Production of anti-rat islet cell surface antibody in guinea pigs.
Anti-rat islet serum was prepared in guinea pigs by multiple subcutaneous inoculations of rat islets homogenates emulsified in complete Freund's adjuvant (CFA). The anti-rat islet serum was cytotoxic against rat spleen cells in the presence of complement and the nonspecific antibodies were observed with homogenates of rat livers and spleens. After absorption, the serum lost the cytotoxicity against the rat spleen cells yet showed specific cytotoxicity against the rat islet cells. The binding capacity of anti-rat islet antibody was determined by the indirect immunofluorescence test using FITC conjugated rabbit anti-guinea pig IgG serum. As the guinea pig anti-rat islet serum contained anti-insulin antibody, the role of this antibody in this cytotoxic activity and surface immunofluorescence was studied. However, the anti-insulin antibody used as the control showed neither cytotoxicity nor surface immunofluorescence. After neutralizing the anti-insulin antibody in the antiserum with insulin, the serum remained cytotoxic to the rat islet cells and a surface immunofluorescence appeared. These data show that specific anti-rat islet cell surface antibody can be produced in guinea pigs by multiple inoculations of rat islets homogenates with CFA.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001323:Autoantibodies; D001681:Biological Assay; D003602:Cytotoxicity, Immunologic; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D008722:Methods; D051381:Rats; D011919:Rats, Inbred Strains; D013154:Spleen
D000906:Antibodies; D001323:Autoantibodies; C024211:islet cell antibody