Autologous CD4 T-cell responses to ectopic class II major histocompatibility complex antigen-expressing single-cell islet cells: an in vitro insight into the pathogenesis of lymphocytic insulitis in nonobese diabetic mice.
We investigated by flow cytometric analysis the expression of class II major histocompatibility complex (MHC) molecules by viable single-cell islet cells (SCICs) prepared from male and female 4- and 10-week-old nonobese diabetic (NOD) mouse islets. With anti-I-Ak monoclonal antibody (specific for I-Ak,f,r,s beta and produced by clone 11-5-2), and fluorescein isothiocyanate-conjugated goat anti-mouse IgG as second-step antibody, we found that SCICs from both sexes aberrantly expressed class II MHC molecules, which was not altered after SCICs were cultured for 24 hr or 120 hr in the presence of 10 ng of recombinant murine interferon gamma per ml. Double-indirect immunofluorescence of male SCICs indicated that the expression of class II MHC molecules was a property of beta cells. Control experiments documented that macrophages and mononuclear cells did not contaminate the SCIC preparations. Coculture experiments with responder splenic CD4 T cells isolated from diabetic NOD mice and stimulator male SCICs indicated a recognition event evidenced by a 12-fold increase in proliferative response. Monoclonal antibodies to class II MHC and CD4 antigens blocked the proliferative response. Results from control autologous and allogeneic mixed lymphocyte reactions suggest that the responder CD4 T cells are autoreactive self-class II MHC restricted. We tentatively conclude that the ability of SCICs from both sexes of NOD mice to express class II MHC molecules as early as 4 weeks of age may represent a mechanism for targeting immune reactions to beta cells and initiate lymphocytic insulitis.
Formby
B
B
Sansum Medical Research Foundation, Laboratory of Immunology, Santa Barbara, CA 93105.
Miller
N
N
D000818:Animals; D000911:Antibodies, Monoclonal; D000938:Antigen-Presenting Cells / Q000276:immunology*; D015704:CD4 Antigens / Q000276:immunology*; D003921:Diabetes Mellitus, Experimental / Q000235:genetics / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000276:immunology*; D007515:Islets of Langerhans / Q000276:immunology*; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D013601:T-Lymphocytes / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D000949:Histocompatibility Antigens Class II