Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells
Nabila Seddiki, Brigitte Santner-Nanan, Jeff Martinson, John Zaunders, Sarah Sasson, Alan Landay, Michael Solomon, Warwick Selby, Stephen I Alexander, Ralph Nanan, Anthony Kelleher, Barbara Fazekas de St Groth, Nabila Seddiki, Brigitte Santner-Nanan, Jeff Martinson, John Zaunders, Sarah Sasson, Alan Landay, Michael Solomon, Warwick Selby, Stephen I Alexander, Ralph Nanan, Anthony Kelleher, Barbara Fazekas de St Groth
Abstract
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 alpha chain of the interleukin-7 receptor, allows an unambiguous flow cytometry-based distinction to be made between CD127(lo) T reg cells and CD127(hi) 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(+)CD127(lo) 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 CD127(lo) phenotype were highly correlated within the CD4(+)CD25(+) population. Moreover, both effector/memory and naive CD25(+)CD127(lo) cells manifested suppressive activity in vitro, whereas CD25(+)CD127(hi) 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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References
- Sakaguchi, S. 2004. Naturally arising CD4+ regulatory T cells for immunologic self-tolerance and negative control of immune responses. Annu. Rev. Immunol. 22:531–562.
- Kriegel, M.A., T. Lohmann, C. Gabler, N. Blank, J.R. Kalden, and H.M. Lorenz. 2004. Defective suppressor function of human CD4+ CD25+ regulatory T cells in autoimmune polyglandular syndrome type II. J. Exp. Med. 199:1285–1291.
- Crispin, J.C., A. Martinez, and J. Alcocer-Varela. 2003. Quantification of regulatory T cells in patients with systemic lupus erythematosus. J. Autoimmun. 21:273–276.
- Cao, D., R. van Vollenhoven, L. Klareskog, C. Trollmo, and V. Malmstrom. 2004. CD25brightCD4+ regulatory T cells are enriched in inflamed joints of patients with chronic rheumatic disease. Arthritis Res. Ther. 6:R335–R346.
- Ehrenstein, M.R., J.G. Evans, A. Singh, S. Moore, G. Warnes, D.A. Isenberg, and C. Mauri. 2004. Compromised function of regulatory T cells in rheumatoid arthritis and reversal by anti-TNFα therapy. J. Exp. Med. 200:277–285.
- Sugiyama, H., R. Gyulai, E. Toichi, E. Garaczi, S. Shimada, S.R. Stevens, T.S. McCormick, and K.D. Cooper. 2005. Dysfunctional blood and target tissue CD4+CD25high regulatory T cells in psoriasis: mechanism underlying unrestrained pathogenic effector T cell proliferation. J. Immunol. 174:164–173.
- Viglietta, V., C. Baecher-Allan, H.L. Weiner, and D.A. Hafler. 2004. Loss of functional suppression by CD4+CD25+ regulatory T cells in patients with multiple sclerosis. J. Exp. Med. 199:971–979.
- Furuno, K., T. Yuge, K. Kusuhara, H. Takada, H. Nishio, V. Khajoee, T. Ohno, and T. Hara. 2004. CD25+CD4+ regulatory T cells in patients with Kawasaki disease. J. Pediatr. 145:385–390.
- Karlsson, M.R., J. Rugtveit, and P. Brandtzaeg. 2004. Allergen-responsive CD4+CD25+ regulatory T cells in children who have outgrown cow's milk allergy. J. Exp. Med. 199:1679–1688.
- Ling, E.M., T. Smith, X.D. Nguyen, C. Pridgeon, M. Dallman, J. Arbery, V.A. Carr, and D.S. Robinson. 2004. Relation of CD4+CD25+ regulatory T-cell suppression of allergen-driven T-cell activation to atopic status and expression of allergic disease. Lancet. 363:608–615.
- Ochs, H.D., S.F. Ziegler, and T.R. Torgerson. 2005. FOXP3 acts as a rheostat of the immune response. Immunol. Rev. 203:156–164.
- Kukreja, A., G. Cost, J. Marker, C. Zhang, Z. Sun, K. Lin-Su, S. Ten, M. Sanz, M. Exley, B. Wilson, et al. 2002. Multiple immuno-regulatory defects in type-1 diabetes. J. Clin. Invest. 109:131–140.
- Huang, Y.M., R. Pirskanen, R. Giscombe, H. Link, and A.K. Lefvert. 2004. Circulating CD4+CD25+ and CD4+CD25− T cells in myasthenia gravis and in relation to thymectomy. Scand. J. Immunol. 59:408–414.
- Putheti, P., A. Pettersson, M. Soderstrom, H. Link, and Y.M. Huang. 2004. Circulating CD4+CD25+ T regulatory cells are not altered in multiple sclerosis and unaffected by disease-modulating drugs. J. Clin. Immunol. 24:155–161.
- van Amelsfort, J.M., K.M. Jacobs, J.W. Bijlsma, F.P. Lafeber, and L.S. Taams. 2004. CD4+CD25+ regulatory T cells in rheumatoid arthritis: differences in the presence, phenotype, and function between peripheral blood and synovial fluid. Arthritis Rheum. 50:2775–2785.
- Baecher-Allan, C., J.A. Brown, G.J. Freeman, and D.A. Hafler. 2001. CD4+CD25high regulatory cells in human peripheral blood. J. Immunol. 167:1245–1253.
- Seddiki, N., B. Santner-Nanan, S.G. Tangye, S.I. Alexander, M. Solomon, S. Lee, R. Nanan, and B. Fazekas de St Groth. 2006. Persistence of naïve CD45RA+ regulatory T cells in adult life. Blood. 107:2830–2838.
- Roncador, G., P.J. Brown, L. Maestre, S. Hue, J.L. Martinez-Torrecuadrada, K.L. Ling, S. Pratap, C. Toms, B.C. Fox, V. Cerundolo, et al. 2005. Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single-cell level. Eur. J. Immunol. 35:1681–1691.
- Ruprecht, C.R., M. Gattorno, F. Ferlito, A. Gregorio, A. Martini, A. Lanzavecchia, and F. Sallusto. 2005. Coexpression of CD25 and CD27 identifies FoxP3+ regulatory T cells in inflamed synovia. J. Exp. Med. 201:1793–1803.
- Sadlack, B., H. Merz, H. Schorle, A. Schimpl, A.C. Feller, and I. Horak. 1993. Ulcerative colitis-like disease in mice with a disrupted interleukin-2 gene. Cell. 75:253–261.
- von Freeden-Jeffry, U., P. Vieira, L.A. Lucian, T. McNeil, S.E. Burdach, and R. Murray. 1995. Lymphopenia in interleukin (IL)-7 gene-deleted mice identifies IL-7 as a nonredundant cytokine. J. Exp. Med. 181:1519–1526.
- Godfrey, W.R., D.J. Spoden, Y.G. Ge, S.R. Baker, B. Liu, B.L. Levine, C.H. June, B.R. Blazar, and S.B. Porter. 2005. Cord blood CD4+CD25+-derived T regulatory cell lines express FoxP3 protein and manifest potent suppressor function. Blood. 105:750–758.
- Valmori, D., A. Merlo, N. Souleimania, C. Hesdorffer, and M. Ayyoub. 2005. A peripheral circulating compartment of natural naive CD4 Tregs. J. Clin. Invest. 115:1953–1962.
- Baecher-Allan, C., E. Wolf, and D.A. Hafler. 2006. MHC class II expression identifies functionally distinct human regulatory T cells. J. Immunol. 176:4622–4631.
- Morgan, M.E., J.H. van Bilsen, A.M. Bakker, B. Heemskerk, M.W. Schilham, F.C. Hartgers, B.G. Elferink, L. van der Zanden, R.R. de Vries, T.W. Huizinga, et al. 2005. Expression of FOXP3 mRNA is not confined to CD4+CD25+ T regulatory cells in humans. Hum. Immunol. 66:13–20.
- Allan, S.E., L. Passerini, R. Bacchetta, N. Crellin, M. Dai, P.C. Orban, S.F. Ziegler, M.G. Roncarolo, and M.K. Levings. 2005. The role of 2 FOXP3 isoforms in the generation of human CD4 Tregs. J. Clin. Invest. 115:3276–3284.
- Gavin, M.A., S.R. Clarke, E. Negrou, A. Gallegos, and A. Rudensky. 2002. Homeostasis and anergy of CD4+CD25+ suppressor T cells in vivo. Nat. Immunol. 3:33–41.
- Cozzo, C., J. Larkin III, and A.J. Caton. 2003. Self-peptides drive the peripheral expansion of CD4+CD25+ regulatory T cells. J. Immunol. 171:5678–5682.
- Cupedo, T., M. Nagasawa, K. Weijer, B. Blom, and H. Spits. 2005. Development and activation of regulatory T cells in the human fetus. Eur. J. Immunol. 35:383–390.
Source: PubMed