Effects of carrier and hapten array on the production of anti-hapten antibodies analyzed by two-dimensional affinity electrophoresis.
Effects of a carrier, bovine serum albumin (BSA) and a hapten array, fluorescein isothiocyanate-conjugated dextran (FITC-DEX), on the production of anti-FITC antibodies in BALB/c mice were analyzed by two-dimensional affinity electrophoresis (2-D AEP). The mice were immunized with FITC-BSA, followed by immunizations with FITC-BSA, saline, BSA, human serum albumin (HSA), fluorescein (Flu), and FITC-DEX. The heterogeneity and quantity of anti-FITC antibodies were increased markedly during the second and third immunization with FITC-BSA. However, the production 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 competition through antigen processing and the clonal expansion of BSA-specific B cells because similar suppression was induced by the immunization with HSA. The hapten array of FITC-DEX generally suppressed the production of anti-FITC antibodies; however, immunization with free hapten of Flu did not affect the production significantly. This may 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 suppression.
The First Department of Biochemistry, Yamaguchi University School of Medicine, Ube, Japan.
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Wang
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Nakamura
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D005260:Female; D019793:Fluorescein; D016650:Fluorescein-5-isothiocyanate; D006241:Haptens / Q000276:immunology*; D006801:Humans; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D012709:Serum Albumin / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000276:immunology*
D000906:Antibodies; D018985:Epitopes, B-Lymphocyte; D006241:Haptens; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D016650:Fluorescein-5-isothiocyanate; D019793:Fluorescein