Humoral immune response to 2,4-dinitrophenyl--keyhole limpet hemocyanin in antigen-free, germ-free and conventional BALB/c mice.
The B cell immune response to 2,4-dinitrophenyl (DNP) keyhole limpet hemocyanin was compared in antigen-free, germ-free and conventional BALB/c mice. The numbers of total and of DNP-specific IgM-, IgG- and IgA-secreting cells in the spleen were determined by enzyme-linked immunosorbent plaque assays after primary, secondary and hyperimmunization. Three days after primary immunization a peak of DNP-specific IgM-secreting cells was seen in conventional mice only. However, this specific response was accompanied by a rise in the total number of IgM-secreting cells. At day 6 after primary immunization the total number and the frequency of DNP-specific IgG-secreting cells were higher in antigen-free mice, compared to germ-free and conventional mice. After secondary immunization in conventional mice only, a considerable bystander IgG response was seen together with the DNP-specific IgG response. At the end of the secondary response 90% of all IgG-secreting cells were DNP specific in antigen-free mice, while the corresponding figure in germ-free and conventional mice was 63% and 14%, respectively. After hyperimmunization, the absolute number of DNP-specific IgG-secreting cells in the spleen was 5-fold and 11-fold higher in antigen-free mice than in germ-free and conventional mice, respectively. Approximately 48% of all IgG-secreting cells were DNP specific in antigen-free mice, while the corresponding figure in germ-free and conventional mice was 17% and 12%, respectively. The results show that the exogenous antigenic load of animals influences the immune response to newly introduced antigens. The higher absolute and relative numbers of antigen-specific IgG-secreting cells after hyperimmunization in antigen-free mice compared to germ-free and conventional mice may provide a better source for antigen-specific B cells that eventually can be used for hybridoma production.
Department of Histology and Cell Biology, University of Groningen, The Netherlands.
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D000821:Animal Feed; D000818:Animals; D000917:Antibody Formation; D000941:Antigens / Q000008:administration & dosage / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005856:Germ-Free Life / Q000276:immunology*; D006433:Hemocyanins / Q000276:immunology*; D007070:Immunoglobulin A / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D013154:Spleen / Q000166:cytology
C042158:2,4-dinitrophenyl keyhole limpet hemocyanin; D000941:Antigens; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D006433:Hemocyanins