Cybulsky A V, Rennke H G, Feintzeig I D, Salant D J
J Clin Invest. 1986 Apr;77(4):1096-107. doi: 10.1172/JCI112408.
In passive Heymann nephritis (PHN) in rats, antibody (anti-Fx1A) reacts in situ with a glomerular epithelial antigen and induces complement (C)-mediated cell-independent proteinuria. To assess the role of the membrane attack complex (MAC), we determined the need for C8 in the pathogenesis of proteinuria in an autologous-phase model of PHN. Isolated rat kidneys, containing nonnephritogenic, non-C-fixing gamma 2 sheep anti-Fx1A (planted antigen), when perfused in vitro with C-fixing guinea pig anti-sheep IgG and a source of C (fresh human plasma 50% vol/vol in buffer containing bovine serum albumin), developed marked proteinuria after 20 min (0.58 +/- 0.08 mg/min X g, n = 8) that increased further to 3.20 +/- 0.93 mg/min X g after 80 min. In contrast, identical kidneys perfused with antibody and heat-inactivated or C8-deficient human plasma and normal kidneys perfused with antibody and fresh plasma excreted only 0.27 +/- 0.03 (n = 6), 0.27 +/- 0.04 (n = 5), and 0.40 +/- 0.05 mg/min X g (n = 6) after 20 min, and 0.13 +/- 0.02, 0.22 +/- 0.03, and 0.32 +/- 0.05 mg/min X g after 80 min, respectively. When C8-deficient plasma was reconstituted with sources of C8 (n = 3), proteinuria was restored to the level observed with fresh normal plasma. Differences in protein excretion could not be explained by quantitative differences in glomerular antigen or antibody content. Extensive ultrastructural damage to glomerular visceral epithelial cells was exclusively seen in antigen-containing kidneys perfused with antibody and C8-replete plasma. Thus, glomerular injury in this model results from an antigen-specific, antibody-directed, C8-dependent reaction involving assembly of the MAC. The ultrastructural findings argue in favor of MAC-induced cytotoxicity of the glomerular visceral epithelial cells.
在大鼠被动型海曼肾炎(PHN)中,抗体(抗Fx1A)与肾小球上皮抗原发生原位反应,并诱导补体(C)介导的非细胞依赖性蛋白尿。为了评估膜攻击复合物(MAC)的作用,我们在PHN的自体期模型中确定了蛋白尿发病机制中C8的必要性。分离的大鼠肾脏含有非致肾炎性、非补体结合性的γ2绵羊抗Fx1A(植入抗原),当在体外与补体结合性豚鼠抗绵羊IgG和补体来源(含牛血清白蛋白的缓冲液中50%体积/体积的新鲜人血浆)灌注时,20分钟后出现明显蛋白尿(0.58±0.08mg/分钟×克,n = 8),80分钟后进一步增加至3.20±0.93mg/分钟×克。相比之下,用抗体和热灭活或C8缺陷型人血浆灌注的相同肾脏,以及用抗体和新鲜血浆灌注的正常肾脏,20分钟后分别仅排泄0.27±0.03(n = 6)、0.27±0.04(n = 5)和0.40±0.05mg/分钟×克(n = 6),80分钟后分别为0.13±0.02、0.22±0.03和0.32±0.05mg/分钟×克。当用C8来源重建C8缺陷型血浆时(n = 3),蛋白尿恢复到新鲜正常血浆观察到的水平。蛋白尿排泄的差异不能用肾小球抗原或抗体含量的定量差异来解释。仅在灌注抗体和C8充足血浆的含抗原肾脏中观察到肾小球脏层上皮细胞广泛的超微结构损伤。因此,该模型中的肾小球损伤是由涉及MAC组装的抗原特异性、抗体导向、C8依赖性反应引起的。超微结构研究结果支持MAC诱导肾小球脏层上皮细胞的细胞毒性作用。