EFFECTS OF CARRIER AND HAPTEN ARRAY ON THE PRODUCTION OF ANTIHAPTEN ANTIBODIES ANALYZED BY 2-DIMENSIONAL AFFINITY ELECTROPHORESIS

Effects of a carrier, bovine serum albumin (BSA) and a hapten array, f luorescein isothiocyanate-conjugated dextran (FITC-DEF;), on the produ ction of anti-FITC antibodies in BALB/c mice were analyzed by two-dime nsional affinity electrophoresis (2-D AEP). The mice were immunized wi th FITC-BSA, followed by immunizations with FITC-BSA, saline, BSA, hum an serum albumin (HSA), fluorescein (Flu), and FITC-DEX. The heterogen eity and quantity of anti-FITC antibodies were increased markedly duri ng the second and third immunization with FITC-BSA. However, the produ ction of anti-FITC antibodies with low affinity to FITC was suppressed by the second and third immunization with the carrier protein of BSA. The suppression of anti-FITC antibodies by immunization with BSA may be due to the major histocompatibility complex (MHC)-mediated competit ion through antigen processing and the clonal expansion of BSA-specifi c B cells because similar suppression was induced by the immunization with PISA. The hapten array of FITC-DEX generally suppressed the produ ction of anti-FITC antibodies; however, immunization with free hapten of Flu did not affect the production significantly. This map indicate that cross-linking of anti-FITC antibodies on the surface of specific B cells via the binding of hapten array is required for the suppressio n.