Adoptive Transfer of Diabetes Into Immunodeficient NOD-scid/scid Mice: Relative Contributions of CD4+ and CD8+ T-Cells From Diabetic Versus Prediabetic NOD.NON-Thy-1a Donors

Precise definition of the role of both CD4 and CD8 T-cell subsets from NOD mice in the adoptive transfer of diabetes has been complicated by the possibility that endogenous T-cells may be recruited. Two newly created NOD congenic stocks, NOD.NON-Thy-1aand NOD/LtSz-scid, have been used as T-cell donors and recipients, respectively, to eliminate contributions from endogenous T-cells and thus to define the requirement for transferred T-cell subsets as a function of underlying diabetes development in the NOD donor. Total T-cells and T-cell subsets prepared from either prediabetic or diabetic NOD.NON-Thy-1a donors were adoptively transferred into 6-wk-old NOD-scid/scid recipients that were monitored for diabetes development. Both flow cytometric and histological analysis of recipient spleen and pancreas after adoptive transfer showed lymphocytes of donor (Thy1.1+) origin exclusively. Total T-cell and enriched CD4+ T-cell preparations from both diabetic and young prediabetic donors transferred diabetes to NOD-scid/scid recipients. However, the mean time to diabetes onset was doubled when CD4+ lymphocytes were isolated from prediabetic versus diabetic donors, and t,+ cells over time. Enriched CD8+ populations alone were unable to transfer disease. More rigorous exclusion of CD8+ cells by means of anti-CD8 MoAb treatment in vivo of the recipients of enriched CD4+ cells demonstrated a significant difference in the diabetogenic potency of CD4+ lymphocytes from diabetic versus nondiabetic donors. Diabetes was adoptively transferred to 58% of the recipients of enriched CD4+ lymphocytes from diabetic donors. In contrast, none of the recipients of enriched CD4+ lymphocytes from young prediabetic donors developed diabetes after MoAb treatment in vivo. The ability of a T-cell population to produce severe insulitis and sialitis in NOD-scid/scid recipients of T-cells closely paralleled its ability to induce diabetes. In an effort to suppress insulitis by suppression of macrophage migration to the islets, NOD-scid/scid mice were treated with silica in conjunction with adoptive transfer of T-cells from diabetic donors. Chronic silica treatment failed to deplete tissue macrophages and did not prevent diabetes development after transfer of unfractionated T-cells. Evidence is discussed indicating that the age-associated differences in ability of CD4+ T-cells to adoptively transfer diabetes in the absence of the CD8+ T-cells subset is a function of prior, chronic exposure of the CD4+ lymphocytes to β-cell antigens in the donor. This study confirms that both CD4+ and CD8+ T-cells are required to initiate β-cell destruction in NOD mice.

[1]  W. Winter,et al.  Flow cytometric enumeration of mononuclear cell populations infiltrating the islets of Langerhans in prediabetic NOD mice: development of a model of autoimmune insulitis for type I diabetes. , 1990, Regional immunology.

[2]  A. Cooke,et al.  The transfer of autoimmune diabetes in NOD mice can be inhibited or accelerated by distinct cell populations present in normal splenocytes taken from young males. , 1990, Journal of autoimmunity.

[3]  T. Rothstein,et al.  Mouse lymphocytes with and without surface immunoglobulin: preparative scale separation in polystyrene tissue culture dishes coated with specifically purified anti-immunoglobulin. , 1977, Journal of immunological methods.

[4]  W. Ogawa,et al.  Morphological Analysis of Selective Destruction of Pancreatic β-cells by Cytotoxic T Lymphocytes in NOD Mice , 1991, Diabetes.

[5]  C. Janeway,et al.  Monoclonal antibodies to murine CD3 epsilon define distinct epitopes, one of which may interact with CD4 during T cell activation. , 1989, Journal of immunology.

[6]  C. Milstein,et al.  Rat × rat hybrid myelomas and a monoclonal anti-Fd portion of mouse IgG , 1979, Nature.

[7]  K. Amano,et al.  Evidence for Initial Involvement of Macrophage in Development of Insulitis in NOD Mice , 1988, Diabetes.

[8]  R. Custer,et al.  Severe combined immunodeficiency (SCID) in the mouse. Pathology, reconstitution, neoplasms. , 1985, The American journal of pathology.

[9]  K. Lafferty,et al.  Pancreatic islet-specific T-cell clones from nonobese diabetic mice. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[10]  B. Charlton,et al.  Administration of Silica Particles or Anti-Lyt2 Antibody Prevents β-Cell Destruction in NOD Mice Given Cyclophosphamide , 1988, Diabetes.

[11]  P. Bedossa,et al.  CD8+ T cell homing to the pancreas in the nonobese diabetic mouse is CD4+ T cell-dependent. , 1991, Journal of immunology.

[12]  Y. Tochino,et al.  Breeding of a non-obese, diabetic strain of mice. , 1980, Jikken dobutsu. Experimental animals.

[13]  Soumitra Ghosh,et al.  Genetic analysis of autoimmune type 1 diabetes mellitus in mice , 1991, Nature.

[14]  K. Lafferty,et al.  The role of CD4+ and CD8+ T cells in the destruction of islet grafts by spontaneously diabetic mice. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[15]  S. Gordon,et al.  Transfer of diabetes in mice prevented by blockade of adhesion-promoting receptor on macrophages , 1990, Nature.

[16]  M. Gefter,et al.  Properties and applications of monoclonal antibodies directed against determinants of the Thy-1 locus. , 1979, Journal of immunology.

[17]  T. Hanafusa,et al.  Predominance of T lymphocytes in pancreatic islets and spleen of pre-diabetic non-obese diabetic (NOD) mice: a longitudinal study. , 1985, Clinical and experimental immunology.

[18]  H. Waldmann,et al.  Therapy with monoclonal antibodies by elimination of T-cell subsets in vivo , 1984, Nature.

[19]  C. Boitard,et al.  Syngeneic transfer of autoimmune diabetes from diabetic NOD mice to healthy neonates. Requirement for both L3T4+ and Lyt-2+ T cells , 1987, The Journal of experimental medicine.

[20]  K. Haskins,et al.  Acceleration of diabetes in young NOD mice with a CD4+ islet-specific T cell clone. , 1990, Science.

[21]  P. Bedossa,et al.  Syngeneic t cell transfer of diabetes into nod newborn mice: in situ studies of the autoimmune steps leading to insulin‐producing cell destruction , 1989, European journal of immunology.

[22]  K. Hamaguchi,et al.  Immunostimulation circumvents diabetes in NOD/Lt mice. , 1989, Journal of autoimmunity.

[23]  E. Leiter,et al.  Defective activation of T suppressor cell function in nonobese diabetic mice. Potential relation to cytokine deficiencies. , 1988, Journal of immunology.

[24]  O. Pankewycz,et al.  Islet‐infiltrating T cell clones from non‐obese diabetic mice that promote or prevent accelerated onset diabetes , 1991, European journal of immunology.

[25]  E. Leiter,et al.  Molecular Mimicry Between Insulin and Retroviral Antigen p73: Development of Cross-Reactive Autoantibodies in Sera of NOD and C57BL/KsJ db/db Mice , 1988, Diabetes.

[26]  A. Monaco,et al.  An improved method for isolation of mouse pancreatic islets. , 1985, Transplantation.

[27]  E. Leiter,et al.  Genetic Control of Diabetogenesis in NOD/Lt Mice: Development and Analysis of Congenic Stocks , 1989, Diabetes.

[28]  L. Wicker,et al.  Both the Lyt-2+ and L3T4+ T cell subsets are required for the transfer of diabetes in nonobese diabetic mice. , 1988, Journal of immunology.

[29]  P. Bedossa,et al.  Adoptive T cell transfer of autoimmune nonobese diabetic mouse diabetes does not require recruitment of host B lymphocytes. , 1988, Journal of immunology.

[30]  J. Reimann,et al.  CD3+ T cells in severe combined immune deficiency (scid) mice. I. Transferred purified CD4+ T cells, but not CD8+ T cells are engrafted in the spleen of congenic scid mice , 1991, European journal of immunology.

[31]  S. Ihm,et al.  Studies on Autoimmunity for Initiation of β-Cell Destruction: VI. Macrophages Essential for Development of β-Cell–Specific Cytotoxic Effectors and Insulitis in NOD Mice , 1990, Diabetes.