Heterogeneity of monoclonal antibodies against pancreatic beta cells.
Sixteen murine hybridoma-secreting monoclonal antibodies against pancreatic islet cell surface antigens (mc-ICSA) have been produced by the cell fusion technique using splenocytes from xenogeneic islet cell- or RIN cell-immunized Balb/c mice. In addition, some mice were autoimmunized by subdiabetogenic doses of the beta cell toxin streptozotocin in combination with complete Freund's adjuvant. Isotyping of the mc-ICSA revealed that 13 of the antibodies belong to the class IgM, and 3 to the subclass IgG1. The specificity of these mc-ICSA has been detected by means of the indirect immunofluorescent technique using primary rat islet cells suspensions, following a procedure of double immunostaining for pancreatic insulin and glucagon. Furthermore, the cross reactivity of these mc-ICSA was studied using endothelial, neuroblastoma and fibroblast cell lines and primary rat splenocytes. One out of the 10 mc-ICSA tested was alpha cell-specific, 2 were beta cell-specific and 7 out of 10 were reactive with both alpha and beta cells. Eleven mc-ICSA showed no cross-reactivity with the 5 other cell types tested. The binding of one mc-ICSA was blocked by 6 of the ICSA-positive human sera which were tested, suggesting that this monoclonal recognizes the same antigenic determinant. The same mc-ICSA diminished the glucose- and arginine-stimulated insulin secretion of isolated rat islets.
Ziegler
B
B
Central Institute of Diabetes Gerhardt Katsch, Karlsburg/GDR.
Witt
S
S
Waterstradt
B
B
Schlosser
M
M
Ziegler
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000916:Antibody Diversity; D001323:Autoantibodies / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D003429:Cross Reactions; D004727:Endothelium / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007340:Insulinoma / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D007515:Islets of Langerhans / Q000166:cytology / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009447:Neuroblastoma / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D051381:Rats; D011919:Rats, Inbred Strains; D013154:Spleen / Q000166:cytology / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; C024211:islet cell antibody