Mannik M, Agodoa L Y, David K A
J Exp Med. 1983 May 1;157(5):1516-28. doi: 10.1084/jem.157.5.1516.
Covalently, cross-linked immune complexes were prepared with multivalent 2-nitro-4-azidophenyl X human serum albumin (NAP X HSA) and antibodies to NAP at five times antigen excess. After purification with gel filtration, affinity chromatography with antigen-agarose column, and addition of the hapten, 9.5% of the antibodies dissociated from the complexes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. After injection of these cross-linked immune complexes into mice, glomeruli stained for the complexes by immunofluorescence microscopy for only a few hours and electron-dense deposits were not detected. In contrast, when the same immune complexes with comparable lattice but without covalent cross-linking were administered to a second group of mice, the initial deposition by immunofluorescence was comparable and then increased to extensive deposits that persisted to 96 h. In this second group of mice extensive electron-dense deposits evolved. These observations supported the conclusion that the immune complexes initially deposited from circulation must undergo rearrangement to persist and to form electron-dense deposits in glomeruli. The covalently cross-linked immune complexes existed in glomeruli only for a short period of time since these complexes could not rearrange.
用多价2-硝基-4-叠氮苯基X人血清白蛋白(NAP X HSA)和针对NAP的抗体,以五倍抗原过量制备共价交联的免疫复合物。经凝胶过滤、抗原琼脂糖柱亲和层析纯化以及添加半抗原后,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析,9.5%的抗体从复合物中解离。将这些交联的免疫复合物注射到小鼠体内后,通过免疫荧光显微镜观察,肾小球中复合物染色仅持续数小时,未检测到电子致密沉积物。相比之下,当将具有可比晶格但无共价交联的相同免疫复合物给予第二组小鼠时,最初通过免疫荧光观察到的沉积物相当,随后增加至广泛的沉积物,且持续至96小时。在这第二组小鼠中出现了广泛的电子致密沉积物。这些观察结果支持了这样的结论,即最初从循环中沉积的免疫复合物必须经过重排才能持续存在并在肾小球中形成电子致密沉积物。共价交联的免疫复合物在肾小球中仅存在很短时间,因为这些复合物无法重排。