Sample records for alpha chain cd25

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

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


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

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

    王燕燕; 蔺辉前; 檀国军; 郭书英; 张建娥


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

  3. Iron modulates the alpha chain of fibrinogen.

    Nielsen, Vance G; Jacobsen, Wayne K


    Iron-bound fibrinogen has been noted to accelerate plasmatic coagulation in patients with divergent conditions involving upregulation of heme oxygenase activity, including hemodialysis, Alzheimer's disease, sickle cell anemia, and chronic migraine. Our goal was to determine if a site of iron-fibrinogen interaction was on the alpha chain. Using thrombelastography, we compared the coagulation kinetic profiles of plasma exposed to 0-10 µM ferric chloride after activation of coagulation with thrombin generated by contact activation of plasma with the plastic sample cup or by exposure to 1 µg/ml of Calloselasma rhodostoma venom (rich in ancrod activity), which causes coagulation via polymerization of alpha chain monomers. Venom mediated coagulation always occurred before thrombin activated thrombus formation, and ferric chloride always diminished the time of onset of coagulation and increased the velocity of clot growth. Iron enhances plasmatic coagulation kinetics by modulating the alpha chain of fibrinogen. PMID:26782808

  4. Alpha decay chain of 292116 nucleus

    Study of superheavy nuclides is of current interest for theoretical physicists as well experimentalists. The present work describes the alpha decay chain of 292116 terminating at 224Pb with corresponding half-lives. Some of these nuclides are reportedly superdeformed but yet stable

  5. Distinct roles for laminin globular domains in laminin alpha1 chain mediated rescue of murine laminin alpha2 chain deficiency.

    Kinga I Gawlik

    Full Text Available BACKGROUND: Laminin alpha2 chain mutations cause congenital muscular dystrophy with dysmyelination neuropathy (MDC1A. Previously, we demonstrated that laminin alpha1 chain ameliorates the disease in mice. Dystroglycan and integrins are major laminin receptors. Unlike laminin alpha2 chain, alpha1 chain binds the receptors by separate domains; laminin globular (LG domains 4 and LG1-3, respectively. Thus, the laminin alpha1 chain is an excellent tool to distinguish between the roles of dystroglycan and integrins in the neuromuscular system. METHODOLOGY/PRINCIPAL FINDINGS: Here, we provide insights into the functions of laminin alpha1LG domains and the division of their roles in MDC1A pathogenesis and rescue. Overexpression of laminin alpha1 chain that lacks the dystroglycan binding LG4-5 domains in alpha2 chain deficient mice resulted in prolonged lifespan and improved health. Importantly, diaphragm and heart muscles were corrected, whereas limb muscles were dystrophic, indicating that different muscles have different requirements for LG4-5 domains. Furthermore, the regenerative capacity of the skeletal muscle did not depend on laminin alpha1LG4-5. However, this domain was crucial for preventing apoptosis in limb muscles, essential for myelination in peripheral nerve and important for basement membrane assembly. CONCLUSIONS/SIGNIFICANCE: These results show that laminin alpha1LG domains and consequently their receptors have disparate functions in the neuromuscular system. Understanding these interactions could contribute to design and optimization of future medical treatment for MDC1A patients.

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

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


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

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

    Shanmugasundaram, Revathi; Selvaraj, Ramesh K


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

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

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

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

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


    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.

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

    Federico Simonetta

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

  11. The proportion of hybrid heterodimers in homozygous or doubly heterozygous beta chain variant hemoglobinopathies associated with alpha chain hemoglobin variants.

    Krauss, J S


    Four alpha genes exist on chromosome 16, but one or more of these genes can be deleted in association with Hemoglobin (Hb)G-Philadelphia in cis to alpha-thalassemia-2 in African-Americans. Therefore, the proportion of HbG-Philadelphia in HbG heterozygotes is trimodal at about 25% for alphaGalpha/alpha alpha, 33% for alphaG-/alpha alpha, and 50% for alphaG-/alpha alpha in patients with HbA. Those who are homozygous or doubly heterozygous for beta chain variants (betaX2 or betaXbetaY) have neither HbA nor the alpha chain variant (alphaX2 betaA2), but have hybrid heterodimers (alphaX2 betaX2). The proportion of hybrid heterodimers here should also be trimodal mirroring alpha gene status. Eleven patients were identified: 4 with Hb SSG, 3 with Hb SCG, and 1 each with Hb OCG, HbSSMontgomery, HbSSChicago, and HbSSBourmedes. Heterodimer proportions were: 43.3 +/- 1.5, 33.5 +/- 2.3, and 15.8 +/- 1.1% for 2, 3, and 4 respective alpha genes which had been studied in 8/11 of the patients (r = 0.98), implying that the prime determinant of the proportion of hybrid heterodimers in this patient group is the number of functional alpha genes. PMID:11045763

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

    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: [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)


    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.

  13. The complete cDNA sequence of laminin alpha 4 and its relationship to the other human laminin alpha chains.

    Richards, A; Al-Imara, L; Pope, F M


    We previously localised the gene (LAMA4) encoding a novel laminin alpha 4 chain to chromosome 6q21. In this study, we describe the complete coding sequence and compare the protein with the other three known human laminin alpha chains. Although closely linked to LAMA2, the LAMA4 product most closely resembles laminin alpha 3, a constituent of laminin 5. Like laminin alpha 3A, the alpha 4 chain is a truncated version of the alpha 1 and alpha 2 chains, with a much reduced short arm. While the alpha 4 molecule is most similar to alpha 3, it shares some features of the C-terminal domains G4 and G5 in common with alpha 2. Unlike the LAMA3 gene, LAMA4 appears to encode only a single transcript, as determined by 5' rapid amplification of cDNA ends. The cDNA sequence encodes 1816 amino acids, which include a 24-residue signal peptide. The gene is expressed in skin, placenta, heart, lung, skeletal muscle, and pancreas. We have also shown that the mRNA can be readily reverse transcribed and amplified from cultured dermal fibroblasts. PMID:8706685

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

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


    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

  15. Localization of laminin alpha4-chain in developing and adult human tissues.

    Petäjäniemi, Noora; Korhonen, Matti; Kortesmaa, Jarkko; Tryggvason, Karl; Sekiguchi, Kiyotoshi; Fujiwara, Hironobu; Sorokin, Lydia; Thornell, Lars-Eric; Wondimu, Zenebech; Assefa, Daniel; Patarroyo, Manuel; Virtanen, Ismo


    Recent studies suggest important functions for laminin-8 (Ln-8; alpha4beta1gamma1) in vascular and blood cell biology, but its distribution in human tissues has remained elusive. We have raised a monoclonal antibody (MAb) FC10, and by enzyme-linked immunoassay (EIA) and Western blotting techniques we show that it recognizes the human Ln alpha4-chain. Immunoreactivity for the Ln alpha4-chain was localized in tissues of mesodermal origin, such as basement membranes (BMs) of endothelia, adipocytes, and skeletal, smooth, and cardiac muscle cells. In addition, the Ln alpha4-chain was found in regions of some epithelial BMs, including epidermis, salivary glands, pancreas, esophageal and gastric glands, intestinal crypts, and some renal medullary tubules. Developmental differences in the distribution of Ln alpha4-chain were detected in skeletal muscle, walls of vessels, and intestinal crypts. Ln alpha4- and Ln alpha2-chains co-localized in BMs of fetal skeletal muscle cells and in some epithelial BMs, e.g., in gastric glands and acini of pancreas. Cultured human pulmonary artery endothelial (HPAE) cells produced Ln alpha4-chain as M(r) 180,000 and 200,000 doublet and rapidly deposited it to the growth substratum. In cell-free extracellular matrices of human kidney and lung, Ln alpha4-chain was found as M(r) 180,000 protein. PMID:12133914

  16. Structure and diversity of the TCR alpha-chain in a teleost fish.

    Partula, S; de Guerra, A; Fellah, J S; Charlemagne, J


    T cell receptor beta-chain genes are well characterized in representatives of most vertebrate phyla, from sharks to mammals, but the molecular structure of complete TCR alpha-chains has not yet been established in cold-blooded vertebrates. We used a PCR approach to isolate cDNAs encoding putative teleost fish (Oncorhynchus mykiss, rainbow trout) TCR alpha-chains. Eight V alpha segments were identified, belonging to six different families, and the best amino acid sequence identity scores for these trout V alpha were all provided by mammalian V alpha or V delta sequences. Twenty-four (60.1 %) of the 39 analyzed V alpha segments belong to the V alpha 2 family, which has limited homology with mammalian V alpha/delta sequences and with the human V pre-B sequence. A total of 32 different J alpha segments were identified from 40 J alpha regions sequenced, suggesting that a large repertoire of J alpha segments is a characteristic of most vertebrates. The structural properties of the TCR alpha-chain complementarity-determining region 3 loop are well conserved between trout and mammals, suggesting that this region has been under continuous selective pressure in jawed vertebrate evolution. The trout C alpha segment has conserved N-terminal and transmembrane domains, but the C alpha intercysteine distance contains only 40 residues, significantly smaller as compared with mammals (49-56 residues). The conserved features of teleost fish TCR beta- and alpha-chains with their mammalian equivalents suggest that TCR-alpha beta receptors were still present in the common Devonian ancestors of modern teleost fish and mammals, about 450 million years ago. PMID:8683116

  17. Molecular characterization of hemoglobin alpha-D chains from Geochelone carbonaria and Geochelone denticulata land turtles.

    Melo, Mônica B; Bordin, Silvana; Duarte, Adriana S S; Ogo, Satie H; Torsoni, Márcio A; Saad, Sara T O; Costa, Fernando F


    In order to help elucidate the evolution of alpha-globins, the complete cDNA and amino acid sequences of Geochelone carbonaria and Geochelone denticulata land turtles alpha-D chains have been described. In G. carbonaria, the cDNA is 539 bp with ATG start codon located at position 46, TGA stop codon at position 469 and AATAAA polyadenylation signal at position 520. In G. denticulata, the cDNA is 536 bp with ATG start codon located at position 46, TGA stop codon at position 469 and AATAAA polyadenylation signal at position 517. Both cDNAs codify 141 amino acid residues, differing from each other in only four amino acid residues. When comparing with human Hb alpha-chain, alterations in important regions can be noted: alpha110 Ala-Gly, alpha114 Pro-Gly, alpha117 Phe-Tyr and alpha122 His-Gln. There is a high homology between the amino acids of these turtles when compared with chicken alpha-D chains, progressively decreasing when compared with human, crocodile, snake, frog and fish alpha-chains. Phylogenetic analysis of alpha-D chains shows that those of turtles are closer to those of birds than to snakes and lizards. PMID:12568815

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

    Lenardo Michael J


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

  19. Deletion of the laminin alpha4 chain leads to impaired microvessel maturation.

    Thyboll, Jill; Kortesmaa, Jarkko; Cao, Renhai; Soininen, Raija; Wang, Ling; Iivanainen, Antti; Sorokin, Lydia; Risling, Mårten; Cao, Yihai; Tryggvason, Karl


    The laminin alpha4 chain, a component of laminin-8 and -9, is expressed in basement membranes, such as those beneath endothelia, the perineurium of peripheral nerves, and around developing muscle fibers. Laminin alpha4-null mice presented with hemorrhages during the embryonic and neonatal period and had extensive bleeding and deterioration of microvessel growth in experimental angiogenesis, as well as mild locomotion defects. Histological examination of newborn mice revealed delayed deposition of type IV collagen and nidogen into capillary basement membranes, and electron microscopy showed discontinuities in the lamina densa. The results demonstrate a central role for the laminin alpha4 chain in microvessel growth and, in the absence of other laminin alpha chains, in the composition of endothelial basement membranes. PMID:11809810

  20. The laminin alpha chains: expression, developmental transitions, and chromosomal locations of alpha1-5, identification of heterotrimeric laminins 8-11, and cloning of a novel alpha3 isoform.

    Miner, J H; Patton, B L; Lentz, S I; Gilbert, D J; Snider, W D; Jenkins, N A; Copeland, N G; Sanes, J R


    Laminin trimers composed of alpha, beta, and gamma chains are major components of basal laminae (BLs) throughout the body. To date, three alpha chains (alpha1-3) have been shown to assemble into at least seven heterotrimers (called laminins 1-7). Genes encoding two additional alpha chains (alpha4 and alpha5) have been cloned, but little is known about their expression, and their protein products have not been identified. Here we generated antisera to recombinant alpha4 and alpha5 and used them to identify authentic proteins in tissue extracts. Immunoprecipitation and immunoblotting showed that alpha4 and alpha5 assemble into four novel laminin heterotrimers (laminins 8-11: alpha4beta1gamma1, alpha4beta2gamma1, alpha5beta1gamma1, and alpha5beta2gamma1, respectively). Using a panel of nucleotide and antibody probes, we surveyed the expression of alpha1-5 in murine tissues. All five chains were expressed in both embryos and adults, but each was distributed in a distinct pattern at both RNA and protein levels. Overall, alpha4 and alpha5 exhibited the broadest patterns of expression, while expression of alpha1 was the most restricted. Immunohistochemical analysis of kidney, lung, and heart showed that the alpha chains were confined to extracellular matrix and, with few exceptions, to BLs. All developing and adult BLs examined contained at least one alpha chain, all alpha chains were present in multiple BLs, and some BLs contained two or three alpha chains. Detailed analysis of developing kidney revealed that some individual BLs, including those of the tubule and glomerulus, changed in laminin chain composition as they matured, expressing up to three different alpha chains and two different beta chains in an elaborate and dynamic progression. Interspecific backcross mapping of the five alpha chain genes revealed that they are distributed on four mouse chromosomes. Finally, we identified a novel full-length alpha3 isoform encoded by the Lama3 gene, which was previously

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

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


    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.

  2. Chondroitin sulphate modification in the alpha4 chain of human recombinant laminin-8 (alpha4beta1gamma1).

    Kortesmaa, Jarkko; Doi, Masayuki; Patarroyo, Manuel; Tryggvason, Karl


    We have produced human laminin-8 (alpha4beta1gamma1) using recombinant technology. Approximately half of the recombinant laminin-8 (rLN-8) molecules were found to have a chondroitin sulphate modification in the alpha4 chain. The substituted and non-substituted forms were separated and tested for cell adhesion activity. Lower cell adhesion promoting activity was seen for the substituted form, but the integrin receptor utilization was similar. We also found the human rLN-8 to behave identically in cell adhesion assays compared to a human/mouse hybrid variant of rLN-8. PMID:12392759

  3. Salt bridge residues between I-Ak dimer of dimers alpha-chains modulate antigen presentation.

    Yadati, S; Nydam, T; Demian, D; Wade, T K; Gabriel, J L; Barisas, B G; Wade, W F


    Class II dimers of dimers are predicted to have functional significance in antigen presentation. The putative contact amino acids of the I-Ak class II dimer of dimers have been identified by molecular modeling based on the DR1 crystal structure (Nydam et al., Int. Immunol. 10, 1237,1998). We have previously reported the role in antigen presentation of dimer of dimers contact amino acids located in the C-terminal domains of the alpha- and beta-chains of class II. Our calculations show that residues Ealpha89 and Ralpha145 in the alpha2-domain form an inter alpha-chain salt bridge between pairs of alphabeta-heterodimers. Other residues, Qalpha92 and Nalpha115, may be involved in close association in that part of the alpha-chain. We investigated the role of these amino acids on class II expression and antigen presentation. Class II composed of an Ealpha89K substituted alpha-chain paired with a wt beta-chain exhibited inhibited antigen presentation and expression of alpha-chain serologic epitopes. In contrast, mutation of Ralpha145E had less affect on antigen presentation and did not affect I-Ak serologic epitopes. Interchanging charges of the salt bridge residues by expressing both Ralpha145E and Ealpha89K on the same chain obviated the large negative effect of the Ealpha89K mutation on antigen presentation but not on the serologic epitopes. Our results are similar for those reported for mutation of DR3's inter-chain salt bridge with the exception that double mutants did not moderate the DR3 defect. Interestingly, the amino acids differences between I-A and DR change the location of the inter-chain salt bridges. In DR1 these residues are located at positions Ealpha88 and Kalpha111; in I-Ak these residues are located at position Ealpha89 and Ralpha145. Inter alpha-chain salt bridges are thus maintained in various class II molecules by amino acids located in different parts of the alpha2-domain. This conservation of structure suggests that considerable functional

  4. cDNA clone for the alpha-chain of human beta-hexosaminidase: deficiency of alpha-chain mRNA in Ashkenazi Tay-Sachs fibroblasts.

    Myerowitz, R; Proia, R L


    We have isolated a cDNA clone containing sequences complementary to mRNA encoding the alpha-chain of the lysosomal enzyme beta-hexosaminidase. RNA from a human lung fibroblast strain, IMR90, was enriched for beta-hexosaminidase messenger by polysome immunoselection with antiserum against beta-hexosaminidase A. This preparation was used to construct cDNA recombinant plasmids by the Okayama-Berg vector primer procedure. After transformation of Escherichia coli, 385 ampicillin-resistant colonies...

  5. Analysis of delocalization of clusters in linear-chain $\\alpha$-cluster states with entanglement entropy

    Kanada-En'yo, Yoshiko


    I investigate entanglement entropy of one dimension (1D) cluster states to discuss the delocalization of clusters in linear-chain $3\\alpha$- and $4\\alpha$-cluster states. In analysis of entanglement entropy of 1D Tohsaki-Horiuchi-Schuck-R\\"opke (THSR) and Brink-Bloch cluster wave functions, I show clear differences in the entanglement entropy between localized cluster wave functions and delocalized cluster wave functions. In order to clarify spatial regions where the entanglement entropy is g...

  6. Physiological covalent regulation of rat liver branched-chain alpha-ketoacid dehydrogenase

    A radiochemical assay was developed for measuring branched-chain alpha-ketoacid dehydrogenase activity of Triton X-100 extracts of freeze-clamped rat liver. The proportion of active (dephosphorylated) enzyme was determined by measuring enzyme activities before and after activation of the complex with a broad-specificity phosphoprotein phosphatase. Hepatic branched-chain alpha-ketoacid dehydrogenase activity in normal male Wistar rats was 97% active but decreased to 33% active after 2 days on low-protein (8%) diet and to 13% active after 4 days on the same diet. Restricting protein intake of lean and obese female Zucker rats also caused inactivation of hepatic branched-chain alpha-ketoacid dehydrogenase complex. Essentially all of the enzyme was in the active state in rats maintained for 14 days on either 30 or 50% protein diets. This was also the case for rats maintained on a commercial chow diet (minimum 23% protein). However, maintaining rats on 20, 8, and 0% protein diets decreased the percentage of the active form of the enzyme to 58, 10, and 7% of the total, respectively. Fasting of chow-fed rats for 48 h had no effect on the activity state of hepatic branched-chain alpha-ketoacid dehydrogenase, i.e., 93% of the enzyme remained in the active state compared to 97% for chow-fed rats. However, hepatic enzyme of rats maintained on 8% protein diet was 10% active before starvation and 83% active after 2 days of starvation. Thus, dietary protein deficiency results in inactivation of hepatic branched-chain alpha-ketoacid dehydrogenase complex, presumably as a consequence of low hepatic levels of branched-chain alpha-ketoacids

  7. Analysis of delocalization of clusters in linear-chain $\\alpha$-cluster states with entanglement entropy

    Kanada-En'yo, Yoshiko


    I investigate entanglement entropy of one dimension (1D) cluster states to discuss the delocalization of clusters in linear-chain $3\\alpha$- and $4\\alpha$-cluster states. In analysis of entanglement entropy of 1D Tohsaki-Horiuchi-Schuck-R\\"opke (THSR) and Brink-Bloch cluster wave functions, I show clear differences in the entanglement entropy between localized cluster wave functions and delocalized cluster wave functions. In order to clarify spatial regions where the entanglement entropy is generated by the delocalization of clusters, I analyze the spatial distribution of entanglement entropy. In the linear-chain $3\\alpha$ cluster state, the delocalization occurs dominantly in a low-density tail region while it is relatively suppressed in an inner region because of Pauli blocking effect between clusters. In the linear-chain 4$\\alpha$ state having a larger system size than the linear-chain $3\\alpha$ state, the delocalization occurs in the whole system. The entanglement entropy is found to be a measure of the d...

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

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


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

  9. Ground state properties of the newly discovered nucleus 265Bh and it's alpha decay chain

    The properties of the nuclei belonging to the newly discovered alpha-decay chain starting from 265Bh have been studied. The axially deformed relativistic mean field (RMF) calculation with the force TMA and NL-Z2 has been performed in the blocked BCS approximation. Some ground state properties such as the binding energies, deformations, spins and parties, as well as Q-values of the alpha decay for this decay chain have been calculated and compared with known experimental data. Good agreement is observed. The single-particle spectrum of the nucleus 265Bh has been studied. (authors)

  10. Promoter and enhancer elements in the rearranged alpha chain gene of the human T cell receptor.

    Luria, S; Gross, G; Horowitz, M; Givol, D


    We cloned and compared the sequence of a rearranged human T cell receptor (TCR) V alpha J alpha gene and its germline counterparts. The only difference in the coding region sequence was confined to the joining region where three nucleotides, TTG, unaccountable by either V alpha or J alpha sequence, were present. By nuclease S1 mapping we identified the mRNA start of the alpha chain 70 nucleotides upstream from the initiator ATG. A 600 bp fragment containing the sequences upstream to the ATG drives the expression of the bacterial chloramphenicol acetyltransferase (CAT) gene. This promoter activity is T cell specific since it can be demonstrated in human T cells but not in B cells or HeLa cells. A 1.1 kb BamHI- HindIII fragment located 5' to the first exon of the C alpha gene was found to enhance transcription from either the heterologous SV40 promoter or the homologous TCR alpha chain promoter. This enhancement activity was independent of the location of the fragment with respect to CAT and was specific to lymphoid cells (either T or B cells) but cannot be demonstrated in HeLa cells. PMID:3501368

  11. Primary structure, developmental expression, and immunolocalization of the murine laminin alpha4 chain.

    Iivanainen, A; Kortesmaa, J; Sahlberg, C; Morita, T; Bergmann, U; Thesleff, I; Tryggvason, K


    The complete primary structure of the mouse laminin alpha4 chain was derived from cDNA clones. The translation product contains a 24-residue signal peptide preceding the mature alpha4 chain of 1,792 residues. Northern analysis on whole mouse embryos revealed that the expression was weak at day 7, but it later increased and peaked at day 15. In adult tissues the strongest expression was observed in lung and cardiac and skeletal muscles. Weak expression was also seen in other adult tissues such as brain, spleen, liver, kidney, and testis. By in situ hybridization of fetal and newborn tissues, expression of the laminin alpha4 chain was mainly localized to mesenchymal cells. Strong expression was seen in the villi and submucosa of the developing intestine, the mesenchymal stroma surrounding the branching lung epithelia, and the external root sheath of vibrissae follicles, as well as in cardiac and skeletal muscle fibers. In the developing kidney, intense but transient expression was associated with the differentiation of epithelial kidney tubules from the nephrogenic mesenchyme. Immunohistologic staining with affinity-purified IgG localized the laminin alpha4 chain primarily to lung septa, heart, and skeletal muscle, capillaries, and perineurium. PMID:9346933

  12. Expression of interleukin-2R alpha and interleukin-2R beta in Hodgkin's disease.

    Tesch, H.; A. Günther; Abts, H.; Jücker, M; Klein, S; Krueger, G. R.; Diehl, V.


    Hodgkin and Reed-Sternberg cells, the putative malignant cells of Hodgkin's disease (HD), carry regularly the CD25 antigen that forms one chain of the interleukin-2 (IL-2) receptor (IL-2R alpha). To analyze the putative role of IL-2R expression in Hodgkin's disease, we have investigated the expression of both IL-2R alpha and IL-2R beta chains in HD-derived cell lines and in primary specimens from patients with HD. Expression of IL-2R alpha and IL-2R beta was detected in all HD-derived cell li...

  13. BstXI RFLP in the human inter-alpha-trypsin inhibitor light chain gene

    Leveillard, T.; Bourguignon, J.; Sesbouee, R.; Hanauer, A.; Salier, J.P.; Diarra-Mehrpour, M.; Martin, J.P.


    The 1.2 kb EcoRI/SmaI fragment of lambdaHuLITI2 was used as probe. lambdaHuLITI2 is a full length cDNA clone coding for human inter-alpha-trypsin inhibitor light chain isolated from immunochemical screening of a lambdagt11 library. Its sequence coding for HI-30 and alpha-1-microglobulin is in agreement. BstXI identifies five invariant bands at 5.0 kb, 2.3 kb, 1.5 kb, 1.1 kb, and 0.7 kb and a diallelic polymorphism with DNA fragments at 2.0 kb or 1.7 kb.

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

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


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

  15. Biotemplated synthesis of metallic nanoparticle chains on an alpha-synuclein fiber scaffold.

    Colby, R; Hulleman, J; Padalkar, S; Rochet, J C; Stanciu, L A


    Biomolecular templates provide an excellent potential tool for bottom-up device fabrication. Self-assembling alpha-synuclein protein fibrils, the formation of which has been linked to Parkinson's disease, have yet to be explored for potential device fabrication. In this paper, alpha-synuclein fibrils were used as a template for palladium (Pd), gold (Au) and copper (Cu) nanoparticle chains synthesis. Deposition over a range of conditions resulted in metal-coated fibers with reproducible average diameters between 50 and 200 nm. Active elemental palladium deposited on the protein fibrils is used as a catalyst for the electroless deposition of Au and Cu. Nanoparticle chains were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray energy dispersive spectrometry (XEDS), and electron energy loss spectrometry (EELS). PMID:18464436

  16. Structure and diversity of the T-cell receptor alpha chain in the Mexican axolotl.

    Fellah, J S; Kerfourn, F; Dumay, A M; Aubet, G; Charlemagne, J


    Polymerase chain reaction was used to isolate cDNA clones encoding putative T-cell receptor (TCR) alpha chains in an amphibian, the Mexican axolotl (Ambystoma mexicanum). Five TCRalpha-V chain-encoding segments were identified, each belonging to a separate family. The best identity scores for these axolotl TCRalpha-V segments were all provided by sequences belonging to the human TCRalpha-V1 family and the mouse TCRalpha-V3 and TCRalpha-V8 families. A total of 14 different TCRA-J segments were identified from 44 TCRA-V/TCRA-J regions sequenced, suggesting that a large repertoire of TCRA-J segments is a characteristic of most vertebrates. The structure of the axolotl CDR3 alpha chain loop is in good agreement with that of mammals, including a majority of small hydrophobic residues at position 92 and of charged, hydrophilic, or polar residues at positions 93 and 94, which are highly variable and correspond to the TCRA-V/J junction. This suggests that some positions of the axolotl CDR3 alpha chain loop are positively selected during T-cell differentiation, particularly around residue 93 that could be selected for its ability to makes contacts with major histocompatibility complex-associated antigenic peptides, as in mammals. The axolotl Calpha domain had the typical structure of mammalian and avian Calpha domains, including the charged residues in the TM segment that are thought to interact with other proteins in the membrane, as well as most of the residues forming the conserved antigen receptor transmembrane motif. PMID:9002443

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

    张明海; 赵建斌


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

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

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


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

  19. 90Y-daclizumab, an anti-CD25 monoclonal antibody, provided responses in 50% of patients with relapsed Hodgkin’s lymphoma

    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.


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

  20. 90Y-daclizumab, an anti-CD25 monoclonal antibody, provided responses in 50% of patients with relapsed Hodgkin's lymphoma.

    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


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

  1. A shortened beta-hexosaminidase alpha-chain in an Italian patient with infantile Tay-Sachs disease.

    Zokaeem, G; Bayleran, J; Kaplan, P; Hechtman, P; Neufeld, E F


    Fibroblasts derived from a beta-hexosaminidase A (HexA)-deficient infant with clinically classic Tay-Sachs disease synthesized a precursor alpha-chain that was smaller than its normal counterpart. Fibroblasts from the infant's parents, who were consanguinous, produced both normal and mutant alpha-chains. The size difference, estimated to be 2-3 kilodaltons on the basis of sodium dodecyl sulfate-polyacrylamide-gel electrophoresis, persisted after removal of oligosaccharides with endo-H and is ...

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

    鞠云飞; 孙立锋; 胡华


    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.

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

    袁海芹; 刁智娟; 周剑锁; 刘彦信; 史娟; 郑德先


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

  4. Dra I polymorphism in the human inter-alpha-tryspin inhibitor heavy chain gene ITIH1

    Leveillard, T.; Salier, J.P.; Sesbouee, R.; Bourguignon, J.; Diarra-Mehrpour, M.; Martin, J.P. (Institut National de la Sante et de la Recherche Medicale, St. Etienne Rouvray (France))


    The 1.0 kb insert of lambda HuHITI-19, used as a probe, codes for the heavy chain H1 of the human inter-alpha-tryspin inhibitor. Dra I (TTT/AAA) identifies one invariant band at 5.8 kb and a diallelic polymorphism with DNA fragments at 4.0 kb or 2.4 kb and 1.6 kb. The allele frequency was studied in 24 healthy caucasians. The ITIH1 gene has been mapped to 3p211-p212 by in situ hybridization. Co-dominant segregation was found in three informative families (11 individuals).

  5. An hypervariable polymorphism detected in the human inter-. alpha. -trypsin inhibitor heavy chain gene ITIH2

    Leveillard, T.; Sesbouee, R.; Bourguignon, J.; Diarra-Mehrpour, M.; Salier, J.P.; Martin, J.P. (Inst. National de la Sante et de la Recherche Medicale, Rouvray (France)); Sirugo, G.; Hanauer, A. (Inst. National de la Sante et de la Recherche Medicale, Strasbourg (France))


    The 112 bp BamHi/Bst YI fragment of lambda HuHITI-9 used as a probe codes for the heavy chain H2 of human inter-{alpha}-trypsin inhibitor. BstXI (CCAN{sub 5}/NTGG) identifies a 5 allele VNTR polymorphism with bands between 2.6 kb and 3.0 kb. DraI, MspI, PstI and TaqI also detect the same polymorphism. The ITH2 gene has been mapped to 10p15 by in situ hybridization.

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

    Pedersen, Anders Elm; Lauritsen, Jens Peter Holst


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

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

    鞠云飞; 孙立锋; 胡华



  8. Murine laminin alpha3A and alpha3B isoform chains are generated by usage of two promoters and alternative splicing.

    Ferrigno, O; Virolle, T; Galliano, M F; Chauvin, N; Ortonne, J P; Meneguzzi, G; Aberdam, D


    We already identified two distinct laminin alpha3A and alpha3B chain isoforms which differ in their amino-terminal ends and display different tissue-specific expression patterns. In this study we have investigated whether these two different isoforms are products of the same laminin alpha3 (lama3) gene and transcribed from one or two separate promoters. Genomic clones were isolated that encompass the sequences upstream to the 5' ends of both the alpha3A and the alpha3B cDNAs. Sequence analysis of the region upstream to the alpha3A open reading frame revealed the presence of a TATA box and potential binding sites for responsive elements. By primer extension analysis, the transcription start site of the alpha3B mRNA isoform was defined. The sequences upstream to the alpha3B mRNA transcription start site do not contain a TATA box near the transcription initiation sites, but AP-1, AP-2, and Sp1 consensus binding site sequences were identified. The genomic regions located immediately upstream of the alpha3A and alpha3B transcription start sites were shown to possess promoter activities in transfection experiments. In the promoter regions, response elements for the acute phase reactant signal and NF-interleukin 6 were found, and their possible relevance in the context of inflammation and wound healing is discussed. Our results demonstrate that the lama3 gene produces the two polypeptides by alternative splicing and contains two promoters, which regulate the production of the two isoforms alpha3A and alpha3B. PMID:9252362

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

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


    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.

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

    Peris Munyaka

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




    We have characterized the structural abnormality of a new alpha chain mutant found in a Kurdish; family. The clinical and hematological investigation of eight individuals have shown that the a variant is associated with a beta degrees-thalassemia mutation (nonsense codon 39). The tryptic peptide map

  12. Murine branched chain alpha-ketoacid dehydrogenase kinase; cDNA cloning, tissue distribution, and temporal expression during embryonic development.

    Doering, C B; Coursey, C; Spangler, W; Danner, D J


    These studies were designed to demonstrate the structural and functional similarity of murine branched chain alpha-ketoacid dehydrogenase and its regulation by the complex-specific kinase. Nucleotide sequence and deduced amino acid sequence for the kinase cDNA demonstrate a highly conserved coding sequence between mouse and human. Tissue-specific expression in adult mice parallels that reported in other mammals. Kinase expression in female liver is influenced by circadian rhythm. Of special interest is the fluctuating expression of this kinase during embryonic development against the continuing increase in the catalytic subunits of this mitochondrial complex during development. The need for regulation of the branched chain alpha-ketoacid dehydrogenase complex by kinase expression during embryogenesis is not understood. However, the similarity of murine branched chain alpha-ketoacid dehydrogenase and its kinase to the human enzyme supports the use of this animal as a model for the human system. PMID:9611264

  13. Chain length dependence of the helix orientation in Langmuir-Blodgett monolayers of alpha-helical diblock copolypeptides

    Nguyen, Le-Thu T; Ardana, Aditya; Vorenkamp, Eltjo J.; ten Brinke, Gerrit; Schouten, Arend J.


    The effect of chain length on the helix orientation of alpha-helical diblock copolypeptides in Langmuir and Langmuir-Blodgett monolayers is reported for the first time. Amphiphilic diblock copolypeptides (PLGA-b-PMLGSLGs) of poly(alpha-L-glutamic acid) (PLGA) and poly(gamma-methyl-L-glutamate-ran-gamma-stearyl-L-glutamate) with 30 mol% of stearyl substituents (PMLGSLG) of various block lengths were studied. The tilt angle between the helices and the substrate-normal decreases upon increasing ...

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

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


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

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

    Yuan-Huan Xiong; Zhen Yuan; Li He


    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.

  16. Mutation in the alpha 5(IV) collagen chain in juvenile-onset Alport syndrome without hearing loss or ocular lesions

    Zhou, J; Hertz, Jens Michael; Tryggvason, K


    A single base mutation was identified in the type IV collagen alpha 5 chain gene (COL4A5) of a Danish kindred with Alport syndrome. The 27-year-old male proband developed hematuria in childhood and terminal renal failure at the age of 25 years. He has no hearing loss or ocular lesions. Electron...

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

    王大伟; 周荣斌; 姚咏明


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

  18. Cloning of the LamA3 gene encoding the alpha 3 chain of the adhesive ligand epiligrin. Expression in wound repair.

    Ryan, M C; Tizard, R; VanDevanter, D R; Carter, W G


    We have isolated cDNA clones encoding the entire 170-kDa chain of epiligrin (alpha 3Ep) and a genomic clone encoding the alpha 3Ep gene (LamA3). Analysis of multiple cDNA clones revealed two distinct transcripts (alpha 3EpA and alpha 3EpB). Sequencing of the alpha 3EpA transcript indicated sequence and structural homology to laminin alpha 1 and alpha 2 chains that extend from domain IIIa through the carboxyl-terminal G domain. The alpha 3EpB transcript encodes a larger amino-terminal domain and contains additional epidermal growth factor repeats and sequences corresponding to domain IV of alpha 1 laminin. Fluorescence in situ hybridization indicated that the LamA3 gene is located on chromosome 18q11.2, a locus distinct from the LamA1 gene (18p11.3). The G domain of the epiligrin alpha 3 chain contains five subdomains that are individually related to the G subdomains reported for Drosophila and vertebrate laminin alpha chains. Sequence divergence within the G domain of alpha 3 epiligrin suggests that it is functionally distinct from laminin, consistent with our previous report showing that epiligrin interacts with different integrin adhesion receptors. Analysis of RNA from human foreskin keratinocytes (HFKs) identified multiple epiligrin transcripts that were down-regulated by viral transformation and differentiation. In contrast, epiligrin expression was up-regulated in wound sites of human skin. PMID:8077230

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

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


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

  20. Alternative splicing produces transcripts coding for alpha and beta chains of a hetero-dimeric phosphagen kinase.

    Ellington, W Ross; Yamashita, Daisuke; Suzuki, Tomohiko


    Glycocyamine kinase (GK) catalyzes the reversible phosphorylation of glycocyamine (guanidinoacetate), a reaction central to cellular energy homeostasis in certain animals. GK is a member of the phosphagen kinase enzyme family and appears to have evolved from creatine kinase (CK) early in the evolution of multi-cellular animals. Prior work has shown that GK from the polychaete Neanthes (Nereis) diversicolor exits as a hetero-dimer in vivo and that the two polypeptide chains (termed alpha and beta) are coded for by unique transcripts. In the present study, we demonstrate that the GK from a congener Nereis virens is also hetero-dimeric and is coded for by alpha and beta transcripts, which are virtually identical to the corresponding forms in N. diversicolor. The GK gene from N. diversicolor was amplified by PCR. Sequencing of the PCR products showed that the alpha and beta chains are the result of alternative splicing of the GK primary mRNA transcript. These results also strongly suggest that this gene underwent an early tandem exon duplication event. Full-length cDNAs for N. virens GKalpha and GKbeta were individually ligated into expression vectors and the resulting constructs used to transform Escherichia coli expression hosts. Regardless of expression conditions, minimal GK activity was observed in both GKalpha and GKbeta constructs. Inclusion bodies for both were harvested, unfolded in urea and alpha chains, beta chains and mixtures of alpha and beta chains were refolded by sequential dialysis. Only modest amounts of GK activity were observed when alpha and beta were refolded individually. In contrast, when refolded the alpha and beta mixture yielded highly active hetero-dimers, as validated by size exclusion chromatography, electrophoresis and mass spectrometry, with a specific activity comparable to that of natural GK. The above evidence suggests that there is a preference for hetero-dimer formation in the GKs from these two polychaetes. The evolution of the

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

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

  2. Alpha chain hemoglobins with electrophoretic mobility similar to that of hemoglobin S in a newborn screening program

    Marcilene Rezende Silva


    Full Text Available OBJECTIVE: To characterize alpha-chain variant hemoglobins with electric mobility similar to that of hemoglobin S in a newborn screening program. METHODS: βS allele and alpha-thalassemia deletions were investigated in 14 children who had undefined hemoglobin at birth and an electrophoretic profile similar to that of hemoglobin S when they were six months old. Gene sequencing and restriction enzymes (DdeI, BsaJI, NlaIV, Bsu36I and TaqI were used to identify hemoglobins. Clinical and hematological data were obtained from children who attended scheduled medical visits. RESULTS: The following alpha chain variants were found: seven children with hemoglobin Hasharon [alpha2 47(CE5 Asp>His, HbA2:c.142G>C], all associated with alpha-thalassemia, five with hemoglobin Ottawa [alpha1 15(A13 Gly>Arg, HBA1:c.46G>C], one with hemoglobin St Luke's [alpha1 95(G2 Pro>Arg, HBA1:c.287C>G] and another one with hemoglobin Etobicoke [alpha212 84(F5 Ser>Arg, HBA212:c.255C>G]. Two associations with hemoglobin S were found: one with hemoglobin Ottawa and one with hemoglobin St Luke's. The mutation underlying hemoglobin Etobicoke was located in a hybrid α212 allele in one child. There was no evidence of clinically relevant hemoglobins detected in this study. CONCLUSION: Apparently these are the first cases of hemoglobin Ottawa, St Luke's, Etobicoke and the α212 gene described in Brazil. The hemoglobins detected in this study may lead to false diagnosis of sickle cell trait or sickle cell disease when only isoelectric focusing is used in neonatal screening. Additional tests are necessary for the correct identification of hemoglobin variants.

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

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


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

  4. Reassembly and reconstitution of separate alpha and beta chains of human leukocyte antigen DR4 molecule isolated from Escherichia coli.

    Kang, J H; Maeng, C Y; Park, J H; Hahm, K S; Han, B D; Kim, K L


    The class II major histocompatibility complex molecules play a major role in presentation of exogenous antigenic peptides to the CD4 positive helper T cell. These are heterodimeric cell surface glycoproteins consisting of alpha- and beta-chains. In the present study, we cloned and expressed the alpha- and beta-chain of HLA-DR4 lacking the transmembrane and cytoplasmic domain separately in Escherichia coli using the pET-5a expression vector system. The expressed alpha- and beta-chains were purified in a denaturing condition by an ion exchange chromatography on Q-Sepharose and a gel filtration chromatography on Sephacryl S-200, respectively. The recombinant proteins were refolded and reassembled by removing the denaturing agent and concomitant reoxidation of the disulfide bond. The refolded heterodimeric rDR4 molecule was resolved by 12.5% SDS-PAGE in a nonreducing condition and confirmed by Western blot using polyclonal antibody against DR-alpha and the monoclonal antibody (L243) for the conformationally correct DR molecule. The rDR4 molecules were reconstituted with a 50-fold molar excess biot-HA (307-319), and the bound peptides to the heterodimer complex were determined by a microplate assay coated with L243 antibody using Extravidin-HRP conjugate. PMID:9163739

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

    薛克营; 金卫国; 王成国; 程立; 杨中卫; 王正艳



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

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


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

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

    彭力; 钟礼立; 黄寒; 厉娟; 梁沫; 李云


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

  8. A polymorphic variant in the human electron transfer flavoprotein alpha-chain (alpha-T171) displays decreased thermal stability and is overrepresented in very-long-chain acyl-CoA dehydrogenase-deficient patients with mild childhood presentation

    Bross, P; Pedersen, P; Nyholm, M;


    -deficient patients homozygous for the K304E mutation (MCAD E304), (iii) a group of patients with elevated urinary excretion of ethylmalonic acid (EMA) possibly due to decreased short-chain acyl-CoA dehydrogenase activity, and (iv) in patients with proven deficiency of very-long-chain acyl-CoA dehydrogenase (VLCAD......). No significant overrepresentations or underrepresentations were found in the first two patient groups, suggesting that the polymorphisms studied are not significant susceptibility factors in either the MCAD E304 or the EMA patient group. However, in the VLCAD deficient patients the alpha-T171 variant...... (decreased thermal stability) was significantly overrepresented. Subgrouping of the VLCAD patients into three phenotypic classes (severe childhood, mild childhood, and adult presentation) revealed that the overrepresentation of the alpha-T171 variant was significant only in patients with mild childhood...

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

    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:


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

    Highlights: → Curcumin inhibits CD4+ T-lymphocyte proliferation. → Curcumin inhibits interleukin-2 (IL-2) synthesis and CD25 expression by CD4+ T-lymphocytes. → Curcumin interferes with IL-2 receptor signaling by inhibiting JAK3 and STAT5 phosphorylation. → IL-2-dependent regulatory T-lymphocyte function and Foxp3 expression is downregulated by curcumin. -- Abstract: Curcumin (diferulomethane) is the principal curcuminoid in the spice tumeric and a potent inhibitor of activation-induced T-lymphocyte proliferation; however, the molecular basis of this immunosuppressive effect has not been well studied. Here we show that micromolar concentrations of curcumin inhibited DNA synthesis by mouse CD4+ T-lymphocytes, as well as interleukin-2 (IL-2) and CD25chain of the high affinity IL-2 receptor) expression in response to antibody-mediated cross-linking of CD3 and CD28. Curcumin acted downstream of protein kinase C activation and intracellular Ca2+ release to inhibit IκB phosphorylation, which is required for nuclear translocation of the transcription factor NFκB. In addition, IL-2-dependent DNA synthesis by mouse CTLL-2 cells, but not constitutive CD25 expression, was impaired in the presence of curcumin, which demonstrated an inhibitory effect on IL-2 receptor (IL-2R) signaling. IL-2-induced phosphorylation of STAT5A and JAK3, but not JAK1, was diminished in the presence of curcumin, indicating inhibition of critical proximal events in IL-2R signaling. In line with the inhibitory action of curcumin on IL-2R signaling, pretreatment of CD4+CD25+ regulatory T-cells with curcumin downregulated suppressor function, as well as forkhead box p3 (Foxp3) expression. We conclude that curcumin inhibits IL-2 signaling by reducing available IL-2 and high affinity IL-2R, as well as interfering with IL-2R signaling.

  11. Association of the human CD3-zeta chain with the alpha beta-T cell receptor/CD3 complex. Clues from a T cell variant with a mutated T cell receptor-alpha chain

    Geisler, C; Schøller, J; Wahi, M A; Rubin, B; Weiss, A


    various components of this multimeric protein complex are not fully understood. In this report, a variant of the human leukemic T cell line Jurkat that synthesized all of the known components of the TCR/CD3 complex but fails to express the TCR/CD3 complex at the cell surface is further characterized. This......The TCR for Ag, on the majority of human T cells, is a disulfide-linked heterodimer composed of TCR-alpha and -beta chains noncovalently associated with the monomorphic CD3 complex composed of the CD3-gamma, -delta, -epsilon, and -zeta chains. The interactions involved in the assembly of the......-CD3-gamma delta epsilon zeta 2). Transfecting a wild-type TCR-alpha gene into J79 reconstituted expression of a complete functionally competent TCR/CD3 complex at the cell surface. The results indicate that the TCR-alpha chain plays a crucial role in the assembly of the CD3-zeta homodimer with the...

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

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


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

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

    Cristina Miyamoto


    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.

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

    Youngjoo Kwon


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

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

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


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

  16. Structural Origins of Nitroxide Side Chain Dynamics on Membrane Protein [alpha]-Helical Sites

    Kroncke, Brett M.; Horanyi, Peter S.; Columbus, Linda (UV)


    Understanding the structure and dynamics of membrane proteins in their native, hydrophobic environment is important to understanding how these proteins function. EPR spectroscopy in combination with site-directed spin labeling (SDSL) can measure dynamics and structure of membrane proteins in their native lipid environment; however, until now the dynamics measured have been qualitative due to limited knowledge of the nitroxide spin label's intramolecular motion in the hydrophobic environment. Although several studies have elucidated the structural origins of EPR line shapes of water-soluble proteins, EPR spectra of nitroxide spin-labeled proteins in detergents or lipids have characteristic differences from their water-soluble counterparts, suggesting significant differences in the underlying molecular motion of the spin label between the two environments. To elucidate these differences, membrane-exposed {alpha}-helical sites of the leucine transporter, LeuT, from Aquifex aeolicus, were investigated using X-ray crystallography, mutational analysis, nitroxide side chain derivatives, and spectral simulations in order to obtain a motional model of the nitroxide. For each crystal structure, the nitroxide ring of a disulfide-linked spin label side chain (R1) is resolved and makes contacts with hydrophobic residues on the protein surface. The spin label at site I204 on LeuT makes a nontraditional hydrogen bond with the ortho-hydrogen on its nearest neighbor F208, whereas the spin label at site F177 makes multiple van der Waals contacts with a hydrophobic pocket formed with an adjacent helix. These results coupled with the spectral effect of mutating the i {+-} 3, 4 residues suggest that the spin label has a greater affinity for its local protein environment in the low dielectric than on a water-soluble protein surface. The simulations of the EPR spectra presented here suggest the spin label oscillates about the terminal bond nearest the ring while maintaining weak

  17. Effect of alpha thalassaemia trait and enhanced gamma chain production on disease severity in beta thalassaemia major and intermedia.

    Gringras, P; Wonke, B; Old, J.; Fitches, A; Valler, D; Kuan, A M; Hoffbrand, V


    One hundred and twenty patients with homozygous beta thalassaemia were selected to determine the clinical effects of certain genetic factors which may modify disease severity. Genetic analysis defined specific beta thalassaemia mutations, the alpha thalassaemia genotype, and the presence of an XmnI restriction enzyme site, associated with increased fetal haemoglobin (HbF) production under certain conditions. Genotypic data with globin chain synthesis were related to the age when regular trans...

  18. Evidence of balanced diversity at the chicken interleukin 4 receptor alpha chain locus

    Podisi Baitsi


    Full Text Available Abstract Background The comparative analysis of genome sequences emerging for several avian species with the fully sequenced chicken genome enables the genome-wide investigation of selective processes in functionally important chicken genes. In particular, because of pathogenic challenges it is expected that genes involved in the chicken immune system are subject to particularly strong adaptive pressure. Signatures of selection detected by inter-species comparison may then be investigated at the population level in global chicken populations to highlight potentially relevant functional polymorphisms. Results Comparative evolutionary analysis of chicken (Gallus gallus and zebra finch (Taeniopygia guttata genes identified interleukin 4 receptor alpha-chain (IL-4Rα, a key cytokine receptor as a candidate with a significant excess of substitutions at nonsynonymous sites, suggestive of adaptive evolution. Resequencing and detailed population genetic analysis of this gene in diverse village chickens from Asia and Africa, commercial broilers, and in outgroup species red jungle fowl (JF, grey JF, Ceylon JF, green JF, grey francolin and bamboo partridge, suggested elevated and balanced diversity across all populations at this gene, acting to preserve different high-frequency alleles at two nonsynonymous sites. Conclusion Haplotype networks indicate that red JF is the primary contributor of diversity at chicken IL-4Rα: the signature of variation observed here may be due to the effects of domestication, admixture and introgression, which produce high diversity. However, this gene is a key cytokine-binding receptor in the immune system, so balancing selection related to the host response to pathogens cannot be excluded.

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

    翟海龙; 赖永榕


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

  20. 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

    潘莉; 翁文骏; 许吕宏; 魏菁; 方建培


    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

  1. Thyroid hormone regulates expression of a transfected human. alpha. -myosin heavy-chain fusion gene in fetal rat heart cells

    Tsika, R.W.; Bahl, J.J.; Morkin, E. (Univ. of Arizona College of Medicine, Tucson (USA)); Leinwand, L.A. (Albert Einstein College of Medicine, Bronx, NY (USA))


    The rat {alpha}-myosin heavy-chain ({alpha}-MHC) gene is regulated by 3,5,3{prime}-triiodo-L-thyronine (T{sub 3}) in ventricular myocardium and is constitutively expressed in atrial tissue. Less is known about regulation of the human gene, but conservation of sequences in the 5{prime}-flanking region between the rat and human {alpha}-MHC genes suggests that the human gene may be regulated similarly. Accordingly, T{sub 3}-responsiveness and tissue-specific expression of human and rat {alpha}-MHC/chloramphenicol acetyltransferase fusion constructs have been compared in rat fetal heart cells, L{sub 6}E{sub 9} myoblasts and myotubes, 3T3 fibroblasts, and HeLa cells. Transient transfection assays revealed a complex series of cis-regulatory elements in the 5{prime}-flanking sequences in the human genes, including a basal promoter element with canonical TATAA and CAAT sequences, two positive regulatory element(s), and two negative regulatory-elements, which markedly diminished both constitutive and T{sub 3}-inducible activity. Interestingly, the human gene seemed to contain a proximal thyroid-hormone response element(s) not found in the rat gene. The authors propose that interactions among the thyroid hormone responsive elements and other cis-acting elements in the human {alpha}-MHC 5{prime}-flanking sequences may be sufficient to explain the characteristic features of expression of this gene in cardiac tissues.

  2. Substitution of a conserved cysteine-996 in a cysteine-rich motif of the laminin {alpha}2-chain in congenital muscular dystrophy with partial deficiency of the protein

    Nissinen, M.; Xu Zhang; Tryggvason, K. [Univ. of Oulu (Finland)


    Congenital muscular dystrophies (CMDs) are autosomal recessive muscle disorders of early onset. Approximately half of CMD patients present laminin {alpha}2-chain (merosin) deficiency in muscle biopsies, and the disease locus has been mapped to the region of the LAMA2 gene (6q22-23) in several families. Recently, two nonsense mutations in the laminin {alpha}2-chain gene were identified in CMD patients exhibiting complete deficiency of the laminin {alpha}2-chain in muscle biopsies. However, a subset of CMD patients with linkage to LAMA2 show only partial absence of the laminin {alpha}2-chain around muscle fibers, by immunocytochemical analysis. In the present study we have identified a homozygous missense mutation in the {alpha}2-chain gene of a consanguineous Turkish family with partial laminin {alpha}2-chain deficiency. The T{r_arrow}C transition at position 3035 in the cDNA sequence results in a Cys996{r_arrow}Arg substitution. The mutation that affects one of the conserved cysteine-rich repeats in the short arm of the laminin {alpha}2-chain should result in normal synthesis of the chain and in formation and secretion of a heterotrimeric laminin molecule. Muscular dysfunction is possibly caused either by abnormal disulfide cross-links and folding of the laminin repeat, leading to the disturbance of an as yet unknown binding function of the laminin {alpha}2-chain and to shorter half-life of the muscle-specific laminin-2 and laminin-4 isoforms, or by increased proteolytic sensitivity, leading to truncation of the short arm. 42 refs., 7 figs.

  3. Branched chain amino acid transaminase and branched chain alpha-ketoacid dehydrogenase activity in the brain, liver and skele­tal muscle of acute hepatic failure rats



    Full Text Available Branched chain amino acid (BCAA transaminase activity increased in both the mitochondrial and supernatant fractions of brain from hepatic failure rats, in which a partial hepatectomy was performed 24h following carbon tetrachloride (CCl4 administration, although the activity of liver and skeletal muscle was the same as in control rats. The elevation of mitochondrial BCAA transaminase activity in liver-injured rats was partly due to increased activity of brain specific Type III isozyme. Branched chain alpha-ketoacid (BCKA dehydrogenase in the brain homogenates was not significantly altered in acute hepatic failure rats, while the liver enzyme activity was markedly diminished. BCKA dehydrogenase activity in the brain homogenates was inhibited by adding ATP to the assay system, and was activated in vitro by preincubating the brain homogenate at 37 degrees C for 15 min. These findings suggest that brain BCAA catabolism is accelerated in acute hepatic failure rats.

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

    E. Bartoloni Bocci


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

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

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


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

  6. Synthesis of beta-hexosaminidase in cell-free translation and in intact fibroblasts: an insoluble precursor alpha chain in a rare form of Tay-Sachs disease.

    Proia, R L; Neufeld, E F


    RNA was isolated from human term placenta or cultured fibroblasts and translated in a rabbit reticulocyte system in the presence of [35S]methionine; the translation products were immunoprecipitated with antisera made against beta-hexosaminidase or its isolated alpha and beta chains and analyzed by polyacrylamide gel electrophoresis. The largest translated alpha and beta chain polypeptides had Mrs of 65,000 and 59,000, respectively. These are approximately equal to 2,000 greater than the Mrs o...

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

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


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

  8. Structure effects in the region of superheavy elements via the $\\alpha$-decay chain of $^{293}$118

    Gupta, Raj K; Kumar, Rajesh; Balasubramaniam, M; Scheid, W; 10.1088/0954-3899/28/11/310


    The $\\alpha$-decay chain of $^{293}$118, first proposed in the Berkeley cold fusion experiment $^{208}$Pb($^{86}$Kr,1n) and now retracted, is calculated by using the preformed cluster model (PCM) of one of us (RKG). Also, the possible branchings of $\\alpha$-particles to heavier cluster decays of all the parents in this chain are calculated for the first time. The calculated Q-values, penetrabilities and preformation factors for $\\alpha$-decays suggest that the $^{285}$114 nucleus with Z=114, N=171 is a magic nucleus, either due to the magicity of Z=114, or of N=172 or of both. The N=172 is proposed to be a magic number in certain relativistic mean-field calculations, but with Z=120. The calculated cluster decays point to new interesting possibilities of $^{14}$C decay of the $^{281}$112 parent, giving rise to a (reasonably) deformed Z=106, N=161, $^{267}$106 daughter (N=162 being now established as the deformed magic shell) or to a doubly magic $^{48}$Ca cluster emitted from any of the parent nucleus in the $...

  9. cDNA sequence coding for the alpha'-chain of the third complement component in the African lungfish.

    Sato, A; Sültmann, H; Mayer, W E; Figueroa, F; Tichy, H; Klein, J


    cDNA clones coding for almost the entire C3 alpha-chain of the African lungfish (Protopterus aethiopicus), a representative of the Sarcopterygii (lobe-finned fishes), were sequenced and characterized. From the sequence it is deduced that the lungfish C3 molecule is probably a disulphide-bonded alpha:beta dimer similar to that of the C3 components of other jawed vertebrates. The deduced sequence contains conserved sites presumably recognized by proteolytic enzymes (e.g. factor I) involved in the activation and inactivation of the component. It also contains the conserved thioester region and the putative site for binding properdin. However, the site for the interaction with complement receptor 2 and factor H are poorly conserved. Either complement receptor 2 and factor H are not present in the lungfish or they bind to different residues at the same or a different site than mammalian complement receptor 2 and factor H. The C3 alpha-chain sequences faithfully reflect the phylogenetic relationships among vertebrate classes and can therefore be used to help to resolve the long-standing controversy concerning the origin of the tetrapods. PMID:10219761

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

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


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

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

    李营营; 冯学斌



  12. Association between genetic polymorphisms in the human interleukin-7 receptor alpha-chain and inhalation allergy

    Shamim, Z; Müller, K; Svejgaard, A; Poulsen, Lars K.; Bodtger, U; Ryder, L P


    Thymic stromal-derived lymphopoietin (TSLP) and interleukin-7 share a common receptor chain, IL-7Ralpha. IL-7 is involved in T-cell homeostasis, and TSLP induces production of pro-allergic cytokines. The gene encoding the IL-7Ralpha chain is polymorphic, and investigation of inhalation allergic p...

  13. Mechanisms of activation of muscle branched-chain alpha-keto acid dehydrogenase during exercise in man

    Van Hall, Gerrit; MacLean, D A; Saltin, B; Wagenmakers, A J

    1. Exercise leads to activation (dephosphorylation) of the branched-chain alpha-keto acid dehydrogenase (BCKADH). Here we investigate the effect of low pre-exercise muscle glycogen content and of branched-chain amino acid (BCAA) ingestion on the activity of BCKADH at rest and after 90 min of one......-leg knee-extensor exercise at 65% maximal one-leg power output in five subjects. 2. Pre-exercise BCAA ingestion (308 mg BCAAs (kg body wt)-1) caused an increased muscle BCAA uptake, a higher intramuscular BCAA concentration and activation of BCKADH both at rest (9 +/- 1 versus 25 +/- 5% for the control and...... BCAA test, respectively) and after exercise (27 +/- 4 versus 54 +/- 7%). 3. At rest the percentage active BCKADH was not different, 6 +/- 2% versus 5 +/- 1%, in the normal and low glycogen content leg (392 +/- 21 and 147 +/- 34 mumol glycosyl units (g dry muscle)-1, respectively). The post...

  14. Urine of patients with early prostate cancer contains lower levels of light chain fragments of inter-alpha-trypsin inhibitor and saposin B but increased expression of an inter-alpha-trypsin inhibitor heavy chain 4 fragment.

    Jayapalan, Jaime J; Ng, Keng L; Shuib, Adawiyah S; Razack, Azad H A; Hashim, Onn H


    The present study was aimed at the identification of proteins that are differentially expressed in the urine of patients with prostate cancer (PCa), those with benign prostatic hyperplasia (BPH) and age-matched healthy male control subjects. Using a combination of 2DE and MS/MS, significantly lower expression of urinary saposin B and two different fragments of inter-alpha-trypsin inhibitor light chain (ITIL) was demonstrated in the PCa patients compared to the controls. However, only one of the ITIL fragments was significantly different between the PCa and BPH patients. When image analysis was performed on urinary proteins that were transferred onto NC membranes and detected using a lectin that binds to O-glycans, a truncated fragment of inter-alpha-trypsin inhibitor heavy chain 4 was the sole protein found to be significantly enhanced in the PCa patients compared to the controls. Together, these urinary peptide fragments might be useful complementary biomarkers to indicate PCa as well as to distinguish it from BPH, although further epidemiological evidence on the specificity and sensitivity of the protein candidates is required. PMID:23417432

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

    王晓娟; 车传珍; 周昱男; 刘仲萍; 黄士昂


    目的 研究患者异基因造血干细胞移植(allo-HSCT)前后骨髓或外周血单个核细胞中CD4+CD25+Foxp3+调节性T淋巴细胞(以下简称CD4+CD25+Foxp3+T细胞)的比率、Foxp3 mRNA和Foxp3蛋白的表达与急性移植物抗宿主病(aGVHD)的关系.方法 选择37例行HIA相合异基因造血干细胞移植(allo-HSCT))的患者作为研究对象.采用流式细胞术检测患者allo-HSCT前7天(-7 d)、移植当天(0 d)、移植后30天(+30d)、+60 d和+90 d时骨髓或外周血单个核细胞中CD4+CD25+Foxp3+T细胞所占的比率;定量聚合酶链反应(PCR)检测Foxp3 mRNA相对表达量;蛋白印迹(Western blot)法检测Foxp3的蛋白表达水平.结果 37例患者存移植后100 d内发生aGVHD 13例(aGVHD组),未发生aGVHD 24例(无aGVHD组).CD4+CD25+Foxp3+T细胞的比率在0 d时最低,与-7、+30、+60、+90d时比较.差异均有统计学意义(P<0.01);与aGVHD组比较,无aGVHD组患者的CD4+CD25+Foxp3+T细胞的比率明显升高,差异有统计学意义(P<0.01).aGVHD组患者移植后Foxp3的表达水平逐渐升高,但明显低于无aGVHD组,尤其在+60和+90 d时的差异有统计学意义(P<0.01).Western blot法检测结果 表明,aGVHD组受者的Foxp3蛋白表达也明显低于无aGVHD组.结论 CD4+CD25+Foxp3+T细胞对防止发生aGVHD具有重要作用,监测患者骨髓或外周血中CD4+CD25+Foxp3+T细胞的比率可以预测aGVHD的发生.%Objective To investigate the expression and significance of CD4+ CD25+ Foxp3+ T Cells, Foxp3 mRNA and Foxp3 protein in the acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods Thirty-seven donor grafts and their respective clinical characteristics were evaluated. Flow cytometric analysis was performed to assess the percentage of CD4+ CD25+ Foxp3+ T cells in the recipients before, and 0, 30, 60, and 90 days after allo-HSCT. The relative expression of Foxp3 mRNA was detected by real time reverse transcription-polymerase chain

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

    马祥; 毛辉; 梁宗安


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

  17. Side-chain-anchored N(alpha)-Fmoc-Tyr-OPfp for bidirectional solid-phase synthesis

    Olsen, Christian A; Jørgensen, Malene; Hansen, Steen H; Witt, Matthias; Jaroszewski, Jerzy W; Franzyk, Henrik


    [reaction: see text] A mild resin-immobilization strategy employing a readily prepared trityl bromide resin for anchoring building blocks via a phenol group has been developed. With N(alpha)-Fmoc-Tyr-OPfp as a starter building block, it was possible to prepare asymmetrically substituted hybrids o...

  18. Affinity purification of human granulocyte macrophage colony-stimulating factor receptor alpha-chain. Demonstration of binding by photoaffinity labeling

    The human granulocyte macrophage colony-stimulating factor (GM-CSF) receptor alpha-chain, a low affinity component of the receptor, was solubilized and affinity-purified from human placenta using biotinylated GM-CSF. Scatchard analysis of 125I-GM-CSF binding to the placental membrane extract disclosed that the GM-CSF receptor had a dissociation constant (Kd) of 0.5-0.8 nM, corresponding to the Kd value of the GM-CSF receptor alpha-chain on the intact placental membrane. Affinity labeling of the solubilized protein using a photoreactive cross-linking agent, N-hydroxysuccinimidyl-4-azidobenzoate (HSAB), demonstrated a single specific band of 70-95 kDa representing a ligand-receptor complex. Approximately 2 g of the placental membrane extract was subjected to a biotinylated GM-CSF-fixed streptavidin-agarose column, resulting in a single major band at 70 kDa on a silver-stained sodium dodecyl sulfate gel. The radioiodination for the purified material disclosed that the purified protein had an approximate molecular mass of 70 kDa and a pI of 6.6. Binding activity of the purified material was demonstrated by photoaffinity labeling using HSAB-125I-GM-CSF, producing a similar specific band at 70-95 kDa as was demonstrated for the crude protein

  19. Affinity purification of human granulocyte macrophage colony-stimulating factor receptor alpha-chain. Demonstration of binding by photoaffinity labeling

    Chiba, S.; Shibuya, K.; Miyazono, K.; Tojo, A.; Oka, Y.; Miyagawa, K.; Takaku, F. (Univ. of Tokyo (Japan))


    The human granulocyte macrophage colony-stimulating factor (GM-CSF) receptor alpha-chain, a low affinity component of the receptor, was solubilized and affinity-purified from human placenta using biotinylated GM-CSF. Scatchard analysis of {sup 125}I-GM-CSF binding to the placental membrane extract disclosed that the GM-CSF receptor had a dissociation constant (Kd) of 0.5-0.8 nM, corresponding to the Kd value of the GM-CSF receptor alpha-chain on the intact placental membrane. Affinity labeling of the solubilized protein using a photoreactive cross-linking agent, N-hydroxysuccinimidyl-4-azidobenzoate (HSAB), demonstrated a single specific band of 70-95 kDa representing a ligand-receptor complex. Approximately 2 g of the placental membrane extract was subjected to a biotinylated GM-CSF-fixed streptavidin-agarose column, resulting in a single major band at 70 kDa on a silver-stained sodium dodecyl sulfate gel. The radioiodination for the purified material disclosed that the purified protein had an approximate molecular mass of 70 kDa and a pI of 6.6. Binding activity of the purified material was demonstrated by photoaffinity labeling using HSAB-{sup 125}I-GM-CSF, producing a similar specific band at 70-95 kDa as was demonstrated for the crude protein.


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


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

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

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


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

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

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


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

  3. 56 Increased Frequency of CD4+ CD25+ FOXP3- in Allergic Conjunctivitis Patients

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


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

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

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


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

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

    Ensaf Khalil Mohammed*, Zeinab Farag Asheiba


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

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

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


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

  7. Cloning of the laminin {alpha}3 chain gene (LAMA3) and identification of a homozygous deletion in a patient with Herlitz junctional epidermolysis bullosa

    Vidal, F.; Ortonne, J.P. [INSERM, Nice (France)]|[Hospital Pasteur, Nice (France); Galliano, M.F. [INSERM, Nice (France)] [and others


    Laminin 5 and laminin 6 are basement membrane proteins synthesized by the basal cells of stratifying squamous epithelia. Altered expression of laminin 5 has been associated with Herlitz junctional epidermolysis bullosa (H-JEB), a severe epidermal blistering disorder inherited as an autosomal recessive disease. We have isolated cDNA clones encoding the {alpha}3 chain of laminin 5 and searched for mutations in the LAMA3 gene in H-JEB patients. In one H-JEB family, an affected individual exhibited drastically reduced immunoreactivity to antibodies directed against the {alpha}3 chain of laminin 5 and an impaired expression of the corresponding mRNA transcripts. RT-PCR analysis of mRNA extracted from the proband`s keratinocytes identified a homozygous single basepair deletion in the transcripts encoding the laminin {alpha}3A and {alpha}3B isoforms. The mutation causes a frameshift and premature termination codon in both alleles of the LAMA3 gene. Inheritance of the clinical H-JEB phenotype was consistent with the segregation of the mutated allele in the family. We also report the identity of the {alpha} chains of laminin 5 and epiligrin and provide evidence that LAMA3 transcripts are distinct from the laminin 6 {alpha} chain mRNA. 35 refs., 5 figs., 1 tab.

  8. Cloning of the laminin alpha 3 chain gene (LAMA3) and identification of a homozygous deletion in a patient with Herlitz junctional epidermolysis bullosa.

    Vidal, F; Baudoin, C; Miquel, C; Galliano, M F; Christiano, A M; Uitto, J; Ortonne, J P; Meneguzzi, G


    Laminin 5 and laminin 6 are basement membrane proteins synthesized by the basal cells of stratifying squamous epithelia. Altered expression of laminin 5 has been associated with Herlitz junctional epidermolysis bullosa (H-JEB), a severe epidermal blistering disorder inherited as an autosomal recessive disease. We have isolated cDNA clones encoding the alpha 3 chain of laminin 5 and searched for mutations in the LAMA3 gene in H-JEB patients. In one H-JEB family, an affected individual exhibited drastically reduced immunoreactivity to antibodies directed against the alpha 3 chain of laminin 5 and an impaired expression of the corresponding mRNA transcripts. RT-PCR analysis of mRNA extracted from the proband's keratinocytes identified a homozygous single basepair deletion in the transcripts encoding the laminin alpha 3A and alpha 3B isoforms. The mutation causes a frameshift and premature termination codon in both alleles of the LAMA3 gene. Inheritance of the clinical H-JEB phenotype was consistent with the segregation of the mutated allele in the family. We also report the identity of the alpha chains of laminin 5 and epiligrin and provide evidence that LAMA3 transcripts are distinct from the laminin 6 alpha chain mRNA. PMID:8586427

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

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


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

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

    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.

  11. 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

    韩文杰; 史艳侠


    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.

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

    Nafiseh Pakravan; Agheel Tabar Molla Hassan; Zuhair Muhammad Hassan


    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".

  13. Structural requirements for assembly of dimeric IgA probed by site-directed mutagenesis of J chain and a cysteine residue of the alpha-chain CH2 domain.

    Krugmann, S; Pleass, R J; Atkin, J D; Woof, J M


    The structural features of J chain required for interaction with IgA in IgA dimer assembly were investigated by coexpression of wild-type and mutant forms of J chain with IgA1 in CHO cells. With wild-type J chain, a mixture of J chain-containing dimers and monomers was secreted. Substitution of Cys14 of J chain with Ser resulted in expression of only monomer IgA covalently associated with J chain. Similarly, mutation of Cys68 to Ser also resulted in expression predominantly of a monomer IgA-J chain species. These results suggest that Cys14 and Cys68 play critical roles in formation of J chain-containing IgA dimers, with each forming a disulfide bridge to an IgA monomer. Substitution of Asn48 with Ala, to prevent attachment of N-linked carbohydrate to J chain, also resulted in markedly reduced dimer assembly, suggesting a requirement for the sugar moiety in J chain function. We also mutated Cys311 on the C alpha2 domain of the IgA heavy chain to Ser. When coexpressed with wild-type J chain, this mutant was still capable of forming dimers, indicating that this residue was not involved in dimerization. Taken together, our results are consistent with an arrangement in which IgA monomers are linked end-to-end with J chain interposed. PMID:9200460

  14. Ground State Properties of New Element Z = 113 and Its Alpha Decay Chain

    TAI Fei; CHEN Ding-Han; XU Chang; REN Zhong-Zhou


    @@ We investigate the ground state properties of the new element 278113 and of the α-decay chain with different models, where the new element Z = 113 has been produced at RIKEN in Japan by cold-fusion reaction [Morita et al.J.Phys.Soc.Jpn.73 (2004) 2593].The experimental decay energies are reproduced by the deformed relativistic mean-field model, by the Skyrme-Hartree-Fock (SHF) model, and by the macroscopic-microscopic model.Theoretical half-lives also reasonably agree with the data.Calculations further show that prolate deformation is important for the ground states of the nuclei in the α-decay chain of 278113.The common points and differences among different models are compared and discussed.

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

    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.

  16. Fetal antigen 2: an amniotic protein identified as the aminopropeptide of the alpha 1 chain of human procollagen type I

    Teisner, B; Rasmussen, H B; Højrup, P;


    Fetal antigen (FA2) was purified from second trimester human amniotic fluid by immunospecific chromatography, gel filtration and reversed-phase chromatography. Gel filtration revealed two molecular forms of FA2 eluting at volumes corresponding to an M(r) of approximately 100 kDa and 30 kDa. SDS...... aminopropeptide of the alpha 1 chain of human procollagen type I as determined by nucleotide sequences. After oxidative procedures normally employed for radio-iodination (iodogen and chloramine-T), FA2 lost its immunoreactivity. An antigen which cross-reacted with polyclonal rabbit anti-human FA2 was demonstrated...... in fetal calf serum. Gel filtration with analysis of fractions by inhibition ELISA showed that the bovine homologue was present in the same molecular forms as those in human amniotic fluid, and immunohistochemical analysis with anti-human FA2 showed that its distribution in bovine skin was identical...

  17. Two RFLPs in human inter-alpha-trypsin inhibitor heavy chain gene ITIH2 on chromosome 10

    Leveillard, T.; Bourguignon, J.; Salier, J.P.; Diarra-Mehrpour, M.; Sesbouee, R.; Martin, J.P. (Institut National de la Sante et de la Recherche Medicale, St. Etienne Rouvray (France))


    The 0.8 kb Eco RI/Bam HI fragment of lambda HuHITI-9 used as a probe codes for human heavy chain H2 of the inter-alpha-trypsin inhibitor. Kpn I (GGTAC/C) identifies one invariant band at 8.5 kb and a diallelic polymorphism with DNA fragments at 26.0 kb or 20.0 kb. Msp I (C/CGG) identifies three invariant bands at 2.35 kb, 2.1 kb and 1.0 kb and a diallelic polymorphism with DNA fragments at 5.0 kb or 2.8 kb and 2.2 kb. The allele frequency for Kpn I and Msp I were determined. The ITIH2 gene has been mapped to 10p15 by in situ hybridization. Co-dominant segregation was found for each polymorphism in two informative families.

  18. Assembly, intracellular processing, and expression at the cell surface of the human alpha beta T cell receptor/CD3 complex. Function of the CD3-zeta chain

    Geisler, C; Kuhlmann, J; Rubin, B


    complex, the role of the CD3 chains for the TCR/CD3 expression have not been experimentally addressed in human T cells. In this study the function of the CD3-zeta chain for the assembly, intracellular processing, and expression of the TCR/CD3 complex in the human leukemic T cell line Jurkat was......The TCR/CD3 complex is a multimeric protein complex composed of a minimum of seven transmembrane chains (TCR alpha beta-CD3 gamma delta epsilon zeta 2). Whereas earlier studies have demonstrated that both the TCR-alpha and -beta chains are required for the cell surface expression of the TCR/CD3...... investigated. The results indicate that: 1) CD3-zeta is required for the cell surface expression of the TCR/CD3 complex; 2) the pentameric form (TCR alpha beta-CD3 gamma delta epsilon) of the TCR/CD3 complex and single TCR chains associated with CD3 (TCR alpha-CD3 gamma delta epsilon and TCR beta-CD3 gamma...

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

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


    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....

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

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


    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....

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

    王娜; 孔宪明; 曹兰芳



  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

    白明辉; 王竞; 涂彩霞; 张蕴颖; 刘敏; 李国艳; 钟良瑞


    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. Creatine, arginine alpha-ketoglutarate, amino acids, and medium-chain triglycerides and endurance and performance.

    Little, Jonathan P; Forbes, Scott C; Candow, Darren G; Cornish, Stephen M; Chilibeck, Philip D


    Creatine (Cr) supplementation increases muscle mass, strength, and power. Arginine a-ketoglutarate (A-AKG) is a precursor for nitric oxide production and has the potential to improve blood flow and nutrient delivery (i.e., Cr) to muscles. This study compared a commercial dietary supplement of Cr, A-AKG, glutamine, taurine, branched-chain amino acids, and medium-chain triglycerides with Cr alone or placebo on exercise performance and body composition. Thirty-five men (approximately 23 yr) were randomized to Cr + A-AKG (0.1 g . kg(-1) . d(-1) Cr + 0.075 g . kg(-1) . d(-1)A-AKG, n = 12), Cr (0.1 g . kg(-1) . d(-1), n = 11), or placebo (1 g . kg(-1) . d(-1) sucrose, n = 12) for 10 d. Body composition, muscle endurance (bench press), and peak and average power (Wingate tests) were measured before and after supplementation. Bench-press repetitions over 3 sets increased with Cr + A-AKG (30.9 +/- 6.6 +/- 34.9 +/- 8.7 reps; p < .01) and Cr (27.6 +/- 5.9 +/- 31.0 +/- 7.6 reps; p < .01), with no change for placebo (26.8 +/- 5.0 +/- 27.1 +/- 6.3 reps). Peak power significantly increased in Cr + A-AKG (741 +/- 112 +/- 794 +/- 92 W; p < .01), with no changes in Cr (722 +/- 138 +/- 730 +/- 144 W) and placebo (696 +/- 63 +/- 705 +/- 77 W). There were no differences in average power between groups over time. Only the Cr-only group increased total body mass (79.9 +/- 13.0 +/- 81.1 +/- 13.8 kg; p < .01), with no significant changes in lean-tissue or fat mass. These results suggest that Cr alone and in combination with A-AKG improves upper body muscle endurance, and Cr + A-AKG supplementation improves peak power output on repeated Wingate tests. PMID:19033611

  4. Radioimmunoassay of inhibin based on synthetic human inhibin alpha-chain peptide

    Polyclonal rabbit antisera were produced against cyclic human inhibin [(Cys6, Tyr7) alpha-(6-30)NH2] peptide, covalently conjugated to bovine serum albumin. The tyrosine residue introduced at position 7 facilitated the oxidative incorporation of radiolabel (125I) to yield a tracer with specific activity of 73.9 Ci/g. These reagents were used to develop a homologous equilibrium radioimmunoassay for human inhibin, with polyethylene glycol, 200 g/L, serving as the separation phase. At a detection limit of 2 micrograms/L (n = 7), immunoactive inhibin was detectable in human pre-ovulatory follicular fluid (128 micrograms/L), seminal plasma (2374 micrograms/L), amniotic fluid (66 micrograms/L), and placental extract (347 micrograms/L). We also demonstrated inhibin immunoreactivity in biological fluids from other mammalian species: macaque, chimpanzee, porcine, and bovine, but not rodent (guinea pig). Although the antisera were raised against a nonbioactive inhibin peptide, immunoglobulins fractionated on Protein A-Sepharose neutralized the bioactivity of human ovarian inhibin. Further characterization of inhibin immuno- and bioactivity was undertaken with immobilized heparin, divalent metal cations, and dye ligands. Only heparin-Sepharose distinguished between immuno- and bioactive inhibin

  5. Th17/CD4+CD25+Treg细胞与自身免疫病

    王琴; 崔向军



  6. Properties of Z=120 nuclei and the \\alpha-decay chains of the (292,304)120 isotopes using relativistic and non-relativistic formalisms

    Ahamad, Shakeb; Patra, S K


    The ground state and first intrinsic excited state of superheavy nuclei with Z=120 and N=160-204 are investigated using both non-relativistic Skyrme-Hartree-Fock and the axially deformed Relativistic Mean Field formalisms. We employ a simple BCS pairing approach for calculating the energy contribution from pairing interaction. The results for isotopic chain of binding energy, quadrupole deformation parameter, two neutron separation energies and some other observables are compared with the FRDM and some recent macroscopic-microscopic calculations. We predict superdeformed ground state solutions for almost all the isotopes. Considering the possibility of magic neutron number, two different mode of \\alpha-decay chains (292)120 and (304)120 are also studied within these frameworks. The Q_{\\alpha}-values and the half-life T^{\\alpha}_{1/2} for these two different mode of decay chains are compared with FRDM and recent macroscopic-microscopic calculations. The calculation is extended for the \\alpha-decay chains of 29...

  7. [Hemoglobin Boumerdès alpha 2(37) (C2) Pro----Arg beta 2: a new variant of the alpha chain associated with hemoglobin S in an Algerian family].

    Dahmane-Arbane, M; Blouquit, Y; Arous, N; Bardakdjian, J; Benamani, M; Riou, J; Benabadji, M; Rosa, J; Galacteros, F


    We report the first case of Hb Boumerdes, an alpha chain variant alpha 2(37) (C2) Pro----Arg beta 2, in an Algerian family. The propositus was also homozygous for the sickle cell gene. The abnormal hybrid Hb alpha 2Boum. beta 2S had an electrophoretic mobility on cellulose acetate pH 8.7 electrophoresis between those of Hb S and Hb A2. Its expression was about 16%. The alpha 2Boum. beta 2A fraction has a mobility between those of Hb F and Hb S. The effects of this mutation on Hb oxygen affinity and deoxy Hb S polymer formation were not studied. The propositus' sickle cell phenotype was benign. PMID:3438164

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

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


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

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

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


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

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

    Ensaf Khalil Mohammed*, Zeinab Farag Asheiba


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

  11. In vivo expression of UDP-N-acetylglucosamine: Alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I (GnT-1) in Aspergillus oryzae and effects on the sugar chain of alpha-amylase.

    Kasajima, Yuya; Yamaguchi, Masako; Hirai, Nobuaki; Ohmachi, Tetsuo; Yoshida, Takashi


    UDP-N-Acetylglucosamine: alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I (GnT-I) is an essential enzyme in the conversion of high mannose type oligosaccharide to the hybrid or complex type. The full length of the rat GnT-I gene was expressed in the filamentous fungus Aspergillus oryzae. A microsomal preparation from a recombinant fungus (strain NG) showed GnT-I activity that transferred N-acetylglucosamine residue to acceptor heptaose, Man(5)GlcNAc(2). The N-linked sugar chain of alpha-amylase secreted by the strain showed a peak of novel retention on high performance liquid chromatography that was same as a reaction product of in vitro GnT-1 assay. The peak of oligosaccharide disappeared on HPLC after beta-N-acetylglucosaminidase treatment. Mass analysis supported the presence of GlcNAcMan(5)GlcNAc(2) as a sugar chain of alpha-amylase from strain NG. Chimera of GnT-I with green fluorescent protein (GFP) showed a dotted pattern of fluorescence in the mycelia, suggesting localization at Golgi vesicles. We concluded that GnT-1 was functionally expressed in A. oryzae cells and that N-acetylglucosamine residue was transferred to N-glycan of alpha-amylase in vivo. A. oryzae is expected to be a potential host for the production of glycoprotein with a genetically altered sugar chain. PMID:17090929

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

    宋丽; 施森; 李敏



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



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

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

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


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

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

    栾樱译; 姚咏明


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

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

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


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

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

    肇静娴; 曾耀英; 李海仙; 曾祥凤; 季煜华; 何贤辉


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

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

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


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

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

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


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

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

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


    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

  1. Developmental alterations in the alpha-fetoprotein sugar chain in maternal serum analyzed by lectin affinity electrophoresis.

    Kawahara N


    Full Text Available Our purpose was to investigate developmental alterations of human alpha-fetoprotein (AFP oligosaccharides in maternal serum by lectin affinity electrophoresis and to compare the AFP glycoforms in maternal serum with those in umbilical cord serum and amniotic fluid. AFP glycoforms were separated by affinity electrophoresis with concanavalin A (Con A, lentil lectin (LCA, erythroagglutinating phytohemagglutinin (E-PHA and Allomyrina dichotoma lectin (allo A and detected by sensitive antibody affinity blotting. In maternal serum, increased proportions of Con A-nonreactive AFP (AFP-C1, LCA strongly-reactive AFP (AFP-L3 and E-PHA-reactive AFP (AFP-P4 and AFP-P5 decreased gradually during the early gestational weeks. Allo A-nonreactive AFP (AFP-A1 and asialo-AFP were found only in amniotic fluids during early gestational weeks. The percentages of these glycoforms at full term were almost the same among those body fluids. Since the glycoforms of maternal serum AFP were close to those of umbilical cord serum AFP, lectin-affinity electrophoretic analysis of maternal serum AFP may be useful for evaluating the developmental state of fetus by examining the nature of AFP sugar chain.

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

    Weiwei Qi


    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.

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

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


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

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

    庞惠勇; 葛恒明; 陈晓芹; 刘小林; 李忠典; 张振宇; 张健


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

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

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


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

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

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


    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.

  7. A homozygous nonsense mutation in the alpha 3 chain gene of laminin 5 (LAMA3) in Herlitz junctional epidermolysis bullosa: prenatal exclusion in a fetus at risk.

    McGrath, J A; Kivirikko, S; Ciatti, S; Moss, C; Dunnill, G S; Eady, R A; Rodeck, C H; Christiano, A M; Uitto, J


    Mutations in the three genes (LAMA3, LAMB3, and LAMC2) that encode the three chains (alpha 3, beta 3, and gamma 2, respectively) of laminin 5, a protein involved in epidermal-dermal adhesion, have been established as the genetic basis for the inherited blistering skin disorder, Herlitz junctional epidermolysis bullosa (H-JEB). In this study, we performed mutational analysis on genomic DNA from a child with H-JEB and identified a nonsense mutation in the alpha 3 chain gene (LAMA3) consisting of a homozygous C-to-T transition resulting in a premature termination codon (CGA-->TGA) on both alleles. The parents were shown to be heterozygous carriers of the same mutation. Direct mutation analysis was used to perform DNA-based prenatal diagnosis from a chorionic villus biopsy at 10 weeks' gestation in a subsequent pregnancy. The fetus was predicted to be genotypically normal with respect to the LAMA3 mutation. PMID:8530087

  8. The interleukin-3 receptor alpha chain is a unique marker for human acute myelogenous leukemia stem cells.

    Jordan, C T; Upchurch, D; Szilvassy, S J; Guzman, M L; Howard, D S; Pettigrew, A L; Meyerrose, T; Rossi, R; Grimes, B; Rizzieri, D A; Luger, S M; Phillips, G L


    Recent studies suggest that the population of malignant cells found in human acute myelogenous leukemia (AML) arises from a rare population of leukemic stem cells (LSCs). LSCs have been documented for nearly all AML subtypes and have been phenotypically described as CD34+/CD38- or CD34+/HLA-DR-. Given the potentially critical role of these primitive cells in perpetuating leukemic disease, we sought to further investigate their molecular and cellular characteristics. Flow cytometric studies using primary AML tissue showed that the interleukin-3 receptor alpha chain (IL-3Ralpha or CD123) was strongly expressed in CD34+/CD38- cells (98 +/- 2% positive) from 16 of 18 primary specimens. Conversely, normal bone marrow derived CD34+/CD38- cells showed virtually no detectable expression of the CD123 antigen. To assess the functional role of IL-3Ralpha positive cells, purified CD34+/CD123+ leukemia cells were transplanted into immune deficient NOD/SCID mice. These experiments showed that CD123+ cells were competent to establish and maintain leukemic populations in vivo. To begin to elucidate a biological role for CD123 in leukemia, primary AML samples were analyzed with respect to signal transduction activity in the MAPK, Akt, and Stat5 pathways. Phosphorylation was not detected in response to IL-3 stimulation, thereby suggesting CD123 is not active in conventional IL-3-mediated signaling. Collectively, these data indicate that CD123 represents a unique marker for primitive leukemic stem cells. Given the strong expression of this receptor on LSCs, we propose that targeting of CD123 may be a promising strategy for the preferential ablation of AML cells. PMID:11021753

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

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


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

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

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


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

  11. The properties of the single chicken MHC classical class II alpha chain ( B-LA) gene indicate an ancient origin for the DR/E-like isotype of class II molecules

    Salomonsen, Jan; Marston, Denise; Avila, David;


    significantly in the peptide-binding alpha(1) domain. The cDNA and genomic DNA sequences from chickens of diverse origins show few alleles, which differ in only four nucleotides and one amino acid. In contrast, significant restriction fragment length polymorphism is detected by Southern blot analysis of genomic...... DNA, suggesting considerable diversity around the gene. Analysis of a large back-cross family indicates that the class II alpha chain locus ( B-LA) is located roughly 5.6 cM from the MHC locus, which encodes the classical class II beta chains. Thus the chicken class II alpha chain gene is like the...

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

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


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

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

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


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

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

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


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

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

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


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

  16. 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

    张淼; 王彤; 陈怀民; 陈彦钦; 邓杨春


    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

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

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


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

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

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


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

  19. 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

    韦月梅; 邹洪才; 崔俊; 孔建忠; 田安国; 葛建英


    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.

  20. Real-time operating mode with DSSSD detector to search for short correlation ER-alpha chains

    Tsyganov, Yury


    Real-time PC based algorithm is developed for DSSSD detector. Complete fusion nuclear reaction natYb+48Ca->217Th is used to test this algorithm at 48Ca beam. Example of successful application of a former algorithm for resistive strip PIPS detector in 249Bk+48Ca nuclear reaction is presented too. Case of alpha-alpha correlations is also under brief consideration.

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

    王露; 张政


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

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

    You Lu

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

  3. Single-cell TCRseq: paired recovery of entire T-cell alpha and beta chain transcripts in T-cell receptors from single-cell RNAseq.

    Redmond, David; Poran, Asaf; Elemento, Olivier


    Accurate characterization of the repertoire of the T-cell receptor (TCR) alpha and beta chains is critical to understanding adaptive immunity. Such characterization has many applications across such fields as vaccine development and response, clone-tracking in cancer, and immunotherapy. Here we present a new methodology called single-cell TCRseq (scTCRseq) for the identification and assembly of full-length rearranged V(D)J T-cell receptor sequences from paired-end single-cell RNA sequencing reads. The method allows accurate identification of the V(D)J rearrangements for each individual T-cell and has the novel ability to recover paired alpha and beta segments. Source code is available at . PMID:27460926

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

    Li JY


    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

  5. BgII reveals two polymorphic sites in the human inter-alpha-trypsin inhibitor heavy chain gene ITI H2

    Leveillard, T.; Diarra-Mehrpour, M.; Salier, J.P.; Sesbouee, R.; Bourguignon, J.; Martin, J.P. (Institut National de la Sante et de la Recherche Medicale, St. Etienne Rouvray (France))


    The 0.8 kb EcoRI/BamHI fragment of lambda HuHITI-9 (1) used as probe codes for human heavy chain H2 of the inter-alpha-trypsin inhibitor. BgII (GCCN4/NGGC) identifies a three allele polymorphism with DNA fragments at 20.0 kb (A) or 11.0 kb (B) or 16.5 kb and 3.5 kb (C). The ITIH2 gene has been mapped to 10p15 by in situ hybridization. Co-dominant segregation was found for each polymorphism in one informative family.

  6. The clinicopathological features of three babies with osteogenesis imperfecta resulting from the substitution of glycine by valine in the pro alpha 1 (I) chain of type I procollagen.

    Cole, W G; Patterson, E; Bonadio, J; Campbell, P E; Fortune, D. W.


    The features of three babies with perinatal lethal osteogenesis imperfecta (OI II) resulting from substitutions of glycine by valine in the triple helical domain of the alpha 1(I) chain of type I collagen were studied. The babies were heterozygous for this substitution at residue 1006 in case 1 (OI35), 973 in case 2 (OI59), and 256 in case 3 (OI7B). OI35 had the most severe clinical form, OI IIC, with premature rupture of membranes, severe antepartum haemorrhage, stillbirth, severe short limb...

  7. CD4+ CD25+调节性T细胞及其分子标记物与支气管哮喘

    马祥; 毛辉; 梁宗安


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

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

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


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

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

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


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

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

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


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

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

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


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

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

    Tang, Qizhi Z; Bluestone, Jeffrey A.


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

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

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


    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.

  14. 银屑病患者外周血CD4+CD25+调节性T细胞水平的研究

    陈丽芳; 陈小敏; 杨秀丽; 史维平; 秦小卫; 郝树媛


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

  15. A homozygous nonsense mutation in the alpha 3 chain gene of laminin 5 (LAMA3) in lethal (Herlitz) junctional epidermolysis bullosa.

    Kivirikko, S; McGrath, J A; Baudoin, C; Aberdam, D; Ciatti, S; Dunnill, M G; McMillan, J R; Eady, R A; Ortonne, J P; Meneguzzi, G


    The inherited mechanobullous disorder, junctional epidermolysis bullosa (JEB), is characterized by extensive blistering and erosions of the skin and mucous membranes. The diagnostic hallmarks of JEB include ultrastructural abnormalities in the hemidesmosomes of the cutaneous basement membrane zone, as well as an absence of staining with antibodies against the anchoring filament protein, laminin 5. Therefore, the three genes encoding alpha 3, beta 3 and gamma 2 chains of laminin 5, known as LAMA3, LAMB3 and LAMC2, are candidate genes for JEB. We have previously demonstrated mutations in the LAMB3 and LAMC2 genes in several families with JEB. We initiated mutation analysis from an affected child by PCR amplification of individual LAMA3 exons, followed by heteroduplex analysis. Nucleotide sequencing of heteroduplexes identified a homozygous nonsense mutation within domain I/II of the alpha 3 chain. These findings provide the first evidence that nonsense mutations within the LAMA3 gene are also involved in the pathogenesis of JEB, and indicate that mutations of all three genes of laminin 5 can result in the JEB phenotype. PMID:7633458

  16. A homozygous nonsense mutation in the {alpha}3 chain gene of laminin 5 (LAMA3) in Herlitz junctional epidermolysis bullosa: Prenatal exclusion in a fetus at risk

    McGrath, J.A. [Jefferson Medical College, Philadelphia, PA (United States)]|[St. Thomas Hospital, London (United Kingdom); Ciatti, S.; Christiano, A.M. [Jefferson Medical College, Philadelphia, PA (United States)] [and others


    Mutations in the three genes (LAMA3, LAMB3, and LAMC2) that encode the three chains ({alpha}3, {Beta}3, and {gamma}2, respectively) of laminin 5, a protein involved in epidermal-dermal adhesion, have been established as the genetic basis for the inherited blistering skin disorder, Herlitz junctional epidermolysis bullosa (H-JEB). In this study, we performed mutational analysis on genomic DNA from a child with H-JEB and identified a nonsense mutation in the {alpha}3 chain gene (LAMA3) consisting of a homozygous C-to-T transition resulting in a premature termination codon (CGA {r_arrow} TGA) on both alleles. The parents were shown to be heterozygous carriers of the same mutation. Direct mutation analysis was used to perform DNA-based prenatal diagnosis from a chorionic villus biopsy at 10 weeks` gestation in a subsequent pregnancy. The fetus was predicted to be genotypically normal with respect to the LAMA3 mutation. 15 refs., 1 fig.

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

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


    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.

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

    Cao Jun


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

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


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

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

    Hong-jun WANG


    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

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

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


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

  2. Alpha-globin loci in homozygous beta-thalassemia intermedia.

    Triadou, P; Lapoumeroulie, C; Girot, R; Labie, D


    Homozygous beta-thalassemia intermediate (TI) differs from thalassemia major (TM) in being less severe clinically. Associated alpha-thalassemia could account for the TI phenotype by reducing the alpha/non-alpha chain imbalance. We have analyzed the alpha loci of 9 TI and 11 TM patients by restriction endonuclease mapping. All the TM and 7 of the TI patients have the normal complement of four alpha-globin genes (alpha alpha/alpha alpha). One TI patient has three alpha-globin genes (alpha alpha/-alpha), and another TI patient has five alpha genes (alpha alpha/alpha alpha alpha). PMID:6305827

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

    徐曼; 蒋小卫; 米粲


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

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

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


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

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

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


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

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

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


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

  7. Diagnosis of. alpha. sub 1 -antitrypsin deficiency by enzymatic amplification of human genomic DNA and direct sequencing of polymerase chain reaction products

    Newton, C.R.; Graham, A.; Powell, S.; Gammack, A.; Riley, J.; Markham, A.F. (ICI Diagnostics, Cheshire (England)); Kalsheker, N. (Univ. of Wales, Cardiff (Wales))


    The authors have compared sequencing of cloned polymerase chain reaction (PCR) products and the direct sequencing of PCR products in the examination of individuals from six families affected with {alpha}{sub 1}-antitrypsin (AAT) deficiency. In families where paternity was in question they confirmed consanguinity by DNA fingerprinting using a panel of locus-specific minisatellite probes. They demonstrate that direct sequencing of PCR amplification products is the method of choice for the absolutely specific diagnosis of AAT deficiency and can distinguish normals, heterozygotes and homozygotes in a single, rapid and facile assay. Furthermore, they demonstrate the reproducibility of the PCR and a rapid DNA isolation procedure. They have also shown that two loci can be simultaneously amplified and that the PCR product from each locus can be independently examined by direct DNA sequencing.

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

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


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

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

    李彩霞; 徐元品; 邹子宏; 周晓鸿


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

  10. Structural and Thermodynamic Basis for Weak Interactions between Dihydrolipoamide Dehydrogenase and Subunit-binding Domain of the Branched-chain [alpha]-Ketoacid Dehydrogenase Complex

    Brautigam, Chad A.; Wynn, R. Max; Chuang, Jacinta L.; Naik, Mandar T.; Young, Brittany B.; Huang, Tai-huang; Chuang, David T. (AS); (UTSMC)


    The purified mammalian branched-chain {alpha}-ketoacid dehydrogenase complex (BCKDC), which catalyzes the oxidative decarboxylation of branched-chain {alpha}-keto acids, is essentially devoid of the constituent dihydrolipoamide dehydrogenase component (E3). The absence of E3 is associated with the low affinity of the subunit-binding domain of human BCKDC (hSBDb) for hE3. In this work, sequence alignments of hSBDb with the E3-binding domain (E3BD) of the mammalian pyruvate dehydrogenase complex show that hSBDb has an arginine at position 118, where E3BD features an asparagine. Substitution of Arg-118 with an asparagine increases the binding affinity of the R118N hSBDb variant (designated hSBDb*) for hE3 by nearly 2 orders of magnitude. The enthalpy of the binding reaction changes from endothermic with the wild-type hSBDb to exothermic with the hSBDb* variant. This higher affinity interaction allowed the determination of the crystal structure of the hE3/hSBDb* complex to 2.4-{angstrom} resolution. The structure showed that the presence of Arg-118 poses a unique, possibly steric and/or electrostatic incompatibility that could impede E3 interactions with the wild-type hSBDb. Compared with the E3/E3BD structure, the hE3/hSBDb* structure has a smaller interfacial area. Solution NMR data corroborated the interactions of hE3 with Arg-118 and Asn-118 in wild-type hSBDb and mutant hSBDb*, respectively. The NMR results also showed that the interface between hSBDb and hE3 does not change significantly from hSBDb to hSBDb*. Taken together, our results represent a starting point for explaining the long standing enigma that the E2b core of the BCKDC binds E3 far more weakly relative to other {alpha}-ketoacid dehydrogenase complexes.

  11. Study of alpha decay chains of superheavy nuclei and magic number beyond Z = 82 and N = 126

    We study various α-decay chains on the basis of the preformed cluster decay model. Our work targets the superheavy elements, which are expected to show extra stability at shell closure. Our computations identify the following combinations of proton and neutron numbers as the most stable nuclei: Z=112, N=161,163; Z=114, N=171,178,179; and Z=124, N=194. We also investigate the alternative of heavy cluster emissions in the decay chain of 301120, instead of α decay. Our study of cluster radioactivity shows that the half-life for 10Be decay in 289114 is larger, indicating enhanced stability at Z=114, N=175. Similar calculations concerning the emission of 14C and 34Si from 301120 find the more stable combinations Z=114, N=173, and Z=106, N=161, respectively. From the same parent, 301120, the emission of a 49−51Ca cluster yielding a Z=100, N=152 daughter is the most probable. (author)

  12. 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

    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: [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)


    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.

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

    Yun, Xiang; SHANG, YUNXIAO; Li, Miao


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

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

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


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

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

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


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

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

    贾丽; 谢晓宝; 邱国强; 钱新瑜; 周民; 肖溶


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

  17. 离体CD4+CD25+调节性T细胞的扩增和混合淋巴细胞反应研究



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

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

    湛洁谊; 卢慧敏; 林穗玲


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

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

    李丽(综述); 李敏(审校)


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

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

    徐林; 徐薇; 蒋正刚; 熊思东


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

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

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


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

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



    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

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

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


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

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

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


    . 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....

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

    Godder Kamar


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

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

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


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

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

    Odeh, Adam N.; Simecka, Jerry W.


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

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

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


    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

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

    鞠云飞; 孙立锋; 胡华; 杨燕; 滕格玲


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

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



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

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

    梁瑞韵; 伍卫; 黄瑾; 江山平; 张蔚


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

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

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

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

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


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

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

    Ebrahim Kouchaki


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

  15. 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

    王俊; 罗英; 颜秉菊


    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

  16. Tumor necrosis factor-alpha-induced activation of RhoA in airway smooth muscle cells: role in the Ca2+ sensitization of myosin light chain20 phosphorylation.

    Hunter, Irene; Cobban, Hannah J; Vandenabeele, Peter; MacEwan, David J; Nixon, Graeme F


    Tumor necrosis factor-alpha (TNF), an inflammatory cytokine, has a potentially important role in the pathogenesis of bronchial asthma and may contribute to airway hyper-responsiveness. Recent evidence has revealed that TNF can increase the Ca(2+) sensitivity of agonist-stimulated myosin light chain(20) (MLC(20)) phosphorylation and contractility in guinea pig airway smooth muscle (ASM). In the present study, the potential intracellular pathways responsible for this TNF-induced Ca(2+) sensitization were investigated. In permeabilized cultured guinea pig ASM cells, recombinant human TNF stimulated an increase in Ca(2+)-activated MLC(20) phosphorylation under Ca(2+) "clamp" conditions. This increased MLC(20) phosphorylation was inhibited by preincubation with the Rho-kinase inhibitor Y27632. TNF also increased the proportion of GTP-bound RhoA, as measured using rhotekin Rho-binding domain, in a time course compatible with a role in the TNF-induced Ca(2+) sensitization. In cultured human ASM cells, recombinant human TNF also activated RhoA with a similar time course. In addition, TNF stimulated phosphorylation of the regulatory subunit of the myosin phosphatase, which was inhibited by Y27632. Although human ASM cells expressed both receptor subtypes, TNF-R1 and TNF-R2, the activation of RhoA was predominantly via stimulation of the TNF-R1, although RhoA did not immunoprecipitate with the TNF-R1. In conclusion, the TNF-induced increase in the Ca(2+) sensitivity of MLC(20) phosphorylation is through stimulation of the TNF-R1 receptor and via a RhoA/Rho-kinase pathway leading to inhibition of the myosin light chain phosphatase. This intracellular mechanism may contribute to TNF-induced airway hyper-responsiveness. PMID:12606782

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

    王子函; 朱继业; 李涛; 冷希圣


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

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

    上官文姬; 沈惠风; 王利民; 叶人诵


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

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

    史彧; 褚莉莉; 张兰芳; 赵德育



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

    陈敏; 樊黎; 吴晓立


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

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

    陈跃华; 朱华芳; 范婷婷


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

  2. Fibrinogen Alpha Chain Precursor and Apolipoprotein A-I in Urine as Biomarkers for Noninvasive Diagnosis of Calcium Oxalate Nephrolithiasis: A Proteomics Study

    Wei Zhu


    Full Text Available Calcium oxalate nephrolithiasis is the most common urological disease, but noninvasive and convenient methods of diagnosis are rarely available. Objective. The present study aimed to identify potential urine biomarkers for noninvasive diagnosis of CaOx nephrolithiasis. Methodology. Urine samples from 72 patients with CaOx nephrolithiasis and 30 healthy controls were collected and proteomics analysis was performed using matrix-assisted laser desorption/ionization-time of flight-mass spectrometer (MALDI-TOF-MS. Results. Thirteen proteins/peptides displayed statistically significant differences. The peptides of m/z 1207.23 and 2773.86 were selected by the genetic algorithm (GA to build a possible diagnostic model. The area under the curve of m/z 1207.23 and 2773.86 was 0.936 and 0.987, respectively. The diagnostic model in distinguishing patients and healthy subjects showed 100% sensitivity and specificity. The peak at m/z 2773.86 was identified as fibrinogen alpha chain (FGA with the sequence G.EGDFLAEGGGVR.G, and the peak at m/z 2773.86 was identified as apolipoprotein A-I (apoA-I with the sequence L.PVLESFKVSFLSALEEYTKKLNTQ. Conclusion. The study results strongly suggested that urinary FGA and apoA-I are highly sensitive and specific biomarkers for noninvasive diagnosis of CaOx nephrolithiasis.

  3. Long-chain fatty acids regulate liver carnitine palmitoyltransferase I gene (L-CPT I) expression through a peroxisome-proliferator-activated receptor alpha (PPARalpha)-independent pathway.

    Louet, J F; Chatelain, F; Decaux, J F; Park, E A; Kohl, C; Pineau, T; Girard, J; Pegorier, J P


    Liver carnitine palmitoyltransferase I (L-CPT I) catalyses the transfer of long-chain fatty acid (LCFA) for translocation across the mitochondrial membrane. Expression of the L-CPT I gene is induced by LCFAs as well as by lipid-lowering compounds such as clofibrate. Previous studies have suggested that the peroxisome-proliferator-activated receptor alpha (PPARalpha) is a common mediator of the transcriptional effects of LCFA and clofibrate. We found that free LCFAs rather than acyl-CoA esters are the signal metabolites responsible for the stimulation of L-CPT I gene expression. Using primary culture of hepatocytes we found that LCFAs failed to stimulate L-CPT I gene expression both in wild-type and PPARalpha-null mice. These results suggest that the PPARalpha-knockout mouse does not represent a suitable model for the regulation of L-CPT I gene expression by LCFAs in the liver. Finally, we determined that clofibrate stimulates L-CPT I through a classical direct repeat 1 (DR1) motif in the promoter of the L-CPT I gene while LCFAs induce L-CPT I via elements in the first intron of the gene. Our results demonstrate that LCFAs can regulate gene expression through PPARalpha-independent pathways and suggest that the regulation of gene expression by dietary lipids is more complex than previously proposed. PMID:11171094

  4. Patients with ovarian carcinoma excrete different altered levels of urine CD59, kininogen-1 and fragments of inter-alpha-trypsin inhibitor heavy chain H4 and albumin

    Hashim Onn H


    Full Text Available Abstract Background Diagnosis of ovarian carcinoma is in urgent need for new complementary biomarkers for early stage detection. Proteins that are aberrantly excreted in the urine of cancer patients are excellent biomarker candidates for development of new noninvasive protocol for early diagnosis and screening purposes. In the present study, urine samples from patients with ovarian carcinoma were analysed by two-dimensional gel electrophoresis and the profiles generated were compared to those similarly obtained from age-matched cancer negative women. Results Significant reduced levels of CD59, kininogen-1 and a 39 kDa fragment of inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4, and enhanced excretion of a 19 kDa fragment of albumin, were detected in the urine of patients with ovarian carcinoma compared to the control subjects. The different altered levels of the proteins were confirmed by Western blotting using antisera and a lectin that bind to the respective proteins. Conclusion CD59, kininogen-1 and fragments of ITIH4 and albumin may be used as complementary biomarkers in the development of new noninvasive protocols for diagnosis and screening of ovarian carcinoma.

  5. Cutting edge: dominant effect of Ile50Val variant of the human IL-4 receptor alpha-chain in IgE synthesis.

    Mitsuyasu, H; Yanagihara, Y; Mao, X Q; Gao, P S; Arinobu, Y; Ihara, K; Takabayashi, A; Hara, T; Enomoto, T; Sasaki, S; Kawai, M; Hamasaki, N; Shirakawa, T; Hopkin, J M; Izuhara, K


    Two variants of the IL-4R alpha-chain (IL-4Ralpha) gene have been recently identified in association with different atopic disorders. To clarify the etiological relationship between the two variants, we analyzed responsiveness to IL-4 of transfectants with four kinds of IL-4Ralpha carrying either Val or Ile at 50 and either Gln or Arg at 551. The substitution of Ile for Val augmented STAT6 activation, proliferation, and transcription activity of the Iepsilon promoter by IL-4, whereas that of Arg for Gln did not change these IL-4 signals. Arg551 was not associated with atopic asthma in the Japanese population. CD23 expression and IgE synthesis by IL-4 were augmented in Ile50-bearing PBMC, compared with those bearing Val50. Taken together, substitution of Arg551 does not enhance the IL-4 signal for generation of germline epsilon transcript, whereas the substitution of Ile50 contributes to enhancement of IgE synthesis. PMID:9973373

  6. Expression of anti-tumor necrosis factor alpha (TNFα) single-chain variable fragment (scFv) in Spirodela punctata plants transformed with Agrobacterium tumefaciens.

    Balaji, Parthasarathy; Satheeshkumar, P K; Venkataraman, Krishnan; Vijayalakshmi, M A


    Therapeutic antibodies against tumor necrosis factor alpha (TNFα) have been considered effective for some of the autoimmune diseases such as rheumatoid arthritis, Crohn's diseases, and so on. But associated limitations of the current therapeutics in terms of cost, availability, and immunogenicity have necessitated the need for alternative candidates. Single-chain variable fragment (scFv) can negate the limitations tagged with the anti-TNFα therapeutics to a greater extent. In the present study, Spirodela punctata plants were transformed with anti-TNFα through in planta transformation using Agrobacterium tumefaciens strain, EHA105. Instead of cefotaxime, garlic extract (1 mg/mL) was used to remove the agrobacterial cells after cocultivation. To the best of our knowledge, this report shows for the first time the application of plant extracts in transgenic plant development. 95% of the plants survived screening under hygromycin. ScFv cDNA integration in the plant genomic DNA was confirmed at the molecular level by PCR. The transgenic protein expression was followed up to 10 months. Expression of scFv was confirmed by immunodot blot. Protein expression levels of up to 6.3% of total soluble protein were observed. β-Glucuronidase and green fluorescent protein expressions were also detected in the antibiotic resistant plants. The paper shows the generation of transgenic Spirodela punctuata plants through in planta transformation. PMID:25786575

  7. Proteomic demonstration of the recurrent presence of inter-alpha-inhibitor H4 heavy-chain during aspergillosis induced in an animal model.

    Desoubeaux, Guillaume; Jourdan, Marie-Lise; Valera, Lionel; Jardin, Bénédicte; Hem, Sonia; Caille, Agnès; Cormier, Bénédicte; Marchand-Adam, Sylvain; Bailly, Éric; Diot, Patrice; Chandenier, Jacques


    Invasive pulmonary aspergillosis remains a matter of great concern in oncology/haematology, intensive care units and organ transplantation departments. Despite the availability of various diagnostic tools with attractive features, new markers of infection are required for better medical care. We therefore looked for potential pulmonary biomarkers of aspergillosis, by carrying out two-dimensional (2D) gel electrophoresis comparing the proteomes of bronchial-alveolar lavage fluids (BALF) from infected rats and from control rats presenting non-specific inflammation, both immunocompromised. A bioinformatic analysis of the 2D-maps revealed significant differences in the abundance of 20 protein spots (ANOVA P-value0.8). One of these proteins, identified by mass spectrometry, was considered of potential interest: inter-alpha-inhibitor H4 heavy-chain (ITIH4), characterised for the first time in this infectious context. Western blotting confirmed its overabundance in all infected BALF, particularly at early stages of murine aspergillosis. Further investigations were carried on rat serum, and confirmed that ITIH4 levels increased during experimental aspergillosis. Preliminary results in human samples strengthened this trend. To our knowledge, this is the first description of the involvement of ITIH4 in aspergillosis. PMID:24360996

  8. Anti-TNF-alpha therapy induces a distinct regulatory T cell population in patients with rheumatoid arthritis via TGF-beta

    Nadkarni, S.; Mauri, C; Ehrenstein, M. R.


    The induction of regulatory T (T reg) cells holds considerable potential as a treatment for autoimmune diseases. We have previously shown that CD4(+)CD25(hi) T reg cells isolated from patients with active rheumatoid arthritis (RA) have a defect in their ability to suppress proinflammatory cytokine production by CD4(+)CD25(-) T cells. This defect, however, was overcome after anti-tumor necrosis factor (TNF)-alpha antibody (infliximab) therapy. Here, we demonstrate that infliximab therapy gives...

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

    HU Zhenping; LI Dazhu; HU Yingfeng; YANG Keping


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

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

    苏虹; 王忱诚; 王保龙; 彭美玲


    目的 探讨在粒细胞集落刺激因子(G-CSF)动员的外周血单个核细胞(PBMC)中诱导产生CD4+CD25+调节性T细胞(Treg)的可行性及其表型和功能.方法 收集G-CSF动员和未动员的BALB/c ♀鼠的PBMC,采用磁活性标记的细胞分选法 (MACS) 分选BALB/c ♀鼠的CD4+CD25-T细胞和CD4+CD25+T细胞及C57BL/6 ♂鼠的CD11c+未成熟树突细胞(iDC),建立细胞培养体系,经iDC体外诱导G-CSF动员和未动员的CD4+CD25-T细胞转换为CD4+CD25+Treg细胞(iTreg).分别应用流式细胞术、混合淋巴细胞反应技术检测诱导后细胞的CD25、Foxp3的表达水平以及抑制功能,比较G-CSF动员与非动员组间iTreg的表型和抑制功能的差异.结果 iDC诱导G-CSF未动员和动员的组间CD25+分子表达率分别为(76.8±4.1)%和(90.4±5.3)%,差异有统计学意义(P<0.01);两组诱导产生的CD25+T细胞的Foxp3表达水平分别为(64.1±2.7)%和(59.5±3.2)%,差异有统计学意义(P<0.05).iDC诱导G-CSF未动员和动员的外周血产生的iCD4+CD25+Treg均表现出免疫抑制功能,其中动员后诱导生成的iTreg的免疫抑制效应明显增强.结论 应用iDC从G-CSF动员的外周血中诱导产生的iCD4+CD25+Treg具有更强的体外抑制效应,为自身免疫性疾病的治疗提供新的思路和手段.%Objective To explore the feasibility of inducing CD4+ CD25 + regulatory T lymphocytes (Tregs) from mononuclear cells of granulocyte-colony stimulating factor (G-CSF) mobilized peripheral blood , and to analyze the phenotype and function of these cells . Methods G-CSF mobilized and un-mobilized peripheral blood mononuclear cells were collected from BALB/c murine. CD4 + CD25 - T cells and CD4 + CD25 + T cells of BALB/c murine, and CDllc+ cells (immature dendritic cell, iDC) of C57BL/6 murine were separated with magnetic activated cell sorting (MACS) respectively. Different cell culture systems were set up to induce CD4+ CD25+ Tregs (iTregs) by iDC from mononuclear cells of G

  11. Expression of mRNAs coding for the alpha 1 chain of type XIII collagen in human fetal tissues: comparison with expression of mRNAs for collagen types I, II, and III


    This paper describes the topographic distribution of the multiple mRNAs coding for a novel human short-chain collagen, the alpha 1 chain of type XIII collagen. To identify the tissues and cells expressing these mRNAs, human fetal tissues of 15-19 gestational wk were studied by Northern and in situ hybridizations. The distribution pattern of the type XIII collagen mRNAs was compared with that of fibrillar collagen types I, II, and III using specific human cDNA probes for each collagen type. No...

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

    李红华; 赵娟; 张景鸿


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

  13. 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

    彭磊磊; 葛圣林; 张成鑫


    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

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

    张莹; 姚咏明; 于燕; 吴瑶; 盛志勇


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

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

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


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

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

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


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

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

    Qian LI; Hua-hao SHEN


    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.

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

    Aleksiewicz Roman


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

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

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


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

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

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


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

  1. Administration of anti-CD25 mAb leads to impaired alpha-galactosylceramide-mediated induction of IFN-gamma production in a murine model

    Rosalia, Rodney Alexander; Štěpánek, Ivan; Polláková, Veronika; Šímová, Jana; Bieblová, Jana; Indrová, Marie; Moravcová, Simona; Přibylová, Hana; Bontkes, H.; Bubeník, Jan; Sparwasser, T.; Reiniš, Milan


    Roč. 218, č. 6 (2013), s. 851-859. ISSN 0171-2985 R&D Projects: GA ČR GA301/07/1410; GA ČR GAP301/10/2174 Institutional research plan: CEZ:AV0Z50520514 Institutional support: RVO:68378050 Keywords : cancer immunotherapy * DEREG mice * interferon γ * natural killer T cells * PC61 monoclonal antibody * T regulatory cells Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.180, year: 2013

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

    葛荣领; 张建华


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

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

    刘春花; 刘恩梅



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

    薛克营; 周咏明; 熊盛道; 熊维宁


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

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

    石涛; 冯学斌; 赵志旭; 张瑾锦


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

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

    祖莹; 李成荣; 郑跃杰; 邓继岿; 付晓玲


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

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

    潘珍珍; 李羚; 郭赟; 贺建


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

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

    湛洁谊; 卢慧敏; 林穗玲


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

  9. A G {r_arrow} A transition at position IVS-11 +1 of the HEX A {alpha}-chain gene in a non-Ashkenazic Mexican Tay-Sachs infant

    Miranda, S.R.P.; Gwon, S.; DeGasperi, R. [New York Univ. Medical Center, NY (United States)] [and others


    Tay-Sachs disease (TSD) is an autosomal recessive storage disorder caused by a deficiency of the lysosomal enzyme, {beta}-N-acetylhexosaminidase A (Hex A), a heteropolymer composed of two polypeptides, {alpha} and {beta}. Mutations in the {alpha}-chain gene render the enzyme defective, resulting in the accumulation of g{sub m2} ganglioside in the nervous system. Deficiency of Hex A was detected in a non-Ashkenazic girl of Mexican origin indicating infantile onset of TSD. Molecular investigation of the {alpha}-chain gene excluded the typical Ashkenazic 4 bp insertion in the exon 11 and the intron 12 splice-junction mutations by Hae III and Dde I restriction analysis, respectively. Single strand conformation polymorphism (SSCP) analysis showed a different pattern in the sample where exon 11 and flanking regions were amplified in the patient DNA as compared to the migration of control DNA. Sequencing of PCR amplified genomic DNA containing exon 11 and flanking intronic regions showed a single base substitution (G {r_arrow} A) at position IVS-11 +1. This mutation creates a recognition site for the restriction enzyme Mbo II. Digestion of exon 11 and flanking regions with Mbo II demonstrated homozygosity of the patient for this mutation and heterozygosity in the mother. mRNA from cultured fibroblasts obtained from a normal control and from the propositus was reverse transcribed. The cDNAs coding for Hex A {alpha}-chain were amplified in four overlapping fragments. In the patient sample it was not possible to amplify the fragment containing the exon 11/intron 11 junction, indicating that this mutation alters normal RNA processing of the Hex A pre-mRNA resulting in the deficiency of Hex A activity.

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

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


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