Detection of autologous antiidiotypic antibody-forming cells by a modified enzyme-linked immunospot (ELISPOT).
We describe here the utilization of a modified enzyme-linked immunospot assay (ELISPOT) in order to detect an autologous antiidiotypic response in mice at the level of single antibody-forming cells (AFC). Severals assays have been routinely used to detect anti-Id producing cells; however, these approaches often produce contrasting data. We present results obtained with the modified ELISPOT, using as a model system the antiidiotypic response in mice after immunization with a vaccine from Streptococcus pneumoniae R36a, expressing the immunodominant epitope phosphorylcholine (PC). The response to PC is mediated by a large fraction of antibodies bearing the public idiotype T15. Mice of different genetics make up were immunized with a single injection of the vaccine. We observed that one mouse strain (D1.LP) out of three was able to mount a significant anti-T15 response during the primary anti-phosphorylcholine response. BALB/c and C57BL/6 mice did not produce significant levels of anti-T15 antibody following a single injection of the antigen. In contrast, BALB/c mice which were repeatedly stimulated showed a specific anti-Id response. Experimental controls were performed using either specific anti-T15 monoclonal antibodies (mAb) or splenocytes from mice immunized with TEPC15 myeloma protein in complete Freund's adjuvant.
Nicoletti
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Institute of Human Anatomy, University of Siena, Italy.
Borghesi-Nicoletti
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D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D000921:Antibody-Producing Cells / Q000276:immunology*; D000942:Antigens, Bacterial / Q000276:immunology*; D001428:Bacterial Vaccines; D016056:Immunodominant Epitopes / Q000276:immunology; D007124:Immunoenzyme Techniques; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008810:Mice, Inbred C57BL / Q000276:immunology; D010767:Phosphorylcholine / Q000276:immunology; D013296:Streptococcus pneumoniae / Q000276:immunology; D014611:Vaccination
D000888:Antibodies, Anti-Idiotypic; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D016056:Immunodominant Epitopes; D007130:Immunoglobulin Idiotypes; D010767:Phosphorylcholine