Human CD25+CD4+ T Suppressor Cell Clones Produce Transforming Growth Factor β, but not Interleukin 10, and Are Distinct from Type 1 T Regulatory Cells

T regulatory (Tr) cells are essential for the induction of peripheral tolerance. Several types of Tr cells exist, including CD4+ T cells which express CD25 constitutively and suppress immune responses via direct cell-to-cell interactions, and type 1 T regulatory (Tr1) cells, which function via secretion of interleukin (IL)-10 and transforming growth factor (TGF)-β. The relationship between CD25+CD4+ T cells and Tr1 cells remains unclear. Here, we demonstrate at the clonal level that Tr1 and CD25+CD4+ T cells are two distinct subsets of regulatory cells with different cytokine production profiles. Furthermore, CD25−CD4+ T cells can be rendered anergic by IL-10 and differentiated into Tr1 cells in the absence of CD25+CD4+ T cells. Cloned human CD25+CD4+ T cell populations are heterogeneous and only a subset of clones continues to express high levels of CD25 and is suppressive. The intensity of CD25, cytotoxic T lymphocyte antigen (CTLA)-4, and glucocorticoid-induced tumor necrosis factor (TNF) receptor expression correlates with the suppressive capacity of the T cell clones. None of the CD25+CD4+ T cell clones with suppressive function produce IL-10, but all produce TGF-β. Suppression mediated by CD25+CD4+ T cell clones is partially dependent on TGF-β, but not on constitutive high expression of CD25. Together these data indicate that naturally occurring human CD25+CD4+ T cells are distinct from IL-10–producing Tr1 cells.

[1]  H. Weiner,et al.  Induction of circulating myelin basic protein and proteolipid protein-specific transforming growth factor-beta1-secreting Th3 T cells by oral administration of myelin in multiple sclerosis patients. , 1996, The Journal of clinical investigation.

[2]  Infectious Tolerance , 2002, The Journal of Experimental Medicine.

[3]  Ana Cumano,et al.  CD25+ CD4+ T Cells Regulate the Expansion of Peripheral CD4 T Cells Through the Production of IL-101 , 2001, The Journal of Immunology.

[4]  M. Roncarolo,et al.  Human Cd25+Cd4+ T Regulatory Cells Suppress Naive and Memory T Cell Proliferation and Can Be Expanded in Vitro without Loss of Function , 2001, The Journal of experimental medicine.

[5]  H. Weiner,et al.  Regulatory T cell clones induced by oral tolerance: suppression of autoimmune encephalomyelitis. , 1994, Science.

[6]  F. Powrie,et al.  Control of intestinal inflammation by regulatory T cells. , 2001, Microbes and infection.

[7]  M. Papiernik Natural CD4+ CD25+ regulatory T cells. Their role in the control of superantigen responses , 2001, Immunological reviews.

[8]  W. Strober,et al.  Cell Contact–Dependent Immunosuppression by Cd4+Cd25+Regulatory T Cells Is Mediated by Cell Surface–Bound Transforming Growth Factor β , 2001, The Journal of experimental medicine.

[9]  David A. Williams,et al.  High-Efficiency Gene Transfer into Normal and Adenosine Deaminase-Deficient T Lymphocytes Is Mediated by Transduction on Recombinant Fibronectin Fragments , 1998, Journal of Virology.

[10]  H. Weiner,et al.  IL‐4 is a differentiation factor for transforming growth factor‐β secreting Th3 cells and oral administration of IL‐4 enhances oral tolerance in experimental allergic encephalomyelitis , 1998, European journal of immunology.

[11]  J. D. de Vries,et al.  Interleukin-10 induces a long-term antigen-specific anergic state in human CD4+ T cells , 1996, The Journal of experimental medicine.

[12]  Hervé Groux,et al.  A CD4+T-cell subset inhibits antigen-specific T-cell responses and prevents colitis , 1997, Nature.

[13]  M. Byrne,et al.  CD4(+)CD25(+) immunoregulatory T cells: gene expression analysis reveals a functional role for the glucocorticoid-induced TNF receptor. , 2002, Immunity.

[14]  R. de Waal Malefyt,et al.  IFN-alpha and IL-10 induce the differentiation of human type 1 T regulatory cells. , 2001, Journal of immunology.

[15]  G. Schuler,et al.  Human CD4+CD25+ Regulatory, Contact-dependent T Cells Induce Interleukin 10–producing, Contact-independent Type 1-like Regulatory T Cells , 2002, The Journal of experimental medicine.

[16]  Fiona Powrie,et al.  An Essential Role for Interleukin 10 in the Function of Regulatory T Cells That Inhibit Intestinal Inflammation , 1999, The Journal of experimental medicine.

[17]  A. Enk,et al.  Identification and Functional Characterization of Human Cd4+Cd25+ T Cells with Regulatory Properties Isolated from Peripheral Blood , 2001, The Journal of experimental medicine.

[18]  T. Mak,et al.  Immunologic Self-Tolerance Maintained by Cd25+Cd4+Regulatory T Cells Constitutively Expressing Cytotoxic T Lymphocyte–Associated Antigen 4 , 2000, The Journal of experimental medicine.

[19]  Fiona Powrie,et al.  Cytotoxic T Lymphocyte–Associated Antigen 4 Plays an Essential Role in the Function of Cd25+Cd4+ Regulatory Cells That Control Intestinal Inflammation , 2000, The Journal of experimental medicine.

[20]  Ethan M. Shevach,et al.  CD4+CD25+ Immunoregulatory T Cells Suppress Polyclonal T Cell Activation In Vitro by Inhibiting Interleukin 2 Production , 1998, The Journal of experimental medicine.

[21]  R. Lechler,et al.  Human CD4(+)CD25(+) cells: a naturally occurring population of regulatory T cells. , 2001, Blood.

[22]  D. Zélénika,et al.  The role of CD4+ T‐cell subsets in determining transplantation rejection or tolerance , 2001, Immunological reviews.

[23]  C. Bordignon,et al.  Peripheral blood lymphocytes as target cells of retroviral vector-mediated gene transfer. , 1994, Blood.

[24]  L. Cosmi,et al.  Phenotype, Localization, and Mechanism of Suppression of CD4+CD25+ Human Thymocytes , 2002, The Journal of experimental medicine.

[25]  M. Toda,et al.  Immunologic self-tolerance maintained by CD25+CD4+ naturally anergic and suppressive T cells: induction of autoimmune disease by breaking their anergic/suppressive state. , 1998, International immunology.

[26]  R. de Waal Malefyt,et al.  IFN-α and IL-10 Induce the Differentiation of Human Type 1 T Regulatory Cells1 , 2001, The Journal of Immunology.

[27]  Ethan M. Shevach,et al.  CD4+CD25+ Regulatory T Cells Can Mediate Suppressor Function in the Absence of Transforming Growth Factor β1 Production and Responsiveness , 2002, The Journal of experimental medicine.

[28]  M. Roncarolo,et al.  Growth and expansion of human T regulatory type 1 cells are independent from TCR activation but require exogenous cytokines , 2002, European journal of immunology.

[29]  M. Salmon,et al.  Human anergic/suppressive CD4+CD25+ T cells: a highly differentiated and apoptosis‐prone population , 2001, European journal of immunology.

[30]  Ethan M. Shevach,et al.  CD4+CD25+ suppressor T cells: more questions than answers , 2002, Nature Reviews Immunology.

[31]  R. Lechler,et al.  Anergic T cells act as suppressor cells in vitro and in vivo , 1999, European journal of immunology.

[32]  G. Schuler,et al.  Ex Vivo Isolation and Characterization of Cd4+Cd25+ T Cells with Regulatory Properties from Human Blood , 2001, The Journal of experimental medicine.

[33]  M. Roncarolo,et al.  Type 1 T regulatory cells , 2001, Immunological reviews.

[34]  D. Mason,et al.  Human CD4+CD25+ thymocytes and peripheral T cells have immune suppressive activity in vitro , 2001, European journal of immunology.

[35]  Ethan M. Shevach,et al.  Suppressor Effector Function of CD4+CD25+ Immunoregulatory T Cells Is Antigen Nonspecific , 2000, The Journal of Immunology.

[36]  H. Weiner,et al.  Induction and mechanism of action of transforming growth factor‐β‐secreting Th3 regulatory cells , 2001, Immunological reviews.

[37]  Sayuri Yamazaki,et al.  Immunologic tolerance maintained by CD25+ CD4+ regulatory T cells: their common role in controlling autoimmunity, tumor immunity, and transplantation tolerance , 2001, Immunological reviews.

[38]  G. Freeman,et al.  CD4+CD25high Regulatory Cells in Human Peripheral Blood1 , 2001, The Journal of Immunology.

[39]  J. Gorski,et al.  Molecular analysis of T cell repertoires. Spectratypes generated by multiplex polymerase chain reaction and evaluated by radioactivity or fluorescence. , 1995, Human immunology.

[40]  J. Shimizu,et al.  Stimulation of CD25+CD4+ regulatory T cells through GITR breaks immunological self-tolerance , 2002, Nature Immunology.