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Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells
Open accessAbstract
Abnormalities in CD4+CD25+Foxp3+ regulatory T (T reg) cells have been implicated in susceptibility to allergic, autoimmune, and immunoinflammatory conditions. However, phenotypic and functional assessment of human T reg cells has been hampered by difficulty in distinguishing between CD25-expressing activated and regulatory T cells. Here, we show that expression of CD127, the α chain of the interleukin-7 receptor, allows an unambiguous flow cytometry–based distinction to be made between CD127lo T reg cells and CD127hi conventional T cells within the CD25+CD45RO+RA− effector/memory and CD45RA+RO− naive compartments in peripheral blood and lymph node. In healthy volunteers, peripheral blood CD25+CD127lo cells comprised 6.35 ± 0.26% of CD4+ T cells, of which 2.05 ± 0.14% expressed the naive subset marker CD45RA. Expression of FoxP3 protein and the CD127lo phenotype were highly correlated within the CD4+CD25+ population. Moreover, both effector/memory and naive CD25+CD127lo cells manifested suppressive activity in vitro, whereas CD25+CD127hi cells did not. Cell surface expression of CD127 therefore allows accurate estimation of T reg cell numbers and isolation of pure populations for in vitro studies and should contribute to our understanding of regulatory abnormalities in immunopathic diseases.
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“In terms of surface phenotype, the best accepted combination of markers is high expression of the IL-2 receptor α chain (CD25) and low expression of the IL-7R α chain (CD127) [134, 385].”
From Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition) (2021) · cites this paper as [385]
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“In combination with CD25 during flow cytometry analysis, CD127 can be used as biomarker for analysis and, because of the expression on the cell membrane, for isolation of Tregs (16, 17).”
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“Secondly, although the low surface expression of CD127 molecule allows more accurate identification of Treg cells [58], we did not check the expression of this molecule.”
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“Expression of CD127, the chain of the IL-7 receptor, has been reported to allow a flow cytometry- based distinction between CD127(lo) Tregs and CD127(hi) conventional T cells within the human CD25CD45RO effector/memory and CD45RA naive compartments in peripheral blood and LN [124, 125].”
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“98% purity), a highly purified human Treg population (36, 37).”
BackgroundRecent advances and current state of immunotherapy in systemic lupus erythematosus
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“Treg.[108] Treg achieve their suppressive effect on other immune cells through cognate interaction or cytokine produc-”
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“At the same time, in comparison to the effect on Tfh, we also studied the effect of treated HIV-1 infection on other key subsets of CD4+ T cells, including CCR5+ memory CD4+ T cells, CD127+ long-term memory cells, CD103+ tissue-resident memory T (Trm) cells, CD161+ cells, which is a surrogate cell surface maker for Th…”
MethodResultInfluentialSelf-citationPawłowski, P., Grubczak, K., Kostecki, J., Iłendo, E., Moniuszko, M., Pawłowska, M., Ręjdak, R., Reszeć, J., & Myśliwiec, J. (2017) · Hormone and Metabolic Research · doi:10.1055/s-0042-122780 · Cited by 17
“More recent studies demonstrated that the combination of low CD127 expression and high CD25 expression identified potently suppressive cells highly enriched with Foxp3s [18, 19].”
BackgroundRegulatory immune cells in kidney disease
Lee, V. W., Wang, Y. M., Wang, Y. P., Zheng, D., Polhill, T. S., Cao, Q., Wu, H., Alexander, I. E., Alexander, S. I., & Harris, D. C. H. (2008) · American Journal of Physiology-Renal Physiology · doi:10.1152/ajprenal.00077.2008 · Cited by 17
“In humans, the recent identification of CD127 expression has been the most definitive marker yet of Tregs (73).”
Self-citationExperimentally induced accumulation of Foxp3+ T cells in upper airway allergy
Skrindo, I., Scheel, C., Johansen, F., & Jahnsen, F. L. (2011) · Clinical & Experimental Allergy · doi:10.1111/j.1365-2222.2011.03710.x · Cited by 16
Human regulatory T cells suppress proliferation of B lymphoma cells
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“CD127 expression is reportedly inversely correlated with the degree of FOXP3 expression in human peripheral blood CD4 + CD25 + T cells.[28,29] CD4 + CD25 + CD127T cells with the low FOXP3 TSDR methylation level are generally considered as a Treg population generated in thymus, whereas FOXP3-expressing CD4 + CD25 + CD1…”
BackgroundDelineation of Homeostatic Immune Signatures Defining Viremic Non-progression in HIV-1 Infection
Singh, A. K., Salwe, S., Padwal, V., Velhal, S., Sutar, J., Bhowmick, S., Mukherjee, S., Nagar, V. S., Patil, P., & Patel, V. V. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.00182 · Cited by 15
“Expression of CD25 (IL-2Rα) and CD127 (IL-7Rα) on T cells was used to enumerate Tregs (CD3+CD4+CD25highCD127low) (31, 32) (Supplementary Figure S1B).”
Immunologic and Genetic Contributors to CD46-Dependent Immune Dysregulation
Meyer, B. J., Kunz, N., Seki, S., Higgins, R., Ghosh, A., Hupfer, R., Baldrich, A., Hirsiger, J. R., Jauch, A., Burgener-Gasser, A.-V., et al. (2023) · Journal of Clinical Immunology · doi:10.1007/s10875-023-01547-y · Cited by 16
“4b), in keeping with literature on Treg surface phenotyping [40, 41].”
ResultHindmarch, D. C., Malashanka, S., Shows, D. M., Clarke, A. S., & Lord, J. D. (2023) · Journal of Crohn s and Colitis · doi:10.1093/ecco-jcc/jjad173 · Cited by 16
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“Consequently, Tregs are now defined as Foxp3 + /CD127low/CD25high/CD4 + cells [13].”
BackgroundRegulatory T cells in renal disease.
Wang, Y. M., Hu, M., Wang, Y., Polhill, T. S., Zhang, G. Y., Wang, Y., Lee, V. W., Harris, D. C. H., & Alexander, S. I. (2008) · PubMed Central · Cited by 16
Self-citationA novel human CD4 + T‐cell inducer subset with potent immunostimulatory properties
Ndhlovu, L. C., Leal, F. E., Eccles-James, I. G., Jha, A. R., Lanteri, M. C., Norris, P. J., Barbour, J. D., Wachter, D. J., Andersson, J., Taskén, K., et al. (2009) · European Journal of Immunology · doi:10.1002/eji.200939258 · Cited by 16
Molecular Regulation ofCellular Immunity by FOXP3
McMurchy, A. N., Nunzio, S. D., Roncarolo, M. G., Bacchetta, R., & Levings, M. K. (2009) · Advances in experimental medicine and biology · doi:10.1007/978-1-4419-1599-3_3 · Cited by 17
REGULATORY T-CELLS IN CHRONIC LYMPHOCYTIC LEUKEMIA
D'Arena, G. F., Rossi, G., Vannata, B., & Deaglio, S. (2012) · Mediterranean Journal of Hematology and Infectious Diseases · doi:10.4084/mjhid.2012.053 · Cited by 16
“Moreover, CD127, that identified the heterodimeric IL-7 receptor, combined with CD4, CD25 and FoxP3, has been shown to better identify Tregs avoiding the contamination of this small cell population (accounting for 1-4% of circulating CD4+ lymphocytes in humans) with activated Tcells.16,17 Tregs and Autoimmunity.”
Jafarinia, M., Mehdipour, F., Hosseini, S. V., Ghahramani, L., Hosseinzadeh, M., & Ghaderi, A. A. (2016) · Tumor Biology · doi:10.1007/s13277-016-5345-y · Cited by 16
Katz, G., Voss, K., Yan, T. F., Kim, Y. C., Kortum, R. L., Scott, D. W., & Snow, A. L. (2018) · Cellular Immunology · doi:10.1016/j.cellimm.2018.02.007 · Cited by 16
“The gating strategy is shown in Figure 1, where Tregs were defined as CD4+ CD25hi CD127lo and Tcons were defined as CD4+ CD25lo CD127hi [22].”
MethodHirabayashi, K., Yanagisawa, R., Saito, S., Higuchi, Y., Koya, T., Sano, K., Koido, S., Okamoto, M., Sugiyama, H., Nakazawa, Y., et al. (2018) · Anticancer Research · doi:10.21873/anticanres.12465 · Cited by 16
Truffault, F., Nazzal, D., Verdier, J., Gradolatto, A., Fadel, É., Roussin, R., Eymard, B., Panse, R. L., & Berrih‐Aknin, S. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.00782 · Cited by 16
“The absence of CD127 is also a hallmark of Treg, yet some CD25hiCD127low CD4 + T cells also contain non-Treg (22, 23).”
BackgroundLee, J. G., Jaeger, K. E., Seki, Y., Lim, Y. W., Cunha, C., Vuchkovska, A., Nelson, A. J., Nikolai-Yogerst, A., Kim, D., Nishimura, M. I., et al. (2021) · Immunology · doi:10.1111/imm.13316 · Cited by 17
Do Treg Speed Up with CARs? Chimeric Antigen Receptor Treg Engineered to Induce Transplant Tolerance
Kaljanac, M., & Abken, H. (2022) · Transplantation · doi:10.1097/tp.0000000000004316 · Cited by 19
“CD127 is a suitable marker in this respect because CD127 inversely correlates with FoxP3 and suppressive function.(69,70) CD4CD25 Treg‚ sorted based on low CD127 expression‚ provide a more potent therapeutic capacity than conventional Treg.”
Ladd, M., Sharma, A., Huang, Q., Wang, A. Y., Xu, L., Genowati, I., Levings, M. K., & Lavoie, P. M. (2010) · Immunology · doi:10.1111/j.1365-2567.2010.03304.x · Cited by 18
“3A), but did express high levels of CD127 (the IL-7 receptor alpha chain), which is absent on Tregs and also characteristic of naïve T cells [27, 28].”
BackgroundDiagnostic Value of Regulatory T Cells: A New Facet of a Much Studied Cell Population
Brown, K., & Wong, W. (2008) · Transplantation · doi:10.1097/tp.0b013e31818f3d2a · Cited by 16
“The low level, or lack of, expression of CD127 has also recently been used to identify Treg (71, 72).”
Li, W., Tenner‐Racz, K., Rácz, P., Janowicz, D. M., Fortney, K. R., Katz, B. P., & Spinola, S. M. (2010) · The Journal of Infectious Diseases · doi:10.1086/652781 · Cited by 16
“Human Treg cells can generally be distinguished from effector T cells on the basis of low surface expression of CD127 (interleukin 7 receptor a) [31, 32].”
BackgroundExpression of IL-7 receptor in human peripheral regulatory T cells
Ciebiada, M., Kasztalska, K., Górska-Ciebiada, M., Barylski, M., & Górski, P. (2012) · Archives of Medical Science · doi:10.5114/aoms.2012.31387 · Cited by 16
He, H., Ni, B., Tian, Y., Tian, Z., Chen, Y., Liu, Z., Yang, X., Lv, Y., & Zhang, Y. (2013) · Immunology · doi:10.1111/imm.12198 · Cited by 16
Immunologic monitoring in kidney transplant recipients
Townamchai, N., Safa, K., & Chandraker, A. K. (2013) · Kidney Research and Clinical Practice · doi:10.1016/j.krcp.2013.04.002 · Cited by 16
“Effector/memory T cells help prevent infection and cancer, but are associated with acute rejection [56]; they can be detected and differentiated from naïve T cells by staining for CD25, CD45RA, CD45RO, and CD62L [57–59].”
BackgroundScaleia, R. L., Morrone, S., Stoppacciaro, A., Scarpino, S., Antonelli, M., Bianchi, E., Nardo, G. D., Oliva, S., Viola, F., Cucchiara, S., et al. (2010) · Journal of Pediatric Gastroenterology and Nutrition · doi:10.1097/mpg.0b013e3181e4d323 · Cited by 16
Zhou, W., Deng, J., Chen, Q., Li, R., Xu, X., Guan, Y., Li, W., Xiong, X., Li, H., Li, J., et al. (2020) · International Journal of Medical Sciences · doi:10.7150/ijms.44088 · Cited by 16
“CD127 is a recently discovered antigen associated with Tregs that is weakly expressed on Tregs, while the self-activated memory T cell CD127 is strongly expressed; therefore, CD4+CD25+CD127 low/- is used to represent Tregs now[4, 5].”
BackgroundPhenotypic Correlations between Monocytes and CD4+ T Cells in Allergic Patients
Moniuszko, M., Kowal, K., Jeznach, M., Rusak, M., Dąbrowska, M., & Bodzenta‐Łukaszyk, A. (2013) · International Archives of Allergy and Immunology · doi:10.1159/000343687 · Cited by 16
Cell exhaustion in HIV-1 infection
Seddiki, N., Brezar, V., & Draenert, R. (2014) · Current Opinion in HIV and AIDS · doi:10.1097/coh.0000000000000087 · Cited by 16
Self-citationGene editing of CD3 epsilon to redirect regulatory T cells for adoptive T cell transfer
Du, W., Noyan, F., McCallion, O., Drosdek, V., Kath, J., Glaser, V., Fuster-García, C., Yang, M.-X., Stein, M., Franke, C., et al. (2025) · Molecular Therapy · doi:10.1016/j.ymthe.2025.01.045 · Cited by 12
Validation of a multicolor staining to monitor phosphoSTAT5 levels in regulatory T-cell subsets
Ehx, G., Hannon, M., Beguin, Y. P., Humblet‐Baron, S., & Baron, F. (2015) · Oncotarget · doi:10.18632/oncotarget.6486 · Cited by 15
“Tregs are also featured by a low expression of CD127, the α chain of the IL-7 receptor [5].”
BackgroundAllergic disease in 8‐year‐old children is preceded by delayed B cell maturation
Strömbeck, A., Nordström, I., Andersson, K., Andersson, H., Johansen, S., Maglio, C., Rabe, H., Adlerberth, I., Wold, A. E., Hesselmar, B., et al. (2017) · Clinical & Experimental Allergy · doi:10.1111/cea.12922 · Cited by 15
“At 8 years of age, additional markers were included as follows: CD24 and CD38 to distinguish immature/transitional (CD24CD38), mature/na€ıve (CD24CD38), memory (CD24CD38) B cells(23) and CD127 to identify CD4CD25CD127 putative Tregs.(24) We also identified proportions of CD45RA and CD45RO putative Tregs.”
MethodInfluentialBrezar, V., Hani, L., Surénaud, M., Hubert, A., Lacabaratz, C., Lelièvre, J., Lévy, Y., & Seddiki, N. (2017) · PLoS Pathogens · doi:10.1371/journal.ppat.1006489 · Cited by 15
Self-citationTabares, P., Berr, S., Langenhorst, D., Sawitzki, B., ten Berge, I. J. M., Tony, H., & Hünig, T. (2017) · European Journal of Immunology · doi:10.1002/eji.201747244 · Cited by 15
Donaubauer, A.-J., Becker, I., Rühle, P. F., Fietkau, R., Gaipl, U. S., & Frey, B. (2019) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/bs.mie.2019.03.003 · Cited by 15
Stroukov, W., Mastronicola, D., Albany, C. J., Catak, Z., Lombardi, G., & Scottà, C. (2023) · Cytometry Part A · doi:10.1002/cyto.a.24720 · Cited by 14
“wherein the very high expression of CD25 combined to the low expression of CD127 describe a well-defined population expressing high levels of the lineage-defining Treg marker FOXP3 [9].”
BackgroundLibetta, C., Esposito, P. B., Gregorini, M., Margiotta, E., Martinelli, C., Borettaz, I., Canevari, M., Rampino, T., Ticozzelli, E., Abelli, M., et al. (2015) · Transplant Immunology · doi:10.1016/j.trim.2015.07.005 · Cited by 15
A flow cytometry based assay for the enumeration of regulatory T cells in whole blood
Hardy, M. Y., Vari, F., Rossetti, T. R., Hart, D. N. J., & Prue, R. L. (2012) · Journal of Immunological Methods · doi:10.1016/j.jim.2012.07.004 · Cited by 15
Łuczyński, W., Stasiak‐Barmuta, A., Juchniewicz, A., Wawrusiewicz‐Kurylonek, N., Iłendo, E., Kos, J., Kretowski, A. J., Górska, M., Chyczewski, L., & Bossowski, A. T. (2010) · Folia Histochemica et Cytobiologica · doi:10.2478/v10042-008-0113-5 · Cited by 15
Dasgupta, A., Mahapatra, M., & Saxena, R. (2014) · Leukemia & lymphoma/Leukemia and lymphoma · doi:10.3109/10428194.2014.966245 · Cited by 15
Dasgupta, A., Mahapatra, M., & Saxena, R. (2012) · Leukemia & lymphoma/Leukemia and lymphoma · doi:10.3109/10428194.2012.730614 · Cited by 15
Therapeutic potential of low‐dose IL‐2 in immune thrombocytopenia: An analysis of 3 cases
Zhang, J., Ruan, Y., Xu, X., Wang, H., Tao, Q., Lu, J., Xia, L., Zhang, Q., Wang, J., Wang, Y., et al. (2017) · Cytometry Part B Clinical Cytometry · doi:10.1002/cyto.b.21601 · Cited by 15
“Interestingly, studies have found that CD4 1 CD25 1 T cells with low expression of CD127 can also be used as reliable markers to define CD4 1 Tregs (10,11).”
BackgroundMetabolic reprogramming augments potency of human pSTAT3–inhibited iTregs to suppress alloreactivity
Walton, K., Fernandez, M. R., Sagatys, E. M., Reff, J., Kim, J., Lee, M. C., Kiluk, J. V., Hui, J. Y. C., McKenna, D. H., Hupp, M., et al. (2020) · JCI Insight · doi:10.1172/jci.insight.136437 · Cited by 15
“The final purity of the iTreg (CD4+, CD127-, CD25+, Foxp3+) (28, 29) product was >90% (Fig.”
BackgroundHey‐Cunningham, A. J., Riaz, A., Fromm, P. D., Kupresanin, F., Markham, R., & McGuire, H. M. (2021) · Reproductive Sciences · doi:10.1007/s43032-021-00658-4 · Cited by 17
“Using multiparametric flow cytometry, through a series of sequential hierarchical gating CD4+ T cells were identified in the blood and/or endometrium of all participants, with Tregs further identified using best practice approach of CD25hi CD127lo CD4+ expression by lymphocytes [40] pro-”
MethodAbadja, F., Alamartine, É., Berthoux, F. C., Mariat, C., Genin, C., & Lambert, C. (2010) · Transplantation · doi:10.1097/tp.0b013e3181bbbd67 · Cited by 15
“9, ACT1, Bb10, BF2, 2A3, BC96, MA251, and 4E3) to detect the CD4 CD25 CD127 regulatory T-cell subset (6,7) was compared.”
Mason, R. D., Rose, R. D., Seddiki, N., Kelleher, A. D., & Kent, S. J. (2008) · Virology · doi:10.1016/j.virol.2008.08.042 · Cited by 15
Self-citation
Metadata
- Authors
- Seddiki, N., Santner-Nanan, B., Martinson, J., Zaunders, J., Sasson, S., Landay, A., Solomon, M., Selby, W., Alexander, S. I., Nanan, R., Kelleher, A., & de St. Groth, B. F.
- Affiliations
- The University of Sydney (AU)
- 4Department of Paediatrics, University of Sydney, Western Clinical School, Penrith, NSW 2751, Australia
- Rush University Medical Center (US)
- St Vincent's Hospital Sydney (AU)
- Royal Prince Alfred Hospital (AU)
- Children's Hospital at Westmead (AU)
- UNSW Sydney (AU)
- 1Centenary Institute of Cancer Medicine and Cell Biology, Faculty of Medicine, University of Sydney, Newtown, NSW 2042, Australia
- ORCID
- Venue
- The Journal of Experimental Medicine
- Publisher
- Rockefeller University Press
- Published
- 2006-07-03
- Volume
- 203
- Issue
- 7
- Pages
- 1693-1700
- ISSN
- 0022-1007 (ISSN-L)
- 1540-9538
- PMC
- PMC2118333
- OpenAlex
- W2168628806
- Cited by
- 1,512
- References
- 30
- Citations by year
- 2026: 18 · 2025: 25 · 2024: 32 · 2023: 32 · 2022: 44 · 2021: 49 · 2020: 58 · 2019: 75 · 2018: 51 · 2017: 64 · 2016: 74 · 2015: 63 · 2014: 87 · 2013: 92 · 2012: 116
- FWCI
- 45.91
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The Journal of Experimental Medicine (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
- PubMed Central (publishedVersion, cc-by-nc-sa)
- PubMed (publishedVersion)
- UNSWorks (University of New South Wales, Sydney, Australia) (submittedVersion, cc-by-nc-nd)
- Funders
- Crohn's and Colitis Foundation of America
- Crohn's and Colitis Foundation
- Medical Research Council
- National Health and Medical Research Council
- National Institute of Allergy and Infectious Diseases (P01AI055793, AI55793, U19AI055793)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Immunology and Microbiology › Immunology › Psoriasis: Treatment and Pathogenesis
- Life Sciences › Immunology and Microbiology › Immunology › IL-33, ST2, and ILC Pathways
- Keywords
- regulatory T cells, CD127, interleukin-2 receptor, Foxp3, CD25, CD4+ T cells, naive T cells, effector memory T cells
- MeSH terms
- Adult, Humans, Infant, Newborn, Lymphocyte Activation, Receptors, Interleukin-2, T-Lymphocyte Subsets, Receptors, Interleukin-7, T-Lymphocytes, Regulatory
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