Jasin H E
Department of Internal Medicine, Southwestern Medical School, Dallas, Texas 75235.
J Clin Invest. 1988 Jan;81(1):6-15. doi: 10.1172/JCI113310.
Incubation of human serum albumin-anti-human serum albumin immune complexes bound to a plastic surface, with human polymorphonuclear leukocytes for 1 h at 37 degrees C resulted in covalent cross-linking of 8.5% +/- 0.5 of the complexes, corresponding to a minimum rate of 700 antibody molecules per cell per minute. Similar results were obtained with IgG-anti-IgG and type II collagen-anticollagen II human antibodies. Cross-linking was defined as the antibody remaining attached to plastic-bound antigen after extraction with 3 M MgCl2 and 0.1 N HCl solutions. The effects of addition of oxygen radical scavengers, heme-enzyme inhibitors, and omission of Cl- indicated that the cross-linking process was mediated by the myeloperoxidase-H2O2-Cl- system. Cross-linking was also obtained with cell lysates, polymorphonuclear granules, and purified human myeloperoxidase in the presence of a steady flux of H2O2 provided by glucose oxidase-glucose. Cross-linking by the cell-free systems was also abolished by sodium azide or omission of chloride ions. Cross-linked immune complexes were also generated by incubation with 20 to 50 microM solutions of freshly distilled hypochlorous acid. Addition of 10 mM hypochlorous acid to soluble IgG resulted in the formation of protein precipitates insoluble in 5 M guanidine, 0.1 N HCl, or boiling 2.3 M sodium dodecyl sulfate-1.4 M 2-mercaptoethanol. The remaining soluble IgG contained fluorescent high molecular aggregates (ex: 360 nm; em: 454 nm). Oxidative cross-linking of antigen-antibody molecules, and of immune complexes to connective tissue macromolecules may play a pathogenic role in acute and chronic inflammatory processes.
将结合在塑料表面的人血清白蛋白 - 抗人血清白蛋白免疫复合物与人类多形核白细胞在37℃孵育1小时，结果8.5%±0.5%的复合物发生共价交联，相当于每个细胞每分钟至少700个抗体分子的交联速率。用IgG - 抗IgG和II型胶原 - 抗II型胶原人类抗体也得到了类似结果。交联定义为用3M MgCl2和0.1N HCl溶液提取后仍附着于塑料结合抗原的抗体。添加氧自由基清除剂、血红素酶抑制剂以及去除Cl-的影响表明，交联过程由髓过氧化物酶 - H2O2 - Cl-系统介导。在葡萄糖氧化酶 - 葡萄糖提供稳定通量的H2O2存在下，细胞裂解物、多形核颗粒和纯化的人髓过氧化物酶也能实现交联。叠氮化钠或去除氯离子也会消除无细胞系统的交联作用。用新鲜蒸馏的20至50微摩尔次氯酸溶液孵育也能产生交联免疫复合物。向可溶性IgG中添加10mM次氯酸会导致形成不溶于5M胍、0.1N HCl或沸腾的2.3M十二烷基硫酸钠 - 1.4M 2 - 巯基乙醇的蛋白质沉淀。剩余的可溶性IgG含有荧光高分子聚集体（激发波长：360nm；发射波长：454nm）。抗原 - 抗体分子以及免疫复合物与结缔组织大分子的氧化交联可能在急性和慢性炎症过程中起致病作用。
