Anti-TMA immunity in mice. I. the appearance of cross-reactive idiotype (s) to the trimethylammonium (TMA) hapten in A/J mice
Alevy, Y.G.; Witherspoon, C.D.; Prange, C.A.; Bellone, C.J.
Journal of Immunology 124(1): 215-221
1980
ISSN/ISBN: 0022-1767 PMID: 6765967 Document Number: 165689
Antibody to the bivalent antigen L-tyrosine-p-azophenyltrimethylammonium (tyr(TMA)), (H-L-tyr(TMA)-NH-(CH2)3)2 (T-S-T) was generated in the A/J mouse strain. Isolated anti-TMA antibody from a single mouse was inoculated into rabbits to produce idiotypic antisera (anti-Id). After adsorption with normal A/J Ig, the antisera was judged idiotype-specific by 2 different assays. In a radioimmunoassay 10-50 ng of cold homologous anti-TMA antibody significantly inhibited the binding between the anti-Id serum and the I125-labeled proband, whereas as much as 20 .mu.g of normal mouse Ig or mouse myeloma protein M104E did not significantly affect this binding. The anti-Id serum inhibited from 48-100% anti-TMA plaque-forming cells (PFC) but there was no significant inhibition of antiazobenzenearsonate (ABA) or sheep red blood cells (SRBC) PFC. In A/J mice the humoral response directed to the TMA determinant conjugated to different protein carriers was examined using the anti-Id serum and was characterized by extensive cross-reactive idiotypes (CRI-TMA). The idiotypic antiserum blocked from 58-82% of the total anti-TMA PFC population depending on the carrier used. Further proof of a restricted response came from hapten inhibition profiles in the PFC assay and also from isoelectric focus (IEF) patterns. Although in individual mice different molar concentrations of free hapten were needed to inhibit 50% of the anti-TMA PFC, a markedly steep inhibition profile was seen for each mouse tested. IEF patterns supported a relatively restricted response, since focused antisera showed distinct bands primarily in the acidic portion of the gel. Many bands appeared similar from mouse to mouse but there were distinct differences among the individuals. The hapten protein conjugate 3-(p-trimethylphenylazo)-N-acetyl-L-tyrosylglycylglycine-human .gamma.-globulin (TMA-S-HGG), which elicited an average of 82% CRI+-TMA PFC in A/J mice, was used to survey several mouse strains that were different in their H-2 haplotypes and allotypes of the Ig-1 locus for the expression of CRI+-TMA PFC. Only strains of the Ig-1e allotype expressed appreciable amounts of the CRI-TMA; Expression appeared independent of the H-2 haplotype. Expression of the CRI-TMA appeared dominant, since in the F1 hybrid CAF1 (responder A/J .times. nonresponder BALB/c) strain 55% of the PFC secreted the CRI-TMA.