsils, spleens, and intestines did not express EBI3. Consistent with these in situ observations, Treg cells purified from blood or tonsils were negative for EBI3 by immunoblotting. Other human T cell subsets, including effector T cells, naive and memory CD4(+) T cells, CD8(+) and gammadelta T cells also did not constitutively express EBI3, which contrasts with IL-35 expression observed in murine CD8(+) and gammadelta T cells. Furthermore, although CD3/CD28 stimulation consistently induced low levels of EBI3 in various CD4(+) T cell subsets, no EBI3 could be detected in CD3/CD28-stimulated Treg cells. RT-PCR analysis showed that, whereas p35 transcripts were detected in both Teff and Treg cells, EBI3 transcripts were detected only in activated Teff cells, but not in resting or activated Treg cells. Thus, in contrast to their murine counterpart, human Treg cells do not express detectable amounts of IL-35.

  2. 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

  3. Correlation of the expression of CD4+CD25+CD127low/- regulatory T cells and transforming growth factor-β with primary rheumatoid arthritis%调节性T细胞和转化生长因子β与初发类风湿性关节炎相关性研究

    Institute of Scientific and Technical Information of China (English)

    罗莉; 罗德梅; 刘盼盼; 董旭南

    2011-01-01

    Objective To investigate the expressions of CD4+ CD25+ CD127low/- regulatory T cells and transforming growth factor-β in the peripheral blood of patients with primary and active rheumatoid arthritis and to explore its mechanism in the pathogenesis of rheumatoid arthritis. Methods The rates of CD4+ CD25+ CD127low/- regulatory T cells in the peripheral blood of 40 primary and active rheumatoid arthritis patients and 35 normal controls were evaluated with flow cytometry. The level of transforming growth factor-β in blood serum was detected with ELISA. All the data were analyzed.Results The positive rate of CD4+ CD25+ T cells in rheumatoid arthritis was significantly lower than that in normal controls(P<0.05). The positive rate of CD4+ CD25+ CD127low/- T cells in rheumatoid arthritis was lower than that in normal controls, but there was no significant difference(P>0.05).There were significant differences between the percentage of CD4+ CD25+ T cells and the percentage of CD4+CD25+CD127low/- T cells in two groups(P<0.05). The level of CD4+ CD25+ T cells was correlated with the level of CD4+ CD25+ CD127low/- T cells in both groups(P<0.05). The level of transforming growth factor-β in rheumatoid arthritis was higher than that in control group(P<0.01).Transforming growth factor-β was not correlated with the level of CD4+ CD25+ CD127low/- regulatory T cells or CD4+ CD25+ T cells in two groups (P> 0.05). The levels of CD4+ CD25+ CD127low/-regulatory T cells, CD4+ CD25+ T cells and transforming growth factor-β were not correlated with erythrocyte sedimentation rate, C-reactive protein, anticyclic citrullinated peptide antibody,antikeratin antibody, or rheumatoid factor level(P>0.05). Conclusion The decrease of CD4+ CD25+CD127low/- regulatory T cells may be correlated with cellular immunity disorder in rheumatoid arthritis. The mechanism of transforming growth factor-β is complex in rheumatoid arthritis.%目的:探讨CD4+CD25+CD127low/-调节性T细

  4. CD4+CD25+调节T细胞与肿瘤免疫治疗策略%CD4+CD25+Regulatory T Cells and Cancer Immunotherapy Strategy

    Institute of Scientific and Technical Information of China (English)

    白平; 王春晖

    2008-01-01

    随着分子生物技术的不断发展,肿瘤的生物治疗在临床应用中的地位日渐突出,其中CD4+CD25+Foxp3+调节T细胞(CD4+CD25+Foxp3+regulatory T cell,Treg)在抑制肿瘤免疫方面起着重要作用,文章就其近年来在肿瘤免疫领域的研究进展作一综述.

  5. Cord Blood Derived CD4+CD25high T Cells Become Functional Regulatory T Cells upon Antigen Encounter

    Science.gov (United States)

    Mayer, Elisabeth; Bannert, Christina; Gruber, Saskia; Klunker, Sven; Spittler, Andreas; Akdis, Cezmi A.

    2012-01-01

    Background: Upon antigen exposure, cord blood derived T cells respond to ubiquitous environmental antigens by high proliferation. To date it remains unclear whether these “excessive” responses relate to different regulatory properties of the putative T regulatory cell (Treg) compartment or even expansion of the Treg compartment itself. Methods: Cord blood (>37 week of gestation) and peripheral blood (healthy controls) were obtained and different Treg cell subsets were isolated. The suppressive potential of Treg populations after antigen exposure was evaluated via functional inhibition assays ([3H]thymidine incorporation assay and CFSE staining) with or without allergen stimulation. The frequency and markers of CD4+CD25highFoxP3+ T cells were characterized by mRNA analysis and flow cytometry. Results: Cord blood derived CD4+CD25high cells did not show substantial suppressor capacity upon TCR activation, in contrast to CD4+CD25high cells freshly purified from adult blood. This could not be explained by a lower frequency of FoxP3+CD4+CD25highcells or FOXP3 mRNA expression. However, after antigen-specific stimulation in vitro, these cells showed strong proliferation and expansion and gained potent suppressive properties. The efficiency of their suppressive capacity can be enhanced in the presence of endotoxins. If T-cells were sorted according to their CD127 expression, a tiny subset of Treg cells (CD4+CD25+CD127low) is highly suppressive even without prior antigen exposure. Conclusion: Cord blood harbors a very small subset of CD4+CD25high Treg cells that requires antigen-stimulation to show expansion and become functional suppressive Tregs. PMID:22272233

  6. Latency-associated peptide identifies a novel CD4+CD25+ regulatory T cell subset with TGFbeta-mediated function and enhanced suppression of experimental autoimmune encephalomyelitis.

    Science.gov (United States)

    Chen, Mei-Ling; Yan, Bo-Shiun; Bando, Yoshio; Kuchroo, Vijay K; Weiner, Howard L

    2008-06-01

    CD4(+)CD25(+) regulatory T cells (Tregs) are essential for maintaining self-tolerance and immune homeostasis. Here we characterize a novel subset of CD4(+)CD25(+) Tregs that express latency-associated peptide (LAP) on their cell surface (CD4(+)CD25(+)LAP(+) cells). CD4(+)CD25(+)LAP(+) cells express elevated levels of Foxp3 and Treg-associated molecules (CTLA4, glucocorticoid-induced TNFR-related gene), secrete TGFbeta, and express both cell surface TGFbeta and surface receptors for TGFbeta. In vitro, the suppressive function of CD4(+)CD25(+)LAP(+) cells is both cell contact and soluble factor dependent; this contrasts with CD4(+)CD25(+)LAP(-) cells, which are mainly cell contact dependent. In a model of experimental autoimmune encephalomyelitis, CD4(+)CD25(+)LAP(+) cells exhibit more potent suppressive activity than CD4(+)CD25(+)LAP(-) cells, and the suppression is TGFbeta dependent. We further show that CD4(+)CD25(+)LAP(+) cells suppress myelin oligodendrocyte glycoprotein-specific immune responses by inducing Foxp3 and by inhibiting IL-17 production. Our findings demonstrate that CD4(+)CD25(+) Tregs are a heterogeneous population and that the CD4(+)CD25(+) subset that expresses LAP functions in a TGFbeta-dependent manner and has greater in vivo suppressive properties. Our work helps elucidate the ambiguity concerning the role of TGFbeta in CD4(+)CD25(+) Treg-mediated suppression and indicates that LAP is an authentic marker able to identify a TGFbeta-expressing CD4(+)CD25(+) Treg subset.

  7. Increased numbers of IL-7 receptor molecules on CD4+CD25-CD107a+ T-cells in patients with autoimmune diseases affecting the central nervous system.

    Directory of Open Access Journals (Sweden)

    Nalini Kumar Vudattu

    Full Text Available BACKGROUND: High content immune profiling in peripheral blood may reflect immune aberrations associated with inflammation in multiple sclerosis (MS and other autoimmune diseases affecting the central nervous system. METHODS AND FINDINGS: Peripheral blood mononuclear cells from 46 patients with multiple sclerosis (MS, 9 patients diagnosed with relapsing remitting MS (RRMS, 13 with secondary progressive multiple sclerosis (SPMS, 9 with other neurological diseases (OND and well as 15 healthy donors (HD were analyzed by 12 color flow cytometry (TCRalphabeta, TCRgammadelta, CD4, CD8alpha, CD8beta, CD45RA, CCR7, CD27, CD28, CD107a, CD127, CD14 in a cross-sectional study to identify variables significantly different between controls (HD and patients (OND, RRMS, SPMS. We analyzed 187 individual immune cell subsets (percentages and the density of the IL-7 receptor alpha chain (CD127 on 59 individual immune phenotypes using a monoclonal anti-IL-7R antibody (clone R34.34 coupled to a single APC molecule in combination with an APC-bead array. A non-parametric analysis of variance (Kruskal-Wallis test was conducted in order to test for differences among the groups in each of the variables. To correct for the multiplicity problem, the FDR correction was applied on the p-values. We identified 19 variables for immune cell subsets (percentages which allowed to segregate healthy individuals and individuals with CNS disorders. We did not observe differences in the relative percentage of IL-7R-positive immune cells in PBMCs. In contrast, we identified significant differences in IL-7 density, measured on a single cell level, in 2/59 variables: increased numbers of CD127 molecules on TCRalphabeta+CD4+CD25 (intermed T-cells and on TCRalphabeta+CD4+CD25-CD107a+ T-cells (mean: 28376 Il-7R binding sites on cells from HD, 48515 in patients with RRMS, 38195 in patients with SPMS and 33692 IL-7 receptor binding sites on cells from patients with OND. CONCLUSION: These data

  8. CD4+CD25+ and CD4+CD25high regulatory T cells in disseminated and localized forms of allergic contact dermatitis: relation to specific cytokines

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    Halina Laudańska

    2011-07-01

    Full Text Available The aim of this study was to evaluate regulatory T lymphocytes (Tregs in the course of allergic contact dermatitis (ACD and to elucidate the role of IL-10 and TGF-b in Tregs activity. Peripheral blood CD4+CD25+ and CD4+CD25high cells were determined by flow cytometry in patients with acute disseminated ACD (‘ad’, n = 36, acute localized ACD (‘al’, n = 26, and disseminated ACD during remission (‘rd’, n = 27 as well as in controls (n = 22. Serum levels of cytokines were measured using ELISA. The mean percentage of CD4+CD25+ and CD4+CD25high cells in patients with ad ACD was significantly higher than in controls (p < 0.01 and the remaining patients (p < 0.05. Both cell populations were significantly elevated in persons with widespread skin lesions (p < 0.05. In ad patients the CD4+CD25+ increased during three weeks of disease, although the significant increase of CD4+CD25high was noted only in the third week. Patients with ad ACD showed a significantly decreased serum level of TGF-b1 as compared with controls and the remaining ACD patients. IL-10 level did not differ between all groups. The elevated population of CD4+CD25high cells in ad ACD patients, and its dependence on the extension of skin lesions, suggest a role of Tregs in regulating the course of ACD. The growing Tregs percentages may indicate their peripheral generation during ACD. The development of lesions despite an increased population of Tregs suggests their functional defect. The role of TGF-b1 in the suppressive activity of Tregs cannot be excluded. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 2, pp. 255–262

  9. CD4+CD25+调节性T细胞在Graves眼病患者外周血中的表达%Detection of CD4+CD25+regulatory T cell in patients with Graves'ophthalmopathy

    Institute of Scientific and Technical Information of China (English)

    辛梦; 王强; 张磊; 韩兆东; 薛海波

    2013-01-01

    AIM: To observe the change law of the ratio of CD4+CD25+regulatory T cell in the peripheral blood of the patients with Graves'ophthalmopathy by detecting the ratio of this cell. METHODS: This experimental study selected 53 GO patients without exophthalmos, 51 GO patients with exophthalmos, and 51 healthy people were collected.The ratio of CD4+CD25+ T cell in peripheral blood was determined by flow cytometry. RESULTS:The ratio of CD4+CD25+T cell in GO patients without exophthalmos was lower than that in healthy group (P<0.01).The ratio of CD4+CD25+T cell in GO patients with exophthalmos was far lower than that in healthy group (P<0.01).Compared with the group of GO patients without exophthalmos, the ratio of CD4+CD25+T cell of GO patients with exophthalmos was also much lower (P<0.01). CONCLUSION:The ratio of CD4+CD25+T cell in the GO patients decreases and there is autoimmune disorder in patients of this disease. Perhaps this is an important mechanism causing immune suppression damage, which provides a new clue for immunological treatment.%目的:通过检测CD4+CD25+调节性T细胞在Graves眼病患者外周血中的表达,以观察该病患者外周血中CD4+CD25+调节性T细胞的变化规律。方法:临床实验研究。采用流式细胞术检测对Graves病不伴有眼征患者组(53例), Graves眼病患者组(51例),正常对照组(51例)的外周静脉血CD4+CD25+Treg细胞比例进行检测。结果:与正常对照组比较, Graves 病不伴有眼征患者组Treg水平下降(P<0.01);与正常对照组比较,Graves眼病患者组外周血Treg水平显著下降( P<0.01);Graves眼病患者组外周血Treg数量明显低于Graves病不伴有眼征患者组(P<0.01)。结论:Graves眼病患者外周血中CD4+CD25+Treg在淋巴细胞中所占的比例降低,存在自身免疫调节紊乱。可能是其机体细胞免疫抑制受损的重要机制,为对该疾病进行免疫学治疗提供了新线索。

  10. The Phenotypic Characterization of Naturally Occurring Regulatory CD4+CD25+T Cells

    Institute of Scientific and Technical Information of China (English)

    Huanfa Yi; Yu Zhen; Lingling Jiang; Jialin Zheng; Yong Zhao

    2006-01-01

    The homeostasis of the immune system depends on the balance between the immune response to an invaded pathogen and the immune tolerance to self antigens. Both central and peripheral tolerances are important mechanisms for the induction and maintenance of T cell tolerance. Recently, much attention has been paid to regulatory T cells (Treg), which play a significant role in maintaining peripheral immune tolerance. So far, there has been no satisfactory advance regarding the surface markers of Treg cells, as none is unique for Treg cells. In this review, we summarize some important molecules expressed in naturally occurring CD4+CD25+ Treg cells(nTreg), including forkhead/winged-helix family transcriptional repressor p3 (Foxp3), the tumor necrosis factor receptor (TNFR) family, CD28/CTLA4 molecules, chemokine receptors, Toll-like receptors (TLRs), membranebound TGF-β and other molecules, such as neuropilin-1, lymphocyte activation gene-3 (LAG)-3 and granzyme.This review provides a collective view on current studies of nTreg cell activation and development related to the expression of molecules and cell phenotype markers, which is important for elucidation of nTreg cell origin,development and function.

  11. Do CD4+ CD25+ immunoregulatory T cells hinder tumor immunotherapy?

    Science.gov (United States)

    Antony, Paul Andrew; Restifo, Nicholas P

    2002-01-01

    After years of banishment from mainstream immunology, the notion that one subset of T cells can exert regulatory effects on other T lymphocytes is back in fashion. Recent work in knockout and transgenic mice has begun to bring molecular definition to our understanding of immunoregulatory CD4+CD25+ T cells (Treg/Th3/Tr1). The identification of the glucocorticoid-induced tumor necrosis factor receptor family-related gene (GITR, also known as TNFRSF18) expressed on T regulatory cells might afford new therapeutic opportunities. Another possible therapeutic intervention could be the blockade of signaling through the molecular pair of tumor necrosis factor-related activation induced cytokine (TRANCE) and receptor activator of NF-kappaB (RANK). Based on the available evidence from experimental mouse tumor models, however, it seems that simply blocking or even eliminating T regulatory function will not be enough to manage established tumors. The challenge for immunotherapists now is to overcome immunosuppression using the knowledge gained through the understanding of T regulatory cell function.

  12. Impairment of Circulating CD4+CD25+GARP+ Regulatory T Cells in Patients with Acute Coronary Syndrome

    Directory of Open Access Journals (Sweden)

    Kai Meng

    2014-02-01

    Full Text Available Background: Atherosclerosis (AS is an inflammatory and immune disease. Regulatory T cells (Tregs suppress the activation of T cells and have been shown to play a protective role during the pathogenesis of AS. However, specific markers for Tregs are lacking. Recently, glycoprotein A repetitions predominant (GARP was discovered as a specific marker of activated Tregs, and we therefore utilized GARP as a specific surface marker for Tregs in the current study. Methods: To assess whether GARP+ Tregs are downregulated in patients with acute coronary syndrome (ACS, we examined CD4+CD25+GARP+ T cell frequencies as well as their associated cytokines and suppressive function. Additionally, we compared GARP expression to that of FOXP3, which may be more sensitive as a marker of activated Tregs in patients with ACS. Results: Patients with ACS demonstrated a significant decrease in circulating CD4+CD25+GARP+ Tregs. Moreover, the suppressive function of Tregs and levels of related cytokines were also impaired in ACS patients compared to those with stable angina (SA or normal coronary artery (NCA. Additionally, after TCR stimulation, peripheral blood mononuclear cells (PBMCs from patients with ACS exhibited a decrease in CD4+CD25+GARP+ Tregs. Conclusions: These fnding indicate that circulating CD4+CD25+GARP+ Tregs are impaired in patients withACS. Thus, targeting GARP may promote the protective function of Tregs in ACS.

  13. Analysis of circulating regulatory T cells (CD4+CD25+CD127-) after cryosurgery in prostate cancer

    Institute of Scientific and Technical Information of China (English)

    Tong-Guo Si; Jun-Ping Wang; Zhi Guo

    2013-01-01

    This study was performed to assess the response of regulatory T cells (Tregs) following cryosurgery in prostate cancer (PCa) patients by measuring their frequency and immune function.Blood was collected prior to and at 4 and 8 weeks after treatment in 30 patients with high-risk PCa who underwent cryosurgery and from 15 healthy volunteers.Circulating CD4+CD25+CD127-Tregs were isolated.Their frequency was detected by flow cytometry,and immune suppressive function was evaluated by measuring the proliferation of CD4+CD25-T cells cocultured with Tregs.The results showed that the percentage of circulating CD4+CD25+CD127-Tregs was increased in PCa patients compared to healthy volunteers (7.6%±0.73% vs.5.8%±-0.54%,P<0.001).The frequency of circulating CD4+CD25+CD127-Tregs was reduced 4 weeks after cryosurgery compared to before surgery (6.3%±0.58% vs.7.6%±0.73%,P<0.001),and the decrease persisted for 8 weeks.However,the suppressive function of Tregs was increased in eight of 12 patients,which might contribute to cancer recurrence.Then the response of circulating rregs is complicated after cryosurgery for PCa,and further studies are warranted.

  14. 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

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

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    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.

  16. 特应性皮炎患者外周血CD4+CD25+调节性T细胞的检测%Detection of peripheral CD4+CD25+ regulatory T (Treg) cells in patients with atopic dermatitis

    Institute of Scientific and Technical Information of China (English)

    曾美; 高谦; 何定阳; 陈丽华; 吴国珍

    2010-01-01

    目的 探讨CD4+CD25+节性T细胞(CD4+CD25+Treg)在特应性皮炎(AD)发病中的作用机制及临床意义.方法 流式细胞仪分析AD患者外周血中CD4+CD25+Treg细胞数量,实时荧光定量PCR检测外周血单核细胞(PBMC)中Foxp3 mRNA水平,ELISA检测血清中IL-2、IL-4、IL-10、IFN-γ水平.结果 AD患者外周血中CD4+CD25+Treg细胞占CD3+T细胞及CD4+T细胞的比例均明显低于正常人对照组(t'=3.775、4.533,P值均<0.01);外周血中CD4+CD25+Treg细胞占CD3+T细胞比例在AD患者急性期明显低于慢性期(t=2.217,P<0.05),而在急性期与亚急性期、亚急性期与慢性期之间差异均无统计学意义(t=1.558、0.49,P值均>0.05).AD患者PBMC中Foxp3 mRNA的水平低于正常人对照组(z=-2.368,P<0.05);其外周血中CD4+CD25+Treg细胞与血清中IL-2和IL-10成正相关(r=0.512、0.494,P值均<0.05),与IL-4和IFN-γ的相关性无统计学意义(r=-0.110、-0.237,P值均>0.05).结论 在AD患者中,外周血中CD4+CD25+Treg细胞数量及Foxp3 mRNA水平均下降,从而可能减少对Th2细胞增生及其细胞因子分泌的抑制,使Th2占优势,参与AD的发病.%Objective To explore the action mechanism and clinical significance of CD4+CD25+ regulatory T (Treg) cells in the development of atopic dermatitis (AD). Methods Peripheral blood mononuclear cells (PBMC) were obtained from 46 patients with AD and 20 normal human controls. Flow cytometry was performed to detect the number of CD4+CD25+ Treg cells, real-time fluorescence PCR assay to measure the Foxp3 mRNA level in PBMC, ELISA to determine the serum levels of IL-2, IL-4, IL-10 and IFN-γ. Results A statistical decrease was observed in the percentages of peripheral CD4+CD25+ Treg cells among CD3+ T cells and CD4+ T cells in AD patients compared with normal controls (t' = 3.775, 4.533, both P< 0.01 ), and in the percentage of peripheral CD4+CD25+ Treg cells among CD3+ T cells in patients with acute AD compared with those with chronic AD

  17. 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.

  18. Circulating subsets and CD4(+)CD25(+) regulatory T cell function in chronic inflammatory demyelinating polyradiculoneuropathy.

    Science.gov (United States)

    Sanvito, Lara; Makowska, Anna; Gregson, Norman; Nemni, Raffaello; Hughes, Richard A C

    2009-01-01

    Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is an inflammatory disease of the peripheral nervous system that is probably autoimmune in origin. Different components of the adaptive and innate immunity may be responsible for the aberrant response towards nerve antigens. To investigate this, we examined lymphocyte subsets and regulatory T cell (Treg) function in the blood of CIDP patients, healthy controls (HC) and subjects with non-immune mediated neuropathies (other neuropathies, ON). We used flow cytometry to determine the frequency of monocytes, B cells, natural killer (NK) and NK-T cells, total and activated CD4(+) and CD8(+) T cells, effector memory and central memory CD4(+) and CD8(+) T cells, and CD4(+)CD25(high)Foxp3(+) Tregs. Treg function was studied after polyclonal stimulation and antigen specific stimulation with myelin protein peptides in CIDP and HC. There was an increased frequency of monocytes (p = 0.02) and decreased frequency of NK cells (p = 0.02) in CIDP compared with HC but not ON. There were no significant differences in other populations. Treg function was impaired in CIDP compared to HC (p = 0.02), whilst T cell proliferation to myelin protein peptides before and after depletion of Tregs was not different between patients and controls. This study shows increased circulating monocytes and reduced NK cells in CIDP. Although Treg frequency was not altered, we confirm that Tregs display a defect of suppressive function. Myelin protein peptides were not the target of the altered peripheral regulation of the immune response. The mechanisms of peripheral immune tolerance in CIDP and their relevance to the pathogenesis deserve further exploration.

  19. 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)

    Xueyan ePeng

    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.

  20. 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.

  1. Rapamycin combined with allogenic immature dendritic cells selectively expands CD4+CD25+Foxp3+ regulatory T cells in rats

    Institute of Scientific and Technical Information of China (English)

    Guo-YingWang; QiZhang; YangYang; Wen-JieChen; WeiLiu; NanJiang; Gui-HuaChen

    2012-01-01

    BACKGROUND: Dendriticcells(DCs)caninitiatetheexpansion of regulatory T cells (Tregs), which play an indispensable role in inducing transplantation tolerance. Some studies have investigated the effect of the immunosuppressant rapamycin (Rapa) on Tregs in vitro. However, the in vivo effect of Rapa combined with immature DCs (iDCs) on Tregs is unknown. This study was undertaken to determine whether allogenic iDCs combined with a short course of Rapa have the ability to selectivelyexpandtheCD4+CD25+Foxp3+ Tregsinarat model. METHODS: Brown Norway rats were injected intravenously with 2×106 Lewis iDCs followed by 1 mg/kg per day Rapa intraperitoneally for 7 consecutive days. On day 8, the levels of CD4+CD25+Foxp3+ Treg cells in peripheral blood and spleen cells were analyzed by flow cytometry. IL-2, IL-4, TGF-β1, and IFN-γ levels in serum were assessed by ELISA. The experimental animals were divided into four groups: control, Rapa-treated, iDC-treated,andcombination-treated. RESULTS: CD4+CD25+Foxp3+ Tregs comprised 7%-8% of CD4+T cells in control rats. Rapa combined with iDCs enhanced this percentage in the peripheral blood and spleen. However, the levels of Tregs did not significantly change after treatment with Rapa or iDCs alone. The levels of CD4+CD25-Foxp3+ T cells and CD4+CD25+Foxp3- T cells in CD4+ T cells did not significantly change in the combined group. The TGF-β1 level in serum from the combined group increased significantly compared with the other groups. CONCLUSIONS: A significantly higher percentage of CD4+CD25+ Foxp3+ Tregs was found in rats treated with allogenic iDCs and a short course of Rapa, along with an increase in the TGF-β1 level in serum. This improved protocol may be a promising therapeutic strategy to increase Tregs, which are beneficial to the induction of peritransplant tolerance.

  2. Autoimmune thyroid disease and CD4~+ CD25~+ regulatory T cell%自身免疫性甲状腺疾病和CD4~+CD25~+调节性T细胞

    Institute of Scientific and Technical Information of China (English)

    侍晓云; 赵勇

    2010-01-01

    自身免疫性甲状腺疾病(AITD)的发生及发展与CD4~+CD25~+调节性T细胞(Treg细胞)的数量和功能密切相关.动物实验证明Treg细胞可抑制AITD的发生.如果清除动物体内的该类细胞,可导致AITD发病或使原有的甲状腺疾病加重,Treg细胞通过抑制效应性T细胞的激活而发挥对AITD的影响作用.无论是胸腺还是外周,不同诱导体系来源的Treg细胞均对AITD有影响作用.%The occurance and development of autoimmune thyroid disease (AITD)is related with changes of quantities and function of CD4~+CD25~+ regulatory T(Treg) cells. Different studies indicated that Treg cells can inhibit the development of AITD. Depletion of Treg cells can improve the incidence of AITD or increase the severity of AITD. Treg cells play roles in inhibiting activation of effector T cell. No matter Treg cells or CD4~+ CD25~+ regulatory T cells generated in the periphery can influence the development of AITD.

  3. Role of CD4 + CD25 + regulatory T cells in the patients with ulcerative colitis%CD4+CD25+T细胞在溃疡性结肠炎患者中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    宋振梅; 王晓娣

    2010-01-01

    目的 研究溃疡性结肠炎(UC)患者外周血CD4+CD25+调节性T细胞和其特异性标志物Foxp3的表达及其在UC不同病期的变化,探讨其在UC发病机制中的意义.方法 研究纳入2007年2月至2008年12月在我院就诊的UC患者40例,其中活动期23例,缓解期17例;同时选择33例肠易激综合征(IBS)患者及32名健康者作为对照.用流式细胞仪检测UC患者、IBS患者和正常对照者的外周血CD4+CD25+T细胞阳性率,用RT-PCR检测外周血单个核细胞(PBMC)中Foxp3 mRNA的表达,用酶联免疫吸附实验检测血清中白细胞介素-10(IL-10)和转化生长因子-β(TGF-β)的浓度.结果 活动期和缓解期UC患者、IBS患者、正常对照者外周血CD4+T细胞总数相比差异无统计学意义(P=0.126).CD4+CD25+T细胞在CD4+T细胞中所占比例在活动期和缓解期UC患者中均明显低于IBS患者和正常对照组(P均<0.01),活动期患者较缓解期患者更低(P<0.001),并且与疾病活动指数(r=-0.660,P<0.001)及红细胞沉降率(r=-0.572,P=0.001)均呈显著负相关,而IBS患者和正常对照者间差异没有统计学意义(P=0.343).PBMC中Foxp3 mRNA的表达在活动期和缓解期UC患者均明显低于IBS患者和正常对照组(P均<0.001).而血清IL-10和TGF-β的浓度在活动期和缓解期UC患者、IBS患者和正常对照者4组间无明显差异(P均>0.05).结论 活动期UC患者外周血CD4+CD25+调节性T细胞明显降低,缓解期UC患者的表达有所升高,并且与临床疾病活动相关,提示这类细胞可能参与了UC疾病的发生发展,Foxp3表达降低可能是导致CD4+CD25+T细胞发育障碍的重要因素.%Objective To investigate the levels of CD4 + CD25 + regulatory T cells and the Foxp3 in the peripheral blood of patients with ulcerative colitis ( UC), and analyze its role in the pathogenesis of UC. Methods From February 2007 to December 2008, 40 UC patients (23 active and 17 remissive), 33 irritable bowel syndrome (IBS

  4. Immunosuppressive effects of rat mesenchymal stem cells:involvement of CD4+CD25+regulator y T cells

    Institute of Scientific and Technical Information of China (English)

    Zhou Ye; Yan Wang; Hai-Yang Xie and Shu-Sen Zheng

    2008-01-01

    BACKGROUND: Recent studies show that mesenchymal stem cells (MSCs) have immunomodulatory properties. They suppress the immune response to alloantigen and modify the proliferation of T cells. CD4+CD25+ regulatory T cells have strong immunomodulatory potential. However, little is known about the effects of rat MSCs (rMSCs) on the development of regulatory T cells. METHODS:MSCs were obtained from bone marrow of male Sprague-Dawley rats, and co-cultured with CD3+ T cells from allogeneic spleen cells. The proportion of CD4+CD25+ regulatory  T  cells  was  analyzed  by  lfow  cytometry.  To further conifrm the immunosuppressive activity of rMSCs, we used MTT assay and lfow cytometry of CD3+ T cells to investigate the proliferative responses of CD3+ T cells to mitogenic stimuli. Enzyme-linked immunosorbent assay was performed to detect alterations of the cytokines TNF-α, TGF-β and IL-10. RESULTS:The proliferation of CD3+ T cells decreased when co-cultured with rMSCs, and the degree of inhibition was concentration-dependent. The percentage of CD4+CD25+ regulatory T cells increased when CD3+ T cells were co-cultured with different concentrations of rMSCs. The levels of pro-inlfammatory cytokine (TNF-α) decreased while anti-inlfammatory (TGF-β, IL-10) cytokines increased in mixed lymphocyte reaction. CONCLUSIONS:rMSCs inhibit allogeneic T cell proliferation in mixed cell cultures. This immunosuppressive effect seems  to  be  mediated  by  inducing  the  generation  of CD4+CD25+ regulatory T cells and soluble factors.

  5. Impairment of circulating CD4+CD25+ regulatory T cells in patients with chronic inflammatory demyelinating polyradiculoneuropathy.

    Science.gov (United States)

    Chi, Li-Jun; Wang, Hua-Bing; Wang, Wei-Zhi

    2008-03-01

    Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is an immune-mediated peripheral nervous system disease. CD4+CD25+ T regulatory cells (Tregs) have been unequivocally shown to be critical in maintaining immune tolerance and preventing auto-immune diseases by suppressing self-reactive T cells. Thus, we hypothesized that the numbers and/or the function of Tregs would be deranged during the progressive or relapse phases of CIDP. The number of Tregs was determined by flow cytometry according to their characteristic CD4+CD25(high) membrane phenotype. Functional characterization of Tregs was analyzed by suppression of proliferation and secretion of cytokines by co-cultured effector CD4+CD25- T cells. FOXP3 message expression level was assessed by quantitative real-time polymerase chain reaction. The results showed significant reduction in both the number and the suppressive function of Tregs in the patients with CIDP compared with healthy controls. Also, Tregs isolated from CIDP patients expressed lower levels of FoxP3 mRNA. During the progressive or the relapsing phases of CIDP, the number of Tregs was reduced, and the suppressive function of them decreased. These findings may be helpful to our understanding of the possible role of Tregs in the pathogenesis of CIDP.

  6. 脓毒症患者CD4+CD25+调节性T细胞检测的临床意义%Clinic significance of Testing CD4+CD25+ Treg cells in patients with sepsis

    Institute of Scientific and Technical Information of China (English)

    邵敏; 刘宝; 王锦权; 陶晓根; 周树生; 金魁; 张翠萍

    2011-01-01

    目的 探讨脓毒症患者CD4+CD25+调节性T细胞(Treg)的水平与机体细胞免疫状态的关系及其检测的临床意义.方法 将脓毒症患者40例按照疾病严重程度分为3组:脓毒症组14例,严重脓毒症组15例,脓毒症休克组11例.所有入选的40例脓毒症患者在入选当天行流式细胞术检测血中CD4+CD25+调节性T细胞的比例和CD3+、CD4+、CD8+和CD4+/CD8+水平.比较脓毒症组、严重脓毒症组和脓毒症休克组血中CD4+CD25+调节性T细胞比例变化和CD3+、CD4+、CD8+和CD4+/CD8+水平变化.同时选取20例健康人为对照组.结果 健康对照组CD4+CD25+调节性T细胞表达率脓毒症休克组最高,严重脓毒症组次之,脓毒症组最低,三组间两两比较差异有统计学意义(P<0.05).脓毒症组CD3+、CD4+和CD4+/CD8+较正常对照组明显升高(P<0.05),CD8+较正常对照组差异不明显(P>0.05).严重脓毒症组和脓毒症休克患者CD3+、CD4+、CD4+/CD8+较正常对照组明显降低(P<0.05),CD8+较正常对照组改变不明显(P>0.05).结论 脓毒症患者外周血CD4+CD25+调节性T细胞表达增加,可能导致CD3+、CD4+、CD4+/CD8+水平降低,免疫抑制加强.提示CD4+CD25+调节性T细胞在脓毒症的免疫发病机制中可能起着重要作用,对评估患者预后有临床价值.%Objective To study the level and significance of CD4+CD25+ Treg cells and CD3+、 CD4+、 CD8+ cells in peripheral blood of patients with sepsis. Methods 40 patients with sepsis Patients were divided into three groups: sepsis group(n=14), severe sepsis group(n=l5), and septic shock group (n=11).The expression of CD4+CD25+ Treg cells and CD3+、 CD4+、 CD8+ cells of 40 patients on the day 1 was detected by flow cytometry. The comparision in the expression of CD4+CD25+ Treg cells and CD3+、 CD4+、 CD8+ cells within groups was made. Twenty healthy individuals served as controls. Results The expression rate. The expression of CD4+CD25+ Treg cells was the highest

  7. 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.

  8. Effects of IL-7 and IL-2 on CD4+CD25-T cells proliferation in vitro%IL-7联合IL-2对脐血CD4+CD25-T细胞体外诱导扩增影响的研究

    Institute of Scientific and Technical Information of China (English)

    仉洁; 郝京生; 李俊甫; 邢龙龙; 刘志文; 钟理; 刘薇

    2014-01-01

    目的:初步探讨IL-7联合IL-2对脐血CD4+CD25-T细胞体外扩增的促进作用,建立稳定的体外培养扩增脐血CD4+CD25-Tregs的培养体系,比较诱导扩增后的CD4+CD25+Tregs和自然分选的CD4+CD25+Tregs对PBMCs功能活性的影响。方法:采用免疫磁珠分选法分选出脐血CD4+CD25-T细胞和CD4+CD25+T细胞;加入不同浓度的细胞因子IL-7结合适当浓度的IL-2作为诱导剂,分析IL-7体外诱导CD4+CD25-T细胞增殖的有效性及最适浓度。我们利用流式细胞术检测体外扩增后的CD4+CD25-T细胞表型变化;MTS法检测体外诱导扩增的CD4+CD25+Tregs及自然分选的CD4+CD25+Tregs分别对成人外周血中单个核细胞增殖的抑制作用;RT-PCR方法分析体外诱导扩增的CD4+CD25+Tregs 及自然分选的CD4+CD25+Tregs FOXP3基因、IL-10基因和TGF-β基因的cDNA表达的变化。结果:经3周体外培养、各组细胞均有明显的扩增。 IL-7诱导组的扩增最强。体外抑制试验显示体外扩增的Tregs对成人外周血中单个核细胞有明显的抑制作用,IL-7联合IL-2诱导CD4+CD25-T细胞生成的CD4+CD25+Tregs具有较自然分选的CD4+CD25+Tregs稍弱的免疫抑制功能,其中IL-7浓度为4 ng/ml,IL-2浓度为2000 U/ml时诱导CD4+CD25-T细胞生成的CD4+CD25+Tregs杀伤活性最强。结论:成功建立了CD4+CD25+Tregs的体外培养体系,联合应用IL-7、大剂量的IL-2和CD3/CD28单抗是体外诱导扩增CD4+CD25-T细胞成为CD4+CD25+Tregs的优选方法,并且扩增倍数高、可持续高表达CD4及CD25细胞表型。%Objective:To explore the promoting effects of IL-7 and IL-2 on CD4+CD25-T cells proliferation in vitro and construct a stable culture system in vitro for CD 4+CD25+regulatory T cells from human umbilical cord blood.To compare the inhibiting effects between induced proliferated CD 4+CD25+Tregs and naturally isolated CD 4+CD25+Tregs

  9. Comparative Study of Regulatory T Cell Function of Human CD25+CD4+ T Cells from Thymocytes, Cord Blood, and Adult Peripheral Blood

    Directory of Open Access Journals (Sweden)

    Wakae Fujimaki

    2008-01-01

    Full Text Available CD25+CD4+ regulatory T cells suppress T cell activation and regulate multiple immune reactions in in vitro and in vivo studies. To define the regulatory function of human CD25+CD4+ T cells at various stages of maturity, we investigated in detail the functional differences of CD25+CD4+ T cells from thymocytes, cord blood (CB, and adult peripheral blood (APB. CB CD25+CD4+ T cells displayed low-FOXP3 protein expression level and had no suppressive activity. In contrast, CD25+CD4+ T cells from thymocytes or APB expressed high expression level of FOXP3 protein associated with significant suppressive activity. Although CB CD25+CD4+ T cells exhibited no suppressive activity, striking suppressive activity was observed following expansion in culture associated with increased FOXP3 expression and a shift from the CD45RA+ to the CD45RA− phenotype. These functional differences in CD25+CD4+ T cells from Thy, CB, and APB hence suggest a pathway of maturation for Treg in the peripheral immune system.

  10. Inhibition of mouse acrosome reaction and sperm-zona pellucida binding by anti-human sperm membrane protein 1 antibody%抗人精子膜蛋白抗体1对小鼠精子顶体反应和精子-透明带结合的抑制作用

    Institute of Scientific and Technical Information of China (English)

    G.Y.Cheng; J.L.Shi; M.Wang; Y.Q.Hu; C.M.Liu; Y.F.Wang; C.Xu

    2007-01-01

    Aim:To investigate the possible functions of human sperm membrane protein (hSMP-1) in the process of fertilization.Methods: A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 (anti-rhSMP-1) antibodies. Sperm acrosome reaction and spermzona pellucida (ZP) binding assay were carried out in 10-week-old BALB/c mice. Results: Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 (P < 0.05). Conclusion: The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.

  11. Effect of Oxymatrine on CD4+CD25+ regulatory T cells and lymphocyte proliferation%氧化苦参碱对CD4+CD25+调节性T细胞(Tr)和淋巴细胞增殖的影响

    Institute of Scientific and Technical Information of China (English)

    李建国; 伍斌; 谢红付; 张江林; 易梅; 李吉

    2008-01-01

    antibody,using flow cytometer to detect the quantity the of CD4+ CD25+ T cells.To utilize Carboxyfluorescein diacetate succinimidyl ester(CFDA-SE)dyed and flow cytometer to examine the fluorescence intensity changes of lymphocytes co-stimulated by polyclonal stimulator Con A and OMT.[Results]500,125 and 31 μ.g/mL OMT has the ability to restrain the proliferation of lymphocyte of mouse depending on its concentration in substrate,but 16,8,4,2μgs/mL OMT improve the proliferation of T lymphocytes of mouse's lymph node,no obvious dependence on its concentration in substrate.Intraperitoneal injection of OMT can improve mouse CD4+ CD36+T cell quantities in peripheral blood obviously,compared OMT groups with HCT and PBS group(P<0.01).[Conclusions]The effects of OMT on the proliferation of lymDhocytes of mouse's lymph node T cells presents the two-way function,OMT can obviously improve the quantity of CD4+ CD25+T cells in the peripheral blood of mouse throush intraperitoneal injection,implying that OMT is a kind of two-way immunoregulator.

  12. 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

  13. Modulation of phenotype and function of human CD4+CD25+ T regulatory lymphocytes mediated by cAMP elevating agents

    Directory of Open Access Journals (Sweden)

    Antonella Riccomi

    2016-09-01

    Full Text Available We have shown that Cholera Toxin (CT and other cyclic AMP (cAMP elevating agents induce up-regulation of the inhibitory molecule CTLA-4 in human resting CD4+ T lymphocytes, which following the treatment acquired suppressive functions. In this study, we evaluated the effect of cAMP elevating agents on human CD4+CD25+ T cells, which include the T regulatory (Treg cells that play a pivotal role in the maintenance of immunological tolerance. We found that cAMP elevating agents induce up-regulation of CTLA-4 in CD4+CD25- and further enhance its expression in CD4+CD25+ T cells. We observed an increase of two isoforms of mRNA coding for the membrane and the soluble CTLA-4 molecules, suggesting that the regulation of CTLA-4 expression by cAMP is at the transcriptional level. In addition, we found that the increase of cAMP in CD4+CD25+ T cells converts the CD4+CD25+Foxp3- T cells in CD4+CD25+Foxp3+ T cells, whereas the increase of cAMP in CD4+CD25- T cells did not up-regulate Foxp3 in the absence of activation stimuli. To investigate the function of these cells, we performed an in vitro suppression assay by culturing CD4+CD25+ T cells untreated or pre-treated with CT with anti-CD3 mAbs-stimulated autologous PBMC. We found that CT enhances the inhibitory function of CD4+CD25+ T cells, CD4+ and CD8+ T cell proliferation and IFNγ production are strongly inhibited by CD4+CD25+ T cells pre-treated with cAMP elevating agents. Furthermore, we found that CD4+CD25+ T lymphocytes pre-treated with cAMP elevating agents induce the up-regulation of CD80 and CD86 co-stimulatory molecules on immature dendritic cells (DCs in the absence of antigenic stimulation, however without leading to full DC maturation. These data show that the increase of intracellular cAMP modulates the phenotype and function of human CD4+CD25+ T cells.

  14. Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells*

    OpenAIRE

    2009-01-01

    Objective: In this paper we compared the two methods of cell sorting (magnetic cell sorting and flow cytometry sorting) for the isolation and function analysis of mouse CD4+ CD25+ regulatory T (Treg) cells, in order to inform further studies in Treg cell function. Methods: We separately used magnetic cell sorting and flow cytometry sorting to identify CD4+ CD25+ Treg cells. After magnetic cell separation, we further used flow cytometry to analyze the purity of CD4+ CD25+ Treg cells, trypan bl...

  15. Reverse effect of CD4+CD25+ regulatory T cells on lupus-like murine%CD4+CD25+调节性T细胞对狼疮样小鼠的干预性研究

    Institute of Scientific and Technical Information of China (English)

    苏虹; 王保龙; 方雪晖; 陈娟; 叶冬青

    2008-01-01

    目的 分析CD4+CD25+调节性T细胞(regulatory T cells,Tregs)对狼疮样小鼠的干预效应,为系统性红斑狼疮(systemic lupus erythematosus,SLE)的免疫干预提供新的思路.方法 采用磁活性标记的细胞分选方法 (magnetic activated cell sorting,MACS)分选BALB/c小鼠CD4+CD25+T细胞,应用流式细胞术(flow cytometry,FCM)检测分选细胞纯度.将6~8 w ♀CB6F1小鼠随机分为3组,包括正常对照组、狼疮鼠模型组(亲代淋巴细胞免疫)、狼疮鼠干预组(亲代淋巴细胞免疫后2 w,输入5×106 Tregs/鼠).分别于诱导后2、4、8、12 w,采用ELISA法检测ANA和抗-dsDNA抗体,并观察肾标本的病理学改变.结果 BALB/c小鼠脾脏单个核细胞经MACS分选后,CD4+CD25+T细胞纯度达98.5%.经亲代淋巴细胞输注后,CB6F1小鼠出现体重降低、皮肤干涩;ANA和抗ds-DNA抗体上升;以及狼疮肾炎的病理学改变.而输入Tregs对已出现自身抗体的狼疮鼠有明显的逆转效应,CB6F1小鼠于输注后抗-dsDNA即开始下降,并伴随其他临床症状和实验室指标的逐渐改善.结论 CD4+CD25+Tregs具有明显的逆转CB6F1鼠狼疮样疾病的效应,将为SLE的免疫干预开辟新途径.

  16. 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.

  17. Evaluation of CD4+ CD25+ FoxP3+ regulatory T cells during treatment of patients with brucellosis.

    Science.gov (United States)

    Hasanjani Roushan, M R; Bayani, M; Soleimani Amiri, S; Mohammadnia-Afrouzi, M; Nouri, H R; Ebrahimpour, S

    2016-01-01

    Cell-mediated immunity (CMI) plays a critical role in the control of brucellosis. Regulatory T cells (Tregs) have a functional character in modulating the balance between host immune response and tolerance, which can eventually lead to chronic infection or relapse. The aim of this study was to assess the alteration of Tregs in cases of brucellosis before and after treatment. Thirty cases of acute brucellosis with the mean age of 41.03±15.15 years (case group) and 30 healthy persons with the mean age of 40.63±13.95 years (control group) were selected and assessed. Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood of all individuals. We analyzed the alteration of Treg cell count using flow cytometry for CD4, CD25, and FoxP3 markers. The level of CD4+ CD25+ FoxP3+ Treg cells was increased in active patients compared with controls (2.5±0.99% vs 1.6±0.84%, p= 0.0004), but it had declined in the treated cases (1.83±0.73%, p=0.02). The level of Tregs was elevated in three relapsed cases. The frequency of Tregs and Treg/Teff (effector T cell) ratio was correlated with inverse serum agglutination test (SAT) and, 2-mercaptoethanol (2-ME) titers as markers of treatment in brucellosis. Based on our findings, we suggest that regulatory cells, such as CD4+ CD25+ FoxP3+ Treg cells, may contribute to the development of infection processes involving immune responses in brucellosis, and evaluation of regulatory T-cell levels may be a potential diagnostic strategy for the treatment outcome in chronic and relapsed cases of brucellosis.

  18. Increase in TGF-β Secreting CD4+CD25+ FOXP3+ T Regulatory Cells in Anergic Lepromatous Leprosy Patients

    Science.gov (United States)

    Saini, Chaman; Ramesh, Venkatesh; Nath, Indira

    2014-01-01

    Background Lepromatous leprosy caused by Mycobacterium leprae is associated with antigen specific T cell unresponsiveness/anergy whose underlying mechanisms are not fully defined. We investigated the role of CD25+FOXP3+ regulatory T cells in both skin lesions and M.leprae stimulated PBMC cultures of 28 each of freshly diagnosed patients with borderline tuberculoid (BT) and lepromatous leprosy (LL) as well as 7 healthy household contacts of leprosy patients and 4 normal skin samples. Methodology/Principle Findings Quantitative reverse transcribed PCR (qPCR), immuno-histochemistry/flowcytometry and ELISA were used respectively for gene expression, phenotype characterization and cytokine levels in PBMC culture supernatants. Both skin lesions as well as in vitro antigen stimulated PBMC showed increased percentage/mean fluorescence intensity of cells and higher gene expression for FOXP3+, TGF-β in lepromatous (p<0.01) as compared to tuberculoid leprosy patients. CD4+CD25+FOXP3+ T cells (Tregs) were increased in unstimulated basal cultures (p<0.0003) and showed further increase in in vitro antigen but not mitogen (phytohemaglutinin) stimulated PBMC (iTreg) in lepromatous as compared to tuberculoid leprosy patients (p<0.002). iTregs of lepromatous patients showed intracellular TGF-β which was further confirmed by increase in TGF-β in culture supernatants (p<0.003). Furthermore, TGF-β in iTreg cells was associated with phosphorylation of STAT5A. TGF-β was seen in CD25+ cells of the CD4+ but not that of CD8+ T cell lineage in leprosy patients. iTregs did not show intracellular IFN-γ or IL-17 in lepromatous leprosy patients. Conclusions/Significance Our results indicate that FOXP3+ iTregs with TGF-β may down regulate T cell responses leading to the antigen specific anergy associated with lepromatous leprosy. PMID:24454972

  19. Reduction of CD4(+)CD25(+) regulatory T-cells in migraine: Is migraine an autoimmune disorder?

    Science.gov (United States)

    Arumugam, Murugesan; Parthasarathy, Varadarajan

    2016-01-15

    Migraine is believed to be a chronic neurological disorder with the exact aetiology being unknown. But, there is a debate on the role of immune dysfunction in migraine pathophysiology. Hence, authors made a debut attempt to explore the link between lymphocyte subset populations and migraine. A significant increase in CD4(+) and decrease in CD8(+) population were observed in migraine patients compared to healthy volunteers. Interestingly, the immunoregulator CD4(+)CD25(+) levels were less in migraine patients compared to the healthy volunteers. The results of the present study indicate that failure of immunoregulation could be implicated in the pathophysiology of migraine.

  20. Dendritic cells control CD4+CD25+ Treg cell suppressor function in vitro through juxtacrine delivery of IL-2.

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    Katarina Kulhankova

    Full Text Available CD4(+CD25(+Foxp3(+ regulatory T cells (Tregs restrict inflammatory responses to self and nonself. Aberrant Treg activity is pathologic: Insufficient Treg activity is implicated in autoimmunity, allergy, and graft-versus-host-disease; overabundant activity is implicated in chronic infection and cancer. Tregs require IL-2 for their expansion and acquisition/execution of suppressor function; however, because Tregs cannot produce IL-2, they depend on IL-2 from an exogenous source. Until now, that IL-2 source had not been established. We asked whether dendritic cells (DCs could supply IL-2 to Tregs and, if so, what was required for that delivery. We used flow cytometry, IL-2 ELISPOT, RT-qPCR, and IL-2 promoter-driven reporter assays to measure intracytoplasmic IL-2, secreted protein, IL-2 message and IL-2 promoter activity in bone marrow-derived (BMDC and splenic DCs. We examined conjugate formation between Tregs, conventional CD4(+ cells, and IL-2-expressing DCs. We measured Treg levels of CD25, Foxp3, and suppressor function after co-culture with IL-2 sufficient and IL-2(-/- DCs. We generated IL-2-mCherry-expressing DCs and used epifluorescence microscopy and flow cytometry to track IL-2 transfer to Tregs and test requirements for transfer. Between 0.7 to 2.4% of DCs constitutively produced IL-2 and diverted IL-2 secretion to Tregs by preferentially forming conjugates with them. Uptake of DC IL-2 by Tregs required cell-cell contact and CD25. Tregs increased levels of CD25 and Foxp3 from baseline and showed greater suppressor function when co-cultured with IL-2-sufficient DCs, but not when co-cultured with IL-2(-/- DCs. Exogenous IL-2, added in excess of 500 U/ml to co-cultures with IL-2(-/- DCs, restored Treg suppressor function. These data support a model of juxtacrine delivery of IL-2 from DCs to Tregs and suggest that a subset of DCs modulates Treg function through controlled, spatial delivery of IL-2. Knowledge of how DCs regulate Tregs should

  1. CD4+ CD25+Treg细胞和Th17细胞与寻常型银屑病发病的相关性

    Institute of Scientific and Technical Information of China (English)

    张玉丽

    2011-01-01

    CD4+ CD25+ Treg细胞和Th17细胞是CD4+T细胞的新亚群,参与自身免疫病、感染和肿瘤等疾病的发生发展.研究表明CD4+ CD25+ Treg细胞和Th17细胞与寻常型银屑病的发病密切相关.通过对CD4+ CD25+ Treg细胞和Th17分化发育和功能发挥过程中的关键调节因子进行阻断或加强,可以上调或下调CD4+ CD25+ Treg细胞和Th17在寻常型银屑病中的表达,以用于寻常型银屑病的预防和诊治.

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

    Science.gov (United States)

    Jun, Cao; Qingshu, Li; Ke, Wei; Ping, Li; Jun, Dong; Jie, Luo; Su, Min

    2015-01-01

    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.

  3. Phenotypes and clinical significance of circulating CD4+CD25+ regulatory T cells (Tregs in patients with acute-on-chronic liver failure (ACLF

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    Yang Jiezuan

    2012-09-01

    Full Text Available Abstract Background CD4+CD25+ regulatory T cells (Tregs play an important role in maintaining immunological tolerance to self and foreign antigens. T cell receptors (TCR reflect the composition and function of T cells. It is not universally agreed that there is a relationship between CD4+CD25+ Treg frequency and the severity of acute-on-chronic liver failure (ACLF. The repertoire of TCR beta chain variable (TCRBV regions of peripheral Tregs in ACLF patients is not well understood. Methods Human PBMCs were separated and sorted into CD4+CD25+ Treg subsets using density gradient centrifugation and magnetic activated cell sorting (MACS. The CD4+CD25high Treg frequency in peripheral blood of ACLF and chronic hepatitis B (CHB patients was measured by flow cytometry. The molecular profiles of TCRBV CDR3 were determined using gene melting spectral pattern (GMSP analysis. TCRBV gene families were cloned and sequenced when the GMSP profiles showed a single-peak. Results CD4+CD25high Treg prevalence in peripheral blood of ACLF patients is increased significantly compared to healthy donors (HDs (P P +CD25high Tregs in ACLF or CHB patients is positively correlated with HBV DNA load. The TCRBV11, BV13.1, BV18, BV20 are the most prevalent TCRBV in CD4+CD25+ Tregs in ACLF and CHB patients. In addition, the CDR3 motifs were relatively conserved in these four TCRBV gene families. Conclusions The CD4+CD25high Tregs prevalence in peripheral blood is indicative of disease severity in ACLF or CHB patients. The relatively conserved TCRBV20 CDR3 motif “TGTGHSPLH” and TCRBV11 CDR3 motif “VYNEQ” may be used in helping diagnosis and treat patients with ACLF.

  4. Lack of Suppressive CD4+CD25+FOXP3+ T Cells in Advanced Stages of Primary Cutaneous T-Cell Lymphoma

    OpenAIRE

    Tiemessen, Machteld M.; Mitchell, Tracey J.; Hendry, Lisa; Whittaker, Sean J; Taams, Leonie S.; John, Susan

    2006-01-01

    Mycosis fungoides and its leukemic variant, Sezary syndrome, are the most common primary cutaneous T-cell lymphomas (CTCLs). In an ex vivo study, we investigated the percentage, phenotype, and suppressive function of CD4+CD25+ regulatory T cells (Tregs) from peripheral blood of CTCL patients. The percentage of Tregs did not differ significantly between patients and controls. Functional assays demonstrated a dichotomy in Treg function: in four out of 10 patients CD4+CD25+ T cells were incapabl...

  5. Cytotoxic T Lymphocyte–Associated Antigen 4 Plays an Essential Role in the Function of Cd25+Cd4+ Regulatory Cells That Control Intestinal Inflammation

    OpenAIRE

    Read, Simon; Malmström, Vivianne; Powrie, Fiona

    2000-01-01

    It is now clear that functionally specialized regulatory T (Treg) cells exist as part of the normal immune repertoire, preventing the development of pathogenic responses to both self- and intestinal antigens. Here, we report that the Treg cells that control intestinal inflammation express the same phenotype (CD25+CD45RBlowCD4+) as those that control autoimmunity. Previous studies have failed to identify how CD25+ Treg cells function in vivo. Our studies reveal that the immune-suppressive func...

  6. DNA Methyltransferase Inhibitor Promotes Human CD4+CD25hFOXP3+ Regulatory T Lymphocyte Induction under Suboptimal TCR Stimulation

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    Chun-Hao Lu

    2016-11-01

    Full Text Available The master transcription factor FOXP3 regulates the differentiation, homeostasis, and suppressor function of CD4+ regulatory T (Treg cells, which are critical in maintaining immune tolerance. Epigenetic regulation of FOXP3 expression has been demonstrated to be important to Treg cell development, but the induction of human Treg cells through epigenetic modification has not been clearly described. We report that the combination of the DNA methyltransferase inhibitor 5-azacytidine (5-Aza and suboptimal T cell receptor (TCR stimulation promoted CD4+CD25hFOXP3+ T cell induction from human CD4+CD25- T cells. 5-Aza treatment enhanced the expression of Treg cell signature genes, CD25, FOXP3, CTLA-4, and GITR, in CD4+CD25h cells. Moreover, 5-Aza-treated CD4+CD25h T cells showed potent suppressive activity in a cell contact-dependent manner and reduced methylation in the Treg-specific demethylated region (TSDR in the FOXP3 gene. The analysis of cytokine production revealed that CD4+CD25- T cells with 5-Aza treatment produced comparable levels of interferon (IFN-γ and transforming growth factor (TGF-β, but less IL-10, and more IL-2 when compared to cells without 5-Aza treatment. The increased IL-2 was indispensible to the enhanced FOXP3 expression in 5-Aza-treated CD4+CD25h cells. Finally, 5-Aza-treated CD4+CD25h T cells could be expanded with IL-2 supplementation alone and maintained FOXP3 expression and suppressor function through the expansion. Our findings demonstrate that DNA demethylation can enhance the induction of human Treg cells and promise to solve one of the challenges with using Treg cells in therapeutic approaches.

  7. Lack of suppressive CD4+CD25+FOXP3+ T cells in advanced stages of primary cutaneous T-cell lymphoma.

    Science.gov (United States)

    Tiemessen, Machteld M; Mitchell, Tracey J; Hendry, Lisa; Whittaker, Sean J; Taams, Leonie S; John, Susan

    2006-10-01

    Mycosis fungoides and its leukemic variant, Sezary syndrome, are the most common primary cutaneous T-cell lymphomas (CTCLs). In an ex vivo study, we investigated the percentage, phenotype, and suppressive function of CD4+CD25+ regulatory T cells (Tregs) from peripheral blood of CTCL patients. The percentage of Tregs did not differ significantly between patients and controls. Functional assays demonstrated a dichotomy in Treg function: in four out of 10 patients CD4+CD25+ T cells were incapable of suppressing autologous CD4+CD25- T-cell proliferation, whereas suppressive function was intact in the other six patients. Suppressive activity of Tregs inversely correlated with the peripheral blood tumor burden. T-plastin gene expression, used as a Sezary cell marker, confirmed that Sezary cells were heterogeneous for CD25 expression. Mixed lymphocyte reactions demonstrated that CD4+CD25- T cells from patients who lacked functional Tregs were susceptible to suppression by Tregs from healthy controls, and had not become suppressive themselves. Furthermore, we found reduced expression of Foxp3 in the CD4+CD25+ Tregs of these patients relative to the other six CTCL patients and controls. Our findings thus indicate a dysfunction of peripheral Tregs in certain CTCL patients, which correlates with tumor burden.

  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. B7-deficient autoreactive T cells are highly susceptible to suppression by CD4(+)CD25(+) regulatory T cells.

    Science.gov (United States)

    May, Kenneth F; Chang, Xing; Zhang, Huiming; Lute, Kenneth D; Zhou, Penghui; Kocak, Ergun; Zheng, Pan; Liu, Yang

    2007-02-01

    CD4(+)CD25(+) regulatory T cells (Tregs) suppress immunity to infections and tumors as well as autoimmunity and graft-vs-host disease. Since Tregs constitutively express CTLA-4 and activated T cells express B7-1 and B7-2, it has been suggested that the interaction between CTLA-4 on Tregs and B7-1/2 on the effector T cells may be required for immune suppression. In this study, we report that autopathogenic T cells from B7-deficient mice cause multiorgan inflammation when adoptively transferred into syngeneic RAG-1-deficient hosts. More importantly, this inflammation is suppressed by adoptive transfer of purified wild-type (WT) CD4(+)CD25(+) T cells. WT Tregs also inhibited lymphoproliferation and acquisition of activation markers by the B7-deficient T cells. An in vitro suppressor assay revealed that WT and B7-deficient T cells are equally susceptible to WT Treg regulation. These results demonstrate that B7-deficient T cells are highly susceptible to immune suppression by WT Tregs and refute the hypothesis that B7-CTLA-4 interaction between effector T cells and Tregs plays an essential role in Treg function.

  10. 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.

  11. Anti-CD25 treatment depletes Treg cells and decreases disease severity in susceptible and resistant mice infected with Paracoccidioides brasiliensis.

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    Maíra Felonato

    Full Text Available Regulatory T (Treg cells are fundamental in the control of immunity and excessive tissue pathology. In paracoccidioidomycosis, an endemic mycosis of Latin America, the immunoregulatory mechanisms that control the progressive and regressive forms of this infection are poorly known. Due to its modulatory activity on Treg cells, we investigated the effects of anti-CD25 treatment over the course of pulmonary infection in resistant (A/J and susceptible (B10.A mice infected with Paracoccidioides brasiliensis. We verified that the resistant A/J mice developed higher numbers and more potent Treg cells than susceptible B10.A mice. Compared to B10.A cells, the CD4(+CD25(+Foxp3(+ Treg cells of A/J mice expressed higher levels of CD25, CTLA4, GITR, Foxp3, LAP and intracellular IL-10 and TGF-β. In both resistant and susceptible mice, anti-CD25 treatment decreased the CD4(+CD25(+Foxp3(+ Treg cell number, impaired indoleamine 2,3-dioxygenase expression and resulted in decreased fungal loads in the lungs, liver and spleen. In A/J mice, anti-CD25 treatment led to an early increase in T cell immunity, demonstrated by the augmented influx of activated CD4(+ and CD8(+ T cells, macrophages and dendritic cells to the lungs. At a later phase, the mild infection was associated with decreased inflammatory reactions and increased Th1/Th2/Th17 cytokine production. In B10.A mice, anti-CD25 treatment did not alter the inflammatory reactions but increased the fungicidal mechanisms and late secretion of Th1/Th2/Th17 cytokines. Importantly, in both mouse strains, the early depletion of CD25(+ cells resulted in less severe tissue pathology and abolished the enhanced mortality observed in susceptible mice. In conclusion, this study is the first to demonstrate that anti-CD25 treatment is beneficial to the progressive and regressive forms of paracoccidioidomycosis, potentially due to the anti-CD25-mediated reduction of Treg cells, as these cells have suppressive effects on the

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

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    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

  13. An experimental study on inhibiting graft rejection following high-risk penetrating keratoplasty by CD25 siRNA nanocarrier in rats

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    Yun-jie SHI

    2015-06-01

    Full Text Available Objective To investigate the effects of CD25 siRNA nanoparticles against immune rejection and prolongation of corneal graft survival time after high-risk corneal grafting in rats. Methods Orthotopic corneal transplantation was performed in SD rats with alkali burned corneas to mimic high-risk rat models. Donor cornea (Wistar rats was grafted into the right cornea of SD recipients on day 14 after alkali burn. The grafted rats were randomly divided into control group (Group A, EntransterTM-control CD25siRNA instillation treatment (Group B, EntransterTM-CD25siRNA instillation treatment (Group C and EntransterTM-CD25siRNA twice instillation treatment (Group D, first administration at 2-hour post-surgery and second on day 7 post-surgery. The recipient eyes were examined using a slit lamp microscope. Then, the mean survival time and rejection index (RI were calculated. The morphologies of grafts were microscopically examined with HE staining, and TEM. CD25 expression after operation was determined by quantitative RT-PCR and immunohistochemistry. Results The survival curves of transplanted cornea showed that the mean survival time in rats of groups C and D was significantly longer than that in groups A and B (P<0.05. No significant difference was found in survival time between group A and group B, and the same between group C and group D. The grafts in groups A and B showed obvious edema and thickening, with irregular arrangement of collagen fibers and infiltration of a large amount of inflammatory cells. Immunohistochemical results showed that expression of CD25 was found in the corneal epithelium, stroma and endothelium in all rats, and higher CD25 expression was observed in groups A and B. Transmission electron microscopy revealed that the degree of stromal fibroblast apoptosis and necrosis in corneal graft was obviously lower in groups C and D than that of groups A and B, with a significant statistical difference. The expression of CD25 m

  14. Antithyroglobulin antibody

    Science.gov (United States)

    Thyroglobulin antibody; Thyroiditis - thyroglobulin antibody; Hypothyroidism - thyroglobulin antibody; Thyroiditis - thyroglobulin antibody; Graves disease - thyroglobulin antibody; Underactive thyroid - thyroglobulin antibody

  15. CD25(+) Bcl6(low) T follicular helper cells provide help to maturing B cells in germinal centers of human tonsil.

    Science.gov (United States)

    Li, Haishan; Pauza, C David

    2015-01-01

    The majority of CXCR5(+) PD1(+) CD4(+) T follicular helper (Tfh) cells (>90%) are CD25(-) Bcl6(hi) , while a small subpopulation (<10%) are CD25(+) Bcl6(low) but do not express FoxP3 and are not T regulatory cells. We purified T:B-cell conjugates from tonsils and found they were enriched for the CD25(+) Bcl6(low) Tfh-cell subpopulation. In response to IL-2, these CD25(+) Tfh cells increased expression of costimulatory molecules ICOS or OX40, upregulated transcription factor cMaf, produced cytokines IL-21, IL-17, and IL-10, and raised the levels of antiapoptotic protein Bcl2. Conjugates formed with CD25(+) BCl6(low) Tfh cells included B cells expressing higher levels of activation-induced cytidine deaminase (AID), memory marker CD45RO, surface IgG or IgA, and MHC class II compared to B-cell conjugates including CD25(-) Bcl6(hi) Tfh cells. While IL-2 suppresses early Tfh-cell differentiation, Tfh-cell recognition of antigen-presenting B cells and signaling through the T-cell receptor likely triggers expression of the high-affinity IL-2 receptor and responses to IL-2 including downregulation of Bcl6. CD25 expression on Tfh cells and local production of IL-2 in tonsil or lymph node may support B helper T-cell function during later stages of B-cell maturation and the development of immune memory.

  16. Ⅱ型胶原蛋白对CIA大鼠外周血CD4+CD25+FOXP3+调节性T细胞的影响

    Institute of Scientific and Technical Information of China (English)

    赵海梅; 刘端勇; 程绍民; 左志琴; 辛增平

    2011-01-01

    目的:观察Ⅱ型胶原蛋白对胶原诱导的关节炎(CIA)大鼠外周血CD4+CD25+FOXP3+调节性T 细胞(Treg)的影响.方法:建立Ⅱ型胶原蛋白诱导的大鼠类风湿性关节炎(CIA)模型.采用流式细胞术分别检测正常对照组、模型组、Ⅱ型胶原蛋白治疗组和雷公藤多苷组大鼠外周血CD4+CD25+FOXP3+Treg的水平.结果:与正常对照组比较, 模型组大鼠外周血CD4+T细胞亚群中CD25+FOXP3+Treg的水 平明显下降(P<0. 05或P<0. 01);与模型组比较, Ⅱ型胶原蛋白治疗组大鼠外周血CD25+FOXP3+Treg显著升高(P<0. 05或P<0. 01);而CD25+FOXP3-T细胞和CD25-FOXP3+ T细胞的水平则明显下降(P<0. 05或P<0. 01).结论:Ⅱ型胶原蛋白治疗CIA大鼠的可能途径是升高外周血CD4+CD25+FOXP3+Treg的水平.

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

    Science.gov (United States)

    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 population were examined by flow cytometry of blood samples at several time-points. Mononuclear cells were isolated from blood and stimulated with birch allergen, ovalbumin or the mitogen PHA, and the levels of IL-1β, IL-6, TNF, IFN-γ, IL-5 and IL-13 were measured. A clinical evaluation regarding the presence of allergen-specific IgE and allergy was performed at 18 and 36 months of age. Results Multivariate discriminant analysis revealed that children who were sensitized at 18 or 36 months of age had higher proportions of FOXP3+CD25high T cells at birth and at 3 days of life than children who remained non-sensitized, whereas allergy was unrelated to the neonatal proportions of these cells. The proportions of CTLA-4+CD25+ T cells were unrelated to both sensitization and allergy. The association between higher proportions of FOXP3+CD25high T cells and sensitization persisted after exclusion of farmer's children. Finally, a farming environment was associated with lower proportions of FOXP3+CD25high T cells in early infancy and to a more prominent T cell memory conversion and cytokine production. Conclusion & Clinical Relevance Our results indicate that high proportions of FOXP3+CD25high T cells in neonates are not protective against later sensitization or development of allergy. PMID:24528482

  18. Characterization of regulatory T cells identified as CD4+CD25highCD39+ in patients with active tuberculosis

    Science.gov (United States)

    Chiacchio, T; Casetti, R; Butera, O; Vanini, V; Carrara, S; Girardi, E; Di Mitri, D; Battistini, L; Martini, F; Borsellino, G; Goletti, D

    2009-01-01

    Forkhead box P3 (FoxP3) is a transcription factor whose expression characterizes regulatory T cells (Treg), but it is also present on activated T cells, thus hindering correct Treg identification. Using classical markers for Treg recognition, discordant results were found in terms of Treg expansion during active tuberculosis (TB) disease. Recently CD39 has been shown to be an accurate marker for Treg detection. The objectives of this study were: (i) to identify Treg expressing CD39 in patients with TB and to compare the results with those obtained by the standard phenotypic markers; (ii) to evaluate if Treg are expanded in vitro by exogenous interleukin (IL)-2 or by antigen-specific stimulation; and (iii) to characterize Treg function on the modulation of antigen-specific responses. We enrolled 13 patients with pulmonary TB and 12 healthy controls. Treg were evaluated by flow cytometry ex vivo and after antigen-specific in vitro stimulation using CD25, FoxP3, CD127 and CD39 markers. Results indicate that CD39+ cells within the CD4+CD25high cells have Treg properties (absence of interferon-γ production and transforming growth factor-β1 release upon stimulation). Ex vivo analysis did not show significant differences between TB patients and controls of Treg by classical or novel markers. In contrast, a significantly higher percentage of Treg was found in TB patients after antigen-specific stimulation both in the presence or absence of IL-2. Depletion of CD39+ Treg increased RD1-specific responses significantly. In conclusion, CD39 is an appropriate marker for Treg identification in TB. These results can be useful for future studies to monitor Mycobacterium tuberculosis-specific response during TB. PMID:19438599

  19. Study on the in vitro expansion of CD4+CD25+ regulatory T cells by mouse dendritic cells%树突状细胞体外扩增小鼠CD4+CD25+调节性T细胞的研究

    Institute of Scientific and Technical Information of China (English)

    罗珊; 李尚为; 靳松; 李小红; 姬巧云

    2013-01-01

    目的 探讨利用小鼠同种异型骨髓源性树突状细胞(BMDC)与CD4+T细胞共培养体外扩增CD4+CD25+调节性T(CD4+CD25+Treg)细胞的方法.方法 将小鼠同种异型imBMDC与CD4+T细胞共培养,通过测定细胞增殖率和CD4+CD25+Treg细胞占CD4+T细胞比例的变化评定该方法体外扩增CD4+CD25+Treg细胞的能力;利用荧光定量RT-PCR检测体外扩增CD4+CD25+Treg细胞FOXP3基因表达及细胞增殖抑制实验检测其细胞功能活性.结果 共培养d7细胞增殖率(5.26±0.286)倍;CD4+CD25+Treg细胞占CD4+T细胞的(33.77±3.69)%,较新鲜分离小鼠脾细胞中CD4+CD25+Treg细胞比例明显增加(P<0.05).体外扩增CD4+CD25+Treg细胞FOXP3基因的表达与新鲜分离CD4+CD25+Treg细胞差异无统计学意义(P>0.05).混合淋巴细胞培养证实体外扩增CD4+CD25+Treg细胞的细胞增殖抑制作用较新鲜分离的CD4+CD25+Treg细胞更强(P<0.05).结论 小鼠同种异型BMDC与CD4+T细胞共培养能有效诱导扩增CD4+CD25+Treg细胞并保持其免疫细胞功能活性.

  20. Deficiency of Mouse CD4+CD25+Foxp3+Regulatory T Cells in Xenogeneic Pig Thymus-Grafted Nude Mice Suffering from Autoimmune Diseases

    Institute of Scientific and Technical Information of China (English)

    Baojun Zhang; Chenming Sun; Yanyan Qu; Aijun Zhang; Jun Liu; Lianjun Zhang; Zeqing Niu; Yong Zhao

    2008-01-01

    Xenogeneic thymus transplantation can efficiently induce specific immune tolerance to donor antigens in athymic recipients.However,many nude mice snffer from autoimmune diseases(AID) for over 10 weeks after xenogeneic thymus transplantation.CD4+CD25+Foxp3+ regulatory T (Treg)cells were recently determined to play a pivotal role in keeping immune tolerance in humans and mice.Thus,we investigated this subpopulation of Treg cells in the periphery of pig thymus-grafted nude mice suffering from AID.Our results showed that the expression of Foxp3, CTLA-4 and GITR on mouse CD4+CD25+T cells and the ratio of CD4+CD25+Foxp3+Treg cells to CD4+T cells were significantly decreased in the periphery of pig thymus-grafted nude mice snfiering from AID,compared with healthy pig or mouse thymus-grafted nude mice.Furthermore,mouse CD4+CD25+T cells in pig thymus-grafted nude mice Sufiering from AID showed more severe deficiency in immunosuppressive function compared with the counterpart in xenogeneic pig or syngeneic thymus-grafted nude mice without AID.Thus,the decreased frequency, altered phenotype and functional deficiency of mouse CD4+CD25+Treg cells in pig thymus-grafted nude mice may contribute to the development of AID in this model.

  1. Studies on the effect and mechanism of CD4+ CD25+ regulatory T calls in Schistosoma japonicum immune evasion%CD4+CD25+调节性T细胞在血吸虫免疫逃避中的作用及机制研究

    Institute of Scientific and Technical Information of China (English)

    唐春莲; 郭思洁; 杨进; 祝青; 刘晓宏

    2014-01-01

    目的 探讨CD4+ CD25+调节性T细胞(Tregs)在日本血吸虫免疫逃避中的作用及其机制.方法 雌性BALB/c小鼠随机分成3组,即正常对照组、感染对照组和抗CD25单克隆抗体(anti-CD25 mAb)组,各感染组每只小鼠均经腹部皮肤感染日本血吸虫尾蚴40条,感染后两周anti-CD25 mAb组每只小鼠经腹腔注射anti-CD25 mAb 300 μg,其它组注射等体积的PBS,感染后5周杀鼠冲虫,计数每只小鼠虫荷.收集脾细胞及培养上清,流式细胞术检测脾淋巴细胞中CD4+ CD25+ Tregs百分比.双抗夹心ELISA法测定脾细胞培养上清中的γ-干扰素(IFN-γ)、IL-4、IL-5、IL-10的含量.结果 Anti-CD25mAb组虫荷(23.17 ±6.94)明显低于感染对照组[(30.17 ±5.85),P=0.047];感染对照组脾淋巴细胞中CD4+ CD25+ Tregs百分比(2.68 ±0.12)%明显高于正常对照组[(1.98±0.33%),P=0.049],而anti-CD25mAb组脾淋巴细胞中CD4+ CD25+ Tregs百分比(1.28±0.30)%明显低于感染对照组(P=0.000);anti-CD25mAb组脾细胞培养上清中IFN-γ的含量(386.87±24.85) pg/mL明显高于感染对照组[(61.32±8.75) pg/mL,P=0.000],其余细胞因子组间无统计学意义.结论 anti-CD25 mAb能部分封闭CD4+ CD25+ Tregs后有利于机体清除日本血吸虫,其机制可能为增强Th1型免疫反应,宿主CD4+ CD25+ Tregs有助于日本血吸虫逃避宿主的免疫攻击.%Objective To explore the effect and mechanism of CD4 + CD25 + Tregs in S.japonicum immune evasion.Methods Female BALB/c mice were randomly divided into normal control group,infected control and anti-CD25 mAb group.Each mouse was infected percutaneously with 40 S.japonicum cercaria.After 2 weeks infection,anti-CD25 mAb group was injected intraperitoneal with 300 μg anti-CD25 mAb each mouse.After 5 weeks infection,all mice were succumbed to measure worm burden.The percent of CD4+ CD25 + Tregs in spleen was measured with flow cytometer.The expression of gamma interferon (IFN-γ),interleukin-4 (IL-4),interleukin-5

  2. EntransterTM纳米载体介导大鼠角膜CD25siRNA转染的效果及安全性评估%The efficiency and safety assessment of EntransterTM nanoparticle carrier for CD25 siRNA transfection in rat cornea

    Institute of Scientific and Technical Information of China (English)

    秦琴; 石韵洁; 赵敏

    2016-01-01

    背景 基因转染是多种眼病基因治疗的有效方法,理想的非病毒载体是研究角膜基因治疗的关键因素,选择高转染率、基因高表达、低毒性的非病毒载体是成功实施基因治疗的前提. 目的 探讨EntransterTM、脂质体非病毒载体对正常SD大鼠角膜转染CD25 siRNA的转染率和安全性,筛选角膜基因转染的最佳载体.方法 应用随机数字表法将80只雄性SPF级成年SD大鼠随机分为EntransterTM-CD25 siRNA 组、脂质体-CD25 siRNA组、单纯CD25 siRNA组和生理盐水组,每组20只,均以右眼作为实验眼.实验眼眼表麻醉后刮除角膜上皮,按照分组不同分别用EntransterTM-CD25 siRNA、脂质体-CD25 siRNA、单纯CD25siRNA和生理盐水各50μl点眼.于点眼后12h、24 h、3d和7d在裂隙灯显微镜下观察大鼠眼表组织反应,检查各组大鼠角膜表面绿色荧光个数.分别于上述时间点处死各组大鼠各4只并获取角膜组织,采用苏木精-伊红染色法行角膜组织病理学检查;采用罗丹明染色行荧光检测,评估各组大鼠角膜基因转染的转染率;采用TUNEL染色法检测实验眼角膜细胞的凋亡情况以评估各种转染载体的安全性;采用免疫荧光技术检测角膜组织中CD11b的表达. 结果 EntransterTM-CD25 siRNA组大鼠角膜表面的荧光染色数量及强度明显高于脂质体-CD25 siRNA组,单纯CD25 siRNA组大鼠角膜荧光染色出现早,但转染后24 h角膜荧光染色消失.角膜组织病理学检查显示,各组大鼠行基因转染后脂质体-CD25 siRNA组大鼠角膜上皮水肿和角膜炎性细胞浸润程度较EntransterTM-CD25 siRNA组、单纯CD25 siRNA组和生理盐水组严重,角膜基质层和内皮层未发现异常,脂质体-CD25 siRNA组大鼠角膜炎性细胞数明显多于EntransterTM-CD25 siRNA组、单纯CD25 siRNA组和生理盐水组,差异均有统计学意义(均P=0.00).TUNEL检测发现,基因转染后12h和3d,脂质体-CD25siRNA组大鼠角

  3. Evaluation of CD4+CD25+ T lymphocyte response time kinetics in patients with chronic Chagas disease after in vitro stimulation with recombinant Trypanosoma cruzi antigens

    Directory of Open Access Journals (Sweden)

    Suellen Carvalho de Moura Braz

    2013-05-01

    Full Text Available Introduction CD4+CD25+ T lymphocytes have been implicated in the regulation of host inflammatory response against Trypanosoma cruzi, and may be involved in the clinical course of the disease. Methods Peripheral blood mononuclear cells from patients with chronic Chagas disease were cultured in the presence of T. cruzi recombinant antigens and assayed for lymphocytes at distinct time points. Results It was possible to differentiate clinical forms of chronic Chagas disease at days 3 and 5 according to presence of CD4+CD25+ T cells in cell cultures. Conclusions Longer periods of cell culture proved to be potentially valuable for prospective evaluations of CD4+CD25+ T lymphocytes in patients with chronic Chagas disease.

  4. Graves病患者外周血FOXP3、GITR及CD25基因的表达%Expression of FOXP3, GITR and CD25 genes in peripheral blood of patients with Graves' disease

    Institute of Scientific and Technical Information of China (English)

    王宏伟; 张莹; 陈福琴

    2009-01-01

    目的 研究Graves病(GD)不同阶段患者外周血叉状头/翅膀状螺旋转录因子(FOXP3)、糖皮质激素诱导肿瘤坏死因子受体(GITR)及IL-2受体α链(CD25)基因的表达变化,探讨其在GD发病机制中的作用.方法 收集GD患者90例,按病情分为GD初诊组30例(男13例,女17例);GD缓解组30例(男10例,女20例);GD复发组30例(男11例,女19例).健康查体者30例(男14例,女16例)作为健康对照组.应用实时荧光定量PCR法检测各组外周血单个核细胞中FOXP3、GITR及CIY25 mRNA含量,同时利用电化学发光的方法测定各组血清甲状腺激素水平及甲状腺过氧化酶抗体(TPOAb)、甲状腺球蛋白抗体(TGAb)的水平.结果 GD各组患者外周血FOXP3mRNA表达均较健康对照组显著降低(P<0.05),GD缓解组FOXP3 mRNA水平较GD初诊组显著升高(P<0.05),复发组FOXP3 mRNA水平虽低于缓解组(P<0.05),但明显高于GD初诊组(P<0.05);GD各组女性患者FOXP3mRNA表达水平显著高于男性患者(P<0.05);Graves初诊及复发组GITR mRNA、CD25mRNA表达水平显著高于对照组(P<0.05).结论 FOXP3、GITR及CD35可能参与了Graves的发生、发展及复发过程.

  5. Study of the Expression of CD4 + CD25 + T Cell and Soluble Cytokines of Peripheral Blood in the Cases of Multiple Sclerosis%多发性硬化患者外周血CD4+CD25+T细胞与可溶性细胞因子表达的相关性研究

    Institute of Scientific and Technical Information of China (English)

    方宇; 张璐; 连亚军; 魏建科; 吴天文; 罗志毅

    2011-01-01

    CD4+ CD25+T细胞和可溶性细胞因子在多发性硬化(multiple sclerosis,MS)发生和发展中起重要作用.本文通过探讨MS患者外周血CD4+ CD25+T细胞数量及可溶性细胞因子与MS病情的关系,为进一步研究MS的发病机制和采取有效治疗措施奠定了基础.%CD4 + CD25+ T cell and soluble cytokines play a critical role during an acute inflammatory attack in multiple sclerosis! MS). In this study, we investigated the relationship between the blood serum levels of CD4 + CD25+ T cell, soluble cytokines and the state of MS. The results would assist in promoting effective therapies and would elucidate pathophysiologic mechanisms relevant to MS.

  6. CD25+CD127+Foxp3- Cells Represent a Major Subpopulation of CD8+ T Cells in the Eye Chambers of Normal Mice

    Science.gov (United States)

    Ziółkowska, Natalia; Ziółkowski, Hubert; Małaczewska, Joanna

    2017-01-01

    The aim of this study has been to determine whether eye chambers constitute part of the normal migratory pathway of naive CD4+ and CD8+ T cells in mouse and if natural CD4+CD25+Foxp3+ and CD8+CD25+Foxp3+ regulatory T cells are present within these eye compartments. To this aim, the cells obtained from aqueous humor (AH) of normal mice were phenotyped in terms of the expression CD4, CD8, CD25, CD127 and transcription factor Foxp3. The mean percentage of CD8+ T cells in the total AH lymphocyte population was as high as 28.69%; the mean percentage of CD8high and CD8low cells in this population was 34.09% and 65.91%, respectively. The presence of cells with the regulatory phenotype, i.e. CD25+Foxp3+ cells, constituted only 0.32% of CD8+ T cell subset. Regarding the expression of CD25, AH CD8+ T cells were an exceptional population in that nearly 85% of these cells expressed this molecule without concomitant Foxp3 expression. Despite having this phenotype, they should not be viewed as activated cells because most of them co-expressed CD127, which indicates that they are naive lymphocytes. With regard to the markers applied in the present research, CD8+CD25+CD127+Foxp3- T cells represent the most numerous subset of AH CD8+ cells. The results suggest that eye chambers in mice are an element in the normal migratory pathway of naive CD8+ T cells. The study presented herein demonstrated only trace presence of CD4+ cells in the eye chambers, as the mean percentage of these cells was just 0.56. Such selective and specific homing of CD8+ and CD4+ cells to the eye chambers is most clearly engaged in the induction and maintenance of ocular immune privilege. PMID:28081241

  7. CD4~+Foxp3~+ regulatory T cells converted by rapamycin from peripheral CD4~+ CD25~-naive T cells display more potent regulatory ability in vitro

    Institute of Scientific and Technical Information of China (English)

    CHEN Jian-fei; GAO Jie; ZHANG Dong; WANG Zi-han; ZHU Ji-ye

    2010-01-01

    Background Rapamycin (RAPA) is a relatively new immunosuppressant drug that functions as a serine/threonine kinase inhibitor to prevent rejection in organ transplantation. RAPA blocks activation of T-effector (Teff) cells by inhibiting the response to interleukin-2. Recently, RAPA was also shown to selectively expand the T-regulator (Treg) cell population. To date, no studies have examined the mechanism by which RAPA converts Teff cells to Treg cells. Methods Peripheral CD4~+CD25~- naive T cells were cultivated with RAPA and B cells as antigen-presenting cells (APCs) in vitro. CD4~+CD25~- T cells were harvested after 6 days and analyzed for expression of forkhead box protein 3 (Foxp3) using flow cytometry. CD4~+CD25~+CD127~- subsets as the converted Tregs were isolated from the mixed lymphocyte reactions (MLR) with CD127 negative selection, followed by CD4 and CD25 positive selection using microbeads and magnetic separation column (MSC). Moreover, mRNA was extracted from converted Tregs and C57BL/6 naive CD4~+CD25~+ T cells and Foxp3 levels were examined by quantitative real-time polymerase chain reaction (rt-PCR). A total of 1×10~5 carboxyfluorescein succinimidyl ester (CFSE)-labeled naive CD4~+CD25~- T cells/well from C57BL/6 mice were cocultured with DBA/2 or C3H maturation of dendritic cells (mDCs) (0.25×10~5/well) in 96-well round-bottom plates for 6 days. Then 1×10~5 or 0.25×10~5 converted Treg cells were added to every well as regulatory cells. Cells were harvested after 6 days of culture and analyzed for proliferation of CFSE-labeled naive CD4~+CD25~- T cells using flow cytometry. Data were analyzed using CellQuest software.Results We found that RAPA can convert peripheral CD4~+CD25~- naive T Cells to CD4~+Foxp3~+ Treg cells using B cells as APCs, and this subtype of Treg can potently suppress Teff proliferation and maintain antigenic specificity. Conclusion Our findings provide evidence that RAPA induces Treg cell conversion from Teff cells and

  8. 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.

  9. CD4+CD25+调节性T细胞对人外周血内皮祖细胞增殖、迁移、黏附的影响%The effect of CD4+CD25+ regulatory T cells on the proliferation,migration and adhesion of endothelial progenitor cells

    Institute of Scientific and Technical Information of China (English)

    谢培益; 苏又苏; 汤海燕; 方红成; 何少林; 李大主

    2011-01-01

    目的:研究CD4+CD25+调节性T细胞对体外培养的人外周血内皮祖细胞(EPCs)增殖、迁移、黏附的影响.方法:密度梯度离心法分离培养人外周血单个核细胞,经FITC-UEA-I和Dil-acLDL双染色鉴定为正在分化的EPCs.进一步采用流式细胞仪检测其表面标志CD34、CD133.磁性细胞分离器(MACS)分离CD4+CD25+调节性T细胞及CD4+CD25-T细胞.将EPCs分别与CD4+CD25+调节性T细胞或CD4+CD25-T细胞共培养36 h.采用MTT比色法、transwell小室、细胞计数法观察EPCs增殖、迁移、黏附能力.结果:与对照组和CD4+CD25-T细胞相比,与CD4+CD25+调节性T细胞共培养EPCs增殖、迁移、黏附能力显著增强,且呈浓度依赖性增强.结论:CD4+CD25+调节性T细胞可显著促进EPCs增殖、迁移、黏附能力,此作用可能是其抗AS作用机制之一.%Objective:To investigat the effect of CD4+CD25+ regulatory T cells (Tregs) on the proliferation,migration and adhesion of endothelial progenitor cells (EPCs) from human circulating blood in vitro. Method : Tregs were isolated from lymphocyte suspensions by magnetic cell sorting column and analyzed by flow cytometry. Human blood mononuclear cells were isolated with Ficoll by density gradient centrifugation. EGM-2MV culture fluid was added, and then cells were plated on dishes coated with human fibronectin. After 7 days, the cells were identified with immunofluorescence and flow cytometry. The cells were cultured alone (control groups), with CD4+CD25- T cells (CD25- groups), or CD4+ CD25+ Tregs (Tregs groups) for 36 hours. The proliferation,migration and adhesion activities of EPCs were determined with MTT assay, transwell assay and adhesive assay, respectively. Result:Compared with control groups and CD25- groups, the proliferative, migratory and adhesive activities of EPCs were significantly enhanced after treated with CD4 + CD25 + Tregs (P<0.05 and P<0. 01). Moreover, the proliferative, migratory and adhesive activities of

  10. Anti-Human Endoglin (hCD105 Immunotoxin—Containing Recombinant Single Chain Ribosome-Inactivating Protein Musarmin 1

    Directory of Open Access Journals (Sweden)

    Begoña Barriuso

    2016-06-01

    Full Text Available Endoglin (CD105 is an accessory component of the TGF-β receptor complex, which is expressed in a number of tissues and over-expressed in the endothelial cells of tumor neovasculature. Targeting endoglin with immunotoxins containing type 2 ribosome-inactivating proteins has proved an effective tool to reduce blood supply to B16 mice tumor xenografts. We prepared anti-endoglin immunotoxin (IT—containing recombinant musarmin 1 (single chain ribosome-inactivating proteins linked to the mouse anti-human CD105 44G4 mouse monoclonal antibody via N-succinimidyl 3-(2-pyridyldithio propionate (SPDP. The immunotoxin specifically killed L929 fibroblast mouse cells transfected with the short form of human endoglin with IC50 values in the range of 5 × 10−10 to 10−9 M.

  11. 耐多药结核患者外周血CD4+CD25+Foxp3+T细胞的变化%Change of the peripheral blood CD4+CD25+Foxp3+T cells in patients with multidrug-resistant tuberculosis

    Institute of Scientific and Technical Information of China (English)

    徐传财; 解卫平; 戴冠群; 谈绮; 闵锐; 王虹

    2011-01-01

    目的:探讨CD4+CD25+Foxp3+T细胞在耐多药结核(multidrug-resistant TB,MDR-TB)发病机制中的作用.方法:随机选择MDR-TB患者31例,药物敏感性结核(drug-susceptible TB,S-TB)33例,健康对照(healthy control,HC)30例,流式细胞仪检测外周血CD4+CD25+Foxp3+T细胞.结果:S-TB患者外周血CD4+CD25+Foxp3+T细胞约占CD4+T细胞的(2.43±0.69)%,显著高于健康人的(1.61±0.46)%,两组之间差异具有显著统计学意义;MDR-TB患者外周血CD4+CD25+Foxp3+T细胞约占CD4+T细胞的(3.14±0.59)%,显著高于S-TB患者的细胞比例,两组之间差异具有显著统计学意义.结论:MDR-TB患者外周血CD4+CD25+Foxp3+T细胞明显增加,揭示其在MDR-TB发病中发挥了重要作用.%Objective :To observe the changes of CD4+CD25+Foxp3+T cells in patients with multidrug-resistant tuberchlosis(TB).Methods:We recruited 31 multidrug-resistant TB (MDR-TB) patients,33 drug-susceptible TB (S-TB) patients and 30 healthy controls.Flow cytometry was used to determined the quantity of CD4+CD25+Foxp3+ir cells in the peripheral blood. Results:The mumber of CD4+CD25+Foxp3+T cell in the peripheral blood of patients with S-TB was(2.43±0.69)% of peripheral CD4+ T cell, it was significantly higher than heathy controls [ ( 1.61±0.46) % ] and lower than patients with MDR-TB [ (3.14±+0.59) % ]. Conclusion: CD4+CD25+Foxp3+T cells increased in MDR-TB patients significantly, this suggests that CD4+CD25+Fox3+T cells play an important role in the pathogenesis of MDR-TB.

  12. Naturally occurring lung CD4(+)CD25(+) T cell regulation of airway allergic responses depends on IL-10 induction of TGF-beta.

    Science.gov (United States)

    Joetham, Anthony; Takeda, Katsuyuki; Takada, Katsuyuki; Taube, Christian; Miyahara, Nobuaki; Matsubara, Shigeki; Matsubara, Satoko; Koya, Toshiyuki; Rha, Yeong-Ho; Dakhama, Azzeddine; Gelfand, Erwin W

    2007-02-01

    Peripheral tolerance to allergens is mediated in large part by the naturally occurring lung CD4(+)CD25(+) T cells, but their effects on allergen-induced airway responsiveness have not been well defined. Intratracheal, but not i.v., administration of naive lung CD4(+)CD25(+) T cells before allergen challenge of sensitized mice, similar to the administration of the combination of rIL-10 and rTGF-beta, resulted in reduced airway hyperresponsiveness (AHR) and inflammation, lower levels of Th2 cytokines, higher levels of IL-10 and TGF-beta, and less severe lung histopathology. Significantly, CD4(+)CD25(+) T cells isolated from IL-10(-/-) mice had no effect on AHR and inflammation, but when incubated with rIL-10 before transfer, suppressed AHR, and inflammation, and was associated with elevated levels of bronchoalveolar lavage TGF-beta levels. By analogy, anti-TGF-beta treatment reduced regulatory T cell activity. These data identify naturally occurring lung CD4(+)CD25(+) T cells as capable of regulating lung allergic responses in an IL-10- and TGF-beta-dependent manner.

  13. Activation of the aryl hydrocarbon receptor reduces the number of precursor and effector T cells, but preserves thymic CD4(+)CD25(+)Foxp3(+) regulatory T cells

    NARCIS (Netherlands)

    Schulz, V.J.; Smit, J.J.; Bol-Schoenmakers, M.; van Duursen, M.B.M.; van den Berg, M.; Pieters, R.H.H.

    2012-01-01

    Aryl hydrocarbon receptor (AhR) activation suppresses immune responses, including allergic sensitization, by increasing the percentage of regulatory (Treg) cells. Furthermore, AhR activation is known to affect thymic precursor T cells. However, the effect of AhR activation on intrathymic CD4(+)CD25(

  14. 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

  15. Identification and monitoring of effector and regulatory T cells during experimental arthritis based on differential expression of CD25 and CD134

    NARCIS (Netherlands)

    E.N.M. Nolte-'t Hoen (Esther); E.P.J. Boot (Elmieke); J.P.A. Wagenaar-Hilbers (Josée); J.H.M. van Bilsen (Jolanda); G.J.A. Arkesteijn (Ger); G. Storm (Gert); L.A. Everse (Linda); W. van Eden (Willem); M.H.M. Wauben (Marca)

    2008-01-01

    textabstractMajor problems in the analysis of CD4+effector cell and regulatory T cell (Treg) populations in an activated immune system are caused by the facts that both cell types can express CD25 and that the discriminatory marker forkhead box p3 can only be analyzed in nonviable (permeabilized) ce

  16. Identification and monitoring of effector and regulatory T cells during experimental arthritis based on differential expression of CD25 and CD134.

    Science.gov (United States)

    Nolte-'t Hoen, Esther N M; Boot, Elmieke P J; Wagenaar-Hilbers, Josée P A; van Bilsen, Jolanda H M; Arkesteijn, Ger J A; Storm, Gert; Everse, Linda A; van Eden, Willem; Wauben, Marca H M

    2008-01-01

    Major problems in the analysis of CD4+ effector cell and regulatory T cell (Treg) populations in an activated immune system are caused by the facts that both cell types can express CD25 and that the discriminatory marker forkhead box p3 can only be analyzed in nonviable (permeabilized) cells. Here, we show that CD134 (OX40) can be used as a discriminatory marker combined with CD25 to isolate and characterize viable CD4+ effector cells and Tregs. Before and during adjuvant arthritis in rats, coexpression of CD134 and CD25 identified activated Tregs consistently, as these T cells proliferated poorly to disease-associated antigens and were suppressive in vitro and in vivo. Depending on the time of isolation and location, CD4+ T cell populations expressing CD134 or CD25 contained effector/memory T cells. Analysis of the function, phenotype, and amount of the CD4+ T cell subsets in different lymph node stations revealed spatiotemporal differences in effector cell and Treg compartments during experimental arthritis.

  17. Identification and monitoring of effector and regulatory T cells during experimental arthritis based on differential expression of CD25 and CD134

    NARCIS (Netherlands)

    Nolte-'t Hoen, E.N.M.; Boot, E.P.J.; Wagenaar-Hilbers, J.P.A.; Bilsen, J.H.M. van; Arkesteijn, G.J.A.; Storm, G.; Everse, L.A.; Eden, W. van; Wauben, M.H.M.

    2008-01-01

    Major problems in the analysis of CD4+ effector cell and regulatory T cell (Treg) populations in an activated immune system are caused by the facts that both cell types can express CD25 and that the discriminatory marker forkhead box p3 can only be analyzed in nonviable (permeabilized) cells. Here,

  18. The dysfunction of CD4(+CD25(+ regulatory T cells contributes to the abortion of mice caused by Toxoplasma gondii excreted-secreted antigens in early pregnancy.

    Directory of Open Access Journals (Sweden)

    Jin-ling Chen

    Full Text Available Toxoplasma gondii is an opportunistic intracellular parasite that is highly prevalent in human and warm-blooded animals throughout the world, leading to potentially severe congenital infections. Although the abortion caused by T. gondii is believed to be dependent on the timing of maternal infection during pregnancy, the mechanism remains unclear. This study was focused on the effects of T. gondii excreted-secreted antigens on pregnant outcomes and CD4(+CD25(+ Foxp3(+ regulatory T cells at different stages of pregnancy. The results showed that in mice the frequency and suppressive function of CD4(+CD25(+ regulatory cells were diminished after injection of T. gondii excreted-secreted antigens at early and intermediate stages of pregnancy. The abortion caused by T. gondii excreted-secreted antigens at early pregnancy could be partly prevented by adoptively transferring of CD4(+CD25(+ cells from the mice injected with T. gondii excreted-secreted antigens at late pregnancy, but not from the mice with the same treatment at early pregnancy. Furthermore, T. gondii excreted-secreted antigens induced apoptosis of CD4(+CD25(+ regulatory cells of mice in early and intermediate stages of pregnancy by down-regulating their Bcl-2 expressions and Bcl-2/Bax ratio. This study provides new insights into the mechanism that T. gondii infection is the high risk factor for abortion in early pregnancy.

  19. Rapamycin, and not cyclosporin A, preserves the highly suppressive CD27+ subset of human CD4+CD25+ regulatory T cells.

    NARCIS (Netherlands)

    Coenen, J.J.A.; Koenen, H.J.P.M.; Rijssen, E. van; Hilbrands, L.B.; Joosten, I.

    2006-01-01

    The immunosuppressive drugs rapamycin and cyclosporin A (CsA) are widely used to prevent allograft rejection. Moreover, they were shown to be instrumental in experimental models of tolerance induction. However, it remains to be elucidated whether these drugs have an effect on the CD4+ CD25+ regulato

  20. 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

  1. Increase in circulating CD4CD25Foxp3 T cells in patients with Philadelphia-negative chronic myeloproliferative neoplasms during treatment with IFN-α

    DEFF Research Database (Denmark)

    Riley, C.H.; Morten Krogh, Jensen; Brimnes, M.K.;

    2011-01-01

    have been reported in both hematologic and solid cancers. We have analyzed the number, phenotype, and function of circulating CD4 CD25Foxp3 T cells in patients with chronic myeloproliferative neoplasms. Surprisingly, we found a marked expansion of this subset of lymphocytes in patients treated with IFN...

  2. Chronic Brucellosis Patients Retain Low Frequency of CD4+ T-Lymphocytes Expressing CD25 and CD28 after Escherichia coli LPS Stimulation of PHA-Cultured PBMCs

    Directory of Open Access Journals (Sweden)

    Panagiotis Skendros

    2008-01-01

    Full Text Available Chronic brucellosis patients display a defective Th1 response to PHA. We have previously shown that heat-killed B. abortus (HKBA can downregulate the PHA-induced increase of CD4+/CD25+ and CD14+/CD80+ cells of brucellosis patients. In the present study, we investigate the effect of E. coli LPS, as a potent stimulant of monocytes and autologous T-lymphocytes, on the PHA-cultured PBMCs of the same groups of patients. Thirteen acute brucellosis (AB patients, 22 chronic brucellosis (CB patients, 11 “cured” subjects, and 15 healthy volunteers were studied. The percentage of CD4+/CD25+ and CD4+/CD28+ T-lymphocytes as well as CD14+/CD80+ monocytes were analyzed by flow cytometry after PBMCs culture with PHA plus E. coli LPS. A significant decrease in the percentage of CD4+/CD25+ and CD4+/CD28+ T-lymphocytes was observed in CB compared to AB. In HKBA cultures, compared to E. coli LPS-cultures, there was a significant reduction of CD4+/CD25+ T-lymphocytes in all groups and CD14+/CD80+ in patients groups. We suggest that Brucella can modulate host immune response, leading to T-cell anergy and chronic infection.

  3. 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

  4. Change of CD4+ CD25+ Regulatory T Cells in Patients with Gastric Cancer before and after Operation%胃癌患者手术前后CD4+CD25+调节性T细胞的变化

    Institute of Scientific and Technical Information of China (English)

    史学菲; 白平; 唐承薇; 王春晖

    2011-01-01

    目的 研究胃癌患者手术前、后调节性T细胞(Treg)及FoxP3表达的变化.方法 采用流式细胞术检测20例胃癌患者术前及其中15例接受了手术者术后1周(简称术后)以及15例因胃部不适接受胃镜检查的自愿者(正常对照组)外周血中Treg数量的变化,用RT-PCR法检测Treg的特异性分子标志物FoxP3的转录水平,同时用免疫组织化学法检测胃癌组织中FoxP3蛋白的表达情况.结果 胃癌患者术前外周血中CD4+T细胞中的CD4+ CD25+比例明显高于正常对照组[(19.39±5.58)%比(9.91±3.23)%,P<0.01],而术后CD4+ CD25+比例较术前明显下降[(13.50±5.93)%比(19.39±5.58)%,P<0.05].胃癌患者术前外周血中FoxP3转录水平明显高于正常对照组(0.86±0.03比0.64±0.02,P<0.01),而术后较术前明显下降(0.73±0.04比0.86±0.03,P<0.05),提示FoxP3转录水平与Treg变化一致.胃癌患者外周血中CD4+T细胞在单个核细胞中的比例与正常对照组相比明显下降(P<0.01),而手术前、后变化不明显.20例胃癌患者中13例胃癌癌细胞的细胞浆中有不同程度的FoxP3蛋白表达(强阳性2例,中阳性6例,弱阳性5例),7例胃癌患者的胃癌细胞中不表达.结论 Treg可能通过免疫抑制作用在胃癌的发生、发展中发挥作用,肿瘤组织本身可能是引起Treg变化的重要始动因素.%Objective To investigate the levels of regulatory T cells (Treg) and FoxP3 gene in patients with gastric cancer before and after operation. Methods Twenty patients with definite diagnosis of gastric cancer and 15 healthy volunteers were selected. The levels of Treg and T cell subsets in peripheral blood were determined by detecting of CD4 and CD25 with immune-fluorescence stain and flow cytometry, the expressions of FoxP3 mRNA in these Treg were detected by RT-PCR technique. The expression of FoxP3 protein in the gastric cancer tissue was measured by immunohistochemistry assay. Results The percentage of Treg cells in

  5. Changes of CD4+CD25+ regulatory T cells in sepsis patients and its relationship with prognosis%脓毒症患者CD4+CD25+调节性T细胞的变化及其与预后的关系

    Institute of Scientific and Technical Information of China (English)

    孙仁华; 李茜; 邱莲女

    2009-01-01

    Objective To evaluate the changes of peripheral blood CD4+CD25+ regulatory T cells and its prognostic value in sepsis patients.MethodThe percentage of CD4+CD25+/CD4+ T cells were measured by flow eytometry present in peripheral blood in 36 sepsis patients on day 1,3 and 5 and 15 healthy people (control group).The changes of CD4+CD25+ regulatory T cells were analyzed in survivors group and non-survivors group respectively.Results There were 19 survivors and 17 non-survivors in sepsis patients.The percentage of CD4+CD25+/CD4+ T cells in survivors group and non-survivors group on day 1 was significantly lower than that in eontrol group [(12.42± 3.26)%,(12.96± 3.00)% vs (16.97 ± 3.66)%,P<0.05].The percentage of CD4+CD25+/CD4+ T cells in survivors group and non-survivors group on day 3 were both signifieanfly higher than that on day 1 [(24.47±4.62)%vs (12.42±3.26)%,(26.61±3.85)%vs (12.96±3.00)%,P<0.05].The percentage of CD4+CD25+/CD4+ T cells in survivors group on day 5[(18.28±4.28)%]was significantly lower than that on day 3 (P<0.05),and the percentage of CD4+CD25+/CD4+ T cells in nonsurvivors group on day 5 were significantly higher than that on day 3(P<0.05).There was no significant difference in the percentage of CD4+CD25+/CD4+ T ceils between survivors group and non-survivors group on day 1,3 respeefively(P > 0.05),but the percentage of CD4+CD25+/CD4+ T cells in survivors group was significantly lower than that in non-survivors group on day 5 (P<0.05).Conclusions The CD4+CD25+ regulatory T cells in sepsis patients decreases following by increase after onset.The persistent increase suggests the emergence of immunoparalysis,which is followed by high monaliy,The CD4+CD25+ regulatory T cells are valuable in evaluation of immune state and prediction the prognosis in sepsis patients.%目的 评价脓毒症患者外周血中CD4+CD25+调节性T细胞的变化趋势及其与预后的相关性.方法 采用流式细胞仪检测36例脓

  6. Study of changes of CD4+CD25+Foxp3+ regulatory T cells of Peripheral Blood in prostate cancer patients and staging of tumor%前列腺癌患者外周血CD4+CD25+Foxp3+调节性T细胞的变化及与肿瘤分期的相关性

    Institute of Scientific and Technical Information of China (English)

    汪铁军; 谢中华; 郑元秀; 王珠美

    2012-01-01

    Abstract Objective To investigate the relationship between the changes of CD4+CD25+Foxp3+ regulatory T cells of peripheral blood in prostate cancer patients and the international staging of tumor. Methods The number of CD4+CD25+Foxp3+ regulatory T cell in peripheral blood of 43 patients with prostate cancer (staging of tumor: II 18, III 10 and IV 8) and 20 healthy examiners were detected by the flow cytometry. The relationship between the changes of CD4+CD25+Foxp3+ regulatory T cells and the staging of tumor was statistically analyzed. Results Compared with that of the healthy control group, the proportion and number of CD4+CD25+Foxp3+ regulatory T cells of peripheral blood in prostate cancer patients were all increased significantly (P0.05). The proportion and number of CD4+CD25+Foxp3+ regulatory T cells in the high-risk group were increased significantly than that in the low-risk group and average-risk group (P<0.01, P<0.05). The proportion and number of CD4+CD25+Foxp3+ regulatory T cells were positively correlated with the staging of tumor and risk level. Conclusion Higher proportion and number of CD4+CD25+Foxp3+ regulatory T cells are found in peripheral blood in prostate cancer patients , and they increase with the staging . The level of CD4+CD25+Foxp3+ regulatory T cells may be useful in evaluation of the patient's condition .%目的 观察前列腺癌(PCa)患者外周血单个核细胞CD4+CD25+Foxp3+调节性T细胞的变化及其与国际肿瘤淋巴结转移(TNM)分期的关系.方法 采用流式细胞术检测20例体检健康者(对照组)及43例PCa患者(患者组,临床TNM分期Ⅰ期7例、Ⅱ期18例、Ⅲ期10例、Ⅳ期8例)外周血单个核细胞CD4+CD25+Foxp3+调节性T细胞数目,并计算CD4+CD25+Foxp3+调节性T细胞占CD4+T淋巴细胞的百分比;分析CD4+CD25+Foxp3+调节性T细胞与肿瘤分期的相关性.结果 与正常对照组比,患者组外周血单核细胞CD4+CD25+Foxp3+Treg占CD4+T淋巴细胞的百分比及CD4+CD25+Foxp3

  7. 外用他克莫司对寻常型银屑病患者CD4+CD25+调节性T细胞的影响%Effect of tacrolimus onitment on CD4+ and CD25+Treg cells in patients with psoriasis vulgaris

    Institute of Scientific and Technical Information of China (English)

    杨晓梅; 何荣国

    2010-01-01

    Objective To explore the effect of tacrolimus onitment on the changes of CD4+ and CD25+ Treg cells in peripheral blood from patients with psoriasis vulgaris. Methods 48 patients with psoria-sis vulgaris were treated with a topical immunomodulator, tacrolimus ointment 0.1%, twice daily for 8 weeks.The proportion of CD4+ and CD25+ Treg cells population in total T cells were evaluated by flowing cytometry before and after treatment. PASI was measured in all the patients. Results After 8-week treatment, the response rates was 91.5%. Before treatment, in the active stage and resting stage of psoriasis vulgaris and healthy controls, the proportion of CD4+ and CD25+ Treg cells population in total T cells was (2.64 ± 0.86) %,(3.98 ± 0.96) %, and (8.46 ± 1.54) %, with a significant difference among the three groups (P<0.01). After treatment, the proportion of CD4+ and CD25+ Treg cells population was obviously increased, but still lower than that in the healthycontrols (P<0.O1). Conclusions CD4+ and CD25+ Treg cells are abnormal in patients with psoriasis vulgaris. Tacrolimus can treat psoriasis vulgaris by upregulating CD4+ and CD25+ Treg cells.%目的 探讨外用他克莫司对寻常型银屑病患者外周血CD4+CD25+调节性T细胞(CD4+CD25+Treg)的影响.方法 给予48例寻常型银屑病患者他克莫司软膏,每日2次外用,用药8周.治疗前和治疗后,用流式细胞仪检测患者外周血CD4+CD25+调节性T细胞的表达水平,并对患者进行PASI评分.结果 治疗8周后,总有效率为91.5%.治疗前,银屑病进行期、静止期患者及正常人外周血CD4+CD25+Treg与CD4+淋巴细胞的百分比为(2.64±0.86)%、(3.98±0.96)%、(8.46±1.54)%,三者比较,差异均有极显著性(P<0.01);治疗后,进行期和静止期患者外周CD4+CD25+Treg与CD4+淋巴细胞的百分比较治疗前均有明显升高(P<0.01),但仍低于正常人,三者比较,差异均有极显著性(P<0.01).结论 寻常型银屑病存在CD4+CD25+调节性T细

  8. Human T cell priming assay: depletion of peripheral blood lymphocytes in CD25(+) cells improves the in vitro detection of weak allergen-specific T cells.

    Science.gov (United States)

    Vocanson, Marc; Achachi, Amine; Mutez, Virginie; Cluzel-Tailhardat, Magalie; Varlet, Béatrice Le; Rozières, Aurore; Fournier, Philippe; Nicolas, Jean-François

    2014-01-01

    To develop an in vitro assay that recapitulates the key event of allergic contact dermatitis (ACD), that is the priming of effector T cells by hapten-presenting dendritic cells, and then allows for the sensitive detection of chemical allergens represents a major challenge. Classical human T cell priming assays (hTCPA) that have been developed in the past, using hapten-loaded monocyte-derived dendritic cells (MDDCs) as antigen-presenting cells and peripheral blood lymphocytes (PBLs) as responding cells, were not efficient to prime T cells to common allergens with moderate/weak sensitizing properties. Recent progress in the understanding of the effector and regulatory mechanisms of ACD have shown that T cell priming requires efficient uptake of allergens by immunogenic DCs and that it is controlled by several subsets of regulatory cells including CD25(+) Tregs. We therefore analyzed various parameters involved in allergen-specific T cell activation in vitro and showed that priming of allergen-specific T cells is hampered by several subsets of immune cells comprising CD1a(neg) DCs, CD25(+) T cells, and CD56(+) regulatory cells.CD4(+)CD25(+)FoxP3(+) Tregs prevented the in vitro T cell priming to moderate/weak allergens, and depletion of human PBLs in CD25(+) cells significantly increased specific T cell proliferation and IFN-γ secretion. CD56(+) cells exerted an additional control of T cell priming since co-depletion of both CD56(+) and CD25(+) cells improved the magnitude of chemical-specific T cell activation. Finally, CD1a(low) MDDCs were able to inhibit T cell activation obtained by allergen-pulsed CD1a(high) MDDC. Moreover, we showed that uptake by DC of allergen-encapsulated nanoparticles significantly increased their activation status and their ability to prompt specific T cell activation. Hence, by combining the different strategies, i.e., depletion of CD25(+) and CD56(+) cells, use of CD1a(high) MDDC, and nanoparticle encapsulation of allergens, it was

  9. CD4~+CD25~+调节性T细胞对巨噬细胞泡沫化的影响%CD4~+ CD25~+ regulatory T cells inhibit macrophage-derived foam cells formation by down-regulating scavenger receptor expression

    Institute of Scientific and Technical Information of China (English)

    林静; 李大主

    2010-01-01

    Objective To investigate whether and how regulatory T cells(Tr) affect macrophages foam-cell formation, and thereby investigate the mechanism of Tr in the development of atherosclerosis. Methods Tr were isolated from lymphocyte suspensions by magnetic cell sorting-column and analyzed by flow cytometry. Macrophages were cultured alone, with CD4~+ CD25~+ T, or CD4~+ CD25~- T cells in the pres-ence of oxLDL for 48 h to transform macrophages into foam cells. Oil red O staining and cellular lipid meas-urement were used to identify macrophage foam cell formation. Semi-quantitative PCR, quantitative real-time PCR and Western blot analysis were carried out to explore the mechanism of Tr-mediated suppression on macrophages foam cell formation. Results Foam cell formation, as identified by oil red O staining, was readily apparent in cells treated with CD4~+ CD25~- T cells and without T cells. After treatment with Tr, a marked decrease(13.9% ± 5.6% ) in foam cell count was observed, compared with untreated cells(13.9% ±5.6% vs 52.9% ± 10.4%, P<0.001 ) or CD4~+ CD25~- T-treated cells(13.9% ± 5.6% vs 53. 1% ± 17.2%, P<0.001 ), 52.9% ± 10.4% and 53.1% ± 17.2%, respectively (P<0.001). The similar effect of Tr was obtained when extracted oil red O and measured by a spectrophotometer. Cholesteryl ester ac-cumulation also used to quantify foam cell formation. Compared with untreated (no T) and CD4~+ CD25~- Tr-treated macrophage cells (CD25~-), the lipids accumulation in CD4~+ CD25~+ Tr-treated macrophage foam cells(CD25~+) was significantly reduced. Total cellular cholesterol and cellular cholesteryl ester was siginifi-cantly reduced in CD25~+ cultures relative to no T[TC(total cholesterol): 57.46 ± 17.92 vs 159.48 ± 16.38, P<0.01 ; CE(esterified cholesterol): 26.68 ± 8.88 vs 102.54 ± 16.67, P<0.001] or CD25~- (TC: 58.50±7.00 vs 150.55±25.11, P<0.01; CE: 26.68±8.88 vs96.90 ± 11.95, P<0.001) cul-tures. Moreover, PCR and Western blot analysis showed that the expression

  10. 急性冠脉综合征患者外周血 CD4+ CD25+ Foxp3+Treg细胞的表达%Peripheral blood CD4+CD25+Foxp3+Treg cells in patients with acute coronary syndrome

    Institute of Scientific and Technical Information of China (English)

    邓穗燕; 李琴; 林丽英; 黄俊

    2014-01-01

    目的::探讨急性冠脉综合征( ACS)患者外周血CD4+CD25+Foxp3+Treg细胞亚群的表达量及其与ACS的关系。方法:采用流式细胞仪( FCM)检测16例ACS患者( ACS组)、12例稳定性心绞痛患者( SA组)和20例健康志愿者( Control组)外周血CD4+CD25+Tre细胞和CD4+CD25+Foxp3+Treg细胞的百分含量;采用酶联免疫吸附试验( ELISA)检测3组研究对象外周血细胞因子TGF ̄β1和IL ̄10水平;并采用透射比浊法检测3组研究对象血浆超敏C ̄反应蛋白( hs ̄CRP )和血糖、血脂水平。结果:3组研究对象外周血CD4+CD25+Treg细胞百分含量差异无统计学意义( P>0.05);ACS组外周血CD4+CD25+Foxp3+Treg细胞水平和血浆TGF ̄β1、IL ̄10浓度较SA组和Control组均明显降低(P0.05)。结论:CD4+CD25+Foxp3+Treg细胞数量和/或功能的下降可能与ACS的发生和发展密切相关,Foxp3标记可能有助于进一步识别ACS相关的Treg细胞。%Objective:To analyze the expression level of CD4+CD25+Foxp3+regulatory T ( Treg) cell subset in peripheral blood of patients with acute coronary syndrome ( ACS) and its correlation with ACS. Methods:Flow cytometry ( FCM) was used to measure the percentage contents of CD4+CD25+ Treg cells and CD4+CD25+Foxp3+Treg cells in the peripheral blood from 16 ACS patients ( ACS group) , 12 patients with stable angina ( SA group) , and 20 healthy volunteers ( Control group) . The levels of peripheral blood cytokines TGF ̄β1 and IL ̄10 in the three groups were measured by enzyme ̄linked immunosorbent assay ( ELISA) . The levels of hypersensitive C ̄reactive protein ( hs ̄CRP ) , blood glucose and blood lipids were measured by transmission turbidimetry. Results:There was no statistical difference in the percentage content of peripheral blood CD4+CD25+Treg cells among the three groups ( P>0. 05 ) . The levels of peripheral CD4+ CD25+ Foxp3+ Treg cells and plasma concentrations of TGF ̄β1 and IL

  11. 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

  12. Interleukin-12 (IL-12p70 promotes induction of highly potent Th1-like CD4+CD25+T regulatory cells that inhibit allograft rejection in unmodified recipients.

    Directory of Open Access Journals (Sweden)

    Nirupama Darshan Verma

    2014-05-01

    Full Text Available In rat models, CD4+CD25+T regulatory cells (Treg play a key role in the induction and maintenance of antigen specific transplant tolerance, especially in DA rats with PVG cardiac allografts(1, 2. We have previously described generation of alloantigen specific Treg (Ts1, by culture of naïve natural CD4+CD25+ Treg (nTreg with specific alloantigen and IL-2 for 4 days. These cells express mRNA for IFN-γ receptor (ifngr and suppress donor but not third party cardiac allograft rejection mediated by alloreactive CD4+T cells at ratios of We induced highly suppressive Th1-like Treg from naïve nTreg in 7 days by culture with alloantigen, first with rIL-2 then with rIL-12p70. These Th1-like Treg delayed specific-donor allograft rejection demonstrating therapeutic potential

  13. Analysis of CD4+CD25+Foxp3+ regulatory T cells in HIV-exposed seronegative persons and HIV-infected persons with different disease progressions.

    Science.gov (United States)

    Li, Lin; Liu, Yongjian; Bao, Zuoyi; Chen, Lili; Wang, Zheng; Li, Tianyi; Li, Hanping; Zhuang, Daomin; Liu, Siyang; Wang, Xiaolin; Li, Jingyun

    2011-02-01

    Regulatory T cells (Tregs) are a subset of T cells that play an important role in the regulation of T-cell function. In a previous study, CD25 was used as a marker of Tregs; however, FoxP3 was recently discovered to be a valuable phenotype of Tregs. In this study, we compared the frequency of Tregs in HIV-1-infected long-term nonprogressors (LTNP), AIDS patients (AP), HIV-exposed seronegative (ES) persons, and healthy controls (HC), by using CD4+CD25+FoxP3+ as a marker of Tregs. The results showed that the frequency of Tregs in AP was significantly higher than in the LTNP, ES, and HC, which suggests that Tregs may play a role in disease progression. Another unique finding in this study is that we found a decrease of Tregs in ES.

  14. 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.

  15. Sublingual Immunotherapy Induces Regulatory Function of IL-10-Expressing CD4+CD25+Foxp3+ T Cells of Cervical Lymph Nodes in Murine Allergic Rhinitis Model

    Directory of Open Access Journals (Sweden)

    Takaya Yamada

    2012-01-01

    Full Text Available Sublingual immunotherapy (SLIT has been considered to be a painless and efficacious therapeutic treatment of allergic rhinitis which is known as type I allergy of nasal mucosa. Nevertheless, its mechanisms need to be further investigated. In this study, we constructed an effective murine model of sublingual immunotherapy in allergic rhinitis, in which mice were sublingually administered with ovalbumin (OVA followed by intraperitoneal sensitization and nasal challenge of OVA. Sublingually treated mice showed significantly decreased specific IgE responses as well as suppressed Th2 immune responses. Sublingual administration of OVA did not alter the frequency of CD4+CD25+ regulatory T cells (Tregs, but led to upregulation of Foxp3- and IL-10-specific mRNAs in the Tregs of cervical lymph nodes (CLN, which strongly suppressed Th2 cytokine production from CD4+CD25− effector T cells in vitro. Furthermore, sublingual administration of plasmids encoding the lymphoid chemokines CCL19 and CCL21-Ser DNA together with OVA suppressed allergic responses. These results suggest that IL-10-expressing CD4+CD25+Foxp3+ Tregs in CLN are involved in the suppression of allergic responses and that CCL19/CCL21 may contribute to it in mice that received SLIT.

  16. 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...... investigated the expression of FoxP3 and regulatory potential of gut-derived CD4(+)CD25(+) T cells cultured from patients with CD and healthy individuals. The FoxP3 expression was analysed by reverse transcriptase polymerase chain reaction (RT-PCR), and the suppressive effect of FoxP3(+)CD4(+)CD25(+) T cells...... on proliferation and cytokine production of autologous CD4(+) T cells was assessed by flow cytometry. Cultured gut-derived T cells with CD4(+)CD25(+) phenotype expressed FoxP3 and were able as the freshly isolated T(regs) from peripheral blood to suppress proliferation and cytokine production of autologous CD4...

  17. Functional and Developmental Analysis of CD4+CD25+ Regulatory T Cells under the Influence of Streptococcal M Protein in Rheumatic Heart Disease

    Directory of Open Access Journals (Sweden)

    Nidhal Abdul-Auhaimen

    2011-06-01

    Full Text Available The purpose of this study was to determine the role of streptococcal M protein in naturally-occurring CD4+CD25+ regulatory T cells (nTregs function and development in rheumatic heart disease in Iraqi patients. Streptococcus pyogenes was isolated for subsequent M protein extraction. Also, peripheral blood nTregs and CD4+ T cells were isolated by using Magnetic Cell Separation System. Tissue culture for isolated cells was performed in the presence and absence of M protein. Cell count was performed, and tumor necrosis factor alpha (TNF-α and interleukin-4 (IL-4 were determined in culture supernatant using ELISA system. There was a significant positive correlation (P0.05, association (r=0.353 between the mean number of nTregs and CD4+ T cells in the presence of M protein. The M protein stimulated CD4+ T cells to produce IL-4 in very little amount (<4 pg/ml in all samples. Compared to the production of IL4, TNF-α was produced in higher concentrations in the culture supernatants. The findings of the study indicate that streptococcal M protein has an important role in increasing the proliferation of CD4+CD25+regulatory T cells and CD4+ T cells. However, CD4+CD25+ regulatory T cells have lower suppressive activity against CD4+ T cells in the presence of M protein

  18. Functional and Developmental Analysis of CD4+CD25+ Regulatory T Cells under the Influence of Streptococcal M Protein in Rheumatic Heart Disease

    Science.gov (United States)

    Abdul-Auhaimena, Nidhal; Al-Kaabi, Zaman I. L

    2011-01-01

    The purpose of this study was to determine the role of streptococcal M protein in naturally-occurring CD4+CD25+ regulatory T cells (nTregs) function and development in rheumatic heart disease in Iraqi patients. Streptococcus pyogenes was isolated for subsequent M protein extraction. Also, peripheral blood nTregs and CD4+ T cells were isolated by using Magnetic Cell Separation System. Tissue culture for isolated cells was performed in the presence and absence of M protein. Cell count was performed, and tumor necrosis factor alpha (TNF-α) and interleukin-4 (IL-4) were determined in culture supernatant using ELISA system. There was a significant positive correlation (P0.05), association (r=0.353) between the mean number of nTregs and CD4+ T cells in the presence of M protein. The M protein stimulated CD4+ T cells to produce IL-4 in very little amount (<4 pg/ml) in all samples. Compared to the production of IL4, TNF-α was produced in higher concentrations in the culture supernatants. The findings of the study indicate that streptococcal M protein has an important role in increasing the proliferation of D4+CD25+regulatory T cells and CD4+ T cells. However, CD4+CD25+ regulatory T cells have lower suppressive activity against CD4+ T cells in the presence of M protein. PMID:23359747

  19. 安子合剂对ACA阳性先兆流产患者CD4+CD25+FOXP3+Treg细胞免疫调节的影响%TheEffect of Anzi Mixture on Immune Regulation of CD4+CD25+FOXP3+Treg Cells in ACA-positive Patients with Threatened Abortion

    Institute of Scientific and Technical Information of China (English)

    朱姝; 陆启滨

    2012-01-01

    目的:观察安子合剂对ACA阳性先兆流产患者CD4+ CD25+ FOXP3+ Treg细胞免疫调节的影响.方法:选取ACA阳性先兆流产患者为治疗组(27例),给服中药安子合剂,125ml,2次/日,连服4周;选取正常孕妇为对照组(15例).检测患者治疗前后及正常孕妇外周血CD4+ CD25+ FOXP3+ Treg细胞的比例,并测定患者治疗前后血清ACA指标、B超检查胚胎发育情况等.结果:治疗组患者治疗前CD4+CD25+FOXP3+Treg细胞的比例显著低于正常妊娠对照组(P<0.01);治疗后,其细胞比例显著高于治疗前(P<0.01).27例患者中20例ACA阳性的转阴率达85.00%,7例ACA定量滴度较治疗前显著下降(P<0.0l).保胎的成功率达92.59%.结论:ACA阳性先兆流产患者外周血CD4+CD25+ FOXP3 +Treg细胞的比例低于正常妊娠妇女,存在免疫调节异常;安子合剂治疗ACA阳性先兆流产的作用机理之一与增加CD4+ CD25+ FOXP3+ Treg细胞的比例、调节妊娠期机体免疫功能有关.%Objective; To observe the effect of Anzi Mixture on immune regulation of CD4+CD25+FOXP3+Treg cells in ACA -positive patients with threatened abortion. Methods: Selecting ACA-positive patients with threatened abortion as treatment group (27cases) , 125ml Anzi Mixture were given, 2 times a day, consecutive administrated for 4 weeks; selecting normal pregnant women as the control group ( 15cases). Detecting the percentage of peripheral blood CD4+CD25+FOXP3+Treg cells in the two groups, detecting the indicators of ACA before and after treatment by Anzi Mixture, B-ultrasound embryo development and so on. Results; Before treatment, the proportion of CD4+CD25+FOXP3+Treg cells in ACA-positive patients with threatened abortion was significantly lower than the control group (P<0. 01) ; after treatment of Anzi Mixture, the proportion of CD4+CD25+FOXP3+ Treg cells was significantly higher than before treatment (P<0. 01 ). The ACA negative rate of the qualitative indicators was 85. 00% , 7 cases of

  20. 3种不同中医证型艾滋病患者外周血CD+4CD+25调节性T细胞的表达%Expression of CD+4CD+25 regulatory T cell in peripheral blood of HIV/AIDS patients with different TCM syndromes

    Institute of Scientific and Technical Information of China (English)

    陈晓蓉; 杨宗国; 沈芳; 王江蓉; 卢洪洲; 杨悦娅

    2011-01-01

    objective: To observe the expression level of CD4+CD25+ regulatory T cell in peripheral blood of HIV/AIDS patients with different TCM syndromes, and to probe the immune negative regulation ability of HIV/AIDS. Methods: Based on the symptoms,signs,tongue manifestations, pulse conditions of AIDS patients, according to the thought of TCM Treatment with syndrome differentiation ,we divided the 97 HIV/AIDS patients into 3 different syndrome type, deficiency of pulmonary and renal syndrome, qi asthenia and blood stasis syndrome, and deficiency of spleen and renal syndrome, in order to observe the expression level of CD4+ CD25+ regulatory T cell in peripheral blood of HIV/AIDS patients with different TCM syndromes and 30cases of normal in control group. Results: The statistical significance of the expression level of CD4+ CD25+ regulatory T cell was observed among the 3 kinds of syndrome and control group (P<0.001). The deficiency of spleen and renal syndrome own the highest expression level of CD4+CD25+ regulatory T cell, followed by deficiency of pulmonary and renal syndrome, and qi asthenia and blood stasis syndrome. Conclusion: The expression level of CD4+CD25+ regulatory T cell of HIV/AIDS patients with different TCM syndromes has being reduced gradually from the deficiency of spleen and renal syndrome, to the deficiency of pulmonary and renal syndrome, and to the qi asthenia and blood stasis syndrome.%目的:观察不同中医证型艾滋病患者CD+4CD+25调节性T细胞(以下简称CD+4CD+25T细胞)的表达水平,探讨不同证型艾滋病免疫负调节能力.方法:根据艾滋病患者的症状、体征、舌象、脉象,按照中医辨证论治思路,分为肺肾不足、气虚血瘀、脾肾亏虚3种证型,设正常对照组30名,收集病例97例,观察不同证型艾滋病患者及正常对照组外周血CD+4CD+25T细胞的表达水平.结果:正常对照组、脾肾亏虚、肺肾不足、气虚血瘀各组患者CD+4CD+25T细胞的表

  1. IKK2dn转染树突状细胞诱导产生的CD4+CD25-T细胞的筛选及鉴定%Screening and function identification of CD4 + CD25 - T cells induced by immature dendritic cells transfected with IKK2dn

    Institute of Scientific and Technical Information of China (English)

    樊彩斌; 张东兴; 温端改; 侯建全; 欧阳骏; 杜科霖

    2012-01-01

    Objective To investigate the method of screening CD4 + CD25 - T cells from regulatory T cells (Treg) induced by recipient-derived immature dendritic cells (imDC) transfected by IKK2dn and loaded with donor antigens,and assess their immunologic function.Methods Rat bone marrow-derived imDC were transfected by IKK2dn and loaded by BN antigen,then cultured with Lewis rats T cells in vitro.From these Treg,CD4 + CD25 T cells were screened by magnetic active cell sorting (MACS),then incubated in secondary mixed lymphocyte reaction ( MLR),with Lewis rat T lymphocytes.Results By MACS,the purity of CD4 + CD25 - T cells was more than 95%.The result of secondary MLR displayed the absorbance value was significantly lower in the CD4 + CD25 - T cells group ( 0.106 ± 0.006 ) than that in BN antigen group (0.189 ± 0.007 ),Adv0-CD4 +T cells group (0.419 ± 0.014) and the third donor group (0.200 ± 0.008) (P <0.05).Conclusion By MACS sereening,we can obtain high purity of CD4 + CD25-T cells from Treg induced by imDC transfected with IKK2dn and loaded by BN rat antigen and recipient Lewis rats T cells.And these T cells can inhibit T cell proliferation with the donor antigen specificity.%目的 探讨从转染IKK2dn并负载供者抗原的未成熟树突状细胞(imDC)诱导产生的调节性T细胞(Treg)中筛选CD4+ CD25 - Treg的方法,并进行鉴定.方法 Lewis大鼠骨髓源性imDC,转染IKK2dn后负载供者BN大鼠抗原,与Lewis大鼠T细胞进行体外混合淋巴细胞反应(MLR)诱导产生Treg,用免疫磁珠法(MACS)筛选出CD4+ CD25 -T细胞,流式细胞仪(FCM)检测细胞纯度.加入CD4+ CD25 -T细胞行再次MLR检测其抑制T细胞增殖的作用.结果 经MACS筛选,CD4+ CD25 -T细胞纯度为(95.78±1.25)%.再次MLR结果显示CD4+ CD25 -T细胞组的吸光度值为(0.106±0.006),低于BN抗原组(0.189±0.007)、Adv0-CD4+T细胞组(0.419±0.014)及第三方供者抗原组(0.200±0.008),差异有统计学意义(P<0.05).结论 转染IKK2dn并负载

  2. Expression of Ets-1 and FOXP3 mRNA in CD4(+)CD25 (+) T regulatory cells from patients with systemic lupus erythematosus.

    Science.gov (United States)

    Xiang, Nan; Li, Xiang-Pei; Li, Xiao-Mei; Wang, Guo-Sheng; Tao, Jin-Hui; Pan, Hai-Feng; Fang, Xuan; Ma, Qian; Yu, Ning

    2014-11-01

    Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease with complex genetic predisposing factors involved. Ets-1 transcription factor plays an important role in the suppressive activity of CD4(+)CD25(+) Treg cells and stable expression of FOXP3. To find its potential role in the pathogenesis of SLE, we investigate the mRNA expression of Ets-1 and FOXP3 mRNA in CD4(+)CD25(+) Treg cells from patients with SLE. Real-time transcription-polymerase chain reaction analysis was used to determine the expression of Ets-1 and FOXP3 mRNA in CD4(+)CD25(+) Treg cells from 36 patients with SLE and 18 sex-and-age-matched healthy controls. The Ets-1 mRNA expression level was decreased in patients with SLE [0.225 (0.135, 0.337)] than healthy controls [0.528 (0.303, 0.681)] (P Ets-1 and FOXP3 expression was also found in new-onset SLE subgroup when compared with healthy controls (P Ets-1 and FOXP3 mRNA was not significantly different in hyperactive and lower active SLE group when compared with inactive SLE group, respectively (P > 0.05). There were no significant differences between SLE with lupus nephritis (LN) and SLE without LN either (P > 0.05). Associations of Ets-1 and FOXP3 mRNA expression levels with major clinical and laboratory parameters of SLE patients were also analyzed. However, no significant association was found. Significant positive correlation was found between Ets-1 and FOXP3 mRNA expression in CD4(+)CD25(+) Treg cells from SLE patients (r = 0.698, P Ets-1 mRNA were decreased in SLE patients and Ets-1 expression was positively correlated with the expression of FOXP3. It indicated that Ets-1 may play an important role in the stable expression of FOXP3 in CD4(+)CD25(+) Treg cells.

  3. Depletion of CD25+CD4+T cells (Tregs) enhances the HBV-specific CD8+ T cell response primed by DNA immunization

    Institute of Scientific and Technical Information of China (English)

    Yoshihiro Furuichi; Hirotake Tokuyama; Satoshi Ueha; Makoto Kurachi; Fuminori Moriyasu; Kazuhiro Kakimi

    2005-01-01

    AIM: Persistent hepatitis B virus (HBV) infection is characterized by a weak CD8+ T cell response to HBV. Immunotherapeutic strategies that overcome tolerance and boost these suboptimal responses may facilitate viral clearance in chronically infected individuals. Therefore, we examined whether CD25+CD4+ regulatory T (Treg) cells might be involved in a inhibition of CD8+T cell priming or in the modulation of the magnitude of the'peak' antiviral CD8+ T cell response primed by DNA immunization. METHODS: B10.D2 mice were immunized once with plasmid pCMV-S. Mice received 500 μg of anti-CD25 mAb injected intraperitoneally 3 d before DNA immunization to deplete CD25+ cells. Induction of HBV-specific CD8+ T ceils in peripheral blood mononuclear cells (PBMCs) was measured by S28-39 peptide loaded DimerX staining and their function was analyzed by intracellular IFN-γ staining.RESULTS: DNA immunization induced HBV-specific CD8+ T cells. At the peak T cell response (d 10), 7.1±2.0% of CD8+ T cells were HBV-specific after DNA immunization, whereas 12.7±3.2% of CD8+ T cells were HBV-specific in Treg-depleted mice, suggesting that DNA immunization induced more antigen-specific CD8+ T cells in the absence of CD25+ Treg cells (n = 6, P<0.05). Similarly, fewer HBVspecific memory T cells were detected in the presence of these cells (1.3±0.4%) in comparison to Treg-depleted mice (2.6±0.9%) on d 30 after DNA immunization (n = 6, P<0.01). Both IFN-γ production and the avidity of the HBV-specific CD8+ T cell response to antigen were higher in HBV-specific CD8+ T cells induced in the absence of Treg cells.CONCLUSION: CD25+ Treg cells suppress priming and/or expansion of antigen-specific CD8+ T cells during DNA immunization and the peak CD8+ T cell response is enhanced by depleting this cell population. Furthermore, Treg cells appear to be involved in the contraction phase of the CD8+ T ceil response and may affect the quality of memory T cell pools. The elimination of Treg

  4. In-vitro inhibition of IFNγ+ iTreg mediated by monoclonal antibodies against cell surface determinants essential for iTreg function

    Directory of Open Access Journals (Sweden)

    Daniel Volker

    2012-08-01

    Full Text Available Abstract Background IFNγ-producing CD4+CD25+Foxp3+ PBL represent a subtype of iTreg that are associated with good long-term graft outcome in renal transplant recipients and suppress alloresponses in-vitro. To study the mechanism of immunosuppression, we attempted to block cell surface receptors and thereby inhibited the function of this iTreg subset in-vitro using monoclonal antibodies and recombinant proteins. Methods PBL of healthy control individuals were stimulated polyclonally in-vitro in the presence of monoclonal antibodies or recombinant proteins against/of CD178, CD152, CD279, CD28, CD95, and HLA-DR. Induction of IFNγ+ iTreg and proliferation of effector cells was determined using four-color fluorescence flow cytometry. Blockade of iTreg function was analyzed using polyclonally stimulated co-cultures with separated CD4+CD25+CD127-IFNγ+ PBL. Results High monoclonal antibody concentrations inhibited the induction of CD4+CD25+Foxp3+IFNγ+ PBL (anti-CD152, anti-CD279, anti-CD95: p +CD25+CD127-IFNγ+ PBL (anti-CD178, anti-CD152, anti-CD279, anti-CD95: p +CD25+Foxp3+IFNγ+ PBL (rCD152 and rCD95: p +CD25+CD127-IFNγ+ PBL showed lower cell proliferation than co-cultures with CD4+CD25+CD127-IFNγ- PBL (p +CD25+CD127-IFNγ- PBL-containing co-cultures in the presence of monoclonal antibody (anti-CD28, anti-CD152, anti-CD279: p +CD25+CD127-IFNγ+ PBL (with the exception anti-CD28 monoclonal antibody: p +CD25+CD127-IFNγ- PBL but do not efficiently block suppressive iTreg function in co-cultures with CD4+CD25+CD127-IFNγ+ PBL. Conclusions CD178, CD152, CD279, CD28, CD95, and HLA-DR determinants are important for induction and suppressive function of IFNγ+ iTreg.

  5. 阻断ICOS/B7h信号的供体特异性输血对移植受体CD4+CD25+调节性T细胞的影响%Influence of donor specific transfusion with impaired ICOS/B7h allorecognition on CD4 + CD25+regulatory T cells of cardiac allograft recipient

    Institute of Scientific and Technical Information of China (English)

    杜峻峰; 李世拥; 于波; 白雪; 左富义

    2010-01-01

    目的 观察阻断ICOS/B7h信号的供体特异性输血(DST)对异基因小鼠心脏移植术后体内CD4+CD25+调节性T细胞(Treg)的影响.方法 按陈氏方法建立小鼠颈部异位心脏移植模型,实验分3组,异基因组及同基因组:供心分别来源于BALB/C和C57BL/6小鼠,受体均为C57BL/6小鼠,未予治疗.治疗组:移植当天给予受体鼠(C57BL/6)尾静脉注射5×106 ICOS-Fc靶定的供体(BALB/C)脾B淋巴细胞,d0~6连续给予受体鼠尾静脉注射ICOS-Fc 200 μg/d.术后统计各组移植物的存活时间,通过流式细胞术检测受体鼠外周血中CD4+CD25+Treg的亚群比例,利用逆转录-聚合酶链反应(RT-PCR)检测移植物中FOXP3的mRNA表达,在混合淋巴细胞反应中检测CD4+CD25+Treg对CD4+CD25-效应T细胞(Teff)的增殖抑制效率.结果 与异基因组比较,治疗组心脏移植物存活时间明显延长[(84.38±29.14)d比(7.00±0.76)d,P<0.01].各组中,治疗组受体外周血中CD4+CD25+Treg亚群比例显著上调[(15.60±5.69)%,P<0.01].与其他两组比较,治疗组心脏移植物中FOXP3 mRNA表达显著上调.以正常鼠为对照,耐受鼠脾脏中获取的CD4+CD25+Treg能够更高效地抑制CD4+CD25-Teff在混合淋巴细胞培养中的增殖效应.结论 通过阻断ICOS/B7h信号的DST可以诱导异基因小鼠心脏移植耐受,CD4+CD25+Treg在耐受的形成与维持中均起着重要作用.%Objective To investigate the possibility of CD4+CD25+ regulatory T cells (Treg) enhancement, which induced by donor specific transfusion combined with blockade of ICOS/B7h costimulation following an allogenic murine heterotopic cardiac transplantation. Methods Recipient mice were assigned to 3 groups. In allogenic group, donor hearts were harvested from BALB/C mice, and allografts were heterotopically transplanted in the neck of C57BL/6 using Chen' s technique, without treatment. In isogeneic group, donor hearts were harvested from C57BL/6 mice, and isografts were heterotopically

  6. 肺积方调节CD4+CD25+Treg细胞及T淋巴细胞增殖与抗Lewis肺癌移植鼠肿瘤生长作用研究%Effects of Feijifang on Levels of CD4+CD25+Treg and T Lymphocytes and Tumor Growth in Mice with Trans-planted Lewis Lung Cancer

    Institute of Scientific and Technical Information of China (English)

    杨国良; 张学进; 胡丹丹

    2013-01-01

    Objective:To investigate the effect of Feijifang combined with chemotherapy on tumor growth and the levels of CD4+CD25+ regulatory T cells(Treg) and T lymphocytes in mice with transplanted Lewis lung cancer and the underlying mechanism of Feijifang modulating immune evasion. Methods:Sixty-four mice with transplanted Lewis lung cancer were randomly divided into control group,Feiji group,comprehensive treatment group(Feijifang combined with diamminedichloroplatinum),and diamminedichloroplatinum (DDP) group,16 mice in each. The proliferation of T lymphocytes in the spleen and thymus was detected by MTT method and the percentage of CD4+CD25+Treg was assessed by flow cytometry. The volume and weight of tumors were recorded and evaluated. Results:The volume and weight of tumors in the DDP,Feiji,and comprehensive treatment groups were less than those in the control group (P0.05). On day 10 and 21,the percentage of CD4+CD25+Treg was lower in the DDP,Feiji,and comprehensive treatment groups as compared with that in the control group (P0.05)。第10、21天DDP组、肺积方组、综合组荷瘤小鼠胸腺CD4+CD25+Treg细胞比例均较对照组降低(P<0.05);DDP组第21天胸腺CD4+CD25+Treg细胞比例较第10天明显升高(P<0.05);综合组脾脏CD4+CD25+Treg细胞比例较低,与肺积方组、DDP组比较差异有统计学意义(P<0.05)。结论:肺积方联合化疗具有抗Lewis肺癌移植小鼠肿瘤生长的作用,该作用可能与其下调CD4+CD25+Treg细胞含量相关。

  7. CD4+CD25 + Foxp3+ T-reg cells play an important role in na(i)ve immune tolerance in rat liver transplantation%CD4+CD25+Foxp3+T-reg细胞在大鼠肝移植天然免疫耐受模型中的作用

    Institute of Scientific and Technical Information of China (English)

    肖江卫; 周彤; 王崇树; 魏寿江; 王城; 李敬东; 彭勇; 金先庆

    2010-01-01

    Objective To study the effect of CD4+ CD25+ Foxp3 + T-reg cells in na(i)ve immune tolerance using a rat liver transplantation model, we monitored the changes of this population cell of CD4+ CD25+ Foxp3+ T-reg cells in liver、thymus、spleen and blood by flow cytometry technique. Methods Na(i)ve immune tolerance model of rat liver transplantation was established (LEW→DA) ,The rejection group (DA→LEW) was used as the control group. CD4+ CD25+ Foxp3+ T-reg cells were respectively extracted and measured in liver、thymus、 spleen and blood. Simultaneously, we recorded the survival rate of rats and liver graft pathological changes in every group. The expression level of Foxp3 mRNA in every group was monitored. Results DA rat in tolerance group could acquire long-time survival without any immunosuppressive drug. Compared with rejection group,CD4+ CD25+ Eoxp3+ T-reg cells in tolerance group were significantly increased, especially in blood. The expression level of Foxp3 mRNA is significantly higher(P2个月),且排斥组大鼠最多只能存活7~10d,免疫耐受组在肝脏、外周血、脾脏和胸腺单核细胞中CD4+CD25+Foxp3+T-reg细胞亚群所占的比例明显高于排斥组(P<0.05),且在移植肝脏中,免疫耐受组的Foxp3 mRNA表达含量明显高于排斥组.结论 在LEW→DA的大鼠免疫耐受模型中,CD4+CD25+Foxp3+T-reg细胞亚群细胞数量和比例明显升高,提示CD4+CD25+Foxp3+T-reg细胞亚群在移植免疫耐受过程中发挥着重要作用,从而为我们临床诱导免疫耐受提供一条新的思路.我们可以通过体外扩增抗原特异性的天然存在的调节性T细胞来抑制效应T细胞的激活和扩增,从而帮助诱导移植免疫耐受和抑制移植物排斥.

  8. Study of expression of CD25 and CXCR3 on T lymphocytes from patients with HCV/HIV co-infection or HIV mono-infection in China%中国HGV/HIV合并感染者或HIV单一感染者T淋巴细胞表面CD25和GXCR3表达的研究

    Institute of Scientific and Technical Information of China (English)

    康辉; 王亚男; 范霞; 姜拥军; 刁莹莹; 尚红

    2006-01-01

    [Objective] To investigate expression and significance of CD25 and CXCR3 on T lymphocytes from peripheral blood mononuclear cells (PBMC) in HCV mono-infection, HIV mono-infection and HCV/HIV co-infection. [Methods] PBMC were isolated from patients with HCV infection (n=21), HIV infection (n=14), HCV/HIV coinfection (n=28) and normal control (n=30). The CD4 and CD8 absolute count, proportion of CD25+CD4+ T cells, and expression of CXCR3 on CD4+ and CD8+ T cells from PBMC were analyzed by flow cytometry. [Results] A lower proportion of CD4+CD25+ T lymphocytes were found in HIV infection and HCV/HIV co-infection (P <0.001), while slight but not significant increased proportion of CD4+CD25+ T lymphocytes were detected in HCV infection. The expression of CXCR3 on CD4+T cells from PBMCs decreased significantly, but the expression of CXCR3 on CD8+T cells increased significantly in HIV and HCV/HIV co-infection (P <0.001). The expression of CXCR3 on CD4+T cells as well as CD8+T cells from PBMCs increased but not significantly in HCV infection. [Conclusion] Results suggested that the expression of CD25 and CXCR3 on T lymphocytes from PBMC were altered and immune regulatory function would be effected in HIV infection in China.%目的探讨T淋巴细胞表面CD25分子和趋化因子受体CXCR3,在丙肝病毒(HCV)单一感染,艾滋病病毒(HIV)单一感染和HCV/HIV合并感染过程中的表达及意义.方法分离HCV感染组(n=21),HIV感染组(n=14),HCV/HIV感染组(n=28)及正常对照组(n=30)人外周血单个核细胞.采用流式细胞术,计数CD4+和CD8+T淋巴细胞数,检测外周血CD4+CD25+T细胞百分含量和CD4+和CD8+T淋巴细胞表面CXCR3的表达水平.结果CD4+T细胞表面CD25的表达在HCV感染组轻度升高,而在HIV感染组及HCV/HIV合并感染组CD25的表达显著降低(P<0.001).HIV感染组及HCV/HIV合并感染组CD4+T细胞表面CXCR3表达显著降低(P<0.001),CD8+T细胞表面CXCR3表达显著升高(P<0.001);HCV感染组CD

  9. Effects of pepper artemisia on CD4+CD25+ regulatory T cells in experimental autoimmune orchitis mice%椒蒿对自身免疫性睾丸炎小鼠CD4+CD25+调节性T细胞的影响

    Institute of Scientific and Technical Information of China (English)

    仵燕; 斯依提尼沙汗·吾满尔; 吴小川; 骆靖华; 玛衣拉·阿不拉克

    2013-01-01

    Objective To study the effects of volatile oil from pepper artemisia leaves on CD4+ CD25+ regulatory T cells in murine autoimmune orchitis mice. Methods Totally 80 male BALB/c mice were randomly divided into 4 groups (normal saline group, model group, olive oil group, and pepper artemisia group). After model making, each group received gavage three days before the surgery, and for 24 days after the surgery (saline group received only the gavage, but no operation). CD4+CD25 + regulatory T cells in the peripheral blood were detected by flow cytometry 3 and 9 weeks after the surgery. Results Three weeks after the surgery, the percentage of peripheral blood CD4+ CD25+ regulatory T cells in model group, olive oil group and pepper artemisia group was significantly lower than that in saline group (P0.05). Nine weeks after the operation, the percentage of peripheral blood CD4+CD25+ regulatory T cells was significantly lower in model group and olive oil group than in saline group (P0.05). Conclusion Pepper artemisia can increase the percentage of CD4+CD25+ regulatory T cells, which exert immunity-regulatory effects on autoimmune orchitis mice.%目的 研究椒蒿挥发油对自身免疫性睾丸炎小鼠CD4+ CD25+调节性T细胞的影响.方法 雄性BALB/c小鼠80只,随机分为4组(生理盐水组、模型组、橄榄油组、椒蒿组),采用睾丸内注射冰醋酸的方法造模,各组都在手术前3天开始灌胃,手术后继续灌胃,共连续灌胃24 d(盐水组只灌胃,不手术).分别于术后3周、9周取外周血,通过流式细胞仪对CD4+ CD25+调节性T细胞进行检测.结果 术后3周,模型组、橄榄油组、椒蒿组小鼠外周血CD4+ CD25+调节性T细胞百分比均低于生理盐水组,差异有统计学意义(P<0.05),而此3组之间差异无统计学意义(P>0.05).术后9周模型组与橄榄油组小鼠外周血CD4+ CD25+调节性T细胞百分比低于生理盐水组,差异有统计学意义(P<0.0S),椒蒿组与生理盐水组比

  10. Research progress of CD4+CD25+ regulatory T cell in bone marrow hematopoietic system diseases%CD4+CD25+调节性T细胞在骨髓造血系统疾病中的研究进展

    Institute of Scientific and Technical Information of China (English)

    刘增慧; 李勇华

    2011-01-01

    CD4 + CD25 + regulatory T cell is a unique T-cell subsets with immune regulation function. Natural CD4+CD25 + regulatory T cell originates from the thymus and acquired CD4+CD25 + regulatory T cell can be induced from CD4* CD25 - T cells in peripheral blood. A specific transcription inhibicory factor-foxp3 , can be expressed in its molecule surface, besides, it also can express other membrane molecules, such as CD4, CD25, CTLA-4 ( CD152), GITR, CD103, CD62L, CD69, CD134, CD71, CD54, CD45RA , TGF-β1 , IL-10, glactin-1 , neuropilin-1 , etc. Its main function is immunosuppression and immune anergy. Recent research finds that it overexpresses in acute myeloid leukemia, chronic lymphocytic leukemia and high-risk myelodysplastic syndrome. However, in aplastic anemia its expression is much lower than normal. Other researchers believe that it has much to do with acute graft versus host disease, in its occurrence and development. In this article, we will review the recent research results of CD4 + CD25 + regulatory T cell in bone marrow hematopoietic system diseases, specifically, in leukemia, acute graft versus host disease, aplastic anemia, myelodysplastic syndrome and multiple myeloma.%CD4+CD25+调节性T细胞(regulatory T cell, Treg)是具有独特免疫调节功能的T细胞亚群. 天然CD4+CD25+ Treg起源于胸腺,外周血中CD4+CD25-T细胞亦可诱导产生.其分子表面表达特异性的转录抑制因子Foxp3,还表达CD4、CD25、CTLA-4(CD152))、GITR、CD103、CD62L、CD69、CD134、CD71、CD54、CD45RA、TGF-β1、IL-10、半乳糖结合蛋白(glactin)-1、神经菌毛素(neuropilin)-1等膜分子,具有免疫抑制和免疫无能作用.近年来研究发现,急性髓细胞白血病(acute myeloid leukemia,AML)、慢性淋巴细胞白血病(chronic lymphocytic leukemia,CLL)、高危骨髓异常增生综合征(myelodysplastic syndrome, MDS)患者Treg表达异常增高,而在再生障碍性贫血(aplastic anemia,AA)患者表达明显降低,其异常表达

  11. HIV慢性感染者Vδ2T淋巴细胞的CD25表达与其凋亡及疾病进展的关系研究%Correlations between CD25 expression on Vδ2 T lymphocytes and the apoptosis of T lymphocytes and disease progression in chronically HIV-infected patients

    Institute of Scientific and Technical Information of China (English)

    许文; 黄磊; 谢杨新; 秦恩强; 石磊; 武晓丽; 杨俊连; 杨欣欣; 赵敏

    2013-01-01

    Objective To investigate the correlations between CD25 expression on Vδ2 T lymphocytes and the frequency and apoptosis of Vδ2 T lymphocytes and disease progression in chronically HIV-infected patients.Methods Thirty-one chronically HIV-infected outpatients treated in our hospital from Mar.2012 to Apr.2013 and 10 healthy controls (HC) were enrolled in the study.Of 31 chronically HIV-infected patients,10 didn't receive highly active antiretroviral therapy (HAART) (without HAART group),10 received 3 months of HAART (3-month HAART group),and 11 received 6 months of HAART (6-month HAART group).The frequency and apoptosis of Vδ2 T lymphocytes and CD25 expression on Vδ2 T lymphocytes were detected by flow cytometry.The changes of the above-mentioned indicators of without HAART group,3-month HAART group and 6-month HAART group were observed respectively,and the correlations between these indicators and CD4+ T lymphocyte counts and viral loads of each group were analyzed respectively.The correlations between CD25 expression and the frequency and apoptosis of Vδ2 T lymphocytes were analyzed as well.Results CD25 expression on Vδ2 T lymphocytes in chronically HIV-infected patients increased significantly as compared with HC.Though 6 months of HAART could down-regulate it,the expression was still higher than that of HC.CD25 expression on Vδ2 T lymphocytes decreased along with the increase in CD4+ T lymphocyte counts,but the correlation was not significant.The correlation between CD25 expression on Vδ2 T lymphocytes and viral loads in the patients not receiving HAART was not significant.The frequency of Vδ2 T lymphocytes was negatively correlated with CD25 expression in chronically HIV-infected patients,but the correlation between the apoptosis of and CD25 expression on Vδ2 T lymphocytes was not significant.Conclusions Vδ2 T lymphocytes are activated in chronically HIV-infected patients,which may be correlated with the decline in the frequency of Vδ2 T lymphocytes

  12. Low circulating CD4(+) CD25(+) Foxp3(+) T regulatory cell levels predict miscarriage risk in newly pregnant women with a history of failure.

    Science.gov (United States)

    Winger, Edward E; Reed, Jane L

    2011-10-01

    PROBLEM  The purpose of this study was to determine whether quantification of peripheral blood Treg cell levels could be used as an indicator of miscarriage risk in newly pregnant women with a history of immunologic reproductive failure. METHOD OF STUDY  Fifty-four pregnant women with a history of immunologic infertility and/or pregnancy loss were retrospectively evaluated (mean age: 36.7 ± 4.9 years, 2.8 ± 2.5 previous miscarriages; 1.5 ± 1.9 previous IVF failures). Twenty-three of these women experienced another first trimester miscarriage, and 31 of these women continued their current pregnancies past 12 weeks ('pregnancy success'). The following immunologic parameters were assessed in the first trimester: NK cell 50:1 cytotoxicity, CD56(+)  16(+)  CD3(-) (NK), CD56(+)  CD3(+) (NKT), TNFα/IL-10, IFNγ/IL-10, CD4(+)  CD25(-) Foxp3(+) , total CD4(+)  Foxp3(+) (CD4(+ ) CD25(+)  Foxp3 plus CD25(- ) Foxp3(+) ), and CD4(+)  CD25(+)  Foxp3(+) levels. RESULTS  Patients with successful ongoing pregnancies experienced a mean (CD4(+)  CD25(+)  Foxp3(+) ) 'Treg' level of 0.72 ± 0.52%, while those that miscarried in the first trimester experienced a mean Treg level of 0.37 ± 0.29% (P = 0.005). Markers not significantly different between the loss and success groups were NK 50:1 cytotoxicity (P = 0.63), CD56(+)  16(+)  3(+) NK cells (P = 0.63), CD56(+)  3(+)   NKT (P = 0.30), TNFα(+) IL-10(+) (P = 0.13), IFNg(+) IL-10(+) (P = 0.63), and CD4(+)  25(-)  Foxp3(+) cells (P = 0.10), although total CD4(+)  Foxp3(+) levels remained significant (P = 0.02) and CD4(+)  25(+) Foxp3(+) showed the most significant difference (P = 0.005). Mean day of blood draw was 49.2 ± 36.1 days pregnant (median 39.0 days). In addition, patients with a low Treg level (0.7%) in the first trimester [44% (15/34) versus 80% (16/20); P = 0.01]. Of the 18 successful pregnancies with sequential Treg

  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. The change of CD4+ CD25+ Treg cells in the collagen-induced arthritis mouse model transplanted with mesenchymal stem cells%骨髓间充质干细胞对胶原诱导的关节炎大鼠CD4+CD25+调节性T细胞的影响

    Institute of Scientific and Technical Information of China (English)

    张明生; 穆永慧; 陈清汉; 朱宇

    2014-01-01

    Objective To observe the immunologic effect of transplantation of mesenchymal stem cells (MSCs) on CD4+ CD25 + Treg cells frequency and function in collagen-induced arthritis (CIA) rat model.Methods Rat CIA model was established by immunizing SD rats with type Ⅱ collagen.MSCs were purified and cultured from the bone marrow of SD rats,then transplanted to CIA model.The frequency of CD4 + CD25 + Treg cells in the spleen was measured by flow cytometry (FCM) respectively after the MSCs transplantation,and the expression of forkhead box P3 (Foxp3),transforming growth factor-β1 (TGF-β1)and interleukin (IL)-10 mRNA was evaluated by real-time quantitative polymerase chain reaction (Real-time PCR).CD4 + CD25 + Treg cells were isolated from the spleen of rats by immunomagnetic beads and the inhibitory effect of CD4 + CD25 + Treg cells on proliferation of CD4 + CD25-Teff cells was estimated by methyl thiazol tetrazolium (MTT) colorimetry.Results The MSCs transplantation relieved the arthritis symptom of the CIA rats.The frequency of CD4 + CD25 + Treg cells in the spleen of MSCs-treated CIA rats was increased significantly (P < 0.05) as compared with the CIA group,and the levels of Foxp3,IL-10 and TGF-β1 mRNA were also significantly increased (P < 0.05).CD4 + CD25 + Treg cells isolated from treatment group showed higher inhibition on the proliferation of CD4 + CD25-Teff cells versus CD4 + CD25 +Treg cells isolated from CIA rats.Conclusion Transplantation of MSCs is effective on preventing the de-velopment of CIA.The increased frequency and reinforced function of CD4+ CD25 + Treg cells may be one of the mechanisms for MCSs transplantation treating CIA in rats.%目的 观察间充质干细胞(MSCs)对Ⅱ型胶原诱导的关节炎(CIA)大鼠Treg细胞数量和功能的影响.方法 以Ⅱ型胶原蛋白免疫SD大鼠建立CIA动物模型;分离培养正常SD大鼠骨髓MSCs,尾静脉移注CIA大鼠;流式细胞仪检测大鼠脾脏CD4 +CD25+T细胞比例变化;实时

  15. 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

    sacrificed weekly from week 1 to 4. Clinical signs of colitis occurred approximately 2 weeks after reconstitution. Disease onset and severity based on histopathology correlated well with the colonic weight:length ratio, fecal consistency score, presence of occult blood in feces, and fecal IL-1beta content....... 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....

  16. 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...... 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...

  17. 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

    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...... course of HIV infection. However, despite HAART during pregnancy, HIV-infected women display different immunologic profiles from HIV-negative women, which may have importance for the induction of fetal-maternal tolerance and in part explain the increased risk of abortion in HIV-infected women....

  18. 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

  19. 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.

  20. 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.

  1. Study of Different Death Patterns of Huaman Peripheral Blood CD4+CD25+T Lymphocytes Induced by Anti-CD137 mAb%共刺激分子CD137单克隆抗体对CD4+CD25+T淋巴细胞死亡方式影响的研究

    Institute of Scientific and Technical Information of China (English)

    尹柯; 周文英; 邬伟明; 黄瑾

    2009-01-01

    目的:初步观察抗共刺激分子CD137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、胀亡现象的影响及其表面特征性标志物FOXp3的表达的变化.方法:采用密度梯度离心法及尼龙棉柱法分离健康志愿者外周血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、检测各分组(抗CD137单克隆抗体2 μg/ml组、IgGl同型抗体对照组)的细胞的自噬率、凋亡率、胀亡率及FOXp3的表达.结果:MACS可成功分离CD4+CD25+T淋巴细胞,纯度可达80.3%~89.5%;抗CD137单克隆抗体可促进人外周血CD4+CD25+T淋巴细胞自噬率及凋亡率增加;各组间的胀亡率改变及FOXp3表达差异无统计学意义.结论:抗CD137单克隆抗体可促进CD4+CD25+T淋巴细胞凋亡和自噬,其作用机制未发现与胀亡显著相关,而且不影响CD4+CD25+T细胞中Foxp3的表达.

  2. Expressions of CD4 + CD25high regulatory T cells, TGF-β1 and COX-2 in the peripheral blood of prostate cancer patients%前列腺癌患者外周血中CD4+CD25high调节性T细胞及调节因子TGF-31和COX-2的表达

    Institute of Scientific and Technical Information of China (English)

    夏桃林; 刘涛; 吴振权; 张海滨; 杨明; 刘绍远; 何灼彬; 李辽源

    2011-01-01

    Objective: To investigate the expressions of CD4+ CD25high regulatory T cells, TGF-β1 and COX-2 in the periphe-ral blood of prostate cancer (Pca) patients, and analyze the role of CD4+ CD25high regulatory T cells in the pathogenesis of Pca and their relationship with TGF-pl and COX-2. Methods: We used flow cytometry to calculate the percentage of CD4+ CD25high regulatory T cells in the CD4 + T cells in the peripheral blood mononuelear cells ( PBMC) from 30 Pca patients (11 localized and 19 non-localized cases) and 20 healthy volunteer controls, determined the expressions of TGF-pl and COX-2 in the serum by ELISA, and analyzed their correlation with the CD4+ CD25high regulatory T cells in the Pca patients as well as the differences between the localized and non- localized cases. Results: CD4+ CD25high regulatory T cells accounted for (18.32 ±7.49) % in the CD4+ T cells in PBMCs from the Pca patients, significantly higher than (7.77 ±1.86) % from the controls (P 0.05 ). The expressions of TGF-pl and COX-2 in the peripheral blood were (215.97 ±55.16) ng/ml and (6.88 ±5.14) ng/ml in the Pca patients, in comparison with (149.75 ±47.11) ng/ml (P 0.05 ) in the controls. Multiple linear regression analysis showed no significant correlation between the expression of CD4 + CD25high regulatory T cells in PBMCs and those of TGF-pl and COX-2 in the peripheral blood of the Pca patients. There were no significant differences between the localized and non-localized Pca groups in the expressions of CD4 + CD25high regulatory T cells, TGF-pl and COX-2 (P > 0.05). Conclusion-. CD4 + CD25high regulatory T cells in in PBMCs are involved in the pathogenesis of Pca. The proliferation of CD4 + CD25 high regulatory T cells is not significantly correlated to the expressions of TGF-pl and COX-2 in the peripheral blood, but maybe to the tumor itself and the local tumor microenvironment.%目的:通过检测前列腺癌患者以及健康志愿者外周血中CD4+ CD25high

  3. 抗肿瘤坏死因子相关凋亡诱导配体受体2嵌合抗体表达载体的构建、表达及其抗肿瘤活性分析%Construction and stable expression of anti-human tumor necrosis factor-related apoptosis-inducing ligand receptor 2 chimeric antibody in CHO cells

    Institute of Scientific and Technical Information of China (English)

    吕付佳; 史娟; 张亚玺; 刘士廉; 刘彦信; 郑德先

    2011-01-01

    目的:构建抗人肿瘤坏死因子相关凋亡诱导配体(TRAIL)受体2(死亡受体5,DR5)的人-鼠嵌合抗体表达载体,获得稳定表达该嵌合抗体的细胞株,并分析嵌合抗体的抗肿瘤活性.方法:采用DNA重组技术,扩增抗人DR5的鼠源单克隆抗体(mAb)AD5-10的重链(HC)、轻链(LC)可变区基因片段,并将其分别插入含有人IgG重链、轻链恒定区基因的真核表达载体RpCI-neo,以重、轻链表达质粒共转染中国仓鼠卵巢细胞(CHO),筛选稳定表达抗人DR5嵌合抗体(hmAD5-10)的重组细胞.采用Western blot和间接ELISA检测嵌合抗体的表达量及其与抗原DR5的结合活性.采用MTS比色法检测嵌合抗体的生物学活性.并对重组细胞株进行无血清培养驯化.结果:获得了2株稳定表达嵌合抗体的重组细胞株CHO-A5和CHO-B11,抗体的表达水平分别为(0.36±0.11)mg/L和(0.16±0.01)mg/L,嵌合抗体与DR5有较好的结合活性,对体外培养的人T淋巴细胞白血病细胞SVT35有显著的杀伤作用.结论:在真核细胞中表达了具有生物学活性的抗DR5的人-鼠嵌合抗体,为其应用于肿瘤治疗研究奠定了基础.%AIM: To establish an human-mouse chimeric antibody against tumor necrosis factor-related apoptosisinducing ligand (TRAIL) receptor 2 (death receptor 5, DR5) in an eukaryotic cell line and analyse ifs tumoricidal activity. METHODS: The cDNAs encoding for the variable regions of heavy chain (VH) and light chain (VL) of AD5-10 were amplified by PCR and inserted into the human lgG heavy and light chain containing expression vector RpClneo, respectively. The recombinant plasmids were co-transfected into HEK293 and/or CHO cells. The production of anti-DR5 human-mouse chimeric antibody (hmAD5-10) and the antibody affinity for DR5 were identified by ELISA and Western blot assay. The tumoricidal activity of hmAD5-10 was demonstrated by MTS assay. The stable expression cells were selected and cultured in serum-free medium

  4. Analysis on CD4 + CD25 + T cell in HCV infection and HCV/HIV co-infection%HCV感染者和HCV/HIV合并感染者免疫调节

    Institute of Scientific and Technical Information of China (English)

    康辉; 尚红; 刁莹莹; 范霞; 代娣; 姜桐军; 耿文清

    2005-01-01

    目的探讨我国单纯丙型肝炎病毒(HCV)感染者和HCV/HIV合并感染者免疫应答的相关机制.方法分离人外周血单个核细胞;流式细胞仪(FACS)检测CD4+T细胞和CD4+CD25+T细胞表达水平.结果CD4+CD25+T细胞占外周血单个核细胞中CD4+T细胞比例(%CD4+CD25+),HCV感染组明显高于健康对照组(19.2%>13.8%,P<0.05),HCV/HIV合并感染组明显低于对照组(6.9%<13.8%,P<0.001),更明显低于HCV感染组(P<0.001).结论HCV感染者体内CD4+CD25+T细胞增殖明显,提示CD4+CD25+T细胞在HCV慢性感染中发挥免疫调节作用.HCV/HIV合并感染时,CD4+CD25+T细胞受损,从而影响免疫调节功能.

  5. The Prospects of the usage of CD+4CD+25 regulatory T cells in cancer immunotherapy%CD+4CD+25调节性T细胞在肿瘤免疫治疗中的价值及应用前景

    Institute of Scientific and Technical Information of China (English)

    付泽娴; 蔡建辉

    2005-01-01

    恶性肿瘤的临床治疗主要是手术治疗、化学治疗和放射治疗.近年来,随着分子生物技术的不断发展,肿瘤的生物治疗在临床应用中的地位日渐突出.但是免疫细胞过继治疗作用的一过性和功能抑制成为影响其临床应用的瓶颈,其中CD+4CD+25调节性T细胞(CD+4CD+25 regulatory T cell ,Treg)在抑制肿瘤免疫方面起着重要作用.本文就其近年来的肿瘤临床研究进展做一综述.

  6. Impact of the CD4 + CD25nt/hiCD127loregulatory T cells on the immune status and disease progression in HIV-1 infected individuals%CD4+CD25 nt/hi CD127lo调节性T细胞对HIV感染者免疫状态及病程进展的影响

    Institute of Scientific and Technical Information of China (English)

    周明奎; 薛以乐; 宫菊丽; 周磊明; 郑晓虹; 盖晶; 沈方伟; 张玮; 宁镇; 岳清; 卢伟; 潘启超; 康来仪; 钟平; 朱文斯; 王盈

    2007-01-01

    目的:探讨具有CD4+CD25nt/hi CD127lo特征的调节性T细胞频率对中国HIV-1感染者免疫状态及病程进展的影响.方法:选取100名未经治疗的HIV-1感染者和4个年龄组的312名健康人对照,采集静脉血,经三重免疫荧光染色,用流式细胞术分析CD4+CD25+Treg表型和CD4+CD25nt/hiCD127loTreg频率并进行CD4+T细胞绝对计数;应用酶联免疫斑点技术(ELISpot)在单细胞水平观察受试者的HIV-1特异性细胞免疫功能;平行检测HIV感染者的病毒载量(NASBA方法).结果:不同病程的HIV-1感染者外周血中CD4+CD25nt/hiCD127loTreg频率存在差异,进展期高于新近感染者(P<0.001),其平均水平显著高于健康人(P<0.001);CD4+CD25nt/hiCD127loTreg频率与HIV感染者CD4+T细胞数量呈显著负相关(r=0.354,P<0.01),与病毒载量呈明显正相关(r=0.287,P<0.01);高频率CD4+CD25nt/hiCD127loTreg HIV-1感染者(进展期)的外周血PBMCs经HIV-1多肽刺激后分泌IFN-γ的CD8+T细胞频率明显低于无症状的新近感染者.结论:初步证实HIV-1感染者外周血中CD4+CD25nt/hiCD127kloTreg细胞频率增加与CD4+T细胞数量降低及病程进展相关;高频率CD4+CD25nt/hiCD127lo Treg细胞对HIV-1感染者的细胞免疫功能具有抑制作用.本结果为进一步阐明HIV持续感染的免疫机制提供了新依据.

  7. 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

  8. Study on the correlation between CD4+CD25+Foxp3+T regulatory cells and NK cells in decidual tissues of patients with preeclampsia%子痫前期患者蜕膜中CD4+CD25+Foxp3+调节性T细胞及NK细胞的相关研究

    Institute of Scientific and Technical Information of China (English)

    王莲莲; 蒋宏颉; 魏军

    2011-01-01

    Objective; To study the correlation between CD4 + CD25 + Foxp3 + T regulatory cells and NK cells in decidual tissues of patients with preeclampsia, explore the related immune changes in pathogenesis of preeclampsia. Methods; 30 cases with preeclampsia (13 cases with mild preeclampsia and 17 cases with severe preeclampsia) and 30 normal pregnant women of late pregnancy were selected, lmmu-nohistochemical method was used to detect and calculate CD4 + CD25 + Foxp3 + T regulatory cells and NK cells in decidual tissues, the average gray value was also detected. Results: The positive expression rate of CD4 + CD25 + Foxp3 + T regulatory cells in decidual tissues in preeclampsia group was significantly lower than that in normal late pregnancy group ( P < 0. 05 ) . The amount of NK cells in decidual tissues in preeclampsia group was significantly higher than that in normal late pregnancy group (P < 0. 05 ) , the average gray value of CD56 immunohis-tochemical staining in NK cells was significantly higher than that in normal late pregnancy group (P < 0. 05) . Conclusion: The amount of CD4 + CDjj + Foxp3 * T regulatory cells in decidual tissues in patients with preeclampsia is significantly lower than that in normal pregnant women of late pregnancy; the amount of NK cells in decidual tissues in patients with preeclampsia is significantly higher than that in normal pregnant women of late pregnancy, which induces maternal - fetal immune unbalance and activity of NK cells, then the killing activity is enhanced, preeclampsia occurs.%目的:对子痫前期(PE)患者蜕膜中CD4+CD25+Foxp3+调节性T细胞(CD4+CD25+Foxp3+Treg)和NK细胞进行相关研究,探讨子痫前期发病机制中相关的免疫学变化.方法:选择子痫前期患者30例(轻度13例、重度17例)、正常晚期妊娠妇女30例.采用免疫组织化学法检测胎盘蜕膜CD4+CD25+Foxp3+调节性T细胞和NK细胞后计数并检测其平均灰度值.结果:1.子痛前期患者蜕膜CD4+CD25+Foxp3+Treg

  9. 初发Graves病患者CD4~+CD25~+Foxp3~+调节性T细胞水平的研究%Study on the Frequencies of CD4~+CD25~+Foxp3~+Regulatory T Cells in the Patients with Graves' Disease

    Institute of Scientific and Technical Information of China (English)

    高桂华; 赵连爽

    2010-01-01

    采用胞内染色技术分别检测45例初发Graves病患者(GD组)和20例健康成人(对照组)CD4~+CD25~+Foxp3~+调节性T细胞水平的变化,并用放射免疫分析法检测TT3、TT4和TSH.结果 显示GD组TT3和TT4水平高于对照组,而TSH水平低于对照组.认为初发GD患者CD4~+CD25~+Forxp3~+调节性T细胞水平明显下降,低于正常人水平,在GD的发病中可能发挥重要作用.

  10. 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.

  11. 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

  12. Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25(+)CD4(+) regulatory cells that control intestinal inflammation.

    Science.gov (United States)

    Read, S; Malmström, V; Powrie, F

    2000-07-17

    It is now clear that functionally specialized regulatory T (Treg) cells exist as part of the normal immune repertoire, preventing the development of pathogenic responses to both self- and intestinal antigens. Here, we report that the Treg cells that control intestinal inflammation express the same phenotype (CD25(+)CD45RB(low)CD4(+)) as those that control autoimmunity. Previous studies have failed to identify how CD25(+) Treg cells function in vivo. Our studies reveal that the immune-suppressive function of these cells in vivo is dependent on signaling via the negative regulator of T cell activation cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), as well as secretion of the immune-suppressive cytokine transforming growth factor beta. Strikingly, constitutive expression of CTLA-4 among CD4(+) cells was restricted primarily to Treg cells, suggesting that CTLA-4 expression by these cells is involved in their immune-suppressive function. These findings raise the possibility that Treg cell function contributes to the immune suppression characteristic of CTLA-4 signaling. Identification of costimulatory molecules involved in the function of Treg cells may facilitate further characterization of these cells and development of new therapeutic strategies for the treatment of inflammatory diseases.

  13. 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.

  14. The role of dendritic cells in the generation of CD4(+) CD25(HI) Foxp3(+) T cells induced by amino acid copolymers.

    Science.gov (United States)

    Kawamoto, Norio; Ohnishi, Hidenori; Kondo, Naomi; Strominger, Jack L

    2013-01-01

    The effects of the amino acid copolymers used in the therapy of experimental autoimmune encephalomyelitis, poly(Y,E,A,K)(n) (Copaxone(®)) and poly(Y,F,A,K)(n), on murine myeloid cells have been investigated. After administration of these copolymers to mice, increases in several splenic myeloid cell populations were observed, including CD11b(+) CD11c(+) dendritic cells. The latter were the major splenic cell type that secreted CCL22 (macrophage-derived chemokine) on stimulation with amino acid copolymers. CCL22 secretion was also stimulated from bone marrow-derived dendritic cells (BMDC) generated with GM-CSF in much larger amounts than from bone marrow-derived macrophages generated with M-CSF. Moreover, CCL22 secretion could also be obtained using BMDC generated from two different types of MHC II(-/-) mice, indicating that an innate immune receptor is involved. Finally, incubation of these BMDC or splenic dendritic cells with naive CD4(+) CD25(-) T cells resulted in formation of CD4(+) CD25(HI) Foxp3 T cells (~25% of which were Foxp3(+)). The number of these regulatory cells was doubled by pretreatment of BMDC with amino acid copolymers.

  15. Clinical Significance of CD4 + CD25 (High) CD127(Low/-) Regulatory T Cells Frequency in Peripheral Blood of Patients with Active Rheumatoid Arthritis%类风湿关节炎外周血CD4+CD25(high)CD127(low/-)T细胞表达的变化及意义

    Institute of Scientific and Technical Information of China (English)

    武加标; 任敏; 汤丽; 李雪飞; 顾巧英; 戚传平; 肖菁; 赵东宝

    2013-01-01

    目的 探讨类风湿关节炎(Rheumatoid arthritis,RA)活动期患者外周血细胞表面CD4+ CD25 (high) CD127(low/-)调节性T细胞数量的变化.方法 用流式细胞术检测32名RA患者,14名膝关节骨性关节炎(osteoarthrosis of knee,OA)患者和13名正常人外周血细胞CD4+ CD25 (high) CD127(low/-)调节性T细胞的百分比,并统计分析其数量变化与有关临床和实验室指标的相关性.结果 RA患者外周血CD4+ CD25 (high) CD127(low/-)调节性T细胞在CD4T细胞中的百分率(4.82±0.49)均显著低于OA组(6.54 ±0.45)和组对照(7.05±0.62) (P <0.05),且与RA活动指数DAS28呈负相关(P<0.05).结论 RA患者外周血CD4+ CD25(high)CD127(low/-)调节性T细胞数量显著降低,提示该细胞亚群可能在调节RA疾病活动中发挥重要作用,并且也可以作为监测RA疾病活动一个重要指标.

  16. The expression of membrane interleukin-2 receptor(CD25) on the surface of peripheral blood mononuclear cells(PBMC) of pulmonary tuberculosis and its clinical significance%肺结核患者PBMC膜白介素-2受体(CD25)表达及其临床意义

    Institute of Scientific and Technical Information of China (English)

    王健; 李朝品; 刘智; 刘炳祥

    2003-01-01

    目的:探讨外周血单个核细胞(PBMC)膜白介素-2受体(CD25)表达在肺结核病鉴别诊断中的应用价值.方法:用生物素-链霉亲和素(BSA)法检测肺结核、支气管肺炎患者T细胞亚群及植物血凝素(PHA)诱导前后CD25表达水平.结果:支气管肺炎患者CD3+、CD4+、CD8+水平分别为(62.32±6.34)%、(47.52±7.16)%、(32.12±6.55)%,CD4+/CD8+ 比值为1.52±0.43,PHA诱导前后CD25水平分别为(4.56±1.52)%、(35.12±7.21)%.空洞型肺结核CD3+、CD4+、CD8+、CD4+/CD8+水平分别为(41.13±5.25)%、(43.38±5.15)%、(36.25±3.46)%和1.15±0.21,非空洞型肺结核CD3+、CD4+、CD8+、CD4+/CD8+水平分别为(46.29±5.60)%、(47.21±4.86)%、(32.36±4.03)%、1.46±0.25,相互比较CD3+、CD4+/CD8+差异均有显著性(P<0.01和P<0.05).空洞型肺结核与非空洞型肺结核患者PHA诱导前后CD25水平分别为(2.13±1.14)%、(27.25±3.50)%和(3.43±1.35)%、(31.14±4.11)%,两者相比差异均有显著性(P<0.01).结论:肺结核病患者体内存在明显的细胞免疫功能紊乱,主要表现为CD25表达水平降低,CD25表达水平与肺结核病的病情似有一定关系,其对肺结核病鉴别诊断具有重要价值.%Objective:To study the value of membrane interleukin-2 receptor(CD25) of peripheral blood mononuclear cells (PBMC) on the differential diagnosis of pulmonary tuberculosis. Methods:The expression of T cell subset and levels of CD25 before and after induction with PHA were detected by biotin-streptavidin(BSA) in patients with pulmonary tuberculosis and bronchopneumonia. Results:The levels of CD3+, CD4+, CD8+, CD4+/CD8+ and CD25 before and after induction in peripheral blood in patients with bronchopneumonia were(62.32±6.34)%,(47.52±7.16)%,(32.12±6.55)%, 1.52±0.43,(4.56±1.52)%,and (35.12±7.21)%,respectively. The levels in pulmonary tuberculosis with cavity were(41.13±5.25)%,(43.38±5.15)%,(36.25±3.46)%, 1.15±0.21,(2.13±1.14)%,and (27.25±3.50)% and in pulmonary tuberculosis

  17. Depressive effect of cigarette smoke extracts on dendritic cells inducing differentiation of CD4+T cells into CD4+CD25+Foxp3+ Tregs%香烟烟雾提取物抑制树突状细胞诱导CD4+T细胞分化为CD4+CD25+Foxp3+调节性T细胞

    Institute of Scientific and Technical Information of China (English)

    梁毅; 周广; 张龙举; 刘积锋; 钟小宁

    2016-01-01

    目的 探讨香烟烟雾提取物(CSE)暴露及抑制CD40-CD40L路径对小鼠髓源性树突状细胞(BMDC)诱导CD4+T细胞分化为CD4+ CD25+ Foxp3+调节性T细胞(Treg)的影响.方法 使用40 ng/mL重组小鼠粒细胞-巨噬细胞集落刺激因子(rmGM-CSF)和10 ng/mL重组小鼠白细胞介素4(rmIL-4)联合诱导无特定病原体级健康雄性BALB/c小鼠骨髓来源的单个核细胞分化为树突状细胞(DC),流式细胞术检测小鼠BMDC表面CD40分子的表达,再采用免疫磁珠分选的方法从BALB/c小鼠脾脏细胞分离出CD4+T细胞.将所培养出的小鼠BMDC与正常对照组小鼠脾脏细胞分选的CD4+T细胞共培养,加入CSE及拮抗性CD40抗体,作用24h;流式细胞术观察各组细胞CD4+ CD25+ Foxp3+ Treg的变化,液相芯片技术检测细胞上清液白细胞介素10(IL-10)、IL-6的水平.结果 BMDC与CD4+T淋巴细胞共培养可以促进CD4+ CD25+ Foxp3+ Treg分化,经CSE刺激后,流式细胞术检测显示小鼠BMDC与CD4+T细胞共培养后分化的CD4+ CD25+ Foxp3+ Treg数量降低,细胞上清液IL-10降低、IL-6的水平升高;而加入拮抗性CD40抗体细胞共培养组CD4+ CD25+ Foxp3+ Treg增加,液相芯片检测细胞上清液IL-10上升、IL-6的水平下降.结论 CSE减少CD4+ CD25+ Foxp3+ Treg数量,体外使用拮抗性抗CD40抗体阻断CD40-CD40L通路可以促进CD4+T细胞分化为CD4+ CD25+ Foxp3+ Treg.

  18. 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

  19. 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

  20. In vitro inhibition of enterobacteria-reactive CD4+CD25- T cells and suppression of immunoinflammatory colitis in mice by the novel immunomodulatory agent VGX-1027

    DEFF Research Database (Denmark)

    Mangano, Katia; Sardesai, Niranjan; D'Alcamo, Maria;

    2008-01-01

    VGX-1027 is an isozaxoline compound that has recently been found to primarily target the function of murine macrophages but not of T cells, inhibiting secretion of tumor necrosis factor (TNF)-alpha in response to different Toll-like receptor agonists in vitro and in vivo. The well-defined role...... inflammatory colitis induced in mice by intracolonic challenge with dinitrobenzene sulfonic acid. The data from the two sets of experiments revealed that VGX-1027 inhibited both proliferation of enterobacterial antigen-reactive CD4+CD25- T cells in vitro and the development of clinical and histological signs...... of colitis in vivo. The beneficial effect in this model was associated with reduced colonic production of proinflammatory cytokines such as interleukin (IL)-1beta, TNF-alpha, IL-12p70 and interferon (IFN)-gamma and lower content of nuclear factor (NF)-kappaB (p65). These findings seem to warrant...

  1. Elevated serum IL-35 and increased expression of IL-35-p35 or -EBI3 in CD4+CD25+ T cells in patients with active tuberculosis

    Science.gov (United States)

    Kong, Bin; Liu, Gan-Bin; Zhang, Jun-Ai; Fu, Xiao-Xia; Xiang, Wen-Yu; Gao, Yu-Chi; Lu, Yuan-Bin; Wu, Xian-Jing; Qiu, Feng; Wang, Wan-Dang; Yi, Lai-Long; Zhong, Ji-Xin; Chen, Zheng W; Xu, Jun-Fa

    2016-01-01

    Despite the recent appreciation of interleukin 35 (IL-35) function in inflammatory diseases, little is known for IL-35 response in patients with active tuberculosis (ATB). In the current study, we demonstrated that ATB patients exhibited increases in serum IL-35 and in mRNA expression of both subunits of IL-35 (p35 and EBI3) in white blood cells and peripheral blood mononuclear cells. Consistently, anti-TB drug treatment led to reduction in serum IL-35 level and p35 or EBI3 expression. TB infection was associated with expression of p35 or EBI3 protein in CD4+ but not CD8+ T cells. Most p35+CD4+ T cells and EBI3+CD4+ T cells expressed Treg-associated marker CD25. Our findings may be important in understanding immune pathogenesis of TB. IL-35 in the blood may potentially serve as a biomarker for immune status and prognosis in TB. PMID:27158354

  2. Elevated serum IL-35 and increased expression of IL-35-p35 or -EBI3 in CD4(+)CD25(+) T cells in patients with active tuberculosis.

    Science.gov (United States)

    Kong, Bin; Liu, Gan-Bin; Zhang, Jun-Ai; Fu, Xiao-Xia; Xiang, Wen-Yu; Gao, Yu-Chi; Lu, Yuan-Bin; Wu, Xian-Jing; Qiu, Feng; Wang, Wan-Dang; Yi, Lai-Long; Zhong, Ji-Xin; Chen, Zheng W; Xu, Jun-Fa

    2016-01-01

    Despite the recent appreciation of interleukin 35 (IL-35) function in inflammatory diseases, little is known for IL-35 response in patients with active tuberculosis (ATB). In the current study, we demonstrated that ATB patients exhibited increases in serum IL-35 and in mRNA expression of both subunits of IL-35 (p35 and EBI3) in white blood cells and peripheral blood mononuclear cells. Consistently, anti-TB drug treatment led to reduction in serum IL-35 level and p35 or EBI3 expression. TB infection was associated with expression of p35 or EBI3 protein in CD4(+) but not CD8(+) T cells. Most p35(+)CD4(+) T cells and EBI3(+)CD4(+) T cells expressed Treg-associated marker CD25. Our findings may be important in understanding immune pathogenesis of TB. IL-35 in the blood may potentially serve as a biomarker for immune status and prognosis in TB.

  3. An increase in CD3+CD4+CD25+ regulatory T cells after administration of umbilical cord-derived mesenchymal stem cells during sepsis.

    Directory of Open Access Journals (Sweden)

    Yu-Hua Chao

    Full Text Available Sepsis remains an important cause of death worldwide, and vigorous immune responses during sepsis could be beneficial for bacterial clearance but at the price of collateral damage to self tissues. Mesenchymal stem cells (MSCs have been found to modulate the immune system and attenuate sepsis. In the present study, MSCs derived from bone marrow and umbilical cord were used and compared. With a cecal ligation and puncture (CLP model, the mechanisms of MSC-mediated immunoregulation during sepsis were studied by determining the changes of circulating inflammation-associated cytokine profiles and peripheral blood mononuclear cells 18 hours after CLP-induced sepsis. In vitro, bone marrow-derived MSCs (BMMSCs and umbilical cord-derived MSCs (UCMSCs showed a similar morphology and surface marker expression. UCMSCs had stronger potential for osteogenesis but lower for adipogenesis than BMMSCs. Compared with rats receiving PBS only after CLP, the percentage of circulating CD3+CD4+CD25+ regulatory T (Treg cells and the ratio of Treg cells/T cells were elevated significantly in rats receiving MSCs. Further experiment regarding Treg cell function demonstrated that the immunosuppressive capacity of Treg cells from rats with CLP-induced sepsis was decreased, but could be restored by administration of MSCs. Compared with rats receiving PBS only after CLP, serum levels of interleukin-6 and tumor necrosis factor-α were significantly lower in rats receiving MSCs after CLP. There were no differences between BMMSCs and UCMSCs. In summary, this work provides the first in vivo evidence that administering BMMSCs or UCMSCs to rats with CLP-induced sepsis could increase circulating CD3+CD4+CD25+ Treg cells and Treg cells/T cells ratio, enhance Treg cell suppressive function, and decrease serum levels of interleukin-6 and tumor necrosis factor-α, suggesting the immunomodulatory association of Treg cells and MSCs during sepsis.

  4. Assessment of the frequency of regulatory T cells (CD4+CD25+CD127-) in children with hemophilia A: relation to factor VIII inhibitors and disease severity.

    Science.gov (United States)

    El-Asrar, Mohamed Abo; Hamed, Ahmed El-Saeed; Darwish, Yasser Wagih; Ismail, Eman Abdel Rahman; Ismail, Noha Ali

    2016-01-01

    A rapidly growing evidence showed that regulatory T cells (Tregs) play a crucial role in tolerance to coagulation factors and may be involved in the pathogenesis of inhibitor formation in patients with hemophilia. We determined the percentage of Tregs (CD4CD25CD127) in 45 children with hemophilia A compared with 45 healthy controls, and assessed their relation to the clinical characteristics of patients and factor VIII (FVIII) inhibitors. Patients were studied stressing on frequency of bleeding attacks, joint pain, history of viral hepatitis, and the received therapy (FVIII precipitate/cryotherapy). FVIII activity and FVIII inhibitors were assessed with flow cytometric analysis of CD4CD25CD127 Tregs. According to residual FVIII activity levels, 30 patients (66.7%) had mild/moderate hemophilia A, whereas 15 (33.3%) patients had severe hemophilia A. The frequency of Tregs was significantly lower among all patients with hemophilia A compared with controls (2.59 ± 1.1 versus 3.73 ± 1.12%; P = 0.002). Tregs were significantly decreased among patients with FVIII inhibitors compared with the inhibitor-negative group (P hemophilia A had lower Tregs levels than those without (P = 0.34 and P = 0.011, respectively). A significant positive correlation was found between the percentage of Tregs and FVIII among hemophilia A patients. ROC curve analysis revealed that the cut-off value of Tregs at 1.91% could differentiate patients with and without FVIII inhibitors, with a sensitivity of 100% and a specificity of 91.3%. We suggest that alteration in the frequency of Tregs in young patients with hemophilia A may contribute to inhibitor formation and disease severity.

  5. Changes of CD4+ CD25+ Regulatory T Cells, FoxP3 in Adjuvant Arthritis Rats with Damage of Pulmonary Function and Effects of Tripterygium Glycosides Tablet

    Directory of Open Access Journals (Sweden)

    Wan Lei

    2012-01-01

    Full Text Available Objective. To observe the effects of tripterygium glycosides tablet (TPT on swelling degree, arthritis index (AI, pulmonary function, cytokines, the expression of regulatory T cells (Treg, and Foxp3 in rats of adjuvant arthritis. Methods. Rats were averagely divided into normal control (NC group, model control (MC group, methotrexate (MTX group, and tripterygium glycosides tablet (TPT group. Except for the rats of normal group, the others were intracutaneously injected with 0.1 mL of Freund’s complete adjuvant in the right hindlimb. NC group and MC group were treated with physiological saline. MTX group and TPT group were treated with MTX, TPT, respectively. Results. The levels of swelling degree, AI, the alveolar inflammation integral, TNF alpha (TNF-, and endothelium-1 (ET-1 in MC group were significantly increased (, and the levels of forced vital capacity (FVC, 25% vital capacity of the peak expiratory flow (FEF25, 50% vital capacity of the peak expiratory flow (FEF50, 75% vital capacity of the peak expiratory flow (FEF75, maximum midexpiratory flow (MMF, peak expiratory flow (PEF, interleukin-10 (IL-10, CD4+ CD25+ Treg, and Foxp3 were decreased (. The scores of alveolitis and ET-1 were decreased with treatment of TPT. The levels of FVC, FEF25, FEF50, FEF75, MMF, PEF, IL-10, and CD4+ CD25+ Treg in peripheral blood were increased. The expressions of Foxp3 protein and mRNA in lung tissue were also increased in TPT group. Conclusions. The paw swelling can be inhibited by TPT, and the inflammatory response in lung tissue was also decreased, which is a significant improvement in pulmonary function. The mechanism is probably associated with upregulating the expression of IL-10, Foxp3, and downregulating the level of TNF-.

  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. 脉冲高答量血液滤过对脓毒症患者CD4+CD25+调节性T细胞的影响%Effect of pulse high volume hemofiltration on CD4+CD25+ Treg cells in patients with sepsis

    Institute of Scientific and Technical Information of China (English)

    邵敏; 刘宝; 王锦权; 陶晓根; 周树生; 金魁; 张翠萍

    2011-01-01

    目的:探讨脓毒症患者CD4+CD25+调节性T细胞(Treg)的水平和意义以及脉冲高容量血液滤过(PHVHF)对它们的影响.方法:将2008年1月~2010年11月在安徽省立医院ICU住院的脓毒症患者40例按照疾病严重程度分为3组:脓毒症组14例,严重脓毒症组15例,脓毒症休克组11例.其中行PHVHF患者25例.40例脓毒症患者在入选当天和d5行流式细胞术检测血中CD4+CD25+Treg细胞的比例和CD3+、CD4+、CD8+和CD4+/CD8+水平.同时选取20例健康人为对照组.结果:健康对照组CD4+CD25+Treg细胞表达率为(0.39±0.23%);脓毒症组为(1.72±0.59%);严重脓毒症组(2.72±0.22%);脓毒性休克组(3.55±0.51%),后3组间两两比较有显著性差异(均P<0.05).脓毒症组CD3+、CD4+和CD4+/CD8+较正常对照组显著升高(P<0.05).严重脓毒症组和脓毒症休克患者CD3+、CD4+、CD4+/CD8+较正常对照组显著降低(均P<0.05).与死亡组比较,生存组PHVHF能更有效地降低CD4+CD25+Treg细胞的异常表达(P<0.05),上调CD3+、CD4+、CD8+和CD4+/CD8+水平.结论:CD4+CD25+Treg细胞在脓毒症的免疫发病机制中可能起着重要作用,随病情加重,免疫抑制加强.PHVHF可调整脓毒症患者T细胞亚群比例,改善其免疫抑制状态,可作为脓毒症免疫调节治疗的手段之一.%Objective:To study the level and significance of CD4+CD25+ Treg cells and CD3+,CD4+,CD8+ cells in peripheral blood of patients with sepsis and evaluate the effects of pulse high volume hemofiltration (PHVHF)on them. Methods:Forty patients with sepsis in ICU of Anhui Provicial Hospital were studied from January 2008 to November 2010.Patients were divided into three groups:sepsis group (n=14),severe sepsis group (n=15)and septic shock group (n=11).The expressions of CD4+CD25+ Treg cells and CD3+,CD4+,CD8+cells of 40 patients on the day 1 and day 5 were detected by flow cytometry and were compared.Twenty five patients receiving PHVHF were divided into two groups:death group (n=7

  8. Study on the relationship between regulatory T cell CD 4+ CD25+ and CD8+ CD28- in the peripheral blood between graves disease%外周血中 CD4+CD25+与 CD8+CD28-调节性 T 细胞与Graves 病的关系研究

    Institute of Scientific and Technical Information of China (English)

    韦巍; 林英辉; 黄小琪

    2015-01-01

    目的:分析Graves病(GD)患者外周血中CD4+ CD25+和CD8+ CD28-调节性 T (Tr)细胞的变化。方法选择该院内分泌科门诊或住院部43例GD患者为研究对象,其中内分泌科门诊确诊的19例GD患者为试验初发组。接受治疗且甲状腺功能恢复正常的24例GD患者为治疗缓解组;选择同期健康体检者20例为健康对照组。采用流式细胞术检测3组外周血中CD4+ CD25+和CD8+ CD28- T r细胞水平。结果试验初发组外周血CD4+CD25+ T r细胞水平为(4.56±4.14)%,明显低于健康对照组的(8.84±4.45)%,差异有统计学意义( P<0.05),CD8+CD28- T r细胞水平为(14.95±5.38)%,与健康对照组的(10.65±6.37)%比较,差异无统计学意义(P>0.05)。治疗缓解组外周血中CD4+CD25+ Tr细胞水平为(6.99±6.35)%,与健康对照组比较,差异无统计学意义( P>0.05),但其CD8+ CD28- T r细胞水平为(20.48±6.07)%,高于健康对照组,差异有统计学意义( P<0.05)。结论 CD4+CD25+ T r细胞水平可能与GD发病有关,CD8+CD28-T r细胞可能与GD病程发展有关。%Objective To analyze the changes of the levels of CD4+CD25+ and CD8+CD28- T cells in the pe‐ripheral blood in patients with graves disease(GD) .Methods A total of 43 patients with GD were selected as objects in this study ,19 cases who were diagnosis in outpatient department were selected into incipience group ,24 cases who were treated and the thyroid function return to normal status were selected into alleviation group ,meanwhile 20 healthy persons were recruited into control group .The levels of CD4+ CD25+ and CD8+ CD28- regulator T cells in the peripheral blood were detected by flow cytometry .Results The CD4+CD25+ regulatory T cell levels in incipience group were (4 .56 ± 4 .14)% ,which was significant lower than(8 .84 ± 4 .45)% in the

  9. TCR usage, gene expression and function of two distinct FOXP3(+)Treg subsets within CD4(+)CD25(hi) T cells identified by expression of CD39 and CD45RO.

    Science.gov (United States)

    Ye, Lingying; Goodall, Jane C; Zhang, Libin; Putintseva, Ekaterina V; Lam, Brian; Jiang, Lei; Liu, Wei; Yin, Jian; Lin, Li; Li, Ting; Wu, Xin; Yeo, Giles; Shugay, Mikhail; Chudakov, Dmitriy M; Gaston, Hill; Xu, Huji

    2016-03-01

    FOXP3+ regulatory T (Treg) cells are indispensable for immune homeostasis, but their study in humans is complicated by heterogeneity within Treg, the difficulty in purifying Tregs using surface marker expression (e.g. CD25) and the transient expression of FOXP3 by activated effector cells. Here, we report that expression of CD39 and CD45RO distinguishes three sub-populations within human CD4(+)CD25(hi) T cells. Initial phenotypic and functional analysis demonstrated that CD4(+)CD25(hi)CD39(+)CD45RO(+) cells had properties consistent with effector Treg, CD4(+)CD25(hi)CD39(-)CD45RO(-) cells were naïve Treg and CD4(+)CD25(hi)CD39(-)CD45RO(+) cells were predominantly non-Treg with effector T-cell function. Differences in these two newly identified Treg subsets were corroborated by studies of gene expression and TCR analysis. To apply this approach, we studied these two newly identified Treg subsets in ankylosing spondylitis, and showed impairment in both effector and naïve Treg. This work highlights the importance of discriminating Treg subsets to enable proper comparisons of immune regulatory capacity in healthy individuals and those with inflammatory disease.

  10. 松果体褪黑素对大鼠胸腺CD4~+CD25~+调节性T细胞发育及其相关基因Foxp3 的影响%Effect of pineal gland and melatonin on CD4~+CD25~+Treg cells development and Foxp3 expression of rat thymus

    Institute of Scientific and Technical Information of China (English)

    林晋; 郑琳; 黄云梅; 刘卉; 魏建恩; 周瑞祥

    2010-01-01

    目的 研究松果体摘除及补充褪黑素对大鼠胸腺CD4~+CD25~+ 调节性T 细胞(Treg 细胞)产生、输出及其相关基因叉状头/翅膀状螺旋转录子(Foxp3)表达的影响.方法 选用雄性清洁级SD 大鼠120只,分为正常组、假手术组、松果体摘除组、松果体摘除+褪黑素低剂量组[腹腔注射7.5mg/(kg.d)]和松果体摘除+褪黑素高剂量组[腹腔注射15mg/(kg.d)],术后4、8周取材.应用流式细胞检测技术、RT-PCR法及免疫组织化学染色法观察并分析松果体摘除及补充外源性褪黑素后胸腺及外周血CD4、CD25双阳性细胞数量及胸腺Foxp3表达的变化.结果 松果体摘除术后无论4周或者8周胸腺CD4~+CD25~+Treg 细胞数量均无明显变化,补充褪黑素后双阳性细胞数量呈时间、剂量依赖性明显增加;松果体摘除术后4周外周血CD4~+CD25~+Treg 细胞数量明显增加,但补充褪黑素后恢复正常,而术后8周各组之间无显著性差异;半定量RT-PCR 及免疫组织化学结果显示,松果体摘除后4周大鼠胸腺Foxp3 表达水平明显升高,补充高低两种剂量褪黑素后均有所下降并达到正常水平,而松果体摘除后8周各组之间Foxp3的表达无明显差异.结论 松果体摘除对胸腺CD4~+CD25~+Treg 细胞的产生无明显影响,但导致大鼠胸腺Foxp3 的转录和翻译明显增加,胸腺CD4~+CD25~+ Treg 细胞的输出增加,而补充外源性褪黑素后能逆转上述异常,长时间作用则其影响逐渐消失.

  11. THE EXPRESSION OF MEMBRANE INTERLEUKIN-2 RECEPTOR(CD25) ON THE SURFACE OF PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC) OF PULMONARY TUBERCULOSIS AND ITS CLINICAL MANIFESTATION%肺结核病患者PBMC膜白介素-2受体(CD25)表达及其临床意义

    Institute of Scientific and Technical Information of China (English)

    王健; 蔡茹; 刘智; 刘炳祥

    2003-01-01

    目的:探讨外周血单个核细胞(PBMC)膜白介素-2受体mIL-2R,(CD25)表达在肺结核病鉴别诊断中的应用价值.方法:用生物素-链霉亲和素(BSA)法检测肺结核、支气管肺炎患者T细胞亚群及植物血凝素(PHA)诱导前后CD25表达水平.结果:支气管肺炎患者CD+3、CD+4、CD+8水平分别为(62.32±6.34)%、(47.52±7.16)%、(32.12±6.55)%,CD+4/CD+8比值为1.52±0.43,PHA诱导前后CD25水平分别为(4.56±1.52)%、(35.12±7.21)%.空洞型肺结核CD+3、CD+4、CD+8、CD+4/CD+8水平分别为(41.13±5.25)%、(43.38±5.15)%、(36.25±3.46)%和1.15±0.21,非空洞型肺结核CD+3、CD+4、CD+8、CD+4/CD+8水平分别为(46.29±5.60)%、(47.21±4.86)%、(32.36±4.03)%、1.46±0.25,相互比较CD+3、CD+3/CD+8差异均有显著性(P<0.01和P<0.05).空洞型肺结核与非空洞型肺结核患者PHA诱导前后CD25水平分别为(2.13±1.14)%、(27.25±3.50)%和(3.43±1.35)%、(31.14±4.11)%,两者相比差异均有显著性(P<0.01).结论:肺结核病患者体内存在明显的细胞免疫功能紊乱 ,主要表现为CD25表达水平降低,CD25表达水平与肺结核病的病情似有一定关系,其对肺结核病鉴别诊断具有重要价值.

  12. CD25+Foxp3+调节性T细胞和Th17细胞在口腔白斑组织中的表达%Expression of CD25+ Foxp3+ Regulatory T Cells and Th17 Cells in Oral Leukoplakia.

    Institute of Scientific and Technical Information of China (English)

    白杨; 王文梅; 王翔; 黄晓峰; 蒋文晖; 刘雅菁

    2011-01-01

    Objective: To investigate the role of Treg and Th17 during the development and progression of the premalignant lesions. Methods: We compared CD25/Foxp3 and IL—17 expression detected by immunohistochemical staining in 42 oral leukoplakia (with or without dysplasia), and compared with 10 normal oral mucosa and 13 oral squamous cell carcinoma. Results: There was increasing trend of expression of CD25+ Foxp3+ Treg cells between the cases of normal oral mucosa , oral leukoplakia with hyperplasia epithelium, oral leukoplakia with dysplasia epithelium and OSCC (P<0. 05). (They were numerated as 2.5±0. 65, 5. 22±1.19, 31.4±16.8 and 42.8±12.67 per high magnification field, respectively). Further, expression of CD25+ Foxp3+ Treg cells was higher in severe dysplasia OLK than in mild/moderate dysplasia OLK. In addition, there was increasing trend of IL— 17 expression which was found between the four groups of normal oral mucosa, oral leukoplakia with hyperplasia epithelium, oral leukoplakia with dysplasia epithelium and OSCC, but no statistical significances. Positive correlation was shown between the density of the subset of co—expression of CD25+ and Foxp3 + T cells and expression of IL—17 (r = 0.318, P<0.05). Conclusion: In assistance with Th17 Cells, Treg cells has played a certain role in the development of oral cancer.%目的:通过观察CD25+Foxp3+和IL-17阳性细胞在口腔黏膜白斑(oral leukoplakia,OLK)和口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)组织中的表达情况,探讨Treg细胞和Th17细胞在口腔癌发生发展过程中的变化.方法:采用免疫组织化学方法检测CD25+Foxp3+、IL-17阳性细胞在10例单纯增生性白斑、32例异常增生性白斑、13例OSCC和10例正常口腔黏膜中的变化情况.结果:正常口腔黏膜组、单纯增生性白斑组、异常增生性白斑组和OSCC组CD25+Foxp3+Treg细胞数量逐渐增高(P<0.05),依次为(2.5±0.65)、(5.22±1.19)、(31.4±16.8)和(42.8±12.67)

  13. Screening of Fungi from Chinese Medical Plants for Anti-Human Immunodeficiency Virus Type 1 Activity

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    In order to isolate anti-human immunodeficiency virus (HIV) agents from natural products, 97 ethanolic extracts of 90 fungi were tested for their inhibitory activity on HIV-1. Most of the extracts tested were relatively non-toxic to human lymphocytic MT-4 cells, but extracts of some fungi exhibited potent anti-HIV activity in an in vitro 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay with a selectivity index greater than 3. Most fungi were isolated from Dendrobium sp. and Taxus sp.

  14. Enzyme-linked immunosorbent assay for detection of streptolysin O antibodies.

    OpenAIRE

    Reitano, M; Pisano, M. A.; Eriquez, L A; D'Amato, R F

    1986-01-01

    An enzyme-linked immumosorbent assay (ELISA), based upon the detection of streptolysin O antibodies in human sera, was developed. Disposable polystyrene tubes, sensitized with streptolysin O antigen, were used as the test vehicles. Corresponding antibodies, present in test sera, were detected by binding of the antibodies to goat anti-human immunoglobulin G conjugated to horseradish peroxidase. Demonstration of bound conjugate was accomplished by monitoring peroxidase activity spectrophotometr...

  15. 不同分期前列腺癌患者外周血CD4+CD25+Foxp3+调节性T细胞的变化及与胰岛素抵抗的相关性%Changes of CD4+ CD25+ Foxp3+ regulatory T cells in the peripheral blood and their correlation with insulin resistance in different stages of prostate cancer

    Institute of Scientific and Technical Information of China (English)

    章步文; 黎钢; 叶津津; 李峥嵘

    2015-01-01

    目的:观察不同分期前列腺癌患者外周血单个核细胞CD4+ CD25+ Foxp3+调节性T细胞的变化及与胰岛素抵抗的关系. 方法:采用流式细胞术检测62例前列腺癌患者(患者组,临床TNM分期Ⅰ期5例、Ⅱ期16例、Ⅲ期21例、Ⅳ期20例)外周血单个核细胞(PBMC)中CD4+ CD25+ Foxp3+调节性T细胞数目,计算CD4+CD25+ Foxp3+调节性T细胞占CD4+T淋巴细胞的百分率;并检测其空腹胰岛素及空腹血糖水平,计算胰岛素抵抗指数(HOMA-IR);采用ELISA法测定外周血胰岛素样生长因子1(IGF-1)水平,分析CD4+ CD25+ Foxp3+调节性T细胞与胰岛素抵抗的相关性,并与42例健康体检者进行对照. 结果:与健康对照组相比,前列腺癌患者HOMA-IR明显升高(6.68±1.66vs3.68±1.42),IGF-1水平明显下降[(96.39 ±21.21) ng/ml vs(164.56 ±30.58) ng/ml],PBMC CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的百分率[(13.88±0.96)%vs(5.33 ±0.65)%]及CD4+CD25+ Foxp3+ Treg绝对值[(3.55±0.29)×107vs(1.99 ±0.78)× 107]明显升高(P<0.05,P<0.01).患者PBMCCD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的百分率及CD4+ CD25+ Foxp3+ Treg绝对数、HOMA-IR均随TNM分期逐渐加重而增加,IGF-1逐渐下降;相关性分析表明:CD4+ CD25+ Foxp3+ Treg/CD4+T及CD4+ CD25+ Foxp3+ Treg绝对数均与HOMA-IR呈明显正相关(r分别为0.689、0.722,P<0.01),与IGF-1呈明显负相关(r分别为-0.896、-0.747,P<0.01). 结论:前列腺癌患者存在不同程度的胰岛素抵抗,且随着疾病程度的加重,外周血CD4+CD25+ Foxp3+调节性T细胞数目和比例及胰岛素抵抗逐渐加重;CD4+ CD25+ Foxp3+调节性T细胞可能通过调节胰岛素抵抗参与其形成和发展.

  16. The influence of allogenetic mesenchymal stem cells transplantation to collagen induced arthritis CD 4 + CD25 + regulatory T cell expression in mice%异基因骨髓间充质干细胞移植对小鼠胶原性关节炎CD4+CD25+调节性T细胞表达的影响

    Institute of Scientific and Technical Information of China (English)

    孙旭; 齐月; 牛广华; 高玉洁; 王柏山; 王思微; 严峰

    2014-01-01

    Objective To study the influence of allogenetic mesenchymal stem cells (MSCs) transplantation on the expression of collagen induced arthritis CD4 + CD25 + regulatory T cell in mice .Methods A total of 40 C57BL/6 (H-2b) mice were selected and randomly divided into normal control group ,model group ,MSCs transplant treatment group and methotrexate treatment of positive control group .Ten mice in each group .Except for mice in the normal control group ,the others were treated with Freund's complete adjuvant + Collagen Ⅱ to induce C57BL/6(H-2b) mice that to make mouse model .Bone marrow mononuclear cells were isolated ,then the MSCs were identified and screened by FAcscaliburTM flow cytometry analyzer .MSCs was transplanted by tail vein injection with the number of cells as 2 × 106 .All the mice in four groups were put to death after transplanting 42 days ,and observed the symptoms and the degree of swelling of the joints .Using flow cytometry to detect the contents of CD4 + CD25 + ,and collected data and made statistical analysis .Results There was no significant difference on the swelling degree of joints between the MSCs transplant treatment group and the normal control group (P> 0 .05) .Compared with methotrexate treatment of positive control group ,there was significant difference on CD4 + CD25 + regulatory T cell expression in the MSCs transplant treatment group (P 0 .05) .Conclusion MSCs transplantion could significantly improve the swelling degree of joints ,and significantly increase the expression of CD4 + CD25 + .Therefore ,MSCs could be used as a new source of replacement cells for transplantation therapy of rheumatoid arthritis .%目的:探讨异基因骨髓间充质干细胞(MSCs)移植对小鼠胶原性关节炎 CD4+、CD25+调节性 T 细胞表达的影响。方法选取40只 C57BL/6(H-2b)小鼠,随机分为正常对照组、Ⅱ型胶原性关节炎(CIA)模型组、MSCs 移植治疗组、甲氨蝶呤

  17. 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

  18. 慢性非细菌性前列腺炎/慢性骨盆疼痛综合征患者外周血CD4+CD25+T调节细胞功能检测及意义%TGF-β1The role of CD4+CD25+ regulatory T cells in the pathogenesis of chronic abacterial prostatitis/ chronic pelvic pain syndrome

    Institute of Scientific and Technical Information of China (English)

    王少刚; 白剑; 席启林; 胡东亮; 刘继红; 叶章群

    2008-01-01

    Objective To explore the role of CD4+CD25+ regulatory T cells in the pathogenesis of chronic abacterial prostatitis/chronic pelvic pain syndrome (CAP/CPPS). Methods Peripheral blood samples were collected from 45 CAP/CPPS patients and 18 healthy age-matched male persons. Peripheral blood mononuclear cells (PBMCs) were isolated. The percentages of CD4+CD25+ and CD4+ CD25high regulatory T cells were detected by flow cytometry. PCR was used to examine the mRNA expression of Foxp3, a transcription factor expressed in the CD4+ CD25+ cells. ELISA was used to examine the plasma level of tumor growth factor (TGF)-β1. Results There were no significant differences in the percentages of peripheral blood CD4+ CD25+ and CD4+ CD25highT cells between the CAP/CPPS patients and normal control group(both P>0.05). The Foxp3 mRNA in the PBMCs of the CAP/CPPS ⅢA and CAP/CPPS ⅢB patients were (0.69±0. 23) and (0.44±0.18) respectively, both significantly lower than that of the control group [(1.37±0.19), P 0.05);CAP/CPPSⅢA、ⅢB组患者外周血单个核细胞(PBMC)中Foxp3mRNA表达水平[(0.69±0.23)、(0.44±0.18)]较对照组(1.37±0.19)明显降低(P=0.035,P=0.014);CAP/CPPSⅢA、ⅢB组患者血清中TGF-β1水平[(18.09±10.45)pg/ml、(14.06±6.22)pg/ml]较对照组[(27.01±13.29)pg/ml]降低(P=0.041,P=0.024).结论 外周血CD44CD25+调节性T细胞数量与CAP/CPPS的发病无明显关联,CD4+CD25+调节性T细胞功能障碍可能参与CAP/CPPS发病机制;Foxp3基因与TGF-β1在CAP/CPPS的发病过程中起重要作用.

  19. T regulatory cells (TREG)(TCD4+CD25+FOXP3+) distribution in the different clinical forms of leprosy and reactional states*

    Science.gov (United States)

    Parente, José Napoleão Tavares; Talhari, Carolina; Schettini, Antônio Pedro Mendes; Massone, Cesare

    2015-01-01

    BACKGROUND Leprosy is characterized histologically by a spectrum of different granulomatous skin lesions, reflecting patients' immune responses to Mycobacterium leprae. Although CD4+CD25+ FoxP3+ T regulatory cells are pivotal in the immuneregulation, presence, frequency, and distribution of Tregs in leprosy, its reactional states have been investigated in few studies. OBJECTIVES This study aimed to verify the frequency and distribution of regulatory T cells in different clinical forms and reactional states of leprosy. METHODS We performed an immunohistochemical study on 96 leprosy cases [Indeterminate (I): 9 patients; tuberculoid tuberculoid: 13 patients; borderline tuberculoid: 26 patients; borderline borderline: 3 patients; borderline lepromatous: 8 patients; lepromatous lepromatous: 27 patients; reversal reaction: 8 patients; and erythema nodosum leprosum: 2 patients]. RESULTS FoxP3-positive cells were present in 100% of the cases with an average density of 2.82% of the infiltrate. Their distribution was not related to granulomatous structures or special locations. There was a statistically significant increment of FoxP3 expression in patients with leprosy reversal reactions when compared with patients presenting with type I leprosy (P= 0.0228); borderline tuberculoid leprosy (P = 0.0351) and lepromatous leprosy (P = 0.0344). CONCLUSIONS These findings suggest that Tregs play a relevant role in the etiopathogenesis of leprosy, mainly in type I leprosy reaction. PMID:25672298

  20. Galectin-9 ameliorates Con A-induced hepatitis by inducing CD4(+CD25(low/int effector T-Cell apoptosis and increasing regulatory T cell number.

    Directory of Open Access Journals (Sweden)

    Kun Lv

    Full Text Available BACKGROUND: T cell-mediated liver damage is a key event in the pathogenesis of many chronic human liver diseases, such as liver transplant rejection, primary biliary cirrhosis, and sclerosing cholangitis. We and other groups have previously reported that galectin-9, one of the β-galactoside binding animal lectins, might be potentially useful in the treatment of T cell-mediated diseases. To evaluate the direct effect of galectin-9 on hepatitis induced by concanavalin A (Con A administration in mice and to clarify the mechanisms involved, we administered galectin-9 into mice, and evaluated its therapeutic effect on Con A-induced hepatitis. METHODOLOGY/PRINCIPAL FINDINGS: Galectin-9 was administrated i.v. to Balb/c mice 30 min before Con A injection. Compared with no treatment, galectin-9 pretreatment significantly reduced serum ALT and AST levels and improved liver histopathology, suggesting an ameliorated hepatitis. This therapeutic effect was not only attributable to a blunted Th1 immune response, but also to an increased number in regulatory T cells, as reflected in a significantly increased apoptosis of CD4(+CD25(low/int effector T cells and in reduced proinflammatory cytokine levels. CONCLUSION/SIGNIFICANCE: Our findings constitute the first preclinical data indicating that interfering with TIM-3/galectin-9 signaling in vivo could ameliorate Con A-induced hepatitis. This strategy may represent a new therapeutic approach in treating human diseases involving T cell activation.

  1. 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.

  2. Partial depletion of CD4(+)CD25(+)Foxp3(+) T regulatory cells significantly increases morbidity during acute phase Toxoplasma gondii infection in resistant BALB/c mice.

    Science.gov (United States)

    Morampudi, Vijay; De Craeye, Stephane; Le Moine, Alain; Detienne, Sophie; Braun, M Y; D'Souza, Sushila

    2011-04-01

    CD4(+)CD25(+)Foxp3(+) T regulatory (Treg) cells, are known to regulate responses to infectious agents. Here we compared disease progression in BALB/c and C57BL/6(B6) mice infected perorally with Toxoplasma gondii for 7 days and examined the affect of partial depletion of Treg cells in these mice. BALB/c mice were seen to be resistant to peroral infection whereas B6 mice were susceptible in terms of mortality. Although the depletion of Treg cells before infection had no effect on the survival of B6 or BALB/c mice, it resulted in increased parasite burdens in BALB/c mice, especially in the lamina propria, but not in B6 mice. Pro-inflammatory cytokines were also increased in Treg cells depleted BALB/c mice as compared to B6 mice. In addition Treg cell depleted BALB/c mice displayed increased ileal histopathology compared to their non-treated counterparts. These findings provide evidence for the contribution of Treg cells, in the resistance of BALB/c mice against peroral T. gondii infection.

  3. 4-1BB Signaling in Conventional T Cells Drives IL-2 Production That Overcomes CD4+CD25+FoxP3+ T Regulatory Cell Suppression.

    Directory of Open Access Journals (Sweden)

    Hampartsoum B Barsoumian

    Full Text Available Costimulation with the recombinant SA-4-1BBL agonist of 4-1BB receptor on conventional CD4+ T cells (Tconvs overcomes the suppression mediated by naturally occurring CD4+CD25+FoxP3+ T regulatory cells (Tregs. The mechanistic basis of this observation has remained largely unknown. Herein we show that Tconvs, but not Tregs, are the direct target of SA-4-1BBL-mediated evasion of Treg suppression. IL-2 produced by Tconvs in response to 4-1BB signaling is both necessary and sufficient for overcoming Treg suppression. Supernatant from Tconvs stimulated with SA-4-1BBL contains high levels of IL-2 and overcomes Treg suppression in ex vivo Tconv:Treg cocultures. Removal of IL-2 from such supernatant restores Treg suppression and repletion of Tconv:Treg cocultures with exogenous recombinant IL-2 overcomes suppression. This study establishes 4-1BB signaling as a key circuit that regulates physical and functional equilibrium between Tregs and Tconvs with important implications for immunotherapy for indications where a fine balance between Tregs and Teffs plays a decisive role.

  4. Galectin-9 Ameliorates Con A-Induced Hepatitis by Inducing CD4+CD25low/int Effector T-Cell Apoptosis and Increasing Regulatory T Cell Number

    Science.gov (United States)

    Zhang, Mengying; Zhong, Min; Suo, Qifeng

    2012-01-01

    Background T cell-mediated liver damage is a key event in the pathogenesis of many chronic human liver diseases, such as liver transplant rejection, primary biliary cirrhosis, and sclerosing cholangitis. We and other groups have previously reported that galectin-9, one of the β-galactoside binding animal lectins, might be potentially useful in the treatment of T cell-mediated diseases. To evaluate the direct effect of galectin-9 on hepatitis induced by concanavalin A (Con A) administration in mice and to clarify the mechanisms involved, we administered galectin-9 into mice, and evaluated its therapeutic effect on Con A-induced hepatitis. Methodology/Principal Findings Galectin-9 was administrated i.v. to Balb/c mice 30 min before Con A injection. Compared with no treatment, galectin-9 pretreatment significantly reduced serum ALT and AST levels and improved liver histopathology, suggesting an ameliorated hepatitis. This therapeutic effect was not only attributable to a blunted Th1 immune response, but also to an increased number in regulatory T cells, as reflected in a significantly increased apoptosis of CD4+CD25low/int effector T cells and in reduced proinflammatory cytokine levels. Conclusion/Significance Our findings constitute the first preclinical data indicating that interfering with TIM-3/galectin-9 signaling in vivo could ameliorate Con A-induced hepatitis. This strategy may represent a new therapeutic approach in treating human diseases involving T cell activation. PMID:23118999

  5. 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.

  6. Cutting edge: trans-signaling via the soluble IL-6R abrogates the induction of FoxP3 in naive CD4+CD25 T cells.

    Science.gov (United States)

    Dominitzki, Sabine; Fantini, Massimo C; Neufert, Clemens; Nikolaev, Alexei; Galle, Peter R; Scheller, Jürgen; Monteleone, Giovanni; Rose-John, Stefan; Neurath, Markus F; Becker, Christoph

    2007-08-15

    Chronic inflammatory diseases may develop when regulatory T cells (Tregs) fail to control the balance between tolerance and immunity. Alternatively, activated immune cells might prevent the induction or activation of Tregs in such diseases. In this study, we demonstrate that trans-signaling into T cells via the soluble IL-6 receptor completely abrogates the de novo induction of adaptive Tregs. Mechanistically, IL-6 trans-signaling augmented the expression of the TGF-beta signaling inhibitor SMAD7. Consequently, SMAD7 overexpression in T cells using newly created transgenic mice rendered CD4(+)CD25(-) T cells resistant to the induction of FoxP3. Finally, IL-6 trans-signaling inhibited Treg-mediated suppression in a murine model of colitis. In summary, IL-6 trans-signaling into T cells emerges as a key pathway for blockade of the development of adaptive Tregs and thus may play a pivotal role in shifting the balance between effector and regulatory T cell numbers in chronic inflammatory and autoimmune diseases.

  7. 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.

  8. Increase in circulating CD4⁺CD25⁺Foxp3⁺ T cells in patients with Philadelphia-negative chronic myeloproliferative neoplasms during treatment with IFN-α

    DEFF Research Database (Denmark)

    Riley, Caroline Hasselbalch; Jensen, Morten Krogh; Brimnes, Marie Klinge;

    2011-01-01

    blood have been reported in both hematologic and solid cancers. We have analyzed the number, phenotype, and function of circulating CD4(+)CD25(+)Foxp3(+) T cells in patients with chronic myeloproliferative neoplasms. Surprisingly, we found a marked expansion of this subset of lymphocytes in patients...

  9. 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

  10. 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

  11. 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.

  12. A Model System for Concurrent Detection of Antigen and Antibody Based on Immunological Fluorescent Method

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    Yuan-Cheng Cao

    2015-01-01

    Full Text Available This paper describes a combined antigen/antibody immunoassay implemented in a 96-well plate using fluorescent spectroscopic method. First, goat anti-human IgG was used to capture human IgG (model antigen; goat anti-human IgG (Cy3 or FITC was used to detect the model antigen; a saturating level of model antigen was then added followed by unlabelled goat anti-human IgG (model antibody; finally, Cy3 labelled rabbit anti-goat IgG was used to detect the model antibody. Two approaches were applied to the concomitant assay to analyze the feasibility. The first approach applied FITC and Cy3 when both targets were present at the same time, resulting in 50 ng/mL of the antibody detection limit and 10 ng/mL of antigen detection limit in the quantitative measurements of target concentration, taking the consideration of FRET efficiency of 68% between donor and acceptor. The sequential approach tended to lower the signal/noise (S/N ratio and the detection of the model antigen (lower than 1 ng/mL had better sensitivity than the model antibody (lower than 50 ng/mL. This combined antigen/antibody method might be useful for combined detection of antigens and antibodies. It will be helpful to screen for both antigen and antibody particularly in the situations of the multiserotype and high-frequency mutant virus infections.

  13. Detection of peripheral blood CD4+ CD25high CD127low regulatory T cells in patients with B-cell non-Hodgkin lymphoma and its clinical significance%B细胞非霍奇金淋巴瘤患者外周血CD4+CD25high CD127low调节性T细胞水平及其临床意义

    Institute of Scientific and Technical Information of China (English)

    白敏; 王列样; 赵志强; 苏文; 苏丽萍; 郑玉萍

    2014-01-01

    目的 探讨外周血CD4+ CD25high CD127low调节性T细胞(Treg细胞)在B细胞非霍奇金淋巴瘤(B-NHL)患者外周血中表达水平及其临床意义.方法 采用流式细胞术检测100例初诊B-NHL患者及50名健康对照者外周血CD4+ CD25high CD127low Treg细胞表达水平,进行统计学分析.结果 健康对照者外周血CD4+ CD25high CD127low Treg细胞中位表达水平为5.00%,初诊B-NHL患者为7.20%,两者之间差异具有统计学意义(P< 0.001).男性患者外周血CD4+ CD25high CD 127low Treg细胞水平高于女性患者(P<0.01),乳酸脱氢酶(LDH)增高患者外周血CD4+ CD25high CD127low Treg细胞水平高于LDH正常患者(P<0.01),Ⅲ~Ⅳ期患者外周血CD4+C D25high CD 127low Treg细胞水平较Ⅰ~Ⅱ期患者增高(P<0.01),有B症状患者外周血CD4+ CD25high CD127low Treg细胞水平高于无B症状患者(P<0.01).而不同年龄、国际预后指数评分、有无大包块患者之间外周血CD4+ CD25high CD 127low Treg细胞水平差异均无统计学意义(均P>0.05).结论 B-NHL患者体内存在免疫抑制,在男性、LDH增高、有B症状及晚期患者中CD4+ CD25high CD127low Treg细胞水平明显增高.检测其水平对于判断B-NHL的预后有一定价值.%Objective To detect peripheral blood level of CD4+ CD25high CD127low regulatory T (Treg) cells in patients with B cell non-Hodgkin lymphoma (B-NHL) and to explore its clinical significance.Methods The levels of peripheral blood CD4+ CD25high CD127low Treg cells of 100 newly diagnosed patients with B-NHL and 50 healthy adults were detected and analyzed by flow cytometry.Results The levels of peripheral blood CD4+ CD25high CD12low Treg cells were 5.00 % and 7.20 % in healthy adults and newly diagnosed B-NHL patients,respectively.The difference was statistically significant (P < 0.001).The level of peripheral blood CD4+ CD25high CD127low Treg cells was significantly higher in the male patients than that in the female (P < 0

  14. QSAR modeling for anti-human African trypanosomiasis activity of substituted 2-Phenylimidazopyridines

    Science.gov (United States)

    Masand, Vijay H.; El-Sayed, Nahed N. E.; Mahajan, Devidas T.; Mercader, Andrew G.; Alafeefy, Ahmed M.; Shibi, I. G.

    2017-02-01

    In the present work, sixty substituted 2-Phenylimidazopyridines previously reported with potent anti-human African trypanosomiasis (HAT) activity were selected to build genetic algorithm (GA) based QSAR models to determine the structural features that have significant correlation with the activity. Multiple QSAR models were built using easily interpretable descriptors that are directly associated with the presence or the absence of a structural scaffold, or a specific atom. All the QSAR models have been thoroughly validated according to the OECD principles. All the QSAR models are statistically very robust (R2 = 0.80-0.87) with high external predictive ability (CCCex = 0.81-0.92). The QSAR analysis reveals that the HAT activity has good correlation with the presence of five membered rings in the molecule.

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

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    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.

  16. 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.

  17. Is there any relationship between PSA and increased peripheral CD4+CD25highFOX3+ Treg in prostate cancer patients?

    Directory of Open Access Journals (Sweden)

    Yigit Akin

    2011-09-01

    Full Text Available Introduction: The aims of this study were fi rst, to determine whether peripheral levels of CD4+CD25highFoxp3+ regulatory T cells (Treg are elevated in Prostate Cancer (PCa patients, and second, to determine the directcorrelation between peripheral Treg and total serum Prostate Specifi c Antigen (PSA levels in these patients.Methods: Peripheral Blood Mononuclear Cells from 56 subjects undergoing diagnostic prostate biopsies (PSA ≥ 2.5 ng/ml were analyzed for Treg numbers. The association between the peripheral Treg and serum PSA values was fi rst determined in the entire population, including people with no prostate pathology and PCa and Benign Prostate Hyperplasia (BPH patients, and second, in nine PCa patients before and after curative prostatectomy.Results: This project was performed in Akdeniz University immunology laboratory and urology out patient clinic from 2008 to 2010. Peripheral Treg frequencies were signifi cantly increased in PCa patients (n = 19, 3.23 ± 1.59 compared with BPH patients (n = 27, 1.66 ± 0.80 and healthy subjects (n = 10, 1.08 ± 0.43 (p < 0.01. The percentage of Treg in BPH patients was also signifi cantly higher than that of healthy subjects (p < 0.01. Importantly, the increase in BPH and PCa patients paralleled the elevation in total serum PSA levels, demonstrating a strong positive correlation (r = 0.75; p < 0.01.Conclusion: These results demonstrate that peripheral Treg densities are correlated with PSA in BPH and PCa patients, suggesting that PSA may have a role in Treg induction and/or maintenance in Treg in these people.

  18. 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.

  19. PD1 blockade reverses the suppression of melanoma antigen-specific CTL by CD4+ CD25(Hi) regulatory T cells.

    Science.gov (United States)

    Wang, Wenshi; Lau, Roy; Yu, Daohai; Zhu, Weiwei; Korman, Alan; Weber, Jeffrey

    2009-09-01

    Regulatory CD4(+)CD25(Hi) T cells (Treg) and programmed death-1 (PD-1) molecule have emerged as pivotal players in immune regulation. However, the underlying mechanisms by which they impact antigen-specific CD8(+) immune responses in cancer patients and how they interact with each other under physiologic conditions remain unclear. Herein, we examined the relationship of PD-1 and its abrogation to the function of Treg in patients with melanoma using short-term in vitro assays to generate melanoma-specific T cells. We identified Treg in the circulation of vaccinated melanoma patients and detected PD-1 expression on vaccine-induced melanoma antigen-specific CTLs, as well as on and within Treg from patients' peripheral blood. Programmed death ligand (PD-L) 1 expression was also detected on patients' Treg. PD-1 blockade promoted the generation of melanoma antigen-specific CTLs and masked their inhibition by Treg. The mechanisms by which PD-1 blockade mediated immune enhancement included direct augmentation of melanoma antigen-specific CTL proliferation, heightening their resistance to inhibition by Treg and direct limitation of the inhibitory ability of Treg. PD-1 blockade reversed the increased expression of PD-1 and PD-L1 on melanoma antigen-specific CTL by Treg, rescued INF-gamma and IL-2 or INF-gamma and tumor necrosis factor-alpha co-expression and expression of IL-7 receptor by melanoma antigen-specific CTL which were diminished by Treg. PD-1 blockade also resulted in down-regulation of intracellular FoxP3 expression by Treg. These data suggest that PD-1 is importantly implicated in the regulation of Treg function in melanoma patients.

  20. PD1 blockade reverses the suppression of melanoma antigen-specific CTL by CD4+CD25Hi regulatory T cells

    Science.gov (United States)

    Lau, Roy; Yu, Daohai; Zhu, Weiwei; Korman, Alan; Weber, Jeffrey

    2009-01-01

    Regulatory CD4+CD25Hi T cells (Treg) and programmed death-1 (PD-1) molecule have emerged as pivotal players in immune regulation. However, the underlying mechanisms by which they impact antigen-specific CD8+ immune responses in cancer patients and how they interact with each other under physiologic conditions remain unclear. Herein, we examined the relationship of PD-1 and its abrogation to the function of Treg in patients with melanoma using short-term in vitro assays to generate melanoma-specific T cells. We identified Treg in the circulation of vaccinated melanoma patients and detected PD-1 expression on vaccine-induced melanoma antigen-specific CTLs, as well as on and within Treg from patients’ peripheral blood. Programmed death ligand (PD-L) 1 expression was also detected on patients’ Treg. PD-1 blockade promoted the generation of melanoma antigen-specific CTLs and masked their inhibition by Treg. The mechanisms by which PD-1 blockade mediated immune enhancement included direct augmentation of melanoma antigen-specific CTL proliferation, heightening their resistance to inhibition by Treg and direct limitation of the inhibitory ability of Treg. PD-1 blockade reversed the increased expression of PD-1 and PD-L1 on melanoma antigen-specific CTL by Treg, rescued INF-γ and IL-2 or INF-γ and tumor necrosis factor-α co-expression and expression of IL-7 receptor by melanoma antigen-specific CTL which were diminished by Treg. PD-1 blockade also resulted in down-regulation of intracellular FoxP3 expression by Treg. These data suggest that PD-1 is importantly implicated in the regulation of Treg function in melanoma patients. PMID:19651643

  1. Mesenchymal stem cells alleviate atherosclerosis by elevating number and function of CD4(+)CD25 (+)FOXP3 (+) regulatory T-cells and inhibiting macrophage foam cell formation.

    Science.gov (United States)

    Wang, Zhi Xiao; Wang, Chong Quan; Li, Xiao Yan; Feng, Gao Ke; Zhu, Hong Ling; Ding, Yan; Jiang, Xue Jun

    2015-02-01

    Atherosclerosis is a chronic inflammatory disease characterized by the formation of plaques inside arteries, leading to narrowing and blockage. Potential therapeutic strategies include expanding the population of regulatory T-cells (Tregs) to enhance atheroprotective immunity, and inhibiting the formation of macrophage foam cells. Here, we studied the effect of bone marrow-derived mesenchymal stem cells (BM-MSCs) on atherosclerotic plaque formation in Apolipoprotein E(-/-) (ApoE-KO) mice, and elucidated the underlying mechanism. BM-MSCs isolated from 4 week-old ApoE-KO mice were evaluated by flow cytometry for expression of MSC-specific markers. Thirty eight week-old ApoE-KO mice were randomly divided into three experimental groups (n = 10 per group): 1. MSC group-received BM-MSCs intravenously; 2. Vehicle group-received DMEM; 3. Control group-did not receive any treatment. Administration of MSCs resulted in a marked decrease in the size of atherosclerotic plaques 3 months after treatment. In addition, the number and function of CD4(+)CD25(+)FOXP3(+) regulatory T-cells (Tregs) in cultured splenocytes, and the expression of FOXP3 at both mRNA and protein levels, was significantly increased in the MSC group. In vitro experiments further indicated that the formation of macrophage foam cells was inhibited by treatment with MSCs, accompanied by a significant downregulation in CD36 and scavenger receptor A (SRA). Our findings suggest that MSCs play an atheroprotective role by enhancing the number and function of Tregs and inhibiting the formation of macrophage foam cells. Hence, administration of MSCs to atherosclerotic patients might have significant clinical benefits.

  2. 骨髓间充质干细胞对肾病综合征大鼠CD4+CD25+调节性T细胞的影响%Effects of bone marrow mesenchymal stem cell transplantation on CD4+CD25+regulatory T cells in rats with primary nephrotic syndrome

    Institute of Scientific and Technical Information of China (English)

    杨焕丹; 张锐锋; 封东进; 朱冰冰; 吕娟

    2014-01-01

    BACKGROUND:Decreased function and reduced number of CD4+CD25+regulatory T cells have been considered the major manifestation of immunity dysfunction in children with primary nephrotic syndrome. Bone marrow mesenchymal stem cells have immunoregulation effects, which up-regulate CD4+CD25+regulatory T cells, inhibit proliferation of lymphocytes, and have been widely used in many immune diseases. OBJECTIVE:To investigate the effects of bone marrow mesenchymal stem celltransplantation on the CD4+CD25+regulatory T cells of peripheral blood in rats with primary nephrotic syndrome. METHODS:Bone marrow mesenchymal stem cells from six Sprague-Dawley rats were isolated, passaged and utilized for cellsuspension preparation. At the third passage, bone marrow mesenchymal stem cells were used for transplantation. The remaining 30 rats were randomly and equal y divided into three groups:normal group, normal saline infusion group, and bone marrow mesenchymal stem cells group. The rat models of primary nephrotic syndrome were established by single injection of adriamycin intravenously through tail vein in the latter two groups. Rats were then treated with bone marrow mesenchymal stem cells (1×10 7 ) (bone marrow mesenchymal stem cells group) or normal saline (normal saline infusion group) through tail vein at the same time after adriamycin administration. The normal group received no treatment. RESULTS AND CONCLUSION:Compared with the normal group, rats in the normal saline infusion group developed nephropathy characterized by ascites, proteinuria, hypoalbuminemia, hypercholastero-lnemia, and progressive renal injury. However, the proteinurine and clinical severity in bone marrow mesenchymal stem cells group were significantly ameliorated after treatment with bone marrow mesenchymal stem cells. CD4+CD25+Treg/CD4+Treg in the peripheral blood in the bone marrow mesenchymal stem cells group and normal saline infusion group were significantly higher than that in the normal group at 28

  3. Influence of Bone Marrow Mesenchymal Stem Cells on CD4 +CD25 +Treg Cells of Rat Heart Failure%骨髓间充质干细胞对鼠心力衰竭CD4+CD25+Treg 细胞的影响

    Institute of Scientific and Technical Information of China (English)

    陶恩学

    2014-01-01

    目的:研究骨髓间充质干细胞在鼠心力衰竭中对CD4+CD25+Treg细胞的影响。方法制作动物模型,雄性SD大鼠随机分为对照组A和模型组,模型组定期用阿霉素腹腔注射,对照组A用同等剂量生理盐水腹腔注射,每周1次,共6次,最后一次注射完成后继续喂养2周,行超声心动图检测左心室收缩末期容积( LVESV )、左室舒张末期容积( LVEDV )、左室射血分数( LVEF);并抽血通过流式细胞仪检测Tregs细胞。在最后一次注射完阿霉素1周后,分离、培养、扩增SD大鼠骨髓间充质干细胞(BMMSCs)。再将上述存活的模型组随机分为干预组,经尾静脉移植骨髓间充质干细胞组动物,对照组B,经尾静脉给予同等剂量生理盐水,分别于移植后第4周抽血通过流式细胞仪检测Tregs细胞,并行超声心动图检测LVESV,LVEDV,LVEF。结果①对照组A,B比较,LVESV,LVEF明显减低(P<0.05),而LVEDV明显升高(P<0.05)。②对照组A,B比较,CD4+CD25+Treg细胞的百分比明显减低( P<0.05)。结论骨髓间充质干细胞移植后可以上调CD4+CD25+Treg细胞,从而抑制扩张型心肌病中的免疫反应,改善心功能。%Objective To study the effect of bone marrow mesenchymal stem cells in the mouse heart failure on CD 4 +CD25 +Treg cells.Methods Male SD rats were randomly divided into control group A (n=10) and model group(n=20),and were treated regularly with intraperitoneal injection of doxorubicin in the model group and the same dose of normal saline in the control group weekly for a total of six times.Left ventricular end systolic volume (LVESV),left ventricular end diastolic volume (LVEDV),left ventricular ejection fraction ( LVEF) and blood Tregs cells were tested by Echocardiography and FlowCytometry respectively after two -week-feeding of the last injection . SD rat bone marrow mesenchymal stem cells (BMMSCs

  4. Antimitochondrial antibody

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/003529.htm Antimitochondrial antibody To use the sharing features on this page, please enable JavaScript. Antimitochondrial antibodies (AMA) are substances ( antibodies ) that form against mitochondria. ...

  5. Requirement of cognate CD4+ T-cell recognition for the regulation of allospecific CTL by human CD4+ CD127- CD25+ FOXP3+ cells generated in MLR.

    Directory of Open Access Journals (Sweden)

    Yuming Yu

    Full Text Available Although immunoregulation of alloreactive human CTLs has been described, the direct influence of CD4(+ Tregs on CD8(+ cytotoxicity and the interactive mechanisms have not been well clarified. Therefore, human CD4(+CD127(-CD25(+FOXP3(+ Tregs were generated in MLR, immunoselected and their allospecific regulatory functions and associated mechanisms were then tested using modified (51Chromium release assays (Micro-CML, MLRs and CFSE-based multi-fluorochrome flow cytometry proliferation assays. It was observed that increased numbers of CD4(+CD127(-CD25(+FOXP3(+ cells were generated after a 7 day MLR. After immunoselection for CD4(+CD127(-CD25(+ cells, they were designated as MLR-Tregs. When added as third component modulators, MLR-Tregs inhibited the alloreactive proliferation of autologous PBMC in a concentration dependent manner. The inhibition was quasi-antigen specific, in that the inhibition was non-specific at higher MLR-Treg modulator doses, but non-specificity disappeared with lower numbers at which specific inhibition was still significant. When tested in micro-CML assays CTL inhibition occurred with PBMC and purified CD8(+ responders. However, antigen specificity of CTL inhibition was observed only with unpurified PBMC responders and not with purified CD8(+ responders or even with CD8(+ responders plus Non-T "APC". However, allospecificity of CTL regulation was restored when autologous purified CD4(+ T cells were added to the CD8(+ responders. Proliferation of CD8(+ cells was suppressed by MLR-Tregs in the presence or absence of IL-2. Inhibition by MLR-Tregs was mediated through down-regulation of intracellular perforin, granzyme B and membrane-bound CD25 molecules on the responding CD8(+ cells. Therefore, it was concluded that human CD4(+CD127(-CD25(+FOXP3(+ MLR-Tregs down-regulate alloreactive cytotoxic responses. Regulatory allospecificity, however, requires the presence of cognate responding CD4(+ T cells. CD8(+ CTL regulatory

  6. Isolation of highly suppressive CD25+FoxP3+ T regulatory cells from G-CSF-mobilized donors with retention of cytotoxic anti-viral CTLs: application for multi-functional immunotherapy post stem cell transplantation.

    Science.gov (United States)

    Samuel, Edward R; Beloki, Lorea; Newton, Katy; Mackinnon, Stephen; Lowdell, Mark W

    2014-01-01

    Previous studies have demonstrated the effective control of cytomegalovirus (CMV) infections post haematopoietic stem cell transplant through the adoptive transfer of donor derived CMV-specific T cells (CMV-T). Strategies for manufacturing CMV immunotherapies has involved a second leukapheresis or blood draw from the donor, which in the unrelated donor setting is not always possible. We have investigated the feasibility of using an aliquot of the original G-CSF-mobilized graft as a starting material for manufacture of CMV-T and examined the activation marker CD25 as a targeted approach for identification and isolation following CMVpp65 peptide stimulation. CD25+ cells isolated from G-CSF-mobilized apheresis revealed a significant increase in the proportion of FoxP3 expression when compared with conventional non-mobilized CD25+ cells and showed a superior suppressive capacity in a T cell proliferation assay, demonstrating the emergence of a population of Tregs not present in non-mobilized apheresis collections. The expansion of CD25+ CMV-T in short-term culture resulted in a mixed population of CD4+ and CD8+ T cells with CMV-specificity that secreted cytotoxic effector molecules and lysed CMVpp65 peptide-loaded phytohaemagglutinin-stimulated blasts. Furthermore CD25 expanded cells retained their suppressive capacity but did not maintain FoxP3 expression or secrete IL-10. In summary our data indicates that CD25 enrichment post CMV stimulation in G-CSF-mobilized PBMCs results in the simultaneous generation of both a functional population of anti-viral T cells and Tregs thus illustrating a potential single therapeutic strategy for the treatment of both GvHD and CMV reactivation following allogeneic haematopoietic stem cell transplantation. The use of G-CSF-mobilized cells as a starting material for cell therapy manufacture represents a feasible approach to alleviating the many problems incurred with successive donations and procurement of cells from unrelated donors

  7. Antibodies to poliovirus detected by immunoradiometric assay with a monoclonal antibody

    Energy Technology Data Exchange (ETDEWEB)

    Spitz, M.; Fossati, C.A.; Schild, G.C.; Spitz, L.; Brasher, M. (National Inst. for Biological Standards and Control, London (UK))

    1982-10-01

    An immunoradiometric assay (IRMA) for the assay of antibodies to poliovirus antigens is described. Dilutions of the test sera or whole (finger prick) blood samples were incubated with the poliovirus antigen bound to a solid phase and the specific antibody was detected by the addition of a mouse anti-human IgG monoclonal antibody (McAb), which was itself revealed by iodinated sheep IgG antimouse F(ab). The authors have shown that this technique is suitable for the estimation of IgG anti-poliovirus antibodies induced in children following polio vaccine. The present study shows that SPRIA provides a simple and inexpensive method for serological studies with poliovirus particularly for use in large-scale surveys.

  8. Effect of acrolein exposure on the percentage of CD4 + CD25 + regulatory T cells and expression of transcription factor Foxp3 in asthmatic rats%丙烯醛暴露对哮喘大鼠CD4+CD25+调节性T细胞及转录因子Foxp3表达的影响

    Institute of Scientific and Technical Information of China (English)

    魏明; 涂玲; 梁颖红; 刘佳; 张俊华; 龚艳杰

    2012-01-01

    Objective This work was intended to investigate the effect of acrolein fog exposure on the ratio of CD4 + CD25 + regulatory T cells(Treg) and expression of transcription factor Foxp3 in asthmatic rats.Methods Sixty 6 - 8 weeks male Wistar rats were randomly divided into 4 groups according to random number table ( 15 rats for each group ) which were control group (animnals were treated with saline ),aerosolized ovalbumin( OVA )exposure group,acrolein exposure group and combined OVA and aerolein fog exposure group,respectively.The rats were exposed to air or/and to saline or OVA aerosol for 6 - 8 weeks respectively.24 h after the last challenge,4 ml of peripheral blood and lung tissue were collected from each rat.The percentage of CD4 + CD25 + T cells was determined by flow cytometry analysis.The concentration of interleukin-4(IL-4) and γ-interferon (IFN-γ)in peripheral blood and lung homogenates were measured by ELISA.The protein expression of Foxp3 in the lung was detected by Western blotting.Results The percentage of CD4 + CD25+ T cells in aerosolized OVA group( (6.23 ± 1.1 1 )% )was significantly lower than that in the normal saline group( ( 9.97 ± 1.23 )% ) ( P < 0.01 ).The percentage of CD4 + CD25 + T cells ((3.26 ±0.84)% )in OVA combined acrolein fog exposure group was remarkably lower than that in the aerosolized OVA exposure group and in the normal saline group( P < 0.01 ).11-4 in both plasma and lung ( (22.57 ±4.34),(0.86 ±0.12) ng/L) was significantly increased in the OVA exposed rats compared with the normal saline group( ( 11.57 ±2.86),(0.31 ±0.10) ng/L) ( P <0.01 ).Further remarkable increase in IL-4 of both plasma and lung tissue was observed in the group exposed to both OVA and acrolein ( ( 34.32 ± 6.21 ),( 1.45 ±0.32)ng/L) compared with the aerosolized OVA exposure group and the normal saline group( P < 0.05 ).γ-IFN of plasma and lung tissue in OVA exposed group ( ( 59.67 ± 20.12 ),( 0.56 ±0.17)ng/L) was significantly

  9. Relationship of CD4+ CD25hi Regulatory T(Treg)Cells to Disease Progression in HIV-infected Patients%HIV/AIDS患者疾病进程与CD4+CD25hi调节性T细胞之间相关性研究

    Institute of Scientific and Technical Information of China (English)

    宋淑静; 冯鑫; 郭晶晶; 刘亚楠; 伦文辉; 魏红山; 刘顺爱

    2009-01-01

    目的 通过分析不同阶段HIV感染者外周血CD4+CD25hi调节性T细胞(CD4+CD25hiregulatory T cells,Treg cells)与外周血免疫状态和病毒载量的相关性,探讨Treg细胞对HIV/AIDS发病进程的影响.方法 采集116例HIV感染者和21例正常人对照外周血,用4色流式细胞术进行CD4+和CD8+T细胞绝对数计数;用3色流式细胞术进行Treg细胞测定;用荧光定量PCR法进行HIVRNA载晕测定.实验数据用回归统计学方法和T检验方法进行分析.结果 HIV感染者外周血Treg细胞频率在HIV感染初期显著下降,之后随着疾病的进程逐渐升高.在CD4+T细胞大于300/μl的患者低于正常对照组,在CD4+T细胞小于100/μl的患者高于正常对照组,差异具有统计学意义.Treg细胞频率与CD4+T淋巴细胞绝对数和CD4+/CD8+之间均呈负相关.其相关系数r和P值各为r=-0.564,P<0.001和r=-0.377,P<0.001;Treg细胞频率与血浆HIV病毒载量呈正相关,其相关系数r=0.514.P<0.001.结论 CD4+CD25hi Treg细胞可能是参与艾滋病免疫发病机理的重要细胞,在HIV感染发病进程的不同阶段具有不同的意义,其确切机制有待进行进一步研究.%Objective Analyzing the relationships between peripheral blood CD4+ CD25hi regulatory T (Treg)cells and peripheral blood immune status or plasma HIV-1 viral load in HIV-infected individuMs,so as to determine whether Treg were related to the progression of HIV-infected disease.Methods116 HIV-infected patients in different stages and 21 healthy control individuals were included in this study.The CD4+ and CD8+ T cell counts were determined by a standard 4-color flow cytometry technique.The Treg cells were examined with 3-color immune staining flow cytometry.The plasma HIV-1 viral load was detected by real time PCR.Results The frequencies of Treg cells decreased in HIV-infected individuals with high CD4+ T cell counts(>300/μl)compared with normal controls.With the progression of disease the frequencies

  10. 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

  11. Increase of Circulating CD4(+)CD25(high)Foxp3(+) 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)

    2010-01-01

    in patients with metastatic renal cell carcinoma on the frequency of CD4(+) CD25(high)Foxp3(+) 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 Treg cells decreased when patients...... had been off IL-2 treatment for only 8 days, but remained higher than pretreatment levels. A functional assay showed that isolated Treg cells were capable of inhibiting proliferation of responder cells. Also, in vitro studies showed that coculture of mature DCs, autologous T cells and IL-2 leads...... to an increase in the number of Treg cells whereas IL-21 does not stimulate the induction of Treg cells. These findings demonstrate that even low doses of IL-2 in combination with DC vaccination are able to expand CD4(+)CD25(+)Foxp3(+) Treg cells in vivo in metastatic renal cell carcinoma patients. Further...

  12. Congenital rubella affecting an infant whose mother had rubella antibodies before conception

    OpenAIRE

    1981-01-01

    A woman who had had high titres of rubella antibodies some months before she became pregnant gave birth to an infant in whom congenital rubella was confirmed at 4 months. Rubella haemagglutination inhibition tests, complement fixation tests, and immunofluorescence tests with anti-human IgG were carried out on sera from the mother. Rubella antibody titres in sera obtained in March 1971, seven and a half months before conception, were equivalent to 400 units, which is usually taken as indicatin...

  13. Clinical significance of detection of CD_4~+CD_(25)~(high)CD_(127)~(low) regulatory T cells in peripheral blood of NHL patients%非霍奇金淋巴瘤患者外周血CD_4~+CD_(25)~(high)CD_(127)~(low)调节性T细胞检测的临床意义

    Institute of Scientific and Technical Information of China (English)

    赵志强; 张巧花; 苏文; 侯淑玲; 贺建霞

    2010-01-01

    目的 观察非霍奇金淋巴瘤(NHL)患者外周血CD_4~+CD_(25)~(high)CD_(127)~(low)调节性T细胞(Treg细胞)亚群水平的变化,初步探讨其临床意义.方法 采用流式细胞术检测65例NHL患者及29名健康人外周血中Treg细胞,并进行统计分析.结果 在65例NHL患者的外周血中Treg细胞的比例为(6.72±1.38)%,高于健康对照组(5.65±0.68)%(P0.05).结论 Treg细胞为免疫抑制细胞,可由肿瘤诱导产生,肿瘤患者外周血中Treg细胞高于健康对照组,且随着患者临床分期的增加,Treg细胞比值升高,Treg细胞比值可以作为临床评估瘤负荷的指标.%Objective To investigate the change of CD_4~+CD_(25)~(high)CD_(127)~(low)> regulatory T cells (Trcg cells) sub-group level in peripheral blood of non-Hodgkin lymphama (NHL) patients, and to explore its clinical significance. Methods Treg cells levels in peripheral blood of lymphoma patients and normal were detected by flow cytometry, followed by statistical analysis. Results In the 65 cases of NHL patients, Treg cells in peripheral blood were (6.72±1.38) %, higher than that in the normal control group (5.65±0.68) % (P 0.05, normal control group (5.65±0.68) %, patients with normal LDH group (6.97±1.20) %, patients with lactate dehydrogenase (6.54±1.02) %]. Conclusion Treg cells induced by tumor and could inhibit the immune cells, Treg cells percentage in peripheral blood of tumor patients is higher than that the normal control group, and increased with the clinical staging, so the percentage of Treg cells may serve as a clinical indicator to evaluate tumor load.

  14. Understanding the conversation between machines: Heidegger's ontology of presence versus Lacan's anti-humanism

    Directory of Open Access Journals (Sweden)

    Katarina Peović Vuković

    2015-12-01

    Full Text Available This paper addresses the question of machine intelligence, a cybernetic problem closely related to the questions of subjectivity, human intelligence and the constitution of human communication. The example of chatterbots and their (movie representations will be used in order to reexamine their similarities and differences, as well as their relation to subjectivity in the broadest sense. Reversing the thesis on the instrumentalisation of the subject by means of a cybernetic framework, which is, as Martin Heidegger claimed, established as a paradigmatic framework of contemporary sciences, the paper will point to the closest relation between the subject and the mechanical or digital quasi-intelligent machine. Jacques Lacan's applied psychoanalysis provides a theoretical framework for this paper since it is in a position to point out the inadequacies of understanding communication as a means of conveying information. Such critique will prove to be fruitful not only for describing communication and the difference between the machinal and the human, but also for indicating the problems of identity politics. Lacan's anti-humanism and ethics of separation appear as tools of contemporary critique of political economy.

  15. Effect of 1,25-(OH) 2-VitD3 combined with spleen ammonia peptide oral freeze dry powder on CD4+CD25+T cells, TGF-β1 and VEGF in patients with asthma%1,25二羟维生素D3联合脾氨肽口服冻干粉对哮喘CD4+CD25+T细胞、TGF-β1及VEGF影响研究

    Institute of Scientific and Technical Information of China (English)

    高志刚; 袁永红; 詹建华; 潘开宇

    2016-01-01

    目的 探讨1,25-二羟维生素D3联合脾氨肽口服冻干粉对哮喘患者CD4+ CD25+T细胞、TGF-β1及VEGF的影响.方法 收集56例支气管哮喘患者,随机分为对照组和实验组,每组各28例,分别给予相应的药物治疗,治疗结束后,对所有患者的静脉血CD4+ CD25+T细胞水平、血清TGF-β1、VEGF水平、ACT评分及肺功能情况进行检测.结果 治疗后,与对照组比较,实验组患者静脉血CD4+CD25+T细胞水平较高(P<0.05);血清TGF-β1水平较高(P<0.05);血清VEGF水平较低(P <0.05);FEV1%、FEV1/FVC值较高(P<0.05);ACT评分较高(P<0.05).结论 1,25-二羟维生素D3联合脾氨肽口服冻干粉能够显著升高支气管哮喘患者静脉血CD4+ CD25+T细胞水平及血清TGF-β1,降低血清VEGF水平,提高免疫力,改善肺功能和治疗效果.

  16. CD4+CD25+调节性T细胞与Th17细胞在寻常型银屑病中的表达%Role of CD4+CD25+ regulatory T cells and Th17 cells in patients with psoriasis vulgaris

    Institute of Scientific and Technical Information of China (English)

    张玉丽; 王逸飞; 张春敏

    2016-01-01

    目的::检测外周血中CD4+CD25+调节性T细胞(简称Treg细胞)与Th17细胞在寻常型银屑病( PV)中的表达。方法:34例寻常型银屑病患者,18例正常人作为对照。采用流式细胞术检测外周血中Treg细胞、FOXP3+Treg细胞及Th17细胞的表达水平。结果: PV患者外周血中Treg细胞比例和FOXP3+Treg细胞比例分别为(3.68±1.22)%和(0.53±0.19)%,低于正常对照组的(6.63±1.00)%和(0.76±0.14)%(P0.05) . Conclusion:The decrease in the expression of Treg cells and the increased response of Th17 cells may be the important factors for the immuno-logical imbalance in PV.

  17. C57BL/6小鼠EAE模型的建立及Foxp3、CD4~+CD25~+调节性T细胞检测%Establishment of experimental allergic encephalomyelitis in C57BL/6 mice and testing of expressions of Foxp3 and quantifies of CD4~+ CD25~+ regulatory T cells

    Institute of Scientific and Technical Information of China (English)

    翁益云; 夏君慧; 鲍剑虹; 张国勇; 张旭

    2010-01-01

    目的 以髓鞘少突胶质细胞糖蛋白细胞外免疫球蛋白样结构域(MOG~(Igd))为抗原免疫C57BL/6小鼠,建立实验性自身免疫性脑脊髓炎(EAE)模型,并检测CD4~+CD25~+T细胞、Foxp3 mRNA等指标的表达.方法 采用分子克隆技术诱导表达MOG~(Igd)-TrxA融合蛋白,纯化后以此融合蛋白为抗原于侧腹壁皮下多点注射免疫C57BL/6小鼠作为实验组(MOG组);同时以硫氧还蛋白(TrxA)、豚鼠脊髓匀浆(GPSCH)、生理盐水免疫小鼠分别作为阴性、阳性及正常对照组.双盲法连续早晚观察30 d,通过评估动物的临床神经功能、组织病理(HE染色和Kluver-Barrera法髓鞘染色、免疫组化法)情况,评价模型的质量.流式细胞仪(FACS)检测小鼠脾脏中CD4~+CD25~+调节性T细胞;real-time PCR检测Foxp3 mRNA的相对表达水平,并分析两者相关性.结果 MOG组及GPSCH组小鼠均呈慢性非缓解型病程,两组在发病时间、达峰时间、临床神经功能评分等方面差异均无统计学意义(P>0.05).TrxA组及正常对照组无一动物发病.模型组(包括MOG组和GPSCH组)动物HE染色可见血管周围炎性细胞浸润,胶质结节形成,髓鞘染色可见斑片状髓鞘脱失,大脑、小脑、脑干和脊髓均有不同程度受累.免疫组化定性和半定量分析显示MOG组和GPSCH组病灶周围炎症浸润处AQP-4表达与TrxA组相比明显增高(P0.05).相关性分析显示,模型组小鼠CD4~+ CD25~+调节性T细胞与Foxp3 mRNA表达水平间存在相关性(r=0.849,P0.05). However, neither in TrxA treated group nor in normal control group did animals exhibit clinical signs of EAE. Histologic sections of the brain and spinal cord taken from affected animals shew perivascular infiltration of mononuclear cells, gliosis, and multifocal demyelination. Lesions scattered throughout the CNS including brainstem, spinal cord, cerebellum, and penventricular white matter. There were significant differences between MOG group and TrxA group

  18. 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

  19. 巨细胞病毒感染致婴幼儿喘息患儿中瘦素、CD4CD25Treg和TLR4的表达

    Institute of Scientific and Technical Information of China (English)

    朱春晖; 陈强; 朱庆雄; 柯江维

    2012-01-01

    Objective The influence of cytomegalovirus infection on the leptin, CD4+CD25+ regulatory T cell and Toll-like receptor.Methods Using PCR to detect the phlegm and serum CMV DNA level of 69 wheezing infants and 28 healthy volunteers and compare the positive rates.detect the leptin level of cytomegalovirus infectious infants with radioimmunoassay in 24 hours after admission, 24 hours after ganciclovir treatment and 12 weeks after the treatment,the CD4+CD25+Treg percentage of CD4+T in peripheral blood and TLR4 expression in monocytes cell surface with flow cytometry.follow-up the wheezing in out - patient clinic or through phone call.Results (1)37 of wheezing infants suffered cytomegalovirus infection,the positive rates was 53.62%,was much higher than that in healthy volunteers group(1/28,3.57%)(p0.05).24h after the treatment:the leptin and TLR4 level in the group accepted ganciclovir treatment, the group refused ganciclovir treatment and group non- cytomegalovirus infection was lower than that before treatment,the level among the three groups has no significant difference(p>0.05); the CD4+CD25+Treg percentage increased after treatment(p0.05).治疗结束24h内:更昔洛韦治疗组、非更昔洛韦治疗组和非巨细胞病毒感染组瘦素水平均较治疗前下降,组间无明显差异(p>0.05),更昔洛韦治疗后CD4+CD25+Treg百分比增高,差异有统计学意义(p<0.05),TLR4水平较治疗前明显下降(p<0.01),且低于同期非更昔洛韦治疗组TLR4水平(p<0.05).治疗后12周:更昔洛韦治疗组瘦素及TLR4水平低于非更昔洛韦治疗组和非巨细胞病毒感染组,CD4+CD25+Treg百分比高于非更昔洛韦治疗组和非巨细胞病毒感染组,差异有统计学意义(p<0.05);非巨细胞病毒感染组瘦素水平、CD4+CD25+Treg百分比及TLR4水平与入院后24小时内及治疗结束24小时内比较无明显差异.(4)更昔洛韦治疗后随访2年患儿喘息次数明显减少(p<0.05).结论(1)巨细胞病毒感染与婴

  20. CONSTRUCTION AND EXPRESSION OF A HUMAN-MOUSE CHIMERIC ANTIBODY AGAINST HUMAN BLADDER CANCER

    Institute of Scientific and Technical Information of China (English)

    白银; 王琰; 周丽君; 俞莉章

    2001-01-01

    To construct and express a human-mouse chimeric antibody against human bladder cancer. Method: The variable region genes of anti-human bladder cancer monoclonal antibody BDI-1 were cloned by RT-PCR. A human-mouse chimeric antibody expression vector was constructed and transfected into CHO cells. The chimeric antibody against bladder cancer was expressed and characterized. Result: Eukaryotic expression vector of the chimeric antibody against human bladder carcinoma was successfully constructed, and was expressed in eukaryotic cells; the expressed chimeric antibody ch-BDI showed same specificity as its parent McAb against human bladder cancer cells. Conclusion: The constructed chimeric antibody was expressed successfully in eukaryotic cells, and the chimeric antibody had desired affinity against human bladder cancer cells.

  1. Relationship between CD4 +CD25highFoxP3 +Treg and the rejection after stem cells transplant for ;Parkinson’s disease%CD4+CD25highFoxP3+Treg与帕金森病干细胞移植后排斥反应间的相关性研究

    Institute of Scientific and Technical Information of China (English)

    赵永波; 沈楠

    2013-01-01

    目的:通过应用骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)移植治疗帕金森病(Parkinson's disease,PD)大鼠,检测大鼠外周血中CD4+CD25highFoxP3+调节性T细胞(regulatory T cell,Treg)比例的变化及相关细胞因子的漂移,观察脑内免疫排斥反应的发生状况。以期探讨干细胞移植治疗PD后,外周血中Treg的改变与脑内免疫排斥反应间的相关性及其在干细胞移植后免疫耐受中的作用。方法 SD大鼠随机分为正常对照组、PD组、(PD+PBS)组、(PD+BMSCs)组。正常对照组大鼠不进行任何处理,为完全空白对照;余下大鼠均通过脑内立体定向注射6-羟基多巴胺(6-hydmxydopamine,6-OHDA)建立PD大鼠模型。成功建模后第3周,(PD+PBS)组大鼠脑内定向注射PBS缓冲液;(PD+BMSCs)组大鼠脑内定向注射应用全骨髓贴壁法培养的大鼠BMSCs悬液;PD组大鼠不做任何处理。PBS缓冲液和BMSCs悬液注射2周后,各组大鼠均应用流式细胞术(lfow cytometry,FCM)检测其外周血中CD4+CD25highFoxP3+Treg占T淋巴细胞比例的变化,并运用酶联免疫吸附测定(ELISA)检测外周血中白介素(IL)-4、IL-10和干扰素-γ(INF-γ)等相关细胞因子的改变。另外,通过免疫组织化学染色法观察大鼠脑内酪氨酸羟化酶(tyrosine hydroxylase,TH)阳性神经元、离子钙接头蛋白分子-1(ionized calcium bindingadaptor molecule-1, iba-1)阳性细胞、主要组织相容性复合体-Ⅱ(major histocompatibility complex-Ⅱ,MHC-Ⅱ)分子的变化。同时,(PD+BMSCs)组大鼠采用EdU染色法监测移植后BMSCs的存活情况。结果脑内立体定向注射6-OHDA 3周后,部分大鼠腹腔注射阿扑吗啡可诱导出旋转行为,表现为向健侧旋转,呈首尾相接状,以210圈/30分钟为标准,建模成功率达65.45%,并且免疫组织化学染色显

  2. 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%Expression and signifiacnce of CD4 +CD25 +regulatory T cells(Treg) in the pathogenesis for the pneumonia patients with leucopenia

    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.

  3. Change of CD4+ CD25+ CD127 low regulatory T cells in peripheral blood of patients with Graves disease treated by 131 I or antithyroid drugs therapy%Graves病131I或抗甲状腺药物治疗前后外周血CD4+CD25+CD127low调节性T细胞的变化

    Institute of Scientific and Technical Information of China (English)

    杨静; 潘天荣; 杜益君; 钟兴

    2016-01-01

    目的:探讨调节性T细胞( Treg)在Graves病( GD)患者外周血中变化,以及131 I或抗甲状腺药物( ATD)治疗后其变化趋势,寻找评价131 I和ATD治疗疗效的新指标。方法健康者40例设为对照组,初诊GD患者40例设为GD组,并将GD组随机分成131 I 治疗组(20例)及 ATD 治疗组(20例)。检测131 I治疗组、ATD治疗组治疗前、治疗后及对照组CD4+CD25+CD127 low Treg 比例、白介素-10( IL-10)、转化生长因子-β1(TGF-β1)水平,通过统计学软件处理相关结果。结果①治疗前GD组Treg比例较对照组明显降低,差异有统计学意义( P<0.01);②131 I治疗组、ATD治疗组治疗后第3个月及第6个月Treg比例较治疗前均升高,差异均有统计学意义( P <0.01);③治疗后第3个月及第6个月, ATD治疗组Treg比例与131 I治疗组差异无统计学意义;④治疗前GD组IL-10、TGF-β1水平较对照组均降低,治疗后6个月,细胞因子水平较治疗前均升高,差异均有统计学意义(P<0.05),各时间点131 I治疗组与ATD治疗组之间,细胞因子差异无统计学意义。结论 GD患者Treg比例和功能显著降低,治疗后部分恢复,因此,对于甲亢患者,Treg可能是评价免疫状态及治疗后病情缓解的指标之一。%Objective To investigate the changes of regulatory T cells ( Treg) in Graves disease ( GD) before and after being treated by 131 I or antithyroid drugs( ATD) ,and to seek for new clinical indicators to evaluate the treat-ment response. Methods The study groups included 40 patients with GD ( GD group) , 20 of whom were treated by 131 I ,others were treated by ATD. Forty healthy donors without history of thyroid or autoimmune disease were en-rolled in control group. The proportions of CD4 +CD25 +CD127low Treg , IL-10 and TGF-β1 were tested before and after treatment respectively. Results ① The significant decrease in the proportion of CD4 +CD25 +CD127low Treg cells in untreated GD patients ( GD group) was

  4. Increase in TGF-β secreting CD4⁺CD25⁺ FOXP3⁺ T regulatory cells in anergic lepromatous leprosy patients.

    Directory of Open Access Journals (Sweden)

    Chaman Saini

    Full Text Available BACKGROUND: Lepromatous leprosy caused by Mycobacterium leprae is associated with antigen specific T cell unresponsiveness/anergy whose underlying mechanisms are not fully defined. We investigated the role of CD25(+FOXP3(+ regulatory T cells in both skin lesions and M.leprae stimulated PBMC cultures of 28 each of freshly diagnosed patients with borderline tuberculoid (BT and lepromatous leprosy (LL as well as 7 healthy household contacts of leprosy patients and 4 normal skin samples. METHODOLOGY/PRINCIPLE FINDINGS: Quantitative reverse transcribed PCR (qPCR, immuno-histochemistry/flowcytometry and ELISA were used respectively for gene expression, phenotype characterization and cytokine levels in PBMC culture supernatants. Both skin lesions as well as in vitro antigen stimulated PBMC showed increased percentage/mean fluorescence intensity of cells and higher gene expression for FOXP3(+, TGF-β in lepromatous (p<0.01 as compared to tuberculoid leprosy patients. CD4(+CD25(+FOXP3(+ T cells (Tregs were increased in unstimulated basal cultures (p<0.0003 and showed further increase in in vitro antigen but not mitogen (phytohemaglutinin stimulated PBMC (iTreg in lepromatous as compared to tuberculoid leprosy patients (p<0.002. iTregs of lepromatous patients showed intracellular TGF-β which was further confirmed by increase in TGF-β in culture supernatants (p<0.003. Furthermore, TGF-β in iTreg cells was associated with phosphorylation of STAT5A. TGF-β was seen in CD25(+ cells of the CD4(+ but not that of CD8(+ T cell lineage in leprosy patients. iTregs did not show intracellular IFN-γ or IL-17 in lepromatous leprosy patients. CONCLUSIONS/SIGNIFICANCE: Our results indicate that FOXP3(+ iTregs with TGF-β may down regulate T cell responses leading to the antigen specific anergy associated with lepromatous leprosy.

  5. Pathophysiological significance of the two-pore domain K+ channel K2P5.1 in splenic CD4+CD25− T cell subset from a chemically-induced murine inflammatory bowel disease model

    Science.gov (United States)

    Nakakura, Sawa; Matsui, Miki; Sato, Aya; Ishii, Mizuki; Endo, Kyoko; Muragishi, Sayaka; Murase, Miki; Kito, Hiroaki; Niguma, Hiroki; Kurokawa, Natsumi; Fujii, Masanori; Araki, Masatake; Araki, Kimi; Ohya, Susumu

    2015-01-01

    The alkaline pH-activated, two-pore domain K+ channel K2P5.1 (also known as TASK2/KCNK5) plays an important role in maintaining the resting membrane potential, and contributes to the control of Ca2+ signaling in several types of cells. Recent studies highlighted the potential role of the K2P5.1 K+ channel in the pathogenesis of autoimmune diseases such as rheumatoid arthritis and multiple sclerosis. The aim of the present study was to elucidate the pathological significance of the K2P5.1 K+ channel in inflammatory bowel disease (IBD). The degrees of colitis, colonic epithelial damage, and colonic inflammation were quantified in the dextran sulfate sodium-induced mouse IBD model by macroscopic and histological scoring systems. The expression and functional activity of K2P5.1 in splenic CD4+ T cells were measured using real-time PCR, Western blot, and fluorescence imaging assays. A significant increase was observed in the expression of K2P5.1 in the splenic CD4+ T cells of the IBD model. Concomitant with this increase, the hyperpolarization response induced by extracellular alkaline pH was significantly larger in the IBD model with the corresponding intracellular Ca2+ rises. The expression of K2P5.1 was higher in CD4+CD25− T cells than in CD4+CD25+ regulatory T cells. The knockout of K2P5.1 in mice significantly suppressed the disease responses implicated in the IBD model. Alternations in intracellular Ca2+ signaling following the dysregulated expression of K2P5.1 were associated with the disease pathogenesis of IBD. The results of the present study suggest that the K2P5.1 K+ channel in CD4+CD25− T cell subset is a potential therapeutic target and biomarker for IBD. PMID:26578971

  6. 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 ...... 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...

  7. Linfócitos T CD4+CD25+ e a regulação do sistema imunológico: perspectivas para o entendimento fisiopatológico da sepse

    OpenAIRE

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

    2012-01-01

    A resposta inflamatória sistêmica representa o evento patogênico central da sepse, subjazendo às manifestações clínicas e aos achados laboratoriais presentes nos enfermos. Inúmeras pesquisas têm demonstrado que os linfócitos T CD4+CD25+ - também conhecidos como células T reguladoras (Treg) - participam dos processos de desenvolvimento da sepse, em virtude de sua capacidade de suprimir a resposta imune. Com base nessas ideias, propôs-se, no presente artigo, a discussão do papel dos linfócitos ...

  8. Identification of anti-HPA-1a allo-antibodies using IgG platelet antibody detection and crossmatch system assay with Galileo Echo.

    Science.gov (United States)

    Di Cristofaro, Julie; Frassati, Coralie; Montagnie, Rolande; Basire, Agnes; Merieux, Yves; Picard, Christophe

    2015-01-01

    Fetal/neonatal allo-immune thrombocytopenia is the most frequent and the most dangerous clinical condition involving anti-human platelet antigens (HPA)-1a allo-antibodies. Anti-HPA-1a allo-immunization requires rapid and accurate diagnosis to determine appropriate treatment. The Capture-P Ready-Screen assay (C-PRS) is a new qualitative immunoassay to detect IgG anti-human leukocyte antigen (HLA) and anti-HPA allo-antibodies. The aim of this study is to assess the identification of anti-HPA-1a allo-antibodies using the C-PRS assay, associated with HLA class I stripping reagents, on the automated benchtop analyzer Galileo Echo. Forty-nine sera were analyzed: without anti-HLA class I or anti-HPA allo-antibodies, with anti-HLA class I allo-antibodies, with anti-HPA-1a allo-antibodies, among which with anti-HLA class I allo-antibodies. None of the samples without allo-antibodies were reactive. Only anti-HLA antibodies, detected by cytotoxicity-dependent complement and not by Luminex, remained positive before and after stripping reagents. Of the 13 samples, anti-HPA-1a allo-antibodies that were correctly identified before and after incubation with HLA assassin reagent were 70% and 85%, respectively. Anti-glycoprotein auto-antibodies and anti-HLA allo-antibodies do not interfere with the detection of anti-HPA-1a antibodies. This preliminary study indicates that further improvement of the test will be helpful in developing a clinically useful assay in the future.

  9. Expressions of folkhead helix transcription factor 3 on CD4+ CD25+ regulatory T lymphocyte in intestinal mucosa in human immunodeficiency virus infected patients%人类免疫缺陷病毒感染者肠黏膜组织中CD4+CD25+调节性T淋巴细胞及叉头状转录因子3的表达

    Institute of Scientific and Technical Information of China (English)

    孙磊; 兰孟东; 郎振为; 王鹏; 李坪; 赵红心; 滕晓英; 周新刚; 张亮; 沈冰

    2011-01-01

    Objective To investigate the changes of CD4+ CD25+ regulatory T lymphocyte (Treg) and expressions of folkhead helix transcription factor 3 (FoxP3) in intestinal mucosa in human immunodeficiency virus (HIV) infected patients. Methods Twenty-one HIV infected patients and 17 control subjects without HIV infection were included in this study. The expression of FoxP3, which was considered as a specific marker of CD4+ CD25 + Treg, was detected in intestinal mucosa specimens from HIV infected patients by immunohistochemistry. Meanwhile, the in situ expression of CD4+ T lymphocyte was also determined by immunohistochemistry. The data were analyzed by t test. Results The positive labeling index of CD4+ T lymphocyte in intestinal mucosa was significantly lower in HIV infected patients compared to the controls (11. 56%±4. 44% vs 43. 49% ±8. 90% ,t=-11. 86,P<0. 01). The positive labeling index of FoxP3 in intestinal mucosa was also significantly lower in HIV infected patients compared to the controls (0.46% ± 0.20% vs 1. 18% ± 0. 44% ,t= - 5. 98,P<0.01). Conclusion The depletion of CD4+ CD25+ Treg is accompanied with the depletion of CD4 + T lymphocyte and the reduction of FoxP3 expression in intestinal mucosa of HIV infected patients.%目的 研究HIV感染者肠黏膜组织中CD4+CD25+调节性T淋巴细胞(Treg)及叉头状转录因子3(FoxP3)的表达.方法 应用免疫组织化学法,检测21例HIV感染者及17例非HIV感染者肠黏膜组织中CD4+CD25+Treg的特异性标志物FoxP3的表达以及CD4+T淋巴细胞的原位表达.数据处理应用t检验.结果 HIV感染者肠黏膜组织中CD4+T淋巴细胞阳性标记指数为11.56%±4.44%,显著低于非HIV感染者的43.49%±8.90%(t=-11.86,P<0.01).HIV感染者肠黏膜组织中FoxP3阳性标记指数为0.46%±0.20%,显著低于非HIV感染者的1.18%±0.44%(t=-5.98,P<0.01).结论 HIV感染者肠黏膜组织中CD4+T淋巴细胞出现消减,CD4+CD25+Treg也出现消减,FoxP3表达下降.

  10. Cytotoxicity of an ebulin l-anti-human CD105 immunotoxin on mouse fibroblasts (L929) and rat myoblasts (L6E9) cells expressing human CD105.

    Science.gov (United States)

    Benítez, Jorge; Ferreras, J Miguel; Muñoz, Raquel; Arias, Yolanda; Iglesias, Rosario; Córdoba-Díaz, Manuel; del Villar, Rosario; Girbés, Tomás

    2005-01-01

    Tumour growth is characterised by the formation of a fine vessel network or neovasculature which nourishes tumour cells. Two kinds of novel anti-angiogenic therapies are based on the prevention of vessels growth and on the destruction of those vessels already formed. We report here on the design and construction of a novel immunotoxin formed with the non-toxic type II ribosome-inactivating protein ebulin l and the mouse anti-human CD105 monoclonal antibody 44G4. The 44G4-ebulin immunotoxin was formed by covalent linking of both proteins with N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and was purified by chromatography on Superdex 200 HiLoad. The analysis of the anti-ribosomal effects in a cell-free translation system indicated that conjugation does not affect the activity of ebulin l. The immunotoxin displays cytotoxicity with nanomolar IC50 values on human CD105+ cells like the mouse fibroblasts L929 cells transfected with the short form of human CD105 and the rat myoblasts L6E9 transfected with the long form of human CD105. In contrast, cells lacking human CD105 were 2-2.5 logs less sensitive to the immunotoxin. Free ebulin displays IC50 values in the range 10(-6) M. Since CD105 is being considered as a potential target for the anti-vascular therapy of tumours, the present immunotoxin could be a promising tool for the anticancer therapy, especially due to the very low in vivo toxicity of ebulin l as compared ricin and other toxins used for immunotoxins.

  11. 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.

  12. Analysis of mRNA expression for genes associated with regulatory T lymphocytes (CD25, FoxP3, CTLA4, and IDO) after experimental infection with bovine viral diarrhea virus of low or high virulence in beef calves.

    Science.gov (United States)

    Palomares, Roberto A; Hurley, David J; Woolums, Amelia R; Parrish, Jacqueline E; Brock, Kenny V

    2014-12-01

    Immunosuppression caused by bovine viral diarrhea virus (BVDV) has been associated with lymphocyte depletion, leukopenia and impairment of leukocyte function; however, no work has been done on the relationship between BVDV and regulatory T lymphocytes (Tregs). The objective of this study was to compare the mRNA expression of genes associated with Tregs (CD25, FoxP3, CTLA4, and IDO), after experimental infection of beef calves with low (LV) or high (HV) virulence BVDV. Thirty BVDV-naïve calves were randomly assigned to three groups. Calves were intra-nasally inoculated with LV (n=10, strain SD-1) or HV (n=10, strain 1373) BVDV or BVDV-free cell culture medium (control, n=10). Quantitative RT-PCR was used to determine the expression of target genes in tracheo-bronchial lymph nodes and spleen on day 5 post-infection. The mRNA expression of CD25 was up-regulated in tracheo-bronchial lymph nodes of LV (Pviral strains, or differences in viral infectivity of the host cells.

  13. An Imbalance between Frequency of CD4+CD25+FOXP3+ Regulatory T Cells and CCR4+ and CCR9+ Circulating Helper T Cells Is Associated with Active Perennial Allergic Conjunctivitis

    Directory of Open Access Journals (Sweden)

    J. Galicia-Carreón

    2013-01-01

    Full Text Available Allergic conjunctivitis (AC is one of the most common eye disorders in ophthalmology. In mice models, it has been suggested that control of allergic conjunctivitis is a delicate balance between Tregs and inflammatory migrating effector cells. Our aim was to evaluate the frequency of Tregs and the frequency of homing receptors expressing cells in peripheral blood mononuclear cells (PBMC from patients with perennial allergic conjunctivitis (PAC. The analyses of phenotypic markers on CD4+ T cells and both soluble or intracellular cytokines were performed by flow cytometry. CD4+CD25+ cells were 15 times more frequent in PBMC from patients than HC; the vast majority of these CD4+CD25+ cells were FOXP3−, and most of CD4+ T cells were CCR4+ and CCR9+ cells. Upon allergen-stimulation, no significant changes were observed in frequency of Treg; however, an increased frequency of CD4+CCR4+CCR9+ cells, CD4+CD103+ cells and CD4+CD108+ cells with increased IL-5, IL-6, and IL-8 production was observed. These findings suggest an immune dysregulation in PAC, characterized by diminished frequency of Tregs and increased frequency of circulating activated CD4+ T cells; upon allergen-stimulation, these cells were expressing cell-surface molecules related to mucosa homing and were able to trigger an inflammatory microenvironment.

  14. Targeting of CD25 and glucocorticoid-induced TNF receptor family-related gene-expressing T cells differentially modulates asthma risk in offspring of asthmatic and normal mother mice.

    Science.gov (United States)

    Hubeau, Cedric; Apostolou, Irina; Kobzik, Lester

    2007-02-01

    Immunological mechanisms leading to increased asthma susceptibility in early life remain obscure. In this study, we examined the effects of neonatal Ab treatments targeting T cell populations on the development of an asthma syndrome. We used a model of increased asthma susceptibility where offspring of asthmatic BALB/c mother mice are more prone (than normal pups) to develop the disease. Neonatal pretreatment of naive pups with mAb directed against the IL-2Ralpha chain (CD25), the costimulatory molecule glucocorticoid-induced TNFR family related gene, and the inhibitory molecule CTLA-4 elicited contrasting effects in offspring depending on the mother's asthma status. Specifically, neonatal CD25(high) T cell depletion stimulated asthma susceptibility in normal offspring whereas it ameliorated the condition of pups born of asthmatic mothers. Conversely, glucocorticoid-induced TNFR family related gene ligation as a primary signal reduced the spleen cellularity and largely abrogated asthma susceptibility in asthma-prone offspring, without inducing disease in normal pups. Striking changes in Th1/Th2 cytokine levels, especially IL-4, followed mAb pretreatment and were consistent with the impact on asthma susceptibility. These results point to major differences in neonatal T cell population and responsiveness related to maternal asthma history. Interventions that temporarily remove and/or inactivate specific T cell subsets may therefore prove useful to attenuate early life asthma susceptibility and prevent the development of Th2-driven allergic airway disease.

  15. Abnormal T-cell reactivity against paternal antigens in spontaneous abortion: adoptive transfer of pregnancy-induced CD4+CD25+ T regulatory cells prevents fetal rejection in a murine abortion model.

    Science.gov (United States)

    Zenclussen, Ana Claudia; Gerlof, Katrin; Zenclussen, Maria Laura; Sollwedel, André; Bertoja, Annarosa Zambon; Ritter, Thomas; Kotsch, Katja; Leber, Joachim; Volk, Hans-Dieter

    2005-03-01

    Mammalian pregnancy is thought to be a state of immunological tolerance. The mechanisms underlying this phenomenon are still poorly understood. Here, we determined whether an inappropriate function of T regulatory (Treg) cells is involved in the pathogenesis of spontaneous abortion. We evaluated spleen and decidual lymphocytes from CBA/J mice undergoing immunological abortion (DBA/2J-mated) or having normal pregnancy (BALB/c-mated) on day 14 of gestation for ex vivo cytokine production after PMA or paternal antigen (alloantigen) stimulation. Treg activity was characterized by quantifying CD4(+)CD25(+) cells, foxp3 expression, and interleukin-10 secretion. Decidual lymphocytes from abortion CBA/J mice contained a significantly higher frequency of interferon-gamma-producing T cells specific for paternal antigens compared to those from normal pregnancy (7.8% versus 2.7%, P abortion mice. Very interestingly, CD4(+)CD25(+) Treg cells from normal pregnant and nonpregnant CBA/J mice could inhibit both proliferation and interferon-gamma secretion of lymphocytes from abortion mice in vitro whereas in vivo prevention of fetal rejection could only be achieved after adoptive transfer of Treg cells from normal pregnant mice. Our data suggest that pregnancy-induced Treg cells play a vital role in maternal tolerance to the allogeneic fetus.

  16. 全身照射预处理供体对异基因大鼠肝移植的作用及对受体内CD4~+CD25~+调节性T细胞的影响%Total body irradiation of the donor in a spontaneous tolerance rat liver transplantation model and the effects on CD4~+ CD25~+ regulatory T cells content

    Institute of Scientific and Technical Information of China (English)

    张业伟; 严栋梁; 卢思聪; 王学浩; 刘允怡

    2009-01-01

    Objective To study the effect of total body irradiation of the donor in a spontaneous tolerance rat liver transplantation model and the role of CD4~+ CD25~+ regulatory T cells on induction of immunotolerance in the recipient.Methods Liver transplantation was performed using male Lewis rats as donors and male DA rats as recipients.These rata were randomly allocated into the following groups:Control group,Homogeneity Liver Transplantation group,Idio-immunotolerance group and Acute Rejection group.After transplantation,survival time rate of each group were observed.Serum ALT,TB level,Foxp3~+ CD4~+ CD25~+ regulatory T cells,expression of GITR on T cell subgroup,histopathology of the hepatic graft on day 14,spleen CTL lytic activity on day 14 were measured.Results In the ldio-immnunotolerance group,the recipients suffered from transient rejection after surgery but acquired immunotolerance and survived long.In the Acute Rejection group,the donors were preconditioned with total body irradiation before liver transplantation.All recipients died between day 17 to 21.Serum ALT and TB increased significantly and the ratio of Foxp3~+ CD4~+ CD25~+ regulatory T cells decreased significantly compared with the Idioimmunotolerance group,the Homogeneity Liver Transplantation group and the Control group.The expression of GITR on CD3~+ CD4~+ T cells in the peripheral blood decreased.the expression of GITR on CD3~+ CD8~+ T cells and CTL lytic activity of the recipients increased by preconditioning of the donors with total body irradiation.Conclusions Preconditioning of the donors with total body irradiation eliminated the passenger lymphocytes of the liver graft,decreased the expression of Foxp3~+ CD4~+ CD25~+ regulatory T cells in peripheral blood,and increased the expression of GITR on CD3~+ CD8~+ T cells,thus affected the course of tolerance and induced acute rejection after liver transplantation.%目的 探讨全身照射(TBI)预处理诱导大鼠肝移植术后急性排

  17. 呼吸道合胞病毒毛细支气管炎患儿外周血CD4+CD25+调节性T细胞与Th17细胞功能变化及意义%Changes and the clinical significance of CD4+ CD25+ regulatory T cells and Th17 cells in peripheral blood of infants with respiratory syncytial virus bronchiolitis

    Institute of Scientific and Technical Information of China (English)

    李宾; 吴福玲; 冯学斌; 孙大康; 崔晴晴; 赵志旭

    2012-01-01

    AIM: To observe the percentages of CD4 + CD25+ regulatory T cells (Tregs) and Thl7 cells and the levels of IL-10, TGF-p and IL-17 in peripheral blood of infants with respiratory syncytial virus ( RSV) bronchiolitis. The relationship between above cells, cytokines and RSV bronchiolitis was determined. METHODS: Thirty-three infants with RSV bronchiolitis, twenty-eight infants with non-RSV pneumonia and twenty-six healthy infants were enrolled. The percentages of Tregs and Thl7 cells in peripheral blood were detected by flow cytometer (FCM), and the levels of IL-10, TGF-pand IL-17 in plasma were determined by ELISA. RESULTS: The percentage of Tregs and the levels of IL-10 and TGF-pin infants with RSV bronchiolitis were significantly lower than those in infants with non-RSV pneumonia and healthy infants ( P < 0. 05), while the percentage of Thl7 cells and the level of IL-17 in infants with RSV bronchiolitis were significantly higher than those in infants with non-RSV pneumonia and healthy infants ( P < 0. 05). CONCLUSION: The imbalance between Tregs and Thl7 cells in peripheral blood of infants with RSV bronchiolitis may be one of the pathogenesis of RSV bronchiolitis.%目的:探讨呼吸道合胞病毒(RSV)毛细支气管炎患儿外周血CD4+ CD25+调节性T细胞和Th17细胞及其分泌细胞因子IL-10、TGF-β、IL-17水平变化与RSV毛细支气管炎发病的关系.方法:收集2010-09/2011-04在滨州医学院附属医院儿科住院的33例RSV毛细支气管炎患儿、28例做为阳性对照的非RSV感染性肺炎患儿(肺炎组)及26例正常对照组的健康体检儿外周血,采用流式细胞术(FCM)检测外周血CD4+ CD25+调节性T细胞、Th17细胞百分率,酶联免疫吸附(ELISA)法检测血浆IL-10、TGF-β、IL-17的水平.结果:RSV毛细支气管炎患儿外周血CD4+ CD25+调节性T细胞、IL-10、TGF-β水平显著低于肺炎患儿及健康体检儿(P<0.05),而Th17、IL-17水平则显著高于肺炎患儿与健康体检儿(P<0

  18. Characterization of immobilization methods of antiviral antibodies in serum for electrochemical biosensors

    Energy Technology Data Exchange (ETDEWEB)

    Huy, Tran Quang, E-mail: huytq@nihe.org.vn [National Institute of Hygiene and Epidemiology (NIHE), No1 Yersin St., Hanoi (Viet Nam); International Training Institute for Materials Science (ITIMS), Hanoi University of Science and Technology (HUST), No1 Dai Co Viet, Hanoi (Viet Nam); Hanh, Nguyen Thi Hong; Van Chung, Pham; Anh, Dang Duc; Nga, Phan Thi [National Institute of Hygiene and Epidemiology (NIHE), No1 Yersin St., Hanoi (Viet Nam); Tuan, Mai Anh, E-mail: tuanma-itims@mail.hut.edu.vn [International Training Institute for Materials Science (ITIMS), Hanoi University of Science and Technology (HUST), No1 Dai Co Viet, Hanoi (Viet Nam)

    2011-06-01

    In this paper, we describes different methods to immobilize Japanese encephalitis virus (JEV) antibodies in human serum onto the interdigitated surface of a microelectrode sensor for optimizing electrochemical detection: (1) direct covalent binding to the silanized surface, (2) binding to the silanized surface via a cross-linker of glutaraldehyde (GA), (3) binding to glutaraldehyde/silanized surface via goat anti-human IgG polyclonal antibody and (4) binding to glutaraldehyde/silanized surface via protein A (PrA). Field emission scanning electron microscopy, Fourier transform infrared spectrometry, and fluorescence microscopy are used to verify the characteristics of antibodies on the interdigitated surface after the serum antibodies immobilization. The analyzed results indicate that the use of protein A is an effective choice for immobilization and orientation of antibodies in serum for electrochemical biosensors. This study provides an advantageous immobilization method of serum containing antiviral antibodies to develop electrochemical biosensors for preliminary screening of viruses in clinical samples from outbreaks.

  19. Fabrication of Anti-human Cardiac Troponin I Immunogold Nanorods for Sensing Acute Myocardial Damage

    Science.gov (United States)

    Guo, Z. R.; Gu, C. R.; Fan, X.; Bian, Z. P.; Wu, H. F.; Yang, D.; Gu, N.; Zhang, J. N.

    2009-12-01

    A facile, rapid, solution-phase method of detecting human cardiac troponin I for sensing myocardial damage has been described using gold nanorods-based biosensors. The sensing is demonstrated by the distinct change of the longitudinal surface plasmon resonance wavelength of the gold nanorods to specific antibody-antigen binding events. For a higher sensitivity, the aspect ratio of gold nanorods is increased up to ca 5.5 by simply adding small amount of HCl in seed-mediated growth solution. Experimental results show that the detecting limit of the present method is 10 ng/mL. Contrast tests reveal that these gold nanorods-based plasmonic biosensors hold much higher sensitivity than that of conventionally spherical gold nanoparticles.

  20. A glucocorticoid amplifies IL-2-induced selective expansion of CD4+CD25+FOXP3+ regulatory T cells in vivo and suppresses graft-versus-host disease after allogeneic lymphocyte transplantation

    Institute of Scientific and Technical Information of China (English)

    Yanhui Xie; Min Wu; Runhua Song; Jiexian Ma; Yi Shi; Wenming Qin; Youxin Jin

    2009-01-01

    Regulatory T (Treg) cells are a subpopulation of T cells that not only prevent autoimmunity, but also control a wide range of T cell-dependent immune responses. Glucocorticoid treatment (dexamethasone, or Dex) has been reported to amplify IL-2-mediated selective in vivo expansion of Treg cells. We simultaneously adminis-tered Dex and IL-2 to the donor in a murine allogeneic lymphocyte transplantation model to expand functional suppressive CD4+CD25+FOXP3+ T cells in the graft and to raise the regulatory T cell/effector T cell (Treg/ Teff) ratio to prevent graft-versus-host disease (GVHD). After combined treatment of the donor with Dex (5 mg/kg/day) and IL-2 (300,000 IU/mouse/day) for 3 days, grafts were subjected to flow cytometric analy-sis, and transplantation was carried out from male C57BL/6 mice to female BALB/c mice aged 8-12 weeks. Results showed that short-term simultaneous administration of Dex and IL-2 markedly expanded functional suppressive CD4+CD25+FOXP3+ T cells in the murine spleen. In this murine allogeneic transplan-tation model, the grafts from donors with Dex and IL-2 pre-treatment led to a longer survival time for the reci-pients than for the control group (median survival time> 60 day vs. 12 day, P = 0.0002). The ratio of Treg/Teff also increased remarkably (0.43 ±0.15 vs. 0.14± 0.01, P=0.01). This study demonstrated that co-stimulation with Dex and IL-2 selectively expanded functional CD4+CD25+FOXP3+ T cells in vivo, and that grafts from donors pre-treated with Dex and IL-2 led to longer survival time and greater suppression of GVHD after allogeneic transplantation. Thus, GVHD can be suppressed by the specific expansion of regulatory T cells with Dex and IL-2 in graft donors.

  1. 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.

  2. Thyroid Antibodies

    Science.gov (United States)

    ... e.g., at regular intervals after thyroid cancer treatment) Thyroid stimulating hormone receptor antibody, Thyroid Stimulating Immunoglobulin TRAb, TSHR Ab, TSI Graves disease When a person has symptoms of hyperthyroidism If a pregnant woman has a known autoimmune ...

  3. An electrochemical quartz crystal impedance study on anti-human immunoglobulin G immobilization in the polymer grown during dopamine oxidation at an Au electrode.

    Science.gov (United States)

    He, Hua; Xie, Qingji; Yao, Shouzhuo

    2005-09-15

    The polymeric film grown during dopamine oxidation at an Au electrode was studied as a novel matrix for immobilizing anti-human immunoglobulin G (IgG) via the electrochemical quartz crystal impedance analysis (EQCIA) method. The growth of the polymeric films at Au electrodes during dopamine oxidation in neutral phosphate buffer (pH 7.4) and the immobilization of anti-human IgG into the polymeric films during their growth have been traced at real time. Lysozyme control experiments suggested that anti-human IgG was electrostatically incorporated into the polymeric film. Also, the porosity of the polymeric films has been discussed by measuring the "wet" and "dry" frequency shifts. Compared with a polypyrrole film immobilized with anti-human IgG, the proposed matrix possessed a larger amount of specific binding sites for human IgG by subsequent immunoreaction tests. The association constant of the anti-human IgG immunoreaction was obtained with satisfactory results.

  4. Design of indirect solid-phase immunosorbent methods for detecting arenavirus antigens and antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Ivanov, A.P.; Rezapkin, G.V.; Dzagurova, T.K.; Tkachenko, E.A.

    1984-05-01

    Specifications have been elaborated for formulating indirect solid-phase enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (SPRIA) methods that employ anti-human and anti-mice G class immunoglobulin (IgG), conjugated with horseradish peroxidase and /sup 125/I for detecting the arenaviruses Junin, Machupo, Tacaribe, Amalpari, Tamiami, Lassa, and LCM (lymphocytic choriomeningitis). These methods make it possible to identify with a high degree of sensitivity arenavirus antigens and antibodies in various kinds of material.

  5. Ascaris lumbricoides-induced suppression of total and specific IgE responses in atopic subjects is interleukin 10-independent and associated with an increase of CD25(+) cells.

    Science.gov (United States)

    Matera, Giovanni; Giancotti, Aida; Scalise, Sonia; Pulicari, Maria Concetta; Maselli, Rosario; Piizzi, Chiara; Pelaia, Girolamo; Tancrè, Valentina; Muto, Valentina; Doldo, Patrizia; Cosco, Vincenzo; Cosimo, Paola; Capicotto, Renata; Quirino, Angela; Scalzo, Rosaria; Liberto, Maria Carla; Parlato, Giuseppe; Focà, Alfredo

    2008-11-01

    Ascaris presence in humans has been associated with high levels of blood eosinophils and serum IgE. This study was designed to address the influence of Ascaris infection on allergic and inflammatory parameters of atopic subjects. A cross-sectional design was used, and atopic individuals to be assessed were divided into 3 groups including Ascaris-infected, anti-Ascaris IgG-positive (seropositive), and control subjects. All subjects enrolled had positive skin test reactivity to at least 1 perennial or seasonal allergen; however, levels of C-reactive protein, C3, and C4 were within normal range values. Eosinophil percentage was not significantly different among the groups studied. Total IgE and specific anti-Ascaris IgE levels in the seropositive group were significantly higher than concentrations found in both control and infected groups. Interleukin (IL)-4 release in Ascaris-infected patients was significantly increased versus seropositives, who were able to produce more IL-4 than controls. The levels of IL-10 were lower in the seropositives as well as infected subjects in comparison with controls. CD25(+) lymphocyte populations were significantly increased in the infected group versus the seropositives as well as the controls. Lung function tests of some selected seropositive subjects were significantly impaired. The same parameters of a representative infected patient were not different from controls. Our data on T helper type 2 cells (Th2) and regulatory T cells (Treg) features, as well as CD25(+) lymphocyte increase, suggest an Ascaris-induced mechanism leading to parasite survival. Moreover, the stable control of both T helper type 1 cells (Th1) and Th2 immunity cascades, paralleled by the absence of overwhelming inflammatory systemic reactions and lack of allergic syndromes, may result in a favorable host condition.

  6. CD4+CD25hiFOXP3+ cells in cord blood of neonates born from filaria infected mother are negatively associated with CD4+Tbet+ and CD4+RORγt+ T cells.

    Directory of Open Access Journals (Sweden)

    Ulysse Ateba-Ngoa

    Full Text Available BACKGROUND: Children who have been exposed in utero to maternal filarial infection are immunologically less responsive to filarial antigens, have less pathology, and are more susceptible to acquire infection than offspring of uninfected mothers. Moreover children from filaria infected mothers have been shown to be less responsive to vaccination as a consequence of an impairment of their immune response. However, it is not well known how in utero exposure to parasite antigens affects cellular immune responses. METHODOLOGY: Here, 30 pregnant women were examined for the presence of microfilaria of Loa loa and Mansonella perstans in peripheral blood. At delivery, cord blood mononuclear cells (CBMC were obtained and the CD4+T cells were phenotyped by expression of the transcription factors Tbet, RORγt, and FOXP3. RESULTS: No significant difference was observed between newborns from infected versus uninfected mothers in the frequencies of total CD4+T cells and CD4+T cells subsets including CD4+Tbet+, CD4+RORγt+ T and CD4+CD25hiFOXP3+ T cells. However, there was a negative association between CD4+CD25hiFOXP3+T cells and CD4+Tbet+ as well as CD4+RORγt+ T cells in the infected group only (B = -0.242, P = 0.002; B = -0.178, P = 0.013 respectively. CONCLUSION: Our results suggest that filarial infection during pregnancy leads to an expansion of functionally active regulatory T cells that keep TH1 and TH17 in check.

  7. Understanding ForteBio's Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant.

    Science.gov (United States)

    Yu, Yao; Mitchell, Scott; Lynaugh, Heather; Brown, Michael; Nobrega, R Paul; Zhi, Xiaoyong; Sun, Tingwan; Caffry, Isabelle; Cao, Yuan; Yang, Rong; Burnina, Irina; Xu, Yingda; Estep, Patricia

    2016-01-01

    Real-time and label-free antibody screening systems are becoming more popular because of the increasing output of purified antibodies and antibody supernatant from many antibody discovery platforms. However, the properties of the biosensor can greatly affect the kinetic and epitope binning results generated by these label-free screening systems. ForteBio human-specific ProA, anti-human IgG quantitation (AHQ), anti-human Fc capture (AHC) sensors, and custom biotinylated-anti-human Fc capture (b-AHFc) sensors were evaluated in terms of loading ability, regeneration, kinetic characterization, and epitope binning with both purified IgG and IgG supernatant. AHC sensors proved unreliable for kinetic or binning assays at times, whereas AHQ sensors showed poor loading and regeneration abilities. ProA sensors worked well with both purified IgG and IgG supernatant. However, the interaction between ProA sensors and the Fab region of the IgG with VH3 germline limited the application of ProA sensors, especially in the epitope binning experiment. In an attempt to generate a biosensor type that would be compatible with a variety of germlines and sample types, we found that the custom b-AHFc sensors appeared to be robust working with both purified IgG and IgG supernatant, with little evidence of sensor-related artifacts.

  8. Antiparietal cell antibody test

    Science.gov (United States)

    APCA; Anti-gastric parietal cell antibody; Atrophic gastritis - anti-gastric parietal cell antibody; Gastric ulcer - anti-gastric parietal cell antibody; Pernicious anemia - anti-gastric parietal cell antibody; ...

  9. 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.

  10. NY-ESO-1-specific circulating CD4+ T cells in ovarian cancer patients are prevalently T(H)1 type cells undetectable in the CD25+ FOXP3+ Treg compartment.

    Science.gov (United States)

    Redjimi, Nassima; Duperrier-Amouriaux, Karine; Raimbaud, Isabelle; Luescher, Immanuel; Dojcinovic, Danijel; Classe, Jean-Marc; Berton-Rigaud, Dominique; Frenel, Jean-Sébastien; Bourbouloux, Emmanuelle; Valmori, Danila; Ayyoub, Maha

    2011-01-01

    Spontaneous CD4(+) T-cell responses to the tumor-specific antigen NY-ESO-1 (ESO) are frequently found in patients with epithelial ovarian cancer (EOC). If these responses are of effector or/and Treg type, however, has remained unclear. Here, we have used functional approaches together with recently developed MHC class II/ESO tetramers to assess the frequency, phenotype and function of ESO-specific cells in circulating lymphocytes from EOC patients. We found that circulating ESO-specific CD4(+) T cells in EOC patients with spontaneous immune responses to the antigen are prevalently T(H)1 type cells secreting IFN-γ but no IL-17 or IL-10 and are not suppressive. We detected tetramer(+) cells ex vivo, at an average frequency of 1:25,000 memory cells, that is, significantly lower than in patients immunized with an ESO vaccine. ESO tetramer(+) cells were mostly effector memory cells at advanced stages of differentiation and were not detected in circulating CD25(+)FOXP3(+)Treg. Thus, spontaneous CD4(+) T-cell responses to ESO in cancer patients are prevalently of T(H)1 type and not Treg. Their relatively low frequency and advanced differentiation stage, however, may limit their efficacy, that may be boosted by immunogenic ESO vaccines.

  11. 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.

  12. Linfócitos T CD4+CD25+ e a regulação do sistema imunológico: perspectivas para o entendimento fisiopatológico da sepse

    Directory of Open Access Journals (Sweden)

    Rodrigo Siqueira-Batista

    2012-09-01

    Full Text Available A resposta inflamatória sistêmica representa o evento patogênico central da sepse, subjazendo às manifestações clínicas e aos achados laboratoriais presentes nos enfermos. Inúmeras pesquisas têm demonstrado que os linfócitos T CD4+CD25+ - também conhecidos como células T reguladoras (Treg - participam dos processos de desenvolvimento da sepse, em virtude de sua capacidade de suprimir a resposta imune. Com base nessas ideias, propôs-se, no presente artigo, a discussão do papel dos linfócitos Treg na sepse, com base na revisão da literatura com estratégia de busca definida (LILACS, PubMed e SciELO, tendo em vista duas abordagens principais: a participação dessas células nos processos de inflamação e imunidade, e as perspectivas de investigação fisiopatológica computacional da condição mórbida.

  13. The ACE inhibitors enalapril and captopril modulate cytokine responses in Balb/c and C57Bl/6 normal mice and increase CD4(+)CD103(+)CD25(negative) splenic T cell numbers.

    Science.gov (United States)

    Albuquerque, Deijanira; Nihei, Jorge; Cardillo, Fabíola; Singh, Ram

    2010-01-01

    Increasing evidence implies beneficial effects of angiotensin-converting enzyme (ACE) inhibitors beyond those of their original indications to control hypertension. One of the most attractive non-hemodynamic properties of ACE inhibitors is their ability to regulate cytokine production. The mechanism(s) underlying the role of ACE inhibitors on cytokine synthesis are not well understood but they have traditionally been attributed to the inhibition of angiotensin (Ang) II formation. In fact, it has been extensively demonstrated that ACE inhibitors decrease Ang II-induced production of proinflammatory cytokines and chemokines. However, it is not well described if inhibition of endogenous Ang II generation by ACE inhibitors modulates systemic cytokine production in mice. To verify that, in this work, we investigated the effects of treatment with the ACE inhibitors enalapril and captopril on cytokine synthesis in C57Bl/6 and Balb/c mice. Our results show that enalapril up regulates IL-10 produced by splenocytes from Balb/c and C57Bl/6 mice and captopril increased it only in Balb/c mice. Furthermore, CD4(+)CD103(+) presented increased IL-10 production after enalapril treatment. Enalapril as well as captopril short-term treatment enhanced IL-2 synthesis in Balb/c mice. Besides, enhanced IL-2 and IL-10 levels correlates with increased CD4(+)CD103(+)CD25(negative) T cells numbers in spleens from enalapril-treated mice.

  14. Adalimumab specifically induces CD3+ CD4+ CD25high Foxp3+ CD127− T-regulatory cells and decreases vascular endothelial growth factor plasma levels in refractory immuno-mediated uveitis: a non-randomized pilot intervention study

    Science.gov (United States)

    Calleja, S; Cordero-Coma, M; Rodriguez, E; Llorente, M; Franco, M; Ruiz de Morales, J G

    2012-01-01

    Aim To explore immunoregulatory and anti-inflammatory pathways specifically targeted by a subcutaneous anti-TNFαdrug—adalimumab—which might be relevant for controlling refractory uveitis. Design Non-randomized pilot intervention study on the effects of adalimumab on Treg populations and plasma VEGF levels in refractory uveitis patients. Inflammatory and immunological parameters were measured in 12 patients before therapy, and 1 and 6 months after therapy, and analyzed in the context of ophthalmological outcomes. The results were compared with those obtained in 10 systemic prednisone-treated uveitis patients. Results After 1 month of treatment, all patients responded, with 67% of adalimumab group and 80% of the corticosteroid group achieving inactivity (P=0.5). Unlike steroid-treated patients, a significant increase in T-regulatory CD4+ CD25high Foxp3+ CD127− cells was observed in adalimumab patients after 1 month of treatment, and maintained after 6 months (P=0.003). A significant adalimumab-specific drop in plasma VEGF was observed after 1 and 6 months of treatment (P=0.019). In every single patient, Tregs but not VEGF correlated with disease activity. Conclusions In refractory uveitis patients treated with adalimumab, clinical efficacy may be mediated through upregulation of Tregs in addition to modulation of VEGF-mediated inflammatory pathways. These biological properties, which were not observed in patients treated with corticosteroids, may reflect the specificity of TNF-αtargeting. PMID:22222264

  15. Antibody-protein A conjugated quantum dots for multiplexed imaging of surface receptors in living cells.

    Science.gov (United States)

    Jin, Takashi; Tiwari, Dhermendra K; Tanaka, Shin-Ichi; Inouye, Yasushi; Yoshizawa, Keiko; Watanabe, Tomonobu M

    2010-11-01

    To use quantum dots (QDs) as fluorescent probes for receptor imaging, QD surface should be modified with biomolecules such as antibodies, peptides, carbohydrates, and small-molecule ligands for receptors. Among these QDs, antibody conjugated QDs are the most promising fluorescent probes. There are many kinds of coupling reactions that can be used for preparing antibody conjugated QDs. Most of the antibody coupling reactions, however, are non-selective and time-consuming. In this paper, we report a facile method for preparing antibody conjugated QDs for surface receptor imaging. We used ProteinA as an adaptor protein for binding of antibody to QDs. By using ProteinA conjugated QDs, various types of antibodies are easily attached to the surface of the QDs via non-covalent binding between the F(c) (fragment crystallization) region of antibody and ProteinA. To show the utility of ProteinA conjugated QDs, HER2 (anti-human epidermal growth factor receptor 2) in KPL-4 human breast cancer cells were stained by using anti-HER2 antibody conjugated ProteinA-QDs. In addition, multiplexed imaging of HER2 and CXCR4 (chemokine receptor) in the KPL-4 cells was performed. The result showed that CXCR4 receptors coexist with HER2 receptors in the membrane surface of KPL-4 cells. ProteinA mediated antibody conjugation to QDs is very useful to prepare fluorescent probes for multiplexed imaging of surface receptors in living cells.

  16. Effect of cell-surface components and metabolites of lactic acid bacteria and probiotic organisms on cytokine production and induction of CD25 expression in human peripheral mononuclear cells.

    Science.gov (United States)

    Ashraf, R; Vasiljevic, T; Smith, S C; Donkor, O N

    2014-05-01

    In the current study, the relative contribution of cell-surface components (CSC) and cell-free supernatants (CFS) in the immuno-modulatory properties of 17 strains of probiotic and lactic acid bacteria (LAB) was assessed. The production of pro- and antiinflammatory cytokines including IL-2, IL-4, IL-10, IL-12 p70, IFN-γ, tumor necrosis factor-α (TNF-α), and transforming growth factor-β was measured at different time points after stimulation of buffy coat derived-peripheral blood mononuclear cells (PBMC) from healthy donors with CSC and CFS of probiotic and LAB. Results showed that CSC of probiotic and LAB strains induced production of T helper 1 and 2 type cytokines. Transforming growth factor-β was stimulated at highest concentrations, followed by IL-10 and TNF-α. The CFS of all tested bacterial strains induced PBMC for significantly high levels of IL-10 secretion compared with unstimulated cells, but the values were less than lipopolysaccharide-stimulated cells. Cytokines due to CFS stimulation showed declined concentration for IL-2, TNF-α, and IL-4, and complete disappearance of IL-12, IFN-γ, and transforming growth factor-β in the cultured medium at 96 h of incubation. Results of cytokine data demonstrate proinflammatory TNF-α immune responses are mainly directed through cell-surface structures of probiotic and LAB, but antiinflammatory immune responses are mediated both by metabolites and cell-surfaces of these bacteria. The induction of CD4(+)CD25(+) regulatory T cells after stimulation of PBMC with CSC and CFS of probiotic and LAB showed regulatory T cell activity appeared to be influenced both by the CSC and metabolites, but was principally triggered by cell surfaces of probiotic and LAB strains.

  17. Early changes of CD4+CD25+Foxp3+ regulatory T cells and Th1/Th2, Tc1/Tc2 profiles in the peripheral blood of rats with controlled hemorrhagic shock and no fluid resuscitation

    Institute of Scientific and Technical Information of China (English)

    ZHANG Qin; LU Yuan-qiang; JIANG Jiu-kun; GU Lin-hui; MOU Han-zhou

    2012-01-01

    Background Hemorrhagic shock induces immune dysfunction.Regulatory T cells (Tregs),T-helper (Th) cells,and cytotoxic T-lymphocytes (CTLs) can execute many crucial actions in immune and inflammatory responses.This study was conducted to investigate the early pathophysiological changes of CD4+CD25+Foxp3+ Treg and Th1/Th2,Tc1/Tc2profiles in the peripheral blood of rats with controlled hemorrhagic shock and no fluid resuscitation. Methods A rat model of controlled hemorrhagic shock with no fluid resuscitation was established.Peripheral blood samples were taken before and four hours after hemorrhagic shock with no fluid resuscitation.Three color flow cytometry was used to detect Tregs,Th1,Th2,Tc1 and Tc2 cells in the samples.Results In the peripheral blood of rats,the percentage of Tregs four hours after hemorrhagic shock was significantly lower than before hemorrhagic shock (P=0.001).The ratios of Th1/Th2 and Tc1/Tc2 were changed from (23.08±8.98)%to (23.91±15.36)%,and from (40.40:1:21.56)% to (65.48±23.88)%,respectively.Conclusions At an eady stage,the advent of hemorrhagic shock is related to an early decrease of Tregs,and a mild shift in the Th1/Th2,Tc1/Tc2 balance toward Th1 and Tc1 dominance.These changes are part of a hyper-inflammatory state of the host,and will deteriorate the maintenance of immune balance.Further influences and detailed mechanisms need to be investigated.

  18. Cross-reactivity of human and bovine antibodies in striped dolphin paraffin wax-embedded tissues.

    Science.gov (United States)

    Jaber, J R; Fernández, A; Herráez, P; Espinosa de los Monteros, A; Ramírez, G A; García, P M; Fernández, T; Arbelo, M; Pérez, J

    2003-11-15

    This study evaluates the cross-reactivity of seven anti-human and one anti-bovine antibodies in formalin-fixed, paraffin-embedded tissue samples of liver and mesenteric lymph nodes of 13 striped dolphins (Stenella coeruleoalba). Four antibodies (CD3, IgG, lysozyme and S100 protein) reacted with striped dolphin lymph nodes in a similar pattern to that observed in the species of origin. The anti-human MHC class II mAb reacted strongly with macrophages and dendritic-like cells of striped dolphins, whereas a small number of lymphocytes were labelled with this antibody. These antibodies were used to study the immunophenotype of the inflammatory infiltrated in non-specific chronic reactive hepatitis (eight cases) and chronic parasite cholangitis (two cases) and normal liver (three cases) of striped dolphins. Non-specific chronic reactive hepatitis was composed of inflammatory infiltration of CD3+ T lymphocytes and IgG+ plasma cells in portal spaces and hepatic sinusoids. Lymphonodular aggregates observed in chronic parasitic cholangitis showed a cellular distribution similar to that found in lymph node cortex, including the presence of S100+ and MHC class II+ dendritic-like cells in lymphoid follicles and interfollicular areas. This result suggests that those inflammatory infiltrates are highly organised to enhance antigen presentation to B and T cells.

  19. Monoclonal Antibody Therapy and Renal Transplantation: Focus on Adverse Effects

    Directory of Open Access Journals (Sweden)

    Gianluigi Zaza

    2014-02-01

    Full Text Available A series of monoclonal antibodies (mAbs are commonly utilized in renal transplantation as induction therapy (a period of intense immunosuppression immediately before and following the implant of the allograft, to treat steroid-resistant acute rejections, to decrease the incidence and mitigate effects of delayed graft function, and to allow immunosuppressive minimization. Additionally, in the last few years, their use has been proposed for the treatment of chronic antibody-mediated rejection, a major cause of late renal allograft loss. Although the exact mechanism of immunosuppression and allograft tolerance with any of the currently used induction agents is not completely defined, the majority of these medications are targeted against specific CD proteins on the T or B cells surface (e.g., CD3, CD25, CD52. Moreover, some of them have different mechanisms of action. In particular, eculizumab, interrupting the complement pathway, is a new promising treatment tool for acute graft complications and for post-transplant hemolytic uremic syndrome. While it is clear their utility in renal transplantation, it is also unquestionable that by using these highly potent immunosuppressive agents, the body loses much of its innate ability to mount an adequate immune response, thereby increasing the risk of severe adverse effects (e.g., infections, malignancies, haematological complications. Therefore, it is extremely important for clinicians involved in renal transplantation to know the potential side effects of monoclonal antibodies in order to plan a correct therapeutic strategy minimizing/avoiding the onset and development of severe clinical complications.

  20. [Antinuclear antibodies].

    Science.gov (United States)

    Cabiedes, Javier; Núñez-Álvarez, Carlos A

    2010-01-01

    Anti-nuclear antibodies (ANA) are immunoglobulin directed against autologous cell nuclear and cytoplasmic components. Besides the autoimmune ANA there are other ANA that can be detected in circulation, like natural and infectious ANA. Because of its high sensibility, detection of the ANA must be done by indirect immunofluorescence (IIF) as screening test and all of those positive samples are convenient to confirm its specificity by ELISA, western blot or other techniques. Positive ANA detected by IIF must be evaluated taking in to account the pattern and titer. The following recommended step is the specificity characterization (reactivity against extractable nuclear antigens [ENA], dsDNA, etc.) which is useful for the diagnosis and follow up of patients with autoimmune diseases, and by such reasoning, its detection must be performed in an orderly and reasonable way using guides or strategies focused to the good use and interpretation of the autoantibodies. The objective of this review is to present a compilation of the literature and our experience in the detection and study of the ANA.

  1. The role of regulatory T lymphocyte in human immunodeficiency syndrome%CD4+CD25+调节性T细胞与人类获得性免疫缺陷病毒感染关系的研究进展

    Institute of Scientific and Technical Information of China (English)

    贺琼; 徐赫; 杜桂书; 邓路瑶; 吴利洋; 徐兰; 王月丹

    2007-01-01

    CD4+CD25+调节性T细胞,是调节性T细胞中功能最重要的一类.它是一类具有特殊免疫调节功能的T细胞亚群.它能够抑制自身免疫病的发生和发展,参与肿瘤免疫的调节,同时在感染和移植免疫中也发挥着极其重要的作用.T细胞的这一亚群具有免疫调节和免疫抑制的特性,新近发现它亦与艾滋病的发生、发展关系密切.HIV进入人体后,CD4+CD25+调节性T细胞抑制了机体的免疫效应但它也同时被感染,最终由于细胞毒的作用而死亡.由于调节性T细胞数量的减少不能有效地发挥其抑制作用,HIV持续的过度活化使得T细胞逐渐耗竭说明在HIV发生、发展的不同阶段Treg细胞可能都发挥了免疫抑制作用.但是却对HIV感染与艾滋病发病的进程产生了不同的效应.此外,CD4+CD25+调节性T细胞还与HIV病毒的持续存在密切相关.本文就CD4+CD25+调节性T细胞与人类获得性免疫缺陷病毒(HIV)感染之间关系进行初步的探讨.

  2. A novel antibody humanization method based on epitopes scanning and molecular dynamics simulation.

    Directory of Open Access Journals (Sweden)

    Ding Zhang

    Full Text Available 1-17-2 is a rat anti-human DEC-205 monoclonal antibody that induces internalization and delivers antigen to dendritic cells (DCs. The potentially clinical application of this antibody is limited by its murine origin. Traditional humanization method such as complementarity determining regions (CDRs graft often leads to a decreased or even lost affinity. Here we have developed a novel antibody humanization method based on computer modeling and bioinformatics analysis. First, we used homology modeling technology to build the precise model of Fab. A novel epitope scanning algorithm was designed to identify antigenic residues in the framework regions (FRs that need to be mutated to human counterpart in the humanization process. Then virtual mutation and molecular dynamics (MD simulation were used to assess the conformational impact imposed by all the mutations. By comparing the root-mean-square deviations (RMSDs of CDRs, we found five key residues whose mutations would destroy the original conformation of CDRs. These residues need to be back-mutated to rescue the antibody binding affinity. Finally we constructed the antibodies in vitro and compared their binding affinity by flow cytometry and surface plasmon resonance (SPR assay. The binding affinity of the refined humanized antibody was similar to that of the original rat antibody. Our results have established a novel method based on epitopes scanning and MD simulation for antibody humanization.

  3. A Unique Report: Development of Super Anti-Human IgG Monoclone with Optical Density Over Than 3

    Directory of Open Access Journals (Sweden)

    Leili Aghebati Maleki

    2013-08-01

    Full Text Available Purpose: Monoclonal antibodies and related conjugates are key reagents used in biomedical researches as well as, in treatment, purification and diagnosis of infectious and non- infectious diseases. Methods: Balb/c mice were immunized with purified human IgG. Spleen cells of the most immune mouse were fused with SP2/0 in the presence of Poly Ethylene Glycol (PEG. Supernatant of hybridoma cells was screened for detection of antibody by ELISA. Then, the sample was assessed for cross-reactivity with IgM & IgA by ELISA and confirmed by immunoblotting. The subclasses of the selected mAbs were determined. The best clone was injected intraperitoneally to some pristane-injected mice. Anti-IgG mAb was purified from the animals' ascitic fluid by Ion exchange chromatography and then, mAb was conjugated with HRP. Results: In the present study, over than 50 clones were obtained that 1 clone had optical density over than 3. We named this clone as supermonoclone which was selected for limiting dilution. The result of the immunoblotting, showed sharp band in IgG position and did not show any band in IgM&IgA position. Conclusion: Based on the findings of this study, the conjugated monoclonal antibody could have application in diagnosis of infectious diseases like Toxoplasmosis, Rubella and IgG class of other infectious and non- infectious diseases.

  4. Detection of specific IgE antibodies in parasite diseases

    Directory of Open Access Journals (Sweden)

    Souza-Atta M.L.B.

    1999-01-01

    Full Text Available Activation of Th1 or Th2 cells is associated with production of specific immunoglobulin isotypes, offering the opportunity to use antibody measurement for evaluation of T cell function. Schistosomiasis and visceral leishmaniasis are diseases associated with Th2 activation. However, an IgE response is not always detected in these patients. In the present study we evaluated specific IgE antibodies to S. mansoni and L. chagasi antigens by ELISA after depletion of serum IgG with protein G immobilized on Sepharose beads or RF-absorbent (purified sheep IgG antibodies anti-human IgG. In schistosomiasis patients, specific IgE to SWAP antigen was demonstrable in only 10 of 21 patients (48% (mean absorbance ± SD = 0.102 ± 0.195 when unabsorbed serum was used. Depletion of IgG with protein G increased the number of specific IgE-positive tests to 13 (62% and the use of RF-absorbent increased the number of positive results to 20 (95% (mean absorbances ± SD = 0.303 ± 0.455 and 0.374 ± 0.477, respectively. Specific IgE anti-L. chagasi antibodies were not detected in unabsorbed serum from visceral leishmaniasis patients. When IgG was depleted with protein G, IgE antibodies were detected in only 3 (11% of 27 patients, and the use of RF-absorbent permitted the detection of this isotype in all 27 visceral leishmaniasis sera tested (mean absorbance ± SD = 0.104 ± 0.03. These data show that the presence of IgG antibodies may prevent the detection of a specific IgE response in these parasite diseases. RF-absorbent, a reagent that blocks IgG-binding sites and also removes rheumatoid factor, was more efficient than protein G for the demonstration of specific IgE antibodies.

  5. Platelet antibody detection by flow cytometry: an effective method to evaluate and give transfusional support in platelet refractoriness

    Directory of Open Access Journals (Sweden)

    Carolina Bonet Bub

    2013-01-01

    Full Text Available BACKGROUND: Immune platelet refractoriness is mainly caused by human leukocyte antigen antibodies (80-90% of cases and, to a lesser extent, by human platelet antigen antibodies. Refractoriness can be diagnosed by laboratory tests and patients should receive compatible platelet transfusions. A fast, effective and low cost antibody-screening method which detects platelet human leukocyte/platelet antigen antibodies is essential in the management of immune platelet refractoriness. OBJECTIVE: The aim of this study was to evaluate the efficiency of the flow cytometry platelet immunofluorescence test to screen for immune platelet refractoriness. METHODS: A group of prospective hematologic patients with clinically suspected platelet refractoriness treated in a referral center in Campinas, SP during July 2006 and July 2011 was enrolled in this study. Platelet antibodies were screened using the flow cytometry platelet immunofluorescence test. Anti-human leukocyte antigen antibodies were detected by commercially available methods. The sensitivity, specificity and predictive values of the immunofluorescence test were determined taking into account that the majority of antiplatelet antibodies presented human leukocyte antigen specificity. RESULTS: Seventy-six samples from 32 female and 38 male patients with a median age of 43.5 years (range: 5-84 years were analyzed. The sensitivity of the test was 86.11% and specificity 75.00% with a positive predictive value of 75.61% and a negative predictive value of 85.71%. The accuracy of the method was 80.26%. CONCLUSION: This study shows that the flow cytometry platelet immunofluorescence test has a high correlation with the anti-human leukocyte antigen antibodies. Despite a few limitations, the method seems to be efficient, fast and feasible as the initial screening for platelet antibody detection and a useful tool to crossmatch platelets for the transfusional support of patients with immune platelet refractoriness.

  6. Selection of antibodies from synthetic antibody libraries.

    Science.gov (United States)

    Harel Inbar, Noa; Benhar, Itai

    2012-10-15

    More than 2 dozen years had passed since the field of antibody engineering was established, with the first reports of bacterial [1-3] and mammalian cells [4] expression of recombinant antibody fragments, and in that time a lot of effort was dedicated to the development of efficient technological means, intended to assist in the creation of therapeutic monoclonal antibodies (mAbs). Research focus was given to two intertwined technological aspects: the selection platform and the recombinant antibody repertoires. In accordance with these areas of interest, it is the goal of this chapter to describe the various selection tools and antibody libraries existing, with emphasis on the later, and their applications. This chapter gives a far from exhaustive, subjective "historic account" of the field, describing the selection platforms, the different formats of antibody repertoires and the applications of both for selecting recombinant antibodies. Several excellent books provide detailed protocols for constructing antibody libraries and selecting antibodies from those libraries [5-13]. Such books may guide a newcomer to the field in the fine details of antibody engineering. We would like to offer advice to the novice: although seemingly simple, effective library construction and antibody isolation provide best benefits in the hands of professionals. It is an art as much as it is science.

  7. GENERATION OF MONOCLONAL ANTIBODY AGAINST HUMAN ANDROGEN RECEPTOR WITH SYNTHETIC PEPTIDE

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Objective: Preparation of anti-human androgen receptor(hAR) monoclonal antibody (McAb). Methods: Four cells lines of hybridoma secreting specific monoclonal antibodies against AR were first established by fusion SP2/0 cell with spleen cell from BALB/c mice immunized with the coupling complex of hAR-KLH. Results: Paraffin-embedded sections of 45 prostate cancers were detected. There was an overall concordance of 91% using Immunohistochemistry between AR polyclonal antibody from Zymed and hAR-N McAb selfmade. Conclusion: The results show that the McAb obtained in this study would be a useful tool to detect the AR status in prostate cancer.

  8. Detection of antibodies to variant antigens on Plasmodium falciparum-infected erythrocytes by flow cytometry

    DEFF Research Database (Denmark)

    Staalsoe, T; Giha, H A; Dodoo, D;

    1999-01-01

    BACKGROUND: Naturally induced antibodies binding to surface antigens of Plasmodium falciparum-infected erythrocytes can be detected by direct agglutination of infected erythrocytes or by indirect immunofluorescence on intact, unfixed, infected erythrocytes. Agglutinating antibodies have previously...... been shown to recognise Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1). This protein is inserted by the parasite into the host cell membrane and mediates the adhesion to the venular endothelium of the host organism in vivo. METHODS: Erythrocytes infected at high parasitaemias...... with ethidium-bromide-labelled mature forms of P. falciparum parasites were sequentially exposed to immune plasma, goat anti-human immunoglobulin (Ig) G, and fluorescein-isothiocyanate-conjugated rabbit anti-goat Ig. Plasma antibodies recognising antigens exposed on the surface of parasitised erythrocytes were...

  9. [The treatment by expanded ex vivo autologous regulatory T-cells CD4+CD25+FoxP3+CD127low restores the balance of immune system in patients with remitting-relapsing multiple sclerosis].

    Science.gov (United States)

    Eliseeva, D D; Lifshits, G V; Lokhonina, A V; Zhdanov, D D; Zavalishin, I A; Bykovskaya, S N

    2016-01-01

    Цель исследования — оценка клинической эффективности и безопасности аутологичных (собственных) Трег-клеток — CD4+CD25+Foxp3+CD127low, выделенных из крови больных ремиттирующим рассеянным склерозом (РРС). Больные аутоиммунными заболеваниями имеют сниженное число и нарушенную супрессорную функцию циркулирующих в крови периферических Трег. С целью коррекции дефекта периферических Трег можно выделить клетки-предшественники, культивировать вне организма (ex vivo) больного и ввести увеличенное количество аутологичных (собственных) Трег в виде вакцины Трег. Материал и методы. Была разработана методика культивирования ex vivo, с помощью которой количество Трег можно увеличить в 30—40 раз в течение 5—7 дней. Выращенные ex vivo клетки состоят более чем из 90% CD4+CD25+Foxp3+CD127low Трег и обладают высоким уровнем супрессорной активности. В пилотные испытания были включены 14 больных РРС, которым подкожно были введены выращенные ex vivo Трег в дозе от 2,8 до 4,5·108 клеток на одну инъекцию. Длительность наблюдения за больными составила 1 год. Результаты и обсуждение. Уровень Трег в крови этих больных повысился в 1,5—2

  10. In-vitro inhibition of IFNγ+ iTreg mediated by monoclonal antibodies against cell surface determinants essential for iTreg function

    OpenAIRE

    Daniel Volker; Sadeghi Mahmoud; Wang Haihao; Opelz Gerhard

    2012-01-01

    Abstract Background IFNγ-producing CD4+CD25+Foxp3+ PBL represent a subtype of iTreg that are associated with good long-term graft outcome in renal transplant recipients and suppress alloresponses in-vitro. To study the mechanism of immunosuppression, we attempted to block cell surface receptors and thereby inhibited the function of this iTreg subset in-vitro using monoclonal antibodies and recombinant proteins. Methods PBL of healthy control individuals were stimulated polyclonally in-vitro i...

  11. The changes of cardiopulmonary function and the correlation with the ratios of regulatory T cells in OA rats%骨关节炎大鼠心、肺功能变化与CD4+ CD25+ Foxp3+ Treg的相关性分析

    Institute of Scientific and Technical Information of China (English)

    程园园; 刘健; 冯云霞; 刘磊; 万磊

    2013-01-01

    Objective:To observe the changes of cardiopulmonary function and expression of CD4 + CD25+ Foxp3+ Treg in Os-teoarthritis ( OA) rats, thus to explore the role that CD4+ CD25 + Foxp3 + Treg played in decline of cardiopulmonary of OA rats. Methods: 20 rats were randomly divided into 2 groups; normal group( NC) and model group( NC) , 10 in each group. Rats in MC group were induced to OA model by injecting papain and L-cystine into right knee joint. Cartilage lesions were scored by the modified Mankin Scale; cardiac function was detected by ultrasonography; pulmonary function was detected by spirometry; interleukin (IL) -17 ,4 were evaluated by ELJSA; the ratios of CD4+ CD25 + Foxp3 + Treg in peripheral blood were measured by Flow cytometry. Results: Compared to NC group, Mankin scores in MC group were significantly increased; part of parameters of cardiopulmonary function were obviously changed, and accounts for 60% and 20% of the selected indexes respectively; expression of IL-17 was significantly ascend, expression of IL-4 and CD4 + CD25+ Foxp3 + Treg were significantly descend ( P < 0. 01 or P < 0. 05 ). Conclusion: While articular cartilage lesions occurred in OA rats, cardiopulmonary function and the expression of peripheral CD4 + CD25 + Foxp3+ Treg were also declined, and significantly correlated with Mankin scores, expression of IL-17,IL-4, suggesting that immune imbalance and aggravated inflammatory due to attenuated regulatory effect of regulatory T cells (Treg) , result in joint, cardiac and pulmonary injury, finally causing cardiopulmonary function declined.%目的:通过观察骨关节炎(Osteoarthritis,OA)大鼠、心肺功能的变化,及其与外周血CD4+CD25+Foxp3+Treg表达量变化,探讨骨关节炎中心、肺功能的异常与CD4+CD25+Foxp3+Treg的关系.方法:20只大鼠随机分为正常对照组(NC)和模型对照组(MC),每组各10只.MC组大鼠采用关节腔注射木瓜蛋白酶(Papain)和L-半胱氨酸(L-cystine)方法制成骨关节炎

  12. Current status of anti-human epidermal growth factor receptor 2 therapies: predicting and overcoming herceptin resistance.

    Science.gov (United States)

    Chung, Alice; Cui, Xiaojiang; Audeh, William; Giuliano, Armando

    2013-08-01

    Human epidermal growth factor receptor 2-overexpressing (HER2+) breast cancer occurs in 20% to 25% of cases and is associated with poor prognosis. Trastuzumab (Herceptin; Genentech, South San Francisco, CA) is a monoclonal antibody targeting the HER2 extracellular domain that has been shown to significantly reduce relapse rates. However, some patients with HER2+ tumors do not respond to Herceptin, and 60% to 85% of patients with HER2+ metastatic breast cancer acquire resistance within a short time period. In this review, we discuss proposed mechanisms of action of trastuzumab and trastuzumab resistance and various drugs that have been developed to overcome drug resistance. We introduce the basal molecular subtype as a predictor of increased risk in HER2+ breast cancer and a possible alternative cause of drug resistance.

  13. Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia

    Science.gov (United States)

    Weng, Ying-Jan; Husebekk, Anne; Skogen, Björn; Kjaer, Mette; Lin, Liang-Tzung; Burnouf, Thierry

    2016-01-01

    Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a severe disease that is caused by maternal alloantibodies generated during pregnancy or at delivery as a result of incompatibility between maternal and fetal human platelet antigens (HPAs) inherited from the father. Antibody-mediated immune suppression using anti-HPA-1a immunoglobulins is thought to be able to prevent FNAIT caused by HPA-1a. A fractionation process to prepare anti-HPA-1a immunoglobulin (Ig) G (IgG) from human plasma was therefore developed. Anti-HPA-1a plasma was obtained from volunteer mothers who underwent alloimmunization against HPA-1a during a previous pregnancy. Plasma was cryoprecipitated and the supernatant treated with caprylic acid and solvent/detergent (S/D), purified by chromatography, nanofiltered, concentrated, and sterile-filtered. The anti-HPA-1a immunoglobulin fraction was characterized for purity and safety. PAK12 and quantitative monoclonal antibody immobilization of platelet antigen (MAIPA) assays were used to detect anti-HPA-1a IgG. Hepatitis C virus (HCV) removal during nanofiltration was assessed by spiking experiments, using cell culture-derived reporter HCV and luciferase analysis. The caprylic acid treatment precipitated non-Ig proteins yielding a 90% pure Ig supernatant. S-HyperCel chromatography of the S/D-treated supernatant followed by HyperCel STAR AX provided high IgG recovery (>80%) and purity (>99.5%), and efficient IgA and IgM removal. Concentrations of complement factors C3 and C4 were HPA-1a throughout the process. Clinical-grade HPA-1a IgG can be prepared using a process compliant with current quality requirements opening perspectives for the prevention of FNAIT. PMID:27627660

  14. Flow cytometric detection of anti-gliadin antibodies.

    Science.gov (United States)

    Presani, G; Perticarari, S; Mangiarotti, M A

    1989-05-12

    A very sensitive solid-phase fluorescent immunoassay to detect anti-alpha-gliadin IgA class antibodies is described. The solid phase consisted of polystyrene carboxylated microspheres, of 5 microns diameter, coated with alpha-gliadin. Serum-specific antibodies bound to the alpha-gliadin were measured by flow cytometry using fluorescein-conjugated anti-human IgA. 41 samples were tested and the results compared with those obtained by a standard method: an enzyme-linked immunosorbent assay (ELISA). A good correlation was found between the two techniques (r = 0.96). The sera of untreated coeliac children showed significantly higher antibody values than the sera of children on a gluten-free diet or healthy control groups. The flow cytometric method was more sensitive when the Kolgomorov/Smirnov test was used to analyse the histograms. This method provides an alternative screening test for coeliac disease and may also be used to confirm borderline results obtained in the ELISA test.

  15. Acetylcholine receptor antibody

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/003576.htm Acetylcholine receptor antibody To use the sharing features on this page, please enable JavaScript. Acetylcholine receptor antibody is a protein found in the blood ...

  16. Antinuclear antibody panel

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/003535.htm Antinuclear antibody panel To use the sharing features on this page, please enable JavaScript. The antinuclear antibody panel is a blood test that looks at ...

  17. Lyme disease antibody

    Science.gov (United States)

    ... JavaScript. The Lyme disease blood test looks for antibodies in the blood to the bacteria that causes ... needed. A laboratory specialist looks for Lyme disease antibodies in the blood sample using the ELISA test . ...

  18. The antibody mining toolbox

    OpenAIRE

    D'Angelo, Sara; Glanville, Jacob; Ferrara, Fortunato; Naranjo, Leslie; Gleasner, Cheryl D.; Shen, Xiaohong; Bradbury, Andrew RM; Kiss, Csaba

    2013-01-01

    In vitro selection has been an essential tool in the development of recombinant antibodies against various antigen targets. Deep sequencing has recently been gaining ground as an alternative and valuable method to analyze such antibody selections. The analysis provides a novel and extremely detailed view of selected antibody populations, and allows the identification of specific antibodies using only sequencing data, potentially eliminating the need for expensive and laborious low-throughput ...

  19. Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia.

    Science.gov (United States)

    Weng, Ying-Jan; Husebekk, Anne; Skogen, Björn; Kjaer, Mette; Lin, Liang-Tzung; Burnouf, Thierry

    2016-01-01

    Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a severe disease that is caused by maternal alloantibodies generated during pregnancy or at delivery as a result of incompatibility between maternal and fetal human platelet antigens (HPAs) inherited from the father. Antibody-mediated immune suppression using anti-HPA-1a immunoglobulins is thought to be able to prevent FNAIT caused by HPA-1a. A fractionation process to prepare anti-HPA-1a immunoglobulin (Ig) G (IgG) from human plasma was therefore developed. Anti-HPA-1a plasma was obtained from volunteer mothers who underwent alloimmunization against HPA-1a during a previous pregnancy. Plasma was cryoprecipitated and the supernatant treated with caprylic acid and solvent/detergent (S/D), purified by chromatography, nanofiltered, concentrated, and sterile-filtered. The anti-HPA-1a immunoglobulin fraction was characterized for purity and safety. PAK12 and quantitative monoclonal antibody immobilization of platelet antigen (MAIPA) assays were used to detect anti-HPA-1a IgG. Hepatitis C virus (HCV) removal during nanofiltration was assessed by spiking experiments, using cell culture-derived reporter HCV and luciferase analysis. The caprylic acid treatment precipitated non-Ig proteins yielding a 90% pure Ig supernatant. S-HyperCel chromatography of the S/D-treated supernatant followed by HyperCel STAR AX provided high IgG recovery (>80%) and purity (>99.5%), and efficient IgA and IgM removal. Concentrations of complement factors C3 and C4 were < 0.5 and < 0.4 mg/dL, respectively. The final IgG could be nanofiltered on Planova 20N under conditions removing more than 3 log HCV infectivity to baseline mock infection level, and concentrated to ca. 30 g/L. Proteolytic activity and thrombin generation were low in the final fraction. The Pak12 and MAIPA assays showed good recovery of anti-HPA-1a throughout the process. Clinical-grade HPA-1a IgG can be prepared using a process compliant with current quality requirements

  20. [VGKC-complex antibodies].

    Science.gov (United States)

    Watanabe, Osamu

    2013-04-01

    Various antibodies are associated with voltage-gated potassium channels (VGKCs). Representative antibodies to VGKCs were first identified by radioimmunoassays using radioisotope-labeled alpha-dendrotoxin-VGKCs solubilized from rabbit brain. These antibodies were detected only in a proportion of patients with acquired neuromyotonia (Isaacs' syndrome). VGKC antibodies were also detected in patients with Morvan's syndrome and in those with a form of autoimmune limbic encephalitis. Recent studies indicated that the "VGKC" antibodies are mainly directed toward associated proteins (for example LGI-1 and CASPR-2) that complex with the VGKCs themselves. The "VGKC" antibodies are now commonly known as VGKC-complex antibodies. In general, LGI-1 antibodies are most commonly detected in patients with limbic encephalitis with syndrome of inappropriate secretion of antidiuretic hormone. CASPR-2 antibodies are present in the majority of patients with Morvan's syndrome. These patients develop combinations of CNS symptoms, autonomic dysfunction, and peripheral nerve hyperexcitability. Furthermore, VGKC-complex antibodies are tightly associated with chronic idiopathic pain. Hyperexcitability of nociceptive pathways has also been implicated. These antibodies may be detected in sera of some patients with neurodegenerative diseases (for example, amyotrophic lateral sclerosis and Creutzfeldt-Jakob disease).

  1. Anti-CD3 antibodies modulate anti-factor VIII immune responses in hemophilia A mice after factor VIII plasmid-mediated gene therapy.

    Science.gov (United States)

    Peng, Baowei; Ye, Peiqing; Rawlings, David J; Ochs, Hans D; Miao, Carol H

    2009-11-12

    One major obstacle in gene therapy is the generation of immune responses directed against transgene product. Five consecutive anti-CD3 treatments concomitant with factor VIII (FVIII) plasmid injection prevented the formation of inhibitory antibodies against FVIII and achieved persistent, therapeutic levels of FVIII gene expression in treated hemophilia A mice. Repeated plasmid gene transfer is applicable in tolerized mice without eliciting immune responses. Anti-CD3 treatment significantly depleted both CD4+ and CD8+ T cells, whereas increased transforming growth factor-beta levels in plasma and the frequency of both CD4+CD25+FoxP3+ and CD4+CD25-Foxp3+ regulatory T cells in the initial few weeks after treatment. Although prior depletion of CD4+CD25+ cells did not abrogate tolerance induction, adoptive transfer of CD4+ cells from tolerized mice at 6 weeks after treatment protected recipient mice from anti-FVIII immune responses. Anti-CD3-treated mice mounted immune responses against both T-dependent and T-independent neo-antigens, indicating that anti-CD3 did not hamper the immune systems in the long term. Concomitant FVIII plasmid + anti-CD3 treatment induced long-term tolerance specific to FVIII via a mechanism involving the increase in transforming growth factor-beta levels and the generation of adaptive FVIII-specific CD4+Foxp3+ regulatory T cells at the periphery. Furthermore, anti-CD3 can reduce the titers of preexisting anti-FVIII inhibitory antibodies in hemophilia A mice.

  2. The possible role of CD4⁺CD25(high)Foxp3⁺/CD4⁺IL-17A⁺ cell imbalance in the autoimmunity of patients with Hashimoto thyroiditis.

    Science.gov (United States)

    Xue, Haibo; Yu, Xiurong; Ma, Lei; Song, Shoujun; Li, Yuanbin; Zhang, Li; Yang, Tingting; Liu, Huan

    2015-12-01

    Hashimoto thyroiditis (HT) is a prototypic organ-specific autoimmune thyroid disease, for which the exact etiology remains unclear. The aim of this study was to investigate dynamic changes in regulatory T cell (Treg) and T helper 17 cell (Th17) populations in patients with HT at different stages of thyroid dysfunction, as well as to analyze the possible correlation between the Treg/Th17 cell axis and autoimmune status in HT. We assessed thyroid function and autoantibody serology both in HT patients and in healthy controls (HCs) and divided HT patients into three subgroups according to thyroid function. We then determined the percentages of Treg and Th17 cells in peripheral blood mononuclear cells and analyzed mRNA expression of the Treg and Th17 cell-defining transcription factors Foxp3 and RORγt. In addition, serum levels of TGF-β and IL-17A were assessed. We found that the percentage of Treg cells, Foxp3 mRNA levels, and the ratio of Treg/Th17 cells were all significantly lower in HT patients, while Th17 cell percentages and RORγt mRNA levels were significantly higher. Interestingly, we also observed significant differences in these measurements between HT patient subgroups. Serum IL-17A levels were markedly increased in HT patients, while serum concentrations of TGF-β were lower, compared to HCs. The ratio of Treg/Th17 cells was negatively correlated with the levels of serum thyroperoxidase antibody, thyroglobulin antibody, and thyrotropin (TSH) in HT patients. Taken together, our data suggest that the balance between Treg and Th17 cells shifts in favor of Th17 cells during clinical progression of HT, which is negatively correlated with levels of thyroid-specific autoantibodies and TSH, implying that Treg/Th17 cell imbalance may contribute to thyroid damage in HT.

  3. Screening, expression, and characterization of an anti-human oxidized low-density lipoprotein single-chain variable fragment.

    Science.gov (United States)

    Kumano-Kuramochi, Miyuki; Fujimura, Takashi; Komba, Shiro; Maeda-Yamamoto, Mari; Machida, Sachiko

    2016-09-01

    Increased levels of oxidized low-density lipoprotein (OxLDL) in the blood circulation are correlated with atherosclerosis. Monoclonal antibody-based detection systems have been reported for OxLDL. We identified novel single-chain variable fragments (scFvs) having affinity for human OxLDL and related ligands. We constructed an scFv library from nonimmunized human spleen mRNA. Two types (γ+κ and μ+λ) of scFv phage libraries were enriched by biopanning, and five scFv clones with affinity for OxLDL were identified. The γκ5 scFv, which showed the highest affinity for OxLDL, was cloned into pET-22b(+) and expressed in Escherichia coli BL21(DE3). γκ5, expressed as an inclusion body in BL21(DE3), was refolded and purified. The specificity and sensitivity of γκ5 were analyzed using enzyme-linked immunosorbent assays (ELISAs). The γκ5 scFv showed affinity for OxLDL and acetylated LDL. The sensitivity of γκ5 to low concentrations (1-2 μg/mL) of OxLDL was higher than that to AcLDL and LDL. Finally, we developed a sandwich ELISA using γκ5 and CTLD14 (a lectin-like OxLDL receptor-1 ligand recognition region), which allowed specific detection of OxLDL at a level below 0.1 μg/mL. Our results indicated that the γκ5 scFv was a promising molecule for the detection of modified LDL at very low concentrations.

  4. Anti-human herpesvirus 6A/B IgG correlates with relapses and progression in multiple sclerosis.

    Directory of Open Access Journals (Sweden)

    Isabel Ortega-Madueño

    Full Text Available To analyze the titers of the IgG and IgM antibodies against human herpesvirus 6A/B (HHV-6A/B in multiple sclerosis (MS patients treated with different disease modified therapies (DMTs along two-years of follow-up.We collected 2163 serum samples from 596 MS; for 301 MS patients a 2-years follow-up was performed. Serum samples of 337 healthy controls were also analyzed. Anti-HHV-6A/B IgG and IgM were analyzed by ELISA (Panbio.We found that 129/187 (69.0% MS patients with a decrease of the anti-HHV-6A/B IgG titers after 2-years with DMTs were free of relapses and progression vs. 46/113 (40.7% of MS patients with an increase of the anti-HHV-6A/B IgG titers (p = 0.0000015; the higher significance was found for natalizumab. Furthermore, we found that anti-HHV-6A/B IgG titers reached their highest value two weeks before the relapse (p = 0.0142, while the anti-HHV-6A/B IgM titers reached their highest value one month before the relapse (p = 0.0344.The measurement of the anti-HHV-6A/B IgG titers could be a good biomarker of clinical response to the different DMTs. The increase of the anti-HHV-6A/B IgG and IgM titers predicts the upcoming clinical relapses. However, further longitudinal studies are needed to validate these results.

  5. The role of CD4+ CD25+ Foxp3+ Treg in the immune tolerance induced by portal vein injection of donor splenocytes%门静脉输注供者脾细胞诱导的供者特异性免疫低反应性

    Institute of Scientific and Technical Information of China (English)

    徐胜元; 何凡; 吴敏; 蔡明; 丁召; 郑翔; 陈知水

    2009-01-01

    目的 探讨经门静脉输注供者脾细胞能否诱导皮肤移植小鼠产生供者特异性的免疫低反应性及其可能机制.方法 取Balb/c小鼠,随机分为空白对照组(经小鼠门静脉输注RPMI 1640培养液)、受者脾细胞组(经小鼠门静脉输注Balb/c小鼠脾细胞)、供者脾细胞组(经小鼠门静脉输注C57BL/6小鼠脾细胞)、空白移植对照组(经小鼠门静脉输注RPMI 1640培养液,7 d后移植C57BL/6小鼠的皮肤)、实验对照组(经小鼠门静脉输注Balb/c小鼠脾细胞,7 d后移植C57BL/6小鼠的皮肤)、实验组(经小鼠门静脉输注C57BL/6小鼠脾细胞,7 d后移植C57BL/6小鼠的皮肤)以及第三方移植组(经小鼠门静脉输注C57BL/6小鼠脾细胞,7 d后移植C3H小鼠的皮肤).记录空白移植对照组、实验对照组、实验组和第三方移植组移植皮肤的存活时间,并观察移植皮肤的病理学变化;脾细胞输注后7 d,分别获取空白对照组、受者脾细胞组和供者脾细胞组小鼠的外周血、脾脏和肝脏,用流式细胞仪测定样本中CD4+CD25+Foxp3+调节性T淋巴细胞(CD4+CD25+Foxp3+Treg细胞)的比例.结果 实验组移植皮肤的存活时间为(19.8±4.6)d,明显长于空白移植对照组、实验对照组和第三方移植组,但仍未达到长期存活.皮肤移植后7 d,空白移植对照组和实验对照组的移植皮肤呈现重度急性排斥反应的病理学改变,而实验组移植皮肤呈现中度急性排斥反应的病理学改变.供者脾细胞组外周血、肝脏和脾脏中CD4+CD25+Foxp3+Treg细胞比例明显高于空白对照组和受者脾细胞组.结论 门静脉输注供者脾细胞可特异性地延长供者皮肤移植物的存活时间,减轻移植物的排斥反应,该效应可能与受者体内的CD4+CD25+Foxp3+Treg细胞增加有关.%Objective To study the roles of CD4+ CD25+ Foxp3+ regulatory T cells (Treg) in the immune tolerance induced by portal vein injection of donor splenocytes. Methods C57

  6. Crystal structure of a human rhinovirus neutralizing antibody complexed with a peptide derived from viral capsid protein VP2.

    OpenAIRE

    1994-01-01

    The three-dimensional structure of the complex between the Fab fragment of an anti-human rhinovirus neutralizing antibody (8F5) and a cross-reactive synthetic peptide from the viral capsid protein VP2 has been determined at 2.5 A resolution by crystallographic methods. The refinement is presently at an R factor of 0.18 and the antigen-binding site and viral peptide are well defined. The peptide antigen adopts a compact fold by two tight turns and interacts through hydrogen bonds, some with io...

  7. Antilaminaribioside and antichitobioside antibodies in inflammatory bowel disease.

    Science.gov (United States)

    Rejchrt, S; Drahosová, M; Kopácová, M; Cyrany, J; Douda, T; Pintér, M; Bures, J

    2008-01-01

    Testing antilaminaribioside (ALCA) and antichitobioside (ACCA) antibodies in 89 Crohn's disease (CD), 31 ulcerative colitis (UC) and 50 controls, mean values were 38.6 and 53.0 ELISA units for CD, 34.0 and 32.6 for UC, 34.5 and 36.4 for controls, respectively. There was no significant difference of ALCA values between CD and UC (p = 0.401), CD and control subjects (p = 0.698) or UC and controls (p = 0.898). ACCA were significantly higher in CD compared with UC (p = 0.011) but not with the controls (p = 0.095). No significant difference of ACCA values between UC and controls (p = 0.107) was found. ALCA and ACCA values significantly correlated in CD (r = 0.548, p ALCA) and 8 % (ACCA), the negative ones (to exclude CD) 25 (ALCA) and 86 % (ACCA). Small and/or large bowel involvement or disease type (i.e. stenosing, perforating or inflammatory) of CD did not differ in the two values. The idea that ALCA and ACCA may be useful either to differentiate between CD, UC and healthy subjects or to stratify CD was not confirmed.

  8. 调节性T细胞对输注树突状细胞后小鼠移植心脏的保护作用%The protective effects of CD4+ CD25+ Treg cell on the allograft after infusion of dendritic cells with low expression of CD40 from donor in mouse heart transplantation

    Institute of Scientific and Technical Information of China (English)

    朱杰昌; 付蔚华; 许翼麟; 朱理玮

    2013-01-01

    Objective To explore the effects of CD4+CD25+Treg cell on the allograft after infusion of dendritic cells (DCs) with low expression of CD40 from donor in mouse heart transplantation.Methods In vitro,mouse bone marrow-derived DCs were infected by CD40-RNAi lentiviral vector,and tolerogenic DCs (Tol-DCs) with low expression of CD40 were prepared.A heterotopic abdominal heart transplantation model was established in mice,and the other three groups that were control group,noninfected DC group and lentivirus infected DC group were designed correspondingly.Cardiac allograft survival time was recorded and pathological grading for acute rejection was assessed on the 7 d after heterotopic abdominal heart transplantation.Concentrations of CD4+CD25+Treg cells in peripheral blood were analyzed before and after transplantation by flow cytometry.Results After 48 h infection of DCs by CD40-RNAi lentiviral vector in vitro,the expression of CD40 mRNA was down-regulated significantly,whose inhibition rate was 80.9%.The expression of CD40 was decreased from 74.37% ±4.08% to 40.07% ± 4.03% (P<0.05) after 48 h infection.Compared with the control group and the noninfected DC group,the cardiac allograft survival time was significantly prolonged in the CD40-RNAi lentivirus infected DC group,which was (14 ± 4) d(P<0.01) ; concentrations of CD4+CD25+Treg cells in peripheral blood were increased both on the 3 d and the 7 d after transplantation (P<0.05) ; the pathological grading for acute rejection was decreased on the 7 d after transplantation (P<0.05).Conclusion The CD4+CD25+Treg cell in peripheral blood was protective to cardiac allograft in prolonging its survival time in mouse heart transplantation.%目的 探讨在输注供者低表达CD40的树突状细胞(dendritic cells,DC)后,调节性T细胞(regulatory T cell,Treg)对小鼠移植心脏的保护作用.方法 以针对小鼠CD40基因的RNA干扰(RNA interference,RNAi)慢病毒载体在体外感染供者骨髓

  9. Expression of recombinant antibodies.

    Science.gov (United States)

    Frenzel, André; Hust, Michael; Schirrmann, Thomas

    2013-01-01

    Recombinant antibodies are highly specific detection probes in research, diagnostics, and have emerged over the last two decades as the fastest growing class of therapeutic proteins. Antibody generation has been dramatically accelerated by in vitro selection systems, particularly phage display. An increasing variety of recombinant production systems have been developed, ranging from Gram-negative and positive bacteria, yeasts and filamentous fungi, insect cell lines, mammalian cells to transgenic plants and animals. Currently, almost all therapeutic antibodies are still produced in mammalian cell lines in order to reduce the risk of immunogenicity due to altered, non-human glycosylation patterns. However, recent developments of glycosylation-engineered yeast, insect cell lines, and transgenic plants are promising to obtain antibodies with "human-like" post-translational modifications. Furthermore, smaller antibody fragments including bispecific antibodies without any glycosylation are successfully produced in bacteria and have advanced to clinical testing. The first therapeutic antibody products from a non-mammalian source can be expected in coming next years. In this review, we focus on current antibody production systems including their usability for different applications.

  10. Therapeutic Recombinant Monoclonal Antibodies

    Science.gov (United States)

    Bakhtiar, Ray

    2012-01-01

    During the last two decades, the rapid growth of biotechnology-derived techniques has led to a myriad of therapeutic recombinant monoclonal antibodies with significant clinical benefits. Recombinant monoclonal antibodies can be obtained from a number of natural sources such as animal cell cultures using recombinant DNA engineering. In contrast to…

  11. Recombinant renewable polyclonal antibodies.

    Science.gov (United States)

    Ferrara, Fortunato; D'Angelo, Sara; Gaiotto, Tiziano; Naranjo, Leslie; Tian, Hongzhao; Gräslund, Susanne; Dobrovetsky, Elena; Hraber, Peter; Lund-Johansen, Fridtjof; Saragozza, Silvia; Sblattero, Daniele; Kiss, Csaba; Bradbury, Andrew R M

    2015-01-01

    Only a small fraction of the antibodies in a traditional polyclonal antibody mixture recognize the target of interest, frequently resulting in undesirable polyreactivity. Here, we show that high-quality recombinant polyclonals, in which hundreds of different antibodies are all directed toward a target of interest, can be easily generated in vitro by combining phage and yeast display. We show that, unlike traditional polyclonals, which are limited resources, recombinant polyclonal antibodies can be amplified over one hundred million-fold without losing representation or functionality. Our protocol was tested on 9 different targets to demonstrate how the strategy allows the selective amplification of antibodies directed toward desirable target specific epitopes, such as those found in one protein but not a closely related one, and the elimination of antibodies recognizing common epitopes, without significant loss of diversity. These recombinant renewable polyclonal antibodies are usable in different assays, and can be generated in high throughput. This approach could potentially be used to develop highly specific recombinant renewable antibodies against all human gene products.

  12. Anti-insulin antibody test

    Science.gov (United States)

    Insulin antibodies - serum; Insulin Ab test; Insulin resistance - insulin antibodies; Diabetes - insulin antibodies ... You appear to have an allergic response to insulin Insulin no longer seems to control your diabetes

  13. The Change of CD4+CD25+ and CD8+CD28- T Regulatory Cells in Rats with the Experimental Colitis Induced by TNBS%调节性T细胞在大鼠TNBS诱发实验性结肠炎中作用的研究

    Institute of Scientific and Technical Information of China (English)

    王智峰; 尤鹏; 刘玉兰; 刘安楠

    2005-01-01

    目的探讨CD4+CD25+和CD8+CD28-调节性T细胞在TNBS(2,4,6-三硝基苯磺酸)诱发的大鼠实验性结肠炎的外周血、脾脏、结肠的改变及其作用.方法建立TNBS实验性结肠炎大鼠模型, 设对照组、建模后第一周和第三周三组,用流式细胞仪检测各组外周血、脾脏和结肠黏膜上皮细胞内单个核细胞中CD4+CD25+和CD8+CD28-调节性T细胞比例的变化.结果成功建立TNBS大鼠实验性结肠炎模型(n=15).结肠部位CD8+CD28-细胞在建模后第一周时较对照组均明显增高[(12.7±5.4)% vs(3.87±3.7)%,P<0.01].在外周血、脾和结肠中CD8+CD28-T细胞比例在建模第三周时均较对照组明显增加[外周血:(14.4±5.5)% vs(6.5±4.6)% ,脾脏:(18.7±4.9)% vs(7.2±5.7)%,结肠:(16.3±4.9)% vs(3.87±3.7)%,P<0.01],其中以结肠部位增加最高,同其它两部位比较有统计学差异(2.8±0.6 vs 1.1±0.4,2.8±0.6 vs 1.5±0.4,P<0.05).在第一周,CD4+CD25+T细胞在结肠部位较对照组增加[(13.6±6.1)vs(5.7±4.2),P<0.05],于第三周回落至对照组水平.结论 CD4+CD25+和CD8+CD28-T细胞在TNBS实验性结肠炎大鼠模型中较正常对照组有明显增加,但增加的部位和时间点有所不同,提示这两种调节性T细胞可能在溃疡性结肠炎发病的不同阶段中起着一定作用,且主要作用于肠道病变部位.

  14. Preparation and Characterization of Covalently Binding of Rat Anti-human IgG Monolayer on Thiol-Modified Gold Surface

    Directory of Open Access Journals (Sweden)

    Lv Zhengjian

    2009-01-01

    Full Text Available Abstract The 16-mercaptohexadecanoic acid (MHA film and rat anti-human IgG protein monolayer were fabricated on gold substrates using self-assembled monolayer (SAM method. The surface properties of the bare gold substrate, the MHA film and the protein monolayer were characterized by contact angle measurements, atomic force microscopy (AFM, grazing incidence X-ray diffraction (GIXRD method and X-ray photoelectron spectroscopy, respectively. The contact angles of the MHA film and the protein monolayer were 18° and 12°, respectively, all being hydrophilic. AFM images show dissimilar topographic nanostructures between different surfaces, and the thickness of the MHA film and the protein monolayer was estimated to be 1.51 and 5.53 nm, respectively. The GIXRD 2θ degrees of the MHA film and the protein monolayer ranged from 0° to 15°, significantly smaller than that of the bare gold surface, but the MHA film and the protein monolayer displayed very different profiles and distributions of their diffraction peaks. Moreover, the spectra of binding energy measured from these different surfaces could be well fitted with either Au4f, S2p or N1s, respectively. Taken together, these results indicate that MHA film and protein monolayer were successfully formed with homogeneous surfaces, and thus demonstrate that the SAM method is a reliable technique for fabricating protein monolayer.

  15. Effect of apoptosis of CD+4 CD25+ regulatory T cells on proliferation as well as secretion of effeetor Tcells and interventional activity of Xuebijing injection in septic rats%脓毒症大鼠调节性T细胞凋亡对效应T细胞增殖和分泌功能的影响及血必净注射液的干预作用

    Institute of Scientific and Technical Information of China (English)

    艾宇航; 姚咏明; 戴新贵

    2009-01-01

    目的 探讨血必净注射液对脓毒症大鼠CD+4CD+25调节性T细胞(Treg)凋亡的影响及与效应T细胞(Teff)增殖及分泌功能的关系.方法 Wister大鼠随机分为对照组、假于术组、肓肠结扎穿孔(CLP)组和血必净治疗组,每组8只;于第3天采用免疫磁珠法分选大鼠脾脏CD+4CD25+Treg和CD+4CD25-T细胞.CD+4CD25+Treg培养12 h后检测凋亡率、叉头翼状螺旋转录因子(Foxp3)和T淋巴细胞毒性相关抗原4(CTLA-4)表达及门细胞介素(IL)-10的分泌.将Treg与CD+4CD25-T细胞共培养,并以刀豆素A刺激后观察Treg对Teff增殖活性和细胞培养上清IL-2/可溶性IL-2受体α(slL-2Rα)含量的影响.结果 CLP组CD+4CD25+Treg细胞凋亡率CLP组(P对照组,而血必净组其抑制率CLP组,sll-2Rα低于CLP组(P<0.01).结论 脓毒症病理过程中CD+4CD25+Treg细胞对Teff抑制效应明显增强,而血必净注射液能诱导Treg细胞凋亡,减轻Treg对Teff的抑制作用.%Objective To investigate the effect of apoptosis of CD+4 CD25+ regulatory T cells (Tregs) on proliferation as well as secretory fullction of effector T ceils (Teff) and potential influence of Xuebijing injection on them in septic rats. Methods A sepsis model was reproduced by cecal ligation puncture (CLP), and Wister rats were randomly divided into the control group (n = 8 ), sham-operated group (n =8), CLP group (n =8), and Xuebijing injection treatment group (n =8). CD+4 CD25+ Tregs in each group were soperated by immunomagnetic beads isolate system on day 3, the apoptosis rate, expression of forkhead/winged helix transcription factor p3 (Foxp3) and cytotoxic T-lymphocyte-associated antigen 4 ( CTLA-4 ) on Tregs were analyzed by flow cytometry, and secretion levels of interleukin (IL) -10 from Tregs were measured by ELISA. Following co-culture of CD+4 CD25 + Treg with CD+4 CD25 - T cells ( 1 : 1 ) for 68 hours, proliferative activity of Teff was determined by MTT, and IL-2/sIL-2Rα levels were measured by ELISA

  16. Monoclonal antibody "gold rush".

    Science.gov (United States)

    Maggon, Krishan

    2007-01-01

    The market, sales and regulatory approval of new human medicines, during the past few years, indicates increasing number and share of new biologics and emergence of new multibillion dollar molecules. The global sale of monoclonal antibodies in 2006 were $20.6 billion. Remicade had annual sales gain of $1 billion during the past 3 years and five brands had similar increase in 2006. Rituxan with 2006 sales of $4.7 billion was the best selling monoclonal antibody and biological product and the 6th among the top selling medicinal brand. It may be the first biologic and monoclonal antibody to reach $10 billion annual sales in the near future. The strong demand from cancer and arthritis patients has surpassed almost all commercial market research reports and sales forecast. Seven monoclonal antibody brands in 2006 had sales exceeding $1 billion. Humanized or fully human monoclonal antibodies with low immunogenicity, enhanced antigen binding and reduced cellular toxicity provide better clinical efficacy. The higher technical and clinical success rate, overcoming of technical hurdles in large scale manufacturing, low cost of market entry and IND filing, use of fully human and humanized monoclonal antibodies has attracted funds and resources towards R&D. Review of industry research pipeline and sales data during the past 3 years indicate a real paradigm shift in industrial R&D from pharmaceutical to biologics and monoclonal antibodies. The antibody bandwagon has been joined by 200 companies with hundreds of new projects and targets and has attracted billions of dollars in R&D investment, acquisitions and licensing deals leading to the current Monoclonal Antibody Gold Rush.

  17. Antiphospholipid antibody syndrome.

    Science.gov (United States)

    Kutteh, William H; Hinote, Candace D

    2014-03-01

    Antiphospholipid antibodies (aPLs) are acquired antibodies directed against negatively charged phospholipids. Obstetric antiphospholipid antibody syndrome (APS) is diagnosed in the presence of certain clinical features in conjunction with positive laboratory findings. Obstetric APS is one of the most commonly identified causes of recurrent pregnancy loss. Thus, obstetric APS is distinguished from APS in other organ systems where the most common manifestation is thrombosis. Several pathophysiologic mechanisms of action of aPLs have been described. This article discusses the diagnostic and obstetric challenges of obstetric APS, proposed pathophysiologic mechanisms of APS during pregnancy, and the management of women during and after pregnancy.

  18. Antibodies against Human Cytomegalovirus in the Pathogenesis of Systemic Sclerosis: A Gene Array Approach.

    Directory of Open Access Journals (Sweden)

    2005-12-01

    Full Text Available BACKGROUND: Systemic sclerosis is an autoimmune disease characterized by immunological abnormalities, vascular damage, and fibroblast proliferation. We have previously shown that a molecular mimicry mechanism links antibodies against the human-cytomegalovirus-derived protein UL94 to the pathogenesis of systemic sclerosis. The UL94 epitope shows homology with NAG-2, a surface molecule highly expressed on endothelial cells. Anti-UL94 peptide antibodies purified from patients' sera induce apoptosis of endothelial cells upon engagement of the NAG-2-integrin complex. METHODS AND FINDINGS: We show here that NAG-2 is expressed on dermal fibroblasts and that anti-UL94 antibodies bind to fibroblasts. We have used the gene array strategy (Affimetrix oligonucleotide microarrays to analyze the transcriptional profile in response to a 4-h and an 8-h treatment with antibodies against the UL94 peptide in endothelial cells and dermal fibroblasts. Exposure of endothelial cells to anti-UL94 antibodies had a profound impact on gene expression, resulting in the upregulation of 1,645 transcripts. Several gene clusters were upregulated including genes encoding adhesion molecules, chemokines, colony-stimulating factors (CSFs, growth factors, and molecules involved in apoptosis. Following antibody stimulation, dermal fibroblasts showed an upregulation of 989 transcripts and acquired a "scleroderma-like" phenotype. Indeed, genes involved in extracellular matrix deposition, growth factors, chemokines, and cytokines were upregulated. We confirmed the microarray results by real-time quantitative polymerase chain reaction and by measuring some of the corresponding proteins with ELISA and Western blotting. CONCLUSION: Our results show that anti-human-cytomegalovirus antibodies may be linked to the pathogenesis of systemic sclerosis not only by inducing endothelial cell activation and apoptosis but also by causing activation of fibroblasts, one of the hallmarks of the disease.

  19. Antibodies against human cytomegalovirus in the pathogenesis of systemic sclerosis: a gene array approach.

    Directory of Open Access Journals (Sweden)

    Claudio Lunardi

    2006-01-01

    Full Text Available BACKGROUND: Systemic sclerosis is an autoimmune disease characterized by immunological abnormalities, vascular damage, and fibroblast proliferation. We have previously shown that a molecular mimicry mechanism links antibodies against the human-cytomegalovirus-derived protein UL94 to the pathogenesis of systemic sclerosis. The UL94 epitope shows homology with NAG-2, a surface molecule highly expressed on endothelial cells. Anti-UL94 peptide antibodies purified from patients' sera induce apoptosis of endothelial cells upon engagement of the NAG-2-integrin complex. METHODS AND FINDINGS: We show here that NAG-2 is expressed on dermal fibroblasts and that anti-UL94 antibodies bind to fibroblasts. We have used the gene array strategy (Affimetrix oligonucleotide microarrays to analyze the transcriptional profile in response to a 4-h and an 8-h treatment with antibodies against the UL94 peptide in endothelial cells and dermal fibroblasts. Exposure of endothelial cells to anti-UL94 antibodies had a profound impact on gene expression, resulting in the upregulation of 1,645 transcripts. Several gene clusters were upregulated including genes encoding adhesion molecules, chemokines, colony-stimulating factors (CSFs, growth factors, and molecules involved in apoptosis. Following antibody stimulation, dermal fibroblasts showed an upregulation of 989 transcripts and acquired a "scleroderma-like" phenotype. Indeed, genes involved in extracellular matrix deposition, growth factors, chemokines, and cytokines were upregulated. We confirmed the microarray results by real-time quantitative polymerase chain reaction and by measuring some of the corresponding proteins with ELISA and Western blotting. CONCLUSION: Our results show that anti-human-cytomegalovirus antibodies may be linked to the pathogenesis of systemic sclerosis not only by inducing endothelial cell activation and apoptosis but also by causing activation of fibroblasts, one of the hallmarks of the disease.

  20. Anti-cartilage antibody.

    Science.gov (United States)

    Greenbury, C L; Skingle, J

    1979-08-01

    Antibody to cartilage has been demonstrated by indirect immunofluorescence on rat trachea in the serum of about 3% of 1126 patients with rheumatoid arthritis. Titres ranged from 1:20 to 1:640. The antibody was not found in 284 patients with primary or secondary osteoarthritis or in 1825 blood donors, nor, with the exception of two weak reactors, in 1314 paraplegic patients. In most cases the antibody appears to be specific for native type II collagen. Using this as an antigen in a haemagglutination test 94% of anti-cartilage sera were positive, whereas among 100 rheumatoid control sera there were only three weak positives. More than 80% of patients with antibody had some erosion of articular cartilage, but there was no correlation with age, sex, duration of disease, nor any recognisable clinical event or change.

  1. Antithyroid microsomal antibody

    Science.gov (United States)

    ... to confirm the cause of thyroid problems, including Hashimoto thyroiditis . The test is also used to find ... positive test may be due to: Granulomatous thyroiditis Hashimoto thyroiditis High levels of these antibodies have also ...

  2. Serum herpes simplex antibodies

    Science.gov (United States)

    ... 2. HSV-1 most often causes cold sores (oral herpes). HSV-2 causes genital herpes. How the Test ... whether a person has ever been infected with oral or genital herpes . It looks for antibodies to herpes simplex virus ...

  3. Quantitation of anti-tetanus and anti-diphtheria antibodies by enzymoimmunoassay: methodology and applications.

    Science.gov (United States)

    Virella, G; Hyman, B

    1991-01-01

    We have developed enzymoimmunoassays (EIA) for the quantitation of antibodies (Ab) to tetanus and diphtheria toxoids (TT, DT) using Immulon I plates coated with the appropriate toxoid. A preparation of human tetanus immunoglobulin with a known concentration of anti-TT Ab was used as calibrator of the anti-TT antibody assay. The assay of anti-DT Ab is calibrated with a pool of human sera whose anti-DT Ab concentration was determined by quantitative immunoelectrophoresis, using a horse anti-DT with known Ab concentration as calibrator. A peroxidase-conjugated anti-human IgG was used in both assays. ABTS was used as substrate, and the reaction was stopped after 1 min incubation with citric acid and the OD measured at 414 nm on a Vmax reader. The assays have been applied to a variety of clinical situations. In patients suspected of having tetanus, the quantitation of antibodies has been helpful in establishing a diagnosis. In patients with a history of hypersensitivity to tetanus toxoid, verification of the levels of anti-TT antibody may prevent unnecessary and potentially harmful immunizations. The assays have also been used for the diagnostic evaluation of the humoral immune response to TT and DT, both in pediatric patients and in immunosuppressed patients. Several non-responders have been detected, and we have recently used the assay to monitor the effects of fish oil administration on the humoral immune response.(ABSTRACT TRUNCATED AT 250 WORDS)

  4. [Evaluation of allergen-specific IgE antibodies by a newly developed mast allergy system].

    Science.gov (United States)

    Nakagawa, T; Iwasaki, E; Baba, M; Matsushita, T; Baba, S; Ito, K; Miyamoto, T

    1989-06-01

    MAST, which stand for multiple antigen simultaneous test, uses enzyme-linked anti-human IgE and chemiluminogenic substate to determine IgE. This system is characterized by simultaneous analysis of multiple allergen items, up to 35, together with total IgE determination. We evaluated usefulness of this MAST system using 191 serum samples obtained from patients with bronchial asthma, allergic rhinitis and/or atopic dermatitis. It was found that there were statistically significant correlations between IgE antibody quantification by MAST and RAST in 24 out of 35 allergen items, with correlation coefficients more than r = 0.60. These included Dermatophagoides farinae and pteronyssinus, Japanese cedar pollen, orchard grass, Alternaria, Candida as inhalant allergens; egg white, milk, soybeans, wheat and rice as food allergens. It was also evaluated how many allergen-specific IgE antibodies could be detected in one serum sample. More than six allergen-specific IgE antibodies were simultaneously detected in 33% of 191 cases, indicating the importance of multiple-allergen analysis. These results indicate the clinical usefulness of the MAST allergy system in detecting IgE antibodies in allergic subjects.

  5. Heparin-Induced Thrombocytopenia Antibody Test

    Science.gov (United States)

    ... Global Sites Search Help? Heparin-induced Thrombocytopenia PF4 Antibody Share this page: Was this page helpful? Also known as: Heparin-PF4 Antibody; HIT Antibody; HIT PF4 Antibody; Heparin Induced Antibody; ...

  6. [New antibodies in cancer treatment].

    Science.gov (United States)

    Pestalozzi, B C; Knuth, A

    2004-09-22

    Since the development of hybridoma technology in 1975 monoclonal antibodies with pre-defined specificity can be produced. Only twenty years later did it become possible to make therapeutic use of monoclonal antibodies in oncology. To this end it was necessary to attach the antigen-binding site of a mouse antibody onto the scaffold of a human antibody molecule. Such chimeric or "humanized" antibodies may be used in passive immunotherapy without eliciting an immune response. Rituximab and trastuzumab are such humanized antibodies. They are used today routinely in the treatment of malignant lymphoma and breast cancer, respectively. These antibodies are usually used in combination with conventional cytostatic anticancer drugs.

  7. Engineering antibodies for cancer therapy.

    Science.gov (United States)

    Boder, Eric T; Jiang, Wei

    2011-01-01

    The advent of modern antibody engineering has led to numerous successes in the application of these proteins for cancer therapy in the 13 years since the first Food and Drug Administration approval, which has stimulated active interest in developing more and better drugs based on these molecules. A wide range of tools for discovering and engineering antibodies has been brought to bear on this challenge in the past two decades. Here, we summarize mechanisms of monoclonal antibody therapeutic activity, challenges to effective antibody-based treatment, existing technologies for antibody engineering, and current concepts for engineering new antibody formats and antibody alternatives as next generation biopharmaceuticals for cancer treatment.

  8. Natural and Man-made Antibody Repertories for Antibody Discovery

    Directory of Open Access Journals (Sweden)

    Juan C eAlmagro

    2012-11-01

    Full Text Available Antibodies are the fastest-growing segment of the biologics market. The success of antibody-based drugs resides in their exquisite specificity, high potency, stability, solubility, safety and relatively inexpensive manufacturing process in comparison with other biologics. We outline here the structural studies and fundamental principles that define how antibodies interact with diverse targets. We also describe the antibody repertoires and affinity maturation mechanisms of human, mice and chickens, plus the use of novel single-domain antibodies in camelids and sharks. These species all utilize diverse evolutionary solutions to generate specific and high affinity antibodies and illustrate the plasticity of natural antibody repertoires. In addition, we discuss the multiple variations of man-made antibody repertoires designed and validated in the last two decades, which have served as tools to explore how the size, diversity and composition of a repertoire impact the antibody discovery process.

  9. Preparation and reactivities of anti-porcine CD44 monoclonal antibodies.

    Science.gov (United States)

    Yang, H; Hutchings, A; Binns, R M

    1993-04-01

    Five monoclonal antibodies (MoAbs) were raised against porcine soluble CD44. The MoAbs recognized the same antigen on the surface of porcine lymphocytes as was recognized by anti-human CD44 MoAb Hermes-1, but identified five different epitopes. They bound to most porcine leucocytes but not to red cells. The epitopes were susceptible to treatment with papain or bromelain, whereas trypsinization of porcine leucocytes only reduced the antigen density. The epitopes seem to be co-expressed among various lymphoid tissues. The MoAbs also cross-reacted to various degrees with leucocytes of humans, dogs, sheep, cattle, goats and horses, suggesting that the corresponding epitopes are differentially conserved among species.

  10. Increased frequency of anti-retina antibodies in asymptomatic patients with chronic t. gondii infection

    Science.gov (United States)

    Cursino, Sylvia Regina Temer; da Costa, Thaís Boccia; Yamamoto, Joyce Hisae; Meireles, Luciana Regina; Silva, Maria Antonieta Longo Galvão; de Andrade Junior, Heitor Franco

    2010-01-01

    PURPOSE: To search for anti-retina antibodies that serve as markers for eye disease in uveitis. MATERIALS AND METHODS: Stored sera from patients with uveitis, ocular toxoplasmosis (n = 30) and non-infectious, immune-mediated uveitis (n = 50) and from asymptomatic individuals who were positive (n = 250) and negative (n = 250) for anti-Toxoplasma antibodies were tested. Serum anti-retina IgG was detected by an optimized ELISA using a solid-phase whole human retina extract, bovine S-antigen or interphotoreceptor retinoid-binding protein. RESULTS: Uveitis patients showed a higher mean reactivity to whole human retina extract, interphotoreceptor retinoid-binding protein and S-antigen in comparison to the asymptomatic population. These findings were independent of the uveitis origin and allowed the determination of the lower anti-retina antibody cut-off for the three antigens. Asymptomatic anti-Toxoplasma serum-positive individuals showed a higher frequency of anti-human whole retina extract antibodies in comparison to asymptomatic anti-Toxoplasma serum-negative patients. The bovine S-antigen and interphotoreceptor retinoid-binding protein ELISAs also showed a higher mean reactivity in the uveitis groups compared to the asymptomatic group, but the observed reactivities were lower and overlapped without discrimination. CONCLUSION: We detected higher levels of anti-retina antibodies in uveitis patients and in a small fraction of asymptomatic patients with chronic toxoplasmosis. The presence of anti-retina antibodies in sera might be a marker of eye disease in asymptomatic patients, especially when whole human retina extract is used in a solid-phase ELISA. PMID:21120306

  11. Antibody affinity maturation

    DEFF Research Database (Denmark)

    Skjødt, Mette Louise

    surface expression of various antibody formats in the generated knockout strain. Functional scFv and scFab fragments were efficiently displayed on yeast whereas impaired chain assembly and heavy chain degradation was observed for display of full-length IgG molecules. To identify the optimal polypeptide...... linker for yeast surface display of scFv and scFab fragments, we compared a series of different Gly-Ser-based linkers in display and antigen binding proficiency. We show that these formats of the model antibody can accommodate linkers of different lengths and that introduction of alanine or glutamate...... fragments by in vivo homologous recombination large combinatorial antibody libraries can easily be generated. We have optimized ordered assembly of three CDR fragments into a gapped vector and observed increased transformation efficiency in a yeast strain carrying a deletion of the SGS1 helicase...

  12. Antibody informatics for drug discovery

    DEFF Research Database (Denmark)

    Shirai, Hiroki; Prades, Catherine; Vita, Randi;

    2014-01-01

    to the antibody science in every project in antibody drug discovery. Recent experimental technologies allow for the rapid generation of large-scale data on antibody sequences, affinity, potency, structures, and biological functions; this should accelerate drug discovery research. Therefore, a robust bioinformatic...... infrastructure for these large data sets has become necessary. In this article, we first identify and discuss the typical obstacles faced during the antibody drug discovery process. We then summarize the current status of three sub-fields of antibody informatics as follows: (i) recent progress in technologies...... for antibody rational design using computational approaches to affinity and stability improvement, as well as ab-initio and homology-based antibody modeling; (ii) resources for antibody sequences, structures, and immune epitopes and open drug discovery resources for development of antibody drugs; and (iii...

  13. Antithyroglobulin Antibodies and Antimicrosomal Antibodies in Various Thyroid Diseases

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Gwon Jun; Hong, Key Sak; Choi, Kang Won; Lee, Kyu; Koh, Chang Soon; Lee, Mun Ho; Park, Sung Hoe; Chi, Je Geun; Lee, Sang Kook [Seoul National University College of Medicine, Seoul (Korea, Republic of)

    1979-03-15

    The authors investigated the incidence of antithyroglobulin antibodies and antibodies and antimicrosomal antibodies measured by tanned red cell hemagglutination method in subjects suffering from various thyroid disorders. 1) In 15 normal patients, neither suffering from any thyroid diseases nor from any other autoimmune disorders, the antithyroglobulin antibodies were all negative, but the antimicrosomal antibody was positive only in one patient (6.7%). 2) The antithyroglobulin antibodies were positive in 31.5% (34 patients) of 108 patients with various thyroid diseases, and the antimicrosomal antibodies were positive in 37.0% (40 patients). 3) of the 25 patients with Graves' diseases, 7 patients (28.0%) showed positive for the antithyroglobulin antibodies, and 9 (36.0%) for the antimicrosomal antibodies. There was no definite differences in clinical and thyroid functions between the groups with positive and negative results. 4) Both antibodies were positive in 16 (88.9%) and 17 (94.4%) patients respectively among 18 patients with Hashimoto's thyroiditis, all of them were diagnosed histologically. 5) Three out of 33 patients with thyroid adenoma showed positive antibodies, and 3 of 16 patients with thyroid carcinoma revealed positive antibodies. 6) TRCH antibodies demonstrated negative results in 2 patients with subacute thyroiditis, but positive in one patient with idiopathic primary myxedema. 7) The number of patients with high titers(>l:802) was 16 for antithyroglobulin antibody, and 62.5% (10 patients) of which was Hashimoto's thyroiditis. Thirteen (65.0) of 20 patients with high titers (>l:802) for antimicrosomal antibody was Hashimoto's thyroiditis. TRCH test is a simple, sensitive method, and has high reliability and reproducibility. The incidences and titers of antithyroglobulin antibody and antimicrosomal antibody are especially high in Hashimoto's thyroiditis.

  14. Prediction of Antibody Epitopes

    DEFF Research Database (Denmark)

    Nielsen, Morten; Marcatili, Paolo

    2015-01-01

    Antibodies recognize their cognate antigens in a precise and effective way. In order to do so, they target regions of the antigenic molecules that have specific features such as large exposed areas, presence of charged or polar atoms, specific secondary structure elements, and lack of similarity...... to self-proteins. Given the sequence or the structure of a protein of interest, several methods exploit such features to predict the residues that are more likely to be recognized by an immunoglobulin.Here, we present two methods (BepiPred and DiscoTope) to predict linear and discontinuous antibody...

  15. Anti-human CD73 monoclonal antibody inhibits metastasis formation in human breast cancer by inducing clustering and internalization of CD73 expressed on the surface of cancer cells

    DEFF Research Database (Denmark)

    Terp, Mikkel G; Olesen, Kristina A; Christensen, Eva Arnspang

    2013-01-01

    -linking of CD73, because both whole IgG anti-CD73 AD2 mAb and Fab' fragments thereof exhibited this effect. Ex vivo treatment of different breast cancer cell lines with anti-CD73 AD2 mAb before i.v. injection into mice inhibited extravasation/colonization of circulating tumor cells and significantly reduced...... internalization and metastasis inhibition. Furthermore, anti-CD73 AD2 mAb inhibited development of metastasis in a spontaneous animal model of human metastatic breast cancer. Our study shows that some anti-CD73 mAbs cause cell-surface clustering of CD73 followed by internalization, thus inhibiting the ability...... another anticancer mechanism of anti-CD73 Abs and show that an anti-CD73 mAb (AD2) inhibits metastasis formation by a mechanism independent of CD73 catalytic activity and inhibition of primary tumor growth. This mechanism involves clustering and internalization of CD73, but does not require cross...

  16. 人去唾液酸糖蛋白受体单克隆抗体制备及其初步应用%Preparation of mouse anti-human asialoglycoprotein receptor monoclonal antibody and its preliminary application

    Institute of Scientific and Technical Information of China (English)

    李军; 陈磊; 孙斌; 张妤; 钱海华; 殷正丰

    2013-01-01

    目的 制备人去唾液酸糖蛋白受体(ASGPR)的小鼠单克隆抗体,并应用该抗体检测ASGPR在细胞系和组织中的表达.方法 对ASGPR H1大亚基进行序列分析,选取ASGPR1全长序列制备免疫原.通过RT PCR合成ASGPR1cDNA,构建ASGPR1表达重组载体,在E.coli BL21中表达后纯化,获得目的抗原.应用传统融合杂交瘤技术制备小鼠单克隆抗体,常规检测单抗亚类和效价,竞争抑制实验鉴定单抗特异性.用流式细胞术和免疫组织化学法分别检测ASGPR在多种肝源性与非肝源性细胞系以及多种肝组织中的表达.结果 制备的单克隆抗体亚型为IgG1,效价达1∶12 800.竞争抑制实验结果显示该单抗具有很好的ASGPR结合特异性,而且识别的肽段位于ASGPR胞外段.流式细胞术检测结果显示,ASGPR不同程度地表达于肝源性细胞系,但不表达于非肝源性细胞系.免疫组织化学检测结果显示,ASGPR专一性表达于正常肝组织和肝癌组织,在肝癌组织的表达与分化程度有关,高分化肝癌中的阳性率高于低分化肝癌(75.0% vs 28.6%,P<0.05).结论 成功制备高特异性人ASGPR小鼠单克隆抗体,适用于用流式细胞术和免疫组织化学法检测ASGPR表达,可用于临床病理鉴定原发性肝癌与转移性肝癌.

  17. Compositions, antibodies, asthma diagnosis methods, and methods for preparing antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Hongjun; Zangar, Richard C.

    2017-01-17

    Methods for preparing an antibody are provided with the method including incorporating 3-bromo-4-hydroxy-benzoic acid into a protein to form an antigen, immunizing a mammalian host with the antigen, and recovering an antibody having an affinity for the antigen from the host. Antibodies having a binding affinity for a monohalotyrosine are provided as well as composition comprising an antibody bound with monohalotyrosine. Compositions comprising a protein having a 3-bromo-4-hydroxy-benzoic acid moiety are also provided. Methods for evaluating the severity of asthma are provide with the methods including analyzing sputum of a patient using an antibody having a binding affinity for monohalotyrosine, and measuring the amount of antibody bound to protein. Methods for determining eosinophil activity in bodily fluid are also provided with the methods including exposing bodily fluid to an antibody having a binding affinity for monohalotyrosine, and measuring the amount of bound antibody to determine the eosinophil activity.

  18. Human germline antibody gene segments encode polyspecific antibodies.

    Science.gov (United States)

    Willis, Jordan R; Briney, Bryan S; DeLuca, Samuel L; Crowe, James E; Meiler, Jens

    2013-04-01

    Structural flexibility in germline gene-encoded antibodies allows promiscuous binding to diverse antigens. The binding affinity and specificity for a particular epitope typically increase as antibody genes acquire somatic mutations in antigen-stimulated B cells. In this work, we investigated whether germline gene-encoded antibodies are optimal for polyspecificity by determining the basis for recognition of diverse antigens by antibodies encoded by three VH gene segments. Panels of somatically mutated antibodies encoded by a common VH gene, but each binding to a different antigen, were computationally redesigned to predict antibodies that could engage multiple antigens at once. The Rosetta multi-state design process predicted antibody sequences for the entire heavy chain variable region, including framework, CDR1, and CDR2 mutations. The predicted sequences matched the germline gene sequences to a remarkable degree, revealing by computational design the residues that are predicted to enable polyspecificity, i.e., binding of many unrelated antigens with a common sequence. The process thereby reverses antibody maturation in silico. In contrast, when designing antibodies to bind a single antigen, a sequence similar to that of the mature antibody sequence was returned, mimicking natural antibody maturation in silico. We demonstrated that the Rosetta computational design algorithm captures important aspects of antibody/antigen recognition. While the hypervariable region CDR3 often mediates much of the specificity of mature antibodies, we identified key positions in the VH gene encoding CDR1, CDR2, and the immunoglobulin framework that are critical contributors for polyspecificity in germline antibodies. Computational design of antibodies capable of binding multiple antigens may allow the rational design of antibodies that retain polyspecificity for diverse epitope binding.

  19. Prediction of antibody persistency from antibody titres to natalizumab

    DEFF Research Database (Denmark)

    Jensen, Poul Erik H; Koch-Henriksen, Nils; Sellebjerg, Finn Thorup;

    2012-01-01

    In a subgroup of patients with multiple sclerosis natalizumab therapy causes generation of anti-natalizumab antibodies that may be transient or persistent. It is recommended to discontinue natalizumab therapy in persistently antibody-positive patients.......In a subgroup of patients with multiple sclerosis natalizumab therapy causes generation of anti-natalizumab antibodies that may be transient or persistent. It is recommended to discontinue natalizumab therapy in persistently antibody-positive patients....

  20. Monoclonal antibodies in myeloma

    DEFF Research Database (Denmark)

    Sondergeld, P.; van de Donk, N. W. C. J.; Richardson, P. G.;

    2015-01-01

    The development of monoclonal antibodies (mAbs) for the treatment of disease goes back to the vision of Paul Ehrlich in the late 19th century; however, the first successful treatment with a mAb was not until 1982, in a lymphoma patient. In multiple myeloma, mAbs are a very recent and exciting add...

  1. Antibody Blood Tests

    Science.gov (United States)

    ... What do I do if I have a negative blood test (or panel) but I’m still having symptoms? While it is rare, it is possible for patients to have a negative antibody test results and still have celiac disease. ...

  2. RBC Antibody Screen

    Science.gov (United States)

    ... test also may be used to help diagnose autoimmune-related hemolytic anemia in conjunction with a DAT. This condition may be caused when a person produces antibodies against his or her own RBC antigens. This can happen with some autoimmune disorders , such as lupus , with diseases such as ...

  3. Acute antibody-directed myostatin inhibition attenuates disuse muscle atrophy and weakness in mice.

    Science.gov (United States)

    Murphy, Kate T; Cobani, Vera; Ryall, James G; Ibebunjo, Chikwendu; Lynch, Gordon S

    2011-04-01

    Counteracting the atrophy of skeletal muscle associated with disuse has significant implications for minimizing the wasting and weakness in plaster casting, joint immobilization, and other forms of limb unloading, with relevance to orthopedics, sports medicine, and plastic and reconstructive surgery. We tested the hypothesis that antibody-directed myostatin inhibition would attenuate the loss of muscle mass and functional capacity in mice during 14 or 21 days of unilateral hindlimb casting. Twelve-week-old C57BL/10 mice were subjected to unilateral hindlimb plaster casting or served as controls. Mice received subcutaneous injections of saline or a mouse chimera of anti-human myostatin antibody (PF-354, 10 mg/kg; n = 6-9) on days 0 and 7 and were tested for muscle function on day 14, or were treated on days 0, 7, and 14 and tested for muscle function on day 21. Hindlimb casting reduced muscle mass, fiber size, and function of isolated soleus and extensor digitorum longus (EDL) muscles (P casting, when wasting and weakness had plateaued (P casting with reductions in muscle size and strength being most apparent during the first 14 days of disuse. These findings highlight the therapeutic potential of antibody-directed myostatin inhibition for disuse atrophy especially within the first 2 wk of disuse.

  4. Plasma functionalization procedure for antibody immobilization for SU-8 based sensor.

    Science.gov (United States)

    Grimaldi, Immacolata Angelica; Testa, Genni; Persichetti, Gianluca; Loffredo, Fausta; Villani, Fulvia; Bernini, Romeo

    2016-12-15

    In this paper, we report the study on a new protocol for the immobilization process of antigen/antibody assay on SU-8 layers by oxygen plasma treatment. Plasma treatments, at different plasma powers and for different duration times, are performed and their effects on immobilization efficiency are studied. The chemical properties and the surface morphology of SU-8 before and after the functionalization and immobilization of (IgG) are then verified by Raman spectroscopy and atomic force microscopy (AFM). An increase of the surface roughness of SU-8 layers is observed after the oxygen plasma treatment and an intensity variation of functional groups is also evidenced. To demonstrate the validity of the process the distribution of IgG immobilized on SU-8 surfaces is detected by fluorescence microscopy measurement after incubation with fluorescein isothiocyanate (FITC)-tagged anti-human IgG. An increase of the amount of the adsorbed protein of about 20% and a good repeatability on antigen/antibody distribution on the surface are detected for IgG on plasma treated substrates. Finally, label free measurements are performed by SU-8 optical ring resonators reaching detection limits of 0.86ngcm(-2). The proposed approach offers a smart protocol for IgG immobilization on SU-8 substrate that can be easily extended to different antigen/antibody assay and polymeric materials for the realization of high performance immunosensors.

  5. Effect of apoptosis of CD4+CD25+ regulatory T lymphocytes on polarization of helper T lymphocytes and potential interventional influence of Xuebijing injection (血必净注射液) in septic rats%脓毒症大鼠调节性T细胞凋亡对辅助性T细胞漂移的影响及血必净注射液的干预作用

    Institute of Scientific and Technical Information of China (English)

    戴新贵; 姚咏明; 艾宇航

    2009-01-01

    Objective To evaluate the influence of apoptosis of CD4+CD25+ regulatory T lymphocyte (Treg) on polarization of helper T lymphocyte (Th) and effect of Xuebijing injection (血必净注射液) in septic rats. Methods A sepsis model was reproduced by cecal ligation and puncture (CLP). Wistar rats were randomly divided into the normal group (n=8),sham operation group (n=8),model group (n=8) and Xuebijing injection treatment group (n=8). CD4+CD25+Tregs in each group were separated by immunomagnetic beads isolation system on day 3,the apoptotic rates,expression of forkhead/winged helix transcription factor p3 (Foxp3) as well as cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) of Treg were analyzed by flow cytometry,and the secretion level of interleukin-10 (IL-10) of Tregs was determined by enzyme linked immunosorbent assay (ELISA) after culturing Tregs for 12 hours. Interferon-γ (IFN-γ),IL-4,and IL-17 levels,which were respectively secreted by Th1,Th2 and Th17,were measured by ELISA in the supernatant after CD4+CD25+Tregs were co-cultured with CD4+CD25-T lymphocytes for 68 hours. Results The apoptosis rate of the normal group was (12.03±0.89)%,which was not significantly different compared with the sham operation group [(9.48±2.17)%]. The apoptosis rate of model group [(5.87±0.44)%] was much lower than that of the normal and sham operation groups,while the Xuebijing injection treatment group [(27.29±2.48)%] had the highest apoptosis rate compared to others (all P0.05). Conclusion Due to the apoptosis of Treg,the suppression function of CD4+CD25+Tregs on CD4+T lymphocyte appears to be abated,and treatment with Xuebijing injection could effectively enhance the apoptosis of Tregs,mediating the response of shifting Th2 to Th1.%目的 探讨脓毒症大鼠CD4+CD25+调节性T细胞(Treg)凋亡对辅助性T细胞(Th)漂移的影响及血必净注射液的干预作用.方法 将Wistar大鼠随机分为正常组、假手术组、模型组、血必净组,每组8只.

  6. What Is Antiphospholipid Antibody Syndrome?

    Science.gov (United States)

    ... page from the NHLBI on Twitter. What Is Antiphospholipid Antibody Syndrome? Antiphospholipid (AN-te-fos-fo-LIP-id) antibody ... weeks or months. This condition is called catastrophic antiphospholipid syndrome (CAPS). People who have APS also are at ...

  7. Red Blood Cell Antibody Identification

    Science.gov (United States)

    ... ID, RBC; RBC Ab ID Formal name: Red Blood Cell Antibody Identification Related tests: Direct Antiglobulin Test ; RBC ... I should know? How is it used? Red blood cell (RBC) antibody identification is used as a follow- ...

  8. Lupus anticoagulants and antiphospholipid antibodies

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/000547.htm Lupus anticoagulants and antiphospholipid antibodies To use the sharing features on this page, please enable JavaScript. Lupus anticoagulants are antibodies against substances in the lining ...

  9. Anti-smooth muscle antibody

    Science.gov (United States)

    ... gov/ency/article/003531.htm Anti-smooth muscle antibody To use the sharing features on this page, please enable JavaScript. Anti-smooth muscle antibody is a blood test that detects the presence ...

  10. Antibody Engineering and Therapeutics Conference

    OpenAIRE

    Larrick, James W; Parren, Paul WHI; Huston, James S; Plückthun, Andreas; Bradbury, Andrew; Tomlinson, Ian M; Chester, Kerry A.; Burton, Dennis R.; Adams, Gregory P.; Weiner, Louis M.; Scott, Jamie K.; Alfenito, Mark R; Veldman, Trudi; Reichert, Janice M.

    2013-01-01

    The Antibody Engineering and Therapeutics conference, which serves as the annual meeting of The Antibody Society, will be held in Huntington Beach, CA from Sunday December 8 through Thursday December 12, 2013. The scientific program will cover the full spectrum of challenges in antibody research and development, and provide updates on recent progress in areas from basic science through approval of antibody therapeutics. Keynote presentations will be given by Leroy Hood (Institute of System Bi...

  11. Structural Characterization of Peptide Antibodies

    DEFF Research Database (Denmark)

    Chailyan, Anna; Marcatili, Paolo

    2015-01-01

    The role of proteins as very effective immunogens for the generation of antibodies is indisputable. Nevertheless, cases in which protein usage for antibody production is not feasible or convenient compelled the creation of a powerful alternative consisting of synthetic peptides. Synthetic peptides...... can be modified to obtain desired properties or conformation, tagged for purification, isotopically labeled for protein quantitation or conjugated to immunogens for antibody production. The antibodies that bind to these peptides represent an invaluable tool for biological research and discovery...

  12. Synthetic peptides for antibody production

    NARCIS (Netherlands)

    Zegers, N.D.

    1995-01-01

    Synthetic peptides are useful tools for the generation of antibodies. The use of antibodies as specific reagents in inununochemical assays is widely applied. In this chapter, the application of synthetic peptides for the generation of antibodies is described. The different steps that lead to the uni

  13. The Presence of Anti-HLA Antibodies before and after Allogeneic Hematopoietic Stem Cells Transplantation from HLA-Mismatched Unrelated Donors

    Directory of Open Access Journals (Sweden)

    Anna Koclega

    2012-01-01

    Full Text Available Although anti-human leukocyte antigen antibodies (anti-HLA Abs are important factors responsible for graft rejection in solid organ transplantation and play a role in post-transfusion complications, their role in allogeneic hematopoietic stem cell transplantation (allo-HSCT has not been finally defined. Enormous polymorphism of HLA-genes, their immunogenicity and heterogeneity of antibodies, as well as the growing number of allo-HSCTs from partially HLA-mismatched donors, increase the probability that anti-HLA antibodies could be important factors responsible for the treatment outcomes. We have examined the incidence of anti-HLA antibodies in a group of 30 allo-HSCT recipients from HLA-mismatched unrelated donors. Anti-HLA Abs were identified in sera collected before and after allo-HSCT. We have used automated DynaChip assay utilizing microchips bearing purified class I and II HLA antigens for detection of anti-HLA Abs. We have detected anit-HLA antibodies against HLA-A, B, C, DR, DQ and DP, but no donor or recipient-specific anti-HLA Abs were detected in the studied group. The preliminary results indicate that anti-HLA antibodies are present before and after allo-HSCT in HLA-mismatched recipients.

  14. Surface modification of polyacrylonitrile fiber for immobilization of antibodies and detection of analyte

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Swati, E-mail: swatijain.iitd@gmail.com [Center for Biomedical Engineering, Indian Institute of Technology, New Delhi, 110016 (India); Chattopadhyay, Sruti, E-mail: srutic@hotmail.com [Center for Biomedical Engineering, Indian Institute of Technology, New Delhi, 110016 (India); Jackeray, Richa, E-mail: richajackeray.iitd@gmail.com [Center for Biomedical Engineering, Indian Institute of Technology, New Delhi, 110016 (India); Singh, Harpal, E-mail: harpal2000@yahoo.com [Center for Biomedical Engineering, Indian Institute of Technology, New Delhi, 110016 (India)

    2009-11-10

    Pendent nitrile groups of multifilamentous polyacrylonitrile (PAN) fibers were reduced to amino groups using lithium aluminum hydride for different time of reduction and amine content was estimated by performing acid-base titrations. Attenuated total reflection-fourier transform infrared spectroscopy (ATR-FTIR) and Differential Scanning Calorimetry (DSC) were used for the characterization of the generated amino groups and thermal properties of the reduced fibers, respectively. The surface morphology of the fibers after reduction and immobilization was characterized using Scanning Electron Microscope (SEM). The newly formed amino groups of the fibers were activated by using glutaraldehyde for the covalent linking of Goat anti-Rabbit IgG-HRP (GAR-HRP) antibody enzyme conjugate. Modified PAN fibers were evaluated as a matrix for sandwich ELISA by using Goat anti-Rabbit antibody (GAR-IgG), Rabbit anti-Goat (RAG-IgG) as analyte and enzyme conjugate GAR-HRP. The fibers reduced for 24 h were able to detect the analyte RAG-IgG at a concentration as low as 3.75 ng mL{sup -1} with 12% skimmed milk as blocking reagent for the optimized concentration of primary antibody GAR-IgG 3 {mu}g mL{sup -1} and peroxidase conjugate GAR-HRP dilution of 8000 fold. The sensitivity, specificity and reproducibility of the developed immunoassay was further established with antibodies present in human blood using Rabbit anti-Human (RAH-IgG) antibody and the corresponding HRP enzyme conjugate. As low as 0.1 {mu}L of human blood was sufficient to perform the assay with the modified fibers.

  15. Antiphospholipid Antibody and Antiphospholipid Syndrome

    Institute of Scientific and Technical Information of China (English)

    吴竞生

    2008-01-01

    @@ Antiphospholipid antibodies (APA) APA is a big category for all kinds of negative charge phospholipid or lecithin - a protein complex autoantibodies or the same antibody, through its recognition of antigen (target protein) different, and phospholipids or lecithin - protein complex combination of various rely on the interference Phospholipid clotting and anti-coagulation factor, and promote endothelial cells, platelets, complement activation and play a role. APA including lupus anticoagulant(LA) and anticardiolipin antibody (ACA), In addition, there are anti-β2 glycoprotein-I (β2-GPI) antibody, anti-prothrombin (a- PT) antibody, anti-lysophosphatidic acid antibody and anti-phosphatidylserine antibody, and so on. APA as the main target of phospholipid-binding protein, including β2-GPI, prothrombin, annexin, protein C (PC) and protein S (PS), plasminogen, and so on.

  16. Engineering antibodies by yeast display.

    Science.gov (United States)

    Boder, Eric T; Raeeszadeh-Sarmazdeh, Maryam; Price, J Vincent

    2012-10-15

    Since its first application to antibody engineering 15 years ago, yeast display technology has been developed into a highly potent tool for both affinity maturing lead molecules and isolating novel antibodies and antibody-like species. Robust approaches to the creation of diversity, construction of yeast libraries, and library screening or selection have been elaborated, improving the quality of engineered molecules and certainty of success in an antibody engineering campaign and positioning yeast display as one of the premier antibody engineering technologies currently in use. Here, we summarize the history of antibody engineering by yeast surface display, approaches used in its application, and a number of examples highlighting the utility of this method for antibody engineering.

  17. A cell sorter with modified bamboo charcoal for the efficient selection of specific antibody-producing hybridomas.

    Science.gov (United States)

    Lin, Chien-Chen; Ni, Mei-Hui; Chang, Yu-Chung; Yeh, Hsiu-Lun; Lin, Feng-Huei

    2010-11-01

    Monoclonal antibodies (mAbs) have been proven useful in research and clinical applications. However, the generation of mAbs by conventional hybridoma technology is time-, cost- and labor-consuming. Here we developed a simplified procedure for efficient generation and selection of antibody-producing hybridomas within 1 h, using a particular cell sorter design, a cytoflow reactor-based cell sorter (CBCS) which consists mainly of the "cytoflow reactor" that comprises two components, a reaction chamber and a glass tubing for air and medium exchange by gravity, and the "sorting material", human EGFR-conjugated bamboo charcoal, for specific B-cell enrichment. The high surface area and porous structure of bamboo charcoal greatly increased cell density and protein production. Moreover, from Raman, FT-IR spectroscopy and IFA analysis, the carboxylation and immobilization of bamboo charcoal can be introduced easily by nitric acid treatment and conjugated handily with human EGFR using EDC/NHS. Other evidences, such as IFA, showed that the specific hybridomas generated in this study could secrete specific anti-human EGFR antibodies. Our design allows the production of mAbs while avoiding time-consuming steps, such as large numbers of limiting dilutions and screening assays, and demonstrates that the CBCS could be a powerful tool for monoclonal antibody production.

  18. A sensitive and specific ELISA using a monoclonal capture antibody for detection of Tamm-Horsfall urinary glycoprotein in serum.

    Science.gov (United States)

    Hunt, J S; Peach, R J; Brünisholz, M C; Lynn, K L; McGiven, A R

    1986-07-11

    An enzyme-linked immunosorbent assay (ELISA) has been established using Nunc polystyrene immunoplates coated with a monoclonal antibody to human Tamm-Horsfall urinary glycoprotein (THP) to detect and measure THP in human serum. Optimal reaction conditions for both the monoclonal capture antibody and the affinity-purified rabbit anti-human THP second antibody were established to produce standard curves which showed linearity between 20-90 ng/ml with a sensitivity of 2-3 ng/ml. The plate-to-plate standard curve mean coefficient of variation (CV) was 5.9 +/- 2.9% on assays performed on the same day while day to day mean CV was 13.3 +/- 2.4%. The specificity of the ELISA was demonstrated by inhibition of binding after preincubation of both urinary THP standards and serum with monoclonal anti-THP antibody. Sera from 195 blood donors tested by the ELISA had a mean concentration of THP antigenic determinants of 260 +/- 105 ng/ml. Results from three control sera run on all plates used in the survey showed mean CV less than 7.6% while no binding was observed with sera from an anephric patient.

  19. How antibodies use complement to regulate antibody responses.

    Science.gov (United States)

    Sörman, Anna; Zhang, Lu; Ding, Zhoujie; Heyman, Birgitta

    2014-10-01

    Antibodies, forming immune complexes with their specific antigen, can cause complete suppression or several 100-fold enhancement of the antibody response. Immune complexes containing IgG and IgM may activate complement and in such situations also complement components will be part of the immune complex. Here, we review experimental data on how antibodies via the complement system upregulate specific antibody responses. Current data suggest that murine IgG1, IgG2a, and IgG2b upregulate antibody responses primarily via Fc-receptors and not via complement. In contrast, IgM and IgG3 act via complement and require the presence of complement receptors 1 and 2 (CR1/2) expressed on both B cells and follicular dendritic cells. Complement plays a crucial role for antibody responses not only to antigen complexed to antibodies, but also to antigen administered alone. Lack of C1q, but not of Factor B or MBL, severely impairs antibody responses suggesting involvement of the classical pathway. In spite of this, normal antibody responses are found in mice lacking several activators of the classical pathway (complement activating natural IgM, serum amyloid P component (SAP), specific intracellular adhesion molecule-grabbing non-integrin R1 (SIGN-R1) or C-reactive protein. Possible explanations to these observations will be discussed.

  20. The antibody Hijikata Tatsumi

    Directory of Open Access Journals (Sweden)

    Éden Peretta

    2012-11-01

    Full Text Available Considered one of the most influential modern dance representatives in Japan, Tatsumi Hijikata’s work was a milestone in the Japanese post-war experimental artistic scene. Heretic son of his time, he staged a fertile mix of artistic and cultural influences, overlapping subversive elements of European arts and philosophy with radical references from pre-modern Japanese culture. In this way he built the foundations of its unstable antibody, its political-artistic project of dissolution of a organism, both physical and social.

  1. Cancer imaging with radiolabeled antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Goldenberg, D.M. (Center for Molecular Medicine and Immunology, Newark, NJ (US))

    1990-01-01

    This book presents a perspective of the use of antibodies to target diagnostic isotopes to tumors. Antibodies with reasonable specificity can be developed against almost any substance. If selective targeting to cancer cells can be achieved, the prospects for a selective therapy are equally intriguing. But the development of cancer detection, or imaging, with radiolabeled antibodies has depended upon advances in a number of different areas, including cancer immunology and immunochemistry for identifying suitable antigen targets and antibodies to these targets, tumor biology for model systems, radiochemistry for he attachment of radionuclides to antibodies, molecular biology for reengineering the antibodies for safer and more effective use in humans, and nuclear medicine for providing the best imaging protocols and instrumentation to detect minute amounts of elevated radioactivity against a background of considerable noise. Accordingly, this book has been organized to address the advances that are being made in many of these areas.

  2. VIRAL ANTIBODIES IN PRESCHOOL CHILDREN

    Directory of Open Access Journals (Sweden)

    S. Saidi

    1974-08-01

    Full Text Available One hundred sera from children 1 - 6 years of age, representative of a large serum collection, were tested for the prevalence of antibodies against different viruses. Hemagglutination-inhibition (HI antibodies were found in 68% for measles; 61 % for rubella; 75'% for influenza A2/Hong Kong/68, 16% for influenza B/Md./59, 0% for group A arboviruses, 10% for group B arboviruses, 3% for phlebotomus fever group and 4% for Congo-Crimean hemorrhagic fever (C-CHF group of arboviruses Poliomyelitis-neutralizing antibodies for type 1, 2 and 3 were 90%; 85% and 84%~ respectively. Antibody to EH virus was detected in 84% of the sera by immuno-fluorescence. None of the sera were positive for hepatitis-B antigen or antibody by immuno-precipitation test. The prevalence of some viral antibodies found in this survey are compared with results obtained from surveys in other parts of the country.

  3. Oriented immobilized anti-LDL antibody carrying poly(hydroxyethyl methacrylate) cryogel for cholesterol removal from human plasma

    Energy Technology Data Exchange (ETDEWEB)

    Bereli, Nilay [Department of Chemistry, Hacettepe University, Beytepe, Ankara (Turkey); Sener, Guelsu [Nanotechnology and Nanomedicine Division, Hacettepe University, Ankara (Turkey); Yavuz, Handan, E-mail: handany@hacettepe.edu.tr [Department of Chemistry, Hacettepe University, Beytepe, Ankara (Turkey); Denizli, Adil [Department of Chemistry, Hacettepe University, Beytepe, Ankara (Turkey)

    2011-07-20

    Low density lipoprotein (LDL) cholesterol is a major ingredient of the plaque that collects in the coronary arteries and causes coronary heart diseases. Among the methods used for the extracorporeal elimination of LDL from intravasal volume, immunoaffinity technique using anti-LDL antibody as a ligand offers superior selectivity and specificity. Proper orientation of the immobilized antibody is the main issue in immunoaffinity techniques. In this study, anti-human {beta}-lipoprotein antibody (anti-LDL antibody) molecules were immobilized and oriented through protein A onto poly(2-hydroxyethyl methacrylate) (PHEMA) cryogel in order to remove LDL from hypercholesterolemic human plasma. PHEMA cryogel was prepared by free radical polymerization initiated with N,N,N',N'-tetramethylene diamine (TEMED). PHEMA cryogel with a swelling degree of 8.89 g H{sub 2}O/g and 67% macro-porosity was characterized by swelling studies, scanning electron microscope (SEM) and blood compatibility tests. All the clotting times were increased when compared with control plasma. The maximum immobilized anti-LDL antibody amount was 63.2 mg/g in the case of random antibody immobilization and 19.6 mg/g in the case of oriented antibody immobilization (protein A loading was 57.0 mg/g). Random and oriented anti-LDL antibody immobilized PHEMA cryogels adsorbed 111 and 129 mg LDL/g cryogel from hypercholesterolemic human plasma, respectively. Up to 80% of the adsorbed LDL was desorbed. The adsorption-desorption cycle was repeated 6 times using the same cryogel. There was no significant loss of LDL adsorption capacity. - Research highlights: {yields} LDL cholesterol is a risk factor in the development of coronary heart diseases. {yields} Antibodies against LDL are used for the selective extracorporeal removal of LDL. {yields} Protein A is used for the oriented immobilization of anti LDL onto PHEMA cryogel. {yields} PHEMA cryogels are biocompatible, exhibit a low pressure drop, lack diffusion

  4. Automated in situ measurement of cell-specific antibody secretion and laser-mediated purification for rapid cloning of highly-secreting producers.

    Science.gov (United States)

    Hanania, Elie G; Fieck, Annabeth; Stevens, Janine; Bodzin, Leon J; Palsson, Bernhard Ø; Koller, Manfred R

    2005-09-30

    Cloning of highly-secreting recombinant cells is critical for biopharmaceutical manufacturing, but faces numerous challenges including the fact that secreted protein does not remain associated with the producing cell. A fundamentally new approach was developed combining in situ capture and measurement of individual cell protein secretion followed by laser-mediated elimination of all non- and poorly-secreting cells, leaving only the highest-secreting cell in a well. Recombinant cells producing humanized antibody were cultured serum-free on a capture matrix, followed by staining with fluorescently-labeled anti-human antibody fragment. A novel, automated, high-throughput instrument (called LEAP) was used to image and locate every cell, quantify the cell-associated and secreted antibody (surrounding each cell), eliminate all undesired cells from a well via targeted laser irradiation, and then track clone outgrowth and stability. Temporarily sparing an island of helper cells around the clone of interest improved cloning efficiency (particularly when using serum-free medium), and helper cells were easily eliminated with the laser after several days. The in situ nature of this process allowed several serial sub-cloning steps to be performed within days of one another, resulting in rapid generation of clonal populations with significantly increased and more stable, homogeneous antibody secretion. Cell lines with specific antibody secretion rates of > 50 pg/cell per day (in static batch culture) were routinely obtained as a result of this cloning approach, often times representing up to 20% of the clones screened.

  5. RE-186 labeled 16.88 IgM and 88BV59 IgG human antibody studies to assess potential for radioimmunotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Breitz, H.; Seiler, C.; Weiden, P. [Virginia Mason Medical Center, Seattle, WA (United States)]|[NeoRx Corp, Seattle, WA (United States)]|[Organon Teknike/Biotechnology Research Institute, Rockville, MD (United States)] [and others

    1994-05-01

    Two studies with Re-186-MAG{sub 2}GABA labeled human antibodies were carried out to assess feasibility for radioimmunotherapy. Antibodies 16.88 and 88BV59 react with different epitopes of CTA 16.88, a tumor associated antigen of colorectal carcinoma. In a phase I dose escalation study, 14 patients received 60 mg/m{sup 2} 16.88 IgM MoAb. The dose of Re-186 ranged from 25 mCi/m{sup 2} to 210 mCi/m{sup 2} divided into 3 weekly infusions. In a pilot study with 88BV59, a human IgG3k MoAb, 20 mg antibody was labeled with 25 mCi/m{sup 2} Re-186 and administered to 4 patients with colon carcinoma. Tumor targeting was seen in 12 of 14 patients with 16.88 and all 4 patients with 88BV59. Retention of antibody at the tumor was longer with 88BV59. One patient developed a rash. No other acute or delayed toxicities were observed. Human anti-human antibody did not develop in any patient. The slower metabolism of the 88BV59 IgG suggests that this form of immunoconjugate merits further investigation for use in radioimmunotherapy.

  6. Antibodies against antibodies: immunogenicity of adalimumab as a model

    NARCIS (Netherlands)

    van Schouwenburg, P.A.

    2012-01-01

    Upon repeated adalimumab exposure part of the patients start to produce ADA. The antibody response is polyclonal and consists mainly of antibodies of IgG1 and IgG4 isotype. In the majority of ADA positive patients ADA are already produced within the first 28 weeks of treatment and in part of the pat