PMID- 2452197 OWN - NLM STAT- MEDLINE DCOM- 19880603 LR - 20061115 IS - 0022-1767 (Print) IS - 0022-1767 (Linking) VI - 140 IP - 9 DP - 1988 May 1 TI - Angiotensin II (AII)-related idiotypic network. II. Heterogeneity and fine specificity of AII internal images analyzed with monoclonal antibodies. PG - 3059-65 AB - Although the structural basis of internal images borne by beta type monoclonal anti-idiotypic antibody (Ab2) begins to be elucidated, there is little information on the repertoire of epitopes which make up the internal images expressed by polyclonal Ab2. We addressed this question by using a two-way approach in the angiotensin II (AII)-related idiotypic network, a system characterized by common occurrence of internal images on rabbit Ab2. First, two sets of internal images were purified in parallel by affinity chromatography on Sepharose 4B covalently linked to either mAb 110 (S4B-110), a mAb specific for a phenylalanine requiring carboxy terminus epitope (Phe8) on AII, or mAb 133 (S4B-133), reactive with a more central epitope also expressed on Phe8 substituted peptide analogs. The respective eluates, EL1 110 and E11 133, exhibited only partially overlapping reactivity, as demonstrated by 1) a different pattern of inhibition by various AII peptide analogues of EL1 110 and E11 133 binding to the same anti-AII antibody (Ab1) (either the homologous polyclonal Ab1 102 or mAb 133), 2) and a distinct profile of EL1 110 and EL1 133 binding to 12 biotinylated monoclonal Ab1 identifying a variety of epitopes on AII. To analyze further the respective distribution of mAb 110 and mAb 133 defined epitopes on Ab2-beta molecules, Ab2 were submitted to sequential affinity chromatography on S4B-110 followed by S4B-133, and the fractionated internal images were characterized by the pattern of binding to the various monoclonal Ab1. It was thus possible to purify two Ab2-beta subpopulations that exclusively imaged the determinant identified by mAb 110 (ii 110) or that identified by mAb 133 (ii 133). A third subpopulation which was successively retained on S4B-110 and S4B-133 expressed both internal images (ii 110 + 133), and was additionally reactive with all the other monoclonal Ab1 tested. In any case, monoclonal Ab1 binding to the different sets of internal images was totally inhibited by an excess of AII. These results indicate that the repertoire of internal epitopes is similar to that of the nominal Ag, but is scattered over distinct subpopulations of Ab2-beta molecules that can be fractionated by affinity chromatography. Some of the latter seem to bear several epitopes and resemble the whole nominal Ag, whereas others appear to image only one determinant. Second, we raised 7 anti-anti-idiotypic mAb (monoclonal Ab3) against affinity-purified Ab2-beta and analyzed their fine specificity for AII.(ABSTRACT TRUNCATED AT 400 WORDS) FAU - Budisavljevic, M AU - Budisavljevic M AD - Unite 64, Institut National de la Sante et de la Recherche Medicale 64, Hopital Tenon, Paris, France. FAU - Geniteau-Legendre, M AU - Geniteau-Legendre M FAU - Baudouin, B AU - Baudouin B FAU - Pontillon, F AU - Pontillon F FAU - Verroust, P J AU - Verroust PJ FAU - Ronco, P M AU - Ronco PM LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - J Immunol JT - Journal of immunology (Baltimore, Md. : 1950) JID - 2985117R RN - 0 (Antibodies, Anti-Idiotypic) RN - 0 (Antibodies, Monoclonal) RN - 0 (Epitopes) RN - 0 (Immunoglobulin Idiotypes) RN - 11128-99-7 (Angiotensin II) SB - IM MH - Angiotensin II/*immunology MH - Animals MH - Antibodies, Anti-Idiotypic/*immunology MH - Antibodies, Monoclonal/*immunology MH - Antibody Specificity MH - Epitopes MH - Immunoglobulin Idiotypes/*immunology MH - Mice MH - Rabbits MH - Structure-Activity Relationship EDAT- 1988/05/01 00:00 MHDA- 1988/05/01 00:01 CRDT- 1988/05/01 00:00 PHST- 1988/05/01 00:00 [pubmed] PHST- 1988/05/01 00:01 [medline] PHST- 1988/05/01 00:00 [entrez] PST - ppublish SO - J Immunol. 1988 May 1;140(9):3059-65.