补体存在时形成的HBsAg-抗-HBs免疫复合物的沉淀性及组成。循环免疫复合物分析模型。

Precipitability and composition of HBsAg-anti-HBs immune complexes formed in the presence of complement. A model of circulating immune complex analysis.

作者信息

Anh-Tuan N, Falus A, Füst G, Merétey K, Medgyesi G A, Hollán S R

出版信息

J Immunol Methods. 1983 Nov 25;64(3):283-94. doi: 10.1016/0022-1759(83)90435-0.

Abstract

Immune complexes (IC) of partially purified HBsAg and human anti-HBs were prepared at different antigen/antibody ratios in the presence of complement in normal human serum (NHS), and under conditions not allowing complement activation in buffers or in NHS containing 10 mM EDTA (NHS-EDTA). Commercial preparations of the radiolabelled antigen and antibody were used. IC formed in NHS were not significantly precipitated even after incubation for 24 h at 4 degrees C, whereas a typical precipitation curve was observed with complexes formed in the absence of complement. Thus, complement activation was found to markedly and permanently inhibit precipitability of HBsAg-anti-HBs immune complexes (HBsAg-IC). HBsAg-IC were precipitated from sera with 3.5% polyethylene glycol (PEG), boiled in sodium dodecyl sulphate (SDS)-urea buffer, and analysed by SDS-polyacrylamide slab gel electrophoresis (SDS-PAGE). With complexes formed in the presence of complement, about one-sixth of the antibody activity was found in high molecular weight fractions corresponding in size to IgG oligomers. By contrast, with complexes formed without complement, no significant amount of antibody was found in these fractions. With blotting technique and radiolabelled anti-human-C3 antibody, it was demonstrated that anti-HBs was covalently bound to C3b fragments in IC formed in the presence of complement and was in the high molecular weight fractions.

摘要

在正常人血清（NHS）中补体存在的情况下，以不同抗原/抗体比例制备部分纯化的乙肝表面抗原（HBsAg）与人抗-HBs的免疫复合物（IC），并在不允许补体激活的条件下，即在缓冲液中或含有10 mM乙二胺四乙酸（EDTA）的NHS（NHS-EDTA）中进行。使用放射性标记抗原和抗体的商业制剂。在NHS中形成的IC即使在4℃孵育24小时后也没有明显沉淀，而在无补体情况下形成的复合物则观察到典型的沉淀曲线。因此，发现补体激活显著且永久性地抑制HBsAg-抗-HBs免疫复合物（HBsAg-IC）的沉淀性。用3.5%聚乙二醇（PEG）从血清中沉淀出HBsAg-IC，在十二烷基硫酸钠（SDS）-尿素缓冲液中煮沸，并通过SDS-聚丙烯酰胺平板凝胶电泳（SDS-PAGE）进行分析。对于在补体存在下形成的复合物，在与IgG寡聚体大小相对应的高分子量组分中发现约六分之一的抗体活性。相比之下，对于在无补体情况下形成的复合物，在这些组分中未发现大量抗体。通过印迹技术和放射性标记的抗人C3抗体，证明抗-HBs在补体存在下形成的IC中与C3b片段共价结合，并存在于高分子量组分中。