Isolation and characterization of human antigen-specific TCRαβ+ CD4-CD8- double-negative regulatory T cells

Down-regulation of immune responses by regulatory T (Treg) cells is an important mechanism involved in the induction of tolerance to allo-antigens (Ags). Recently, a novel subset of Ag-specific T-cell receptor (TCR)alpha beta+ CD4(-)CD8- (double-negative [DN]) Treg cells has been found to be able to prevent the rejection of skin and heart allografts by specifically inhibiting the function of antigraft-specific CD8+ T cells. Here we demonstrate that peripheral DN Treg cells are present in humans, where they constitute about 1% of total CD3+ T cells, and consist of both naive and Ag-experienced cells. Similar to murine DN Treg cells, human DN Treg cells are able to acquire peptide-HLA-A2 complexes from antigen-presenting cells by cell contact-dependent mechanisms. Furthermore, such acquired peptide-HLA complexes appear to be functionally active, in that CD8+ T cells specific for the HLA-A2-restricted self-peptide, Melan-A, became sensitive to apoptosis by neighboring DN T cells after acquisition of Melan-A-HLA-A2 complexes and revealed a reduced proliferative response. These results demonstrate for the first time that a sizable population of peripheral DN Treg cells, which are able to suppress Ag-specific T cells, exists in humans. DN Treg cells may serve to limit clonal expansion of allo-Ag-specific T cells after transplantation.

[1]  Mark J. Smyth,et al.  Functional significance of the perforin/granzyme cell death pathway , 2002, Nature Reviews Immunology.

[2]  S. Distler,et al.  Characterization and function of CD3+ CD4- CD8- TcR-alpha beta bearing cells infiltrating the lung during the immune response. , 1992, Regional immunology.

[3]  M. Mannie,et al.  Antigen presentation by T cells: T cell receptor ligation promotes antigen acquisition from professional antigen‐presenting cells , 1997, European journal of immunology.

[4]  Andreas Radbruch,et al.  Two Subsets of Naive T Helper Cells with Distinct T Cell Receptor Excision Circle Content in Human Adult Peripheral Blood , 2002, The Journal of experimental medicine.

[5]  R. Andreesen,et al.  Large-scale in vitro expansion of polyclonal human CD4+CD25high regulatory T cells , 2004 .

[6]  A. Hayday.  [gamma][delta] cells: a right time and a right place for a conserved third way of protection. , 2000, Annual review of immunology.

[7]  T. Strom,et al.  Tracking the Immunoregulatory Mechanisms Active During Allograft Tolerance1 , 2002, The Journal of Immunology.

[8]  Li Zhang,et al.  Identification of a previously unknown antigen-specific regulatory T cell and its mechanism of suppression , 2000, Nature Medicine.

[9]  F. Rieux-Laucat,et al.  Autoimmune lymphoproliferative syndrome with somatic Fas mutations. , 2004, The New England journal of medicine.

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

[11]  M. Roncarolo,et al.  The role of different subsets of T regulatory cells in controlling autoimmunity. , 2000, Current opinion in immunology.

[12]  D. Richman,et al.  Memory CD8+ T cells vary in differentiation phenotype in different persistent virus infections , 2002, Nature Medicine.

[13]  S. Quaratino,et al.  Definition of unique traits of human CD4− CD8−αβ T cells , 1993 .

[14]  M. Mannie,et al.  Class II MHC/peptide complexes are released from APC and are acquired by T cell responders during specific antigen recognition. , 1999, Journal of immunology.

[15]  C. Fathman,et al.  CD4+CD25+ regulatory T cells preserve graft-versus-tumor activity while inhibiting graft-versus-host disease after bone marrow transplantation , 2003, Nature Medicine.

[16]  A. Mackensen,et al.  Survival and Tumor Localization of Adoptively Transferred Melan-A-Specific T Cells in Melanoma Patients 1 , 2003, The Journal of Immunology.

[17]  Li Zhang,et al.  Inhibition of Graft-Versus-Host Disease by Double-Negative Regulatory T Cells1 , 2003, The Journal of Immunology.

[18]  E. Jaffe,et al.  A novel lymphoproliferative/autoimmune syndrome resembling murine lpr/gld disease. , 1992, The Journal of clinical investigation.

[19]  J. Bluestone,et al.  Suppressor T cells – they’re back and critical for regulation of autoimmunity! , 2001, Immunological reviews.

[20]  D. Speiser,et al.  Ex Vivo IFN-γ Secretion by Circulating CD8 T Lymphocytes: Implications of a Novel Approach for T Cell Monitoring in Infectious and Malignant Diseases1 , 2001, The Journal of Immunology.

[21]  Warren Strober,et al.  Dominant interfering fas gene mutations impair apoptosis in a human autoimmune lymphoproliferative syndrome , 1995, Cell.

[22]  R. Lechler,et al.  Anergic T cells as suppressor cells in vitro. , 1994, Science.

[23]  A. Naji,et al.  Thymic selection of CD4+CD25+ regulatory T cells induced by an agonist self-peptide , 2001, Nature Immunology.

[24]  Li Zhang,et al.  Donor-lymphocyte infusion induces transplantation tolerance by activating systemic and graft-infiltrating double-negative regulatory T cells. , 2002, Blood.

[25]  G. Salles,et al.  Human γδ T Cells Express a Higher TCR/CD3 Complex Density Than αβ T Cells , 2001 .

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

[27]  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.

[28]  G. Carcelain,et al.  Direct evidence to support the immunosurveillance concept in a human regressive melanoma. , 1994, The Journal of clinical investigation.

[29]  M. Takiguchi,et al.  Differentiation of Human CD8+ T Cells from a Memory to Memory/Effector Phenotype1 , 2002, The Journal of Immunology.

[30]  E. Brooks,et al.  Novel T-lymphocyte population in combined immunodeficiency with features of graft-versus-host disease. , 1989, The New England journal of medicine.

[31]  G. Tsokos,et al.  T cell receptor alpha/beta expressing double-negative (CD4-/CD8-) and CD4+ T helper cells in humans augment the production of pathogenic anti-DNA autoantibodies associated with lupus nephritis. , 1989, Journal of immunology.

[32]  M. Jackson,et al.  TCR-Mediated internalization of peptide-MHC complexes acquired by T cells. , 1999, Science.

[33]  B. Seliger,et al.  High Frequency of Functionally Active Melan-A–Specific T Cells in a Patient with Progressive Immunoproteasome-Deficient Melanoma , 2004, Cancer Research.

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

[35]  J. Allison,et al.  Cloned natural suppressor cell lines express the CD3+CD4-CD8- surface phenotype and the alpha, beta heterodimer of the T cell antigen receptor. , 1989, Journal of immunology.

[36]  B. Diamond,et al.  Human Double-Negative T Cells in Systemic Lupus Erythematosus Provide Help for IgG and Are Restricted by CD1c1 , 2000, The Journal of Immunology.

[37]  K. Young,et al.  Infusion of in vitro-generated DN T regulatory cells induces permanent cardiac allograft survival in mice. , 2003, Transplantation proceedings.

[38]  D. Voehringer,et al.  Frequent expression of the natural killer cell receptor KLRG1 in human cord blood T cells: correlation with replicative history , 2004, European journal of immunology.

[39]  J. Reimann,et al.  Double‐Negative (CD4‐CD8‐), TCRαβ‐Expressing, Peripheral T Cells , 1991, Scandinavian journal of immunology.

[40]  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.

[41]  R. Lechler,et al.  Antigen presentation by mouse CD4+ T cells involving acquired MHC class II:peptide complexes: another mechanism to limit clonal expansion? , 2003, Blood.

[42]  F. Sallusto,et al.  Two subsets of memory T lymphocytes with distinct homing potentials and effector functions , 1999, Nature.

[43]  C. Benoist,et al.  Damage control, rather than unresponsiveness, effected by protective DX5+ T cells in autoimmune diabetes , 2001, Nature Immunology.

[44]  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.

[45]  E. Shevach.  Regulatory T cells in autoimmmunity*. , 2000, Annual review of immunology.

[46]  L. Chess,et al.  The specific regulation of immune responses by CD8+ T cells restricted by the MHC class Ib molecule, Qa-1. , 2000, Annual review of immunology.

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

[48]  J. Priatel,et al.  TCR/Self-Antigen Interactions Drive Double-Negative T Cell Peripheral Expansion and Differentiation into Suppressor Cells1 , 2001, The Journal of Immunology.

[49]  M. Toda,et al.  Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. , 1995, Journal of immunology.

[50]  Li Zhang,et al.  The nature and mechanisms of DN regulatory T-cell mediated suppression. , 2002, Human immunology.

[51]  M. Akiyama,et al.  Evidence for in vivo clonal proliferation of unique population of blood CD4-/CD8- T cells bearing T-cell receptor alpha and beta chains in two normal men. , 1992, Blood.

[52]  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.