Budisavljevic M, Geniteau-Legendre M, Baudouin B, Pontillon F, Verroust P J, Ronco P M
Unité 64, Institut National de la Santé et de la Recherche Médicale 64, Hôpital Tenon, Paris, France.
J Immunol. 1988 May 1;140(9):3059-65.
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)
尽管β型单克隆抗独特型抗体（Ab2）所携带的内影像的结构基础已开始得到阐明，但关于构成多克隆Ab2所表达内影像的表位库的信息却很少。我们在血管紧张素II（AII）相关的独特型网络中采用双向方法解决了这个问题，该系统的特征是兔Ab2上经常出现内影像。首先，通过在共价连接到mAb 110（S4B - 110）的琼脂糖4B上进行亲和层析，平行纯化两组内影像，mAb 110是针对AII上需要苯丙氨酸的羧基末端表位（Phe8）的单克隆抗体，或者是与Phe8取代肽类似物上也表达的更中心表位反应的mAb 133（S4B - 133）。各自的洗脱液，EL1 110和E11 133，表现出仅部分重叠的反应性，如以下所示：1）各种AII肽类似物对EL1 110和E11 133与同一抗AII抗体（Ab1）（同源多克隆Ab1 102或mAb 133）结合的抑制模式不同；2）EL1 110和EL1 133与12种生物素化单克隆Ab1结合的独特图谱，这些Ab1识别AII上的多种表位。为了进一步分析mAb 110和mAb 133定义的表位在Ab2 - β分子上的各自分布，将Ab2依次在S4B - 110上进行亲和层析，然后在S4B - 133上进行亲和层析，并且通过与各种单克隆Ab1的结合模式来表征分级分离的内影像。因此，可以纯化出两个Ab2 - β亚群，它们分别仅成像由mAb 110鉴定的决定簇（ii 110）或由mAb 133鉴定的决定簇（ii 133）。第三个亚群先后保留在S4B - 110和S4B - 133上，表达两种内影像（ii 110 + 133），并且还与所有其他测试的单克隆Ab1反应。在任何情况下，过量的AII都能完全抑制单克隆Ab1与不同组内影像的结合。这些结果表明，内表位库与名义抗原的表位库相似，但分散在可以通过亲和层析分级分离的Ab2 - β分子的不同亚群中。后者中的一些似乎带有几个表位，类似于整个名义抗原，而其他一些似乎仅成像一个决定簇。其次，我们针对亲和纯化的Ab2 - β产生了7种抗抗独特型单克隆抗体（单克隆Ab3），并分析了它们对AII的精细特异性。（摘要截断于400字）
