Hartigan-O'Connor DJ, Poon C, Sinclair E, McCune JMHuman CD4+ regulatory T cells express lower levels of the IL-7 receptor alpha chain (CD127), allowing consistent identification and sorting of live cells. J Immunol Methods 319:41-52

Division of Experimental Medicine, Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
Journal of Immunological Methods (Impact Factor: 1.82). 02/2007; 319(1-2):41-52. DOI: 10.1016/j.jim.2006.10.008
Source: PubMed


Although quantitative identification and viable enrichment of natural regulatory T cells (T-regs) in humans are problematic, such steps would greatly facilitate the analysis of these cells in disease states. In an attempt to identify markers that are sensitive and specific for human T-regs, we analyzed the expression of fourteen intracellular and cell surface markers on human CD4(+) cells. Many markers were partially selective for CD25(hi) T-regs, but consistent and specific discrimination of functional T-regs was only made possible by focus on CD127, the alpha chain of the IL-7 receptor. Although most CD4(+) human T cells express CD127, T-regs exhibiting suppressive activity in vitro display distinctly lower surface expression of this marker, irrespective of their level of CD25 expression. Sorted cells with the surface phenotype CD4(+)CD25(+)CD127(low) had higher levels of intracellular FOXP3 and CTLA-4 and, as determined by functional assays, were suppressive, hypoproliferative, and poorly responsive to TCR signaling. The CD4(+)CD25(+)CD127(low) phenotype was also found to be characteristic of T-regs found in mice and in rhesus macaques. This surface phenotype should allow for quantitative studies of regulatory T cells in disease states as well as for enrichment of live regulatory T cells for functional analyses and/or expansion in vitro.

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    • "Therefore, many studies have focused on finding more specific Treg surface markers, and demonstrated that surface expression of CD127, the α-chain of the IL-7 receptor, in combination with CD25 can distinguish human regulatory from conventional CD4 + T cells [5] [6]. In particular, CD127 expression inversely correlates with that of FoxP3 and the suppressive function of human CD4 + Treg cells, so that low expression of CD127 is currently one of the most important features used to define Tregs [7] [8]. "
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    ABSTRACT: Regulatory T cells (Tregs), defined as CD4+CD25+highFoxP3+CD127- cells, could promote tolerance in renal transplantation (Tx). In an open-label, randomized, controlled trial 62 de-novo Tx recipients received induction with basiliximab and cyclosporine A (CsA) for the first month after Tx and then were assigned to treatment with sirolimus (SRL) or CsA and followed-up for 2years. The primary endpoint was to evaluate the effects of induction and maintenance treatments on circulating Tregs, while the secondary endpoint was the assessment of Treg renal infiltration and the relationship between Treg count and clinical outcomes. There were no significant differences in either circulating or tissue Treg number between the two groups. At 1month post-Tx, all patients presented a profound Treg depletion, followed by a significant increase in Tregs that resulted stable during the follow-up. The same trend was also observed for non-activated Tregs (CD69-) and for other immunocompetent cells (CD4+ and CD8+ T cells, B cells and NK cells). Moreover, the Treg count did not correlate either with renal function or with acute rejection and graft loss. Initial immunosuppression is crucial to regulate circulating Tregs, regardless of subsequent immunosuppressive maintenance regimens. Strategies aiming to promote tolerance should consider the effects of different induction regimens. Copyright © 2015. Published by Elsevier B.V.
    Transplant Immunology 07/2015; 33(2). DOI:10.1016/j.trim.2015.07.005 · 1.46 Impact Factor
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    • "As activated human T cells can transiently express FoxP3 and CD25, differentiation of Tregs from activated effector T cells by only using these two markers may suffer from inaccuracies. CD127 is a newly described surface marker that allows distinguishing regulatory T cells from other CD25+ cells [40]. For TH17 identification, we chose two experimental approaches: determination the mRNA expression of the TH17 specific transcription factor RORγT by qPCR, and FACS analyses of IL-17 production, which has revealed as a very reliable method to identify TH17 cells [41]. "
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    ABSTRACT: Chronic low back pain (CLBP) is a leading cause of disability and costs in health care systems worldwide. Despite extensive research, the exact pathogenesis of CLBP, particularly the individual risk of chronification remains unclear. To investigate a possible role of the adaptive immune system in the pathophysiology of CLBP, we analyzed T cell related cytokine profiles, T cell related mRNA expression patterns and the distribution of T cell subsets in 37 patients suffering from nonspecific CLBP before and after multimodal therapy in comparison to 25 healthy controls. Serum patterns of marker cytokines were analyzed by Luminex technology, mRNA expression of cytokines and specific transcription factors was measured by real-time PCR, and distribution of TH1-, TH2-, TH17- and regulatory T cell (Tregs) subsets was determined by multicolor flow cytometry. We found that CLBP patients exhibit an increased number of anti-inflammatory Tregs, while pro-inflammatory TH17 cells are decreased, resulting in an altered TH17/Treg ratio. Accordingly, FoxP3 and TGF-β-mRNA expression was elevated, while expression of IL-23 was reduced. Serum cytokine analyses proved to be unsuitable to monitor the adaptive immune response in CLBP patients. We further show that even after successful therapy with lasting reduction of pain, T cell subset patterns remained altered after a follow-up period of 6 months. These findings suggest an involvement of TH17/Treg cells in the pathogenesis of CLBP and emphasize the importance of these cells in the crosstalk of pain and immune response. Trial Registration German Clinical Trial Register: Registration Trial DRKS00005954.
    PLoS ONE 08/2014; 9(8):e104883. DOI:10.1371/journal.pone.0104883 · 3.23 Impact Factor
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    • "In the present study, we investigated the phenotypic profile of membrane molecules associated with functional activation and inhibition of TREG and effector T cells. Using a highly specific strategy for TREG cell identification based on the surface markers CD25+CD127Ø/low∅ (5-7,25), we could not find any abnormality in TREG cell frequency, but we originally identified a series of peculiar qualitative differences in the composition of surface molecules of TREG cells in SLE patients. Most TREG cells consistently expressed CD95, approximately half were CD45RO+, and one-third were HLA-DR+ in SLE patients and healthy controls. "
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    ABSTRACT: Regulatory T (TREG) cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE). TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25+/highCD127Ø/lowFoxP3+, and effector T cells were defined as CD25+CD127+FoxP3Ø. The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4+TREG and CD28+TREG cells and an increased frequency of CD40L+TREG cells. There was a decrease in the TREG/effector-T ratio for GITR+, HLA-DR+, OX40+, and CD45RO+ cells, and an increased ratio of TREG/effector-T CD40L+ cells in patients with SLE. In addition, CD40L+TREG cell frequency correlated with the SLE disease activity index (P=0.0163). In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease.
    Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas / Sociedade Brasileira de Biofisica ... [et al.] 08/2014; 47(8):662-9. DOI:10.1590/1414-431X20143483 · 1.01 Impact Factor
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