Zalman L S, Wood L M, Frank M M, Müller-Eberhard H J
J Exp Med. 1987 Feb 1;165(2):572-7. doi: 10.1084/jem.165.2.572.
The affected E of two patients with paroxysmal nocturnal hemoglobinuria (PNH) were enriched by lysing the unaffected, normal E with anti-human decay-accelerating factor (DAF) and guinea pig serum. The membranes of the unlysed, DAF-deficient cells (PNH-E) were dissolved and examined by SDS-PAGE and immunoblotting using an antiserum to homologous restriction factor (HRF). Whereas the 65 kD complement regulatory protein was readily detectable in the normal controls, it was completely lacking in both samples of PNH-E membranes. Functional studies likewise indicated the absence of HRF activity from PNH-E. When radiolabeled, isolated HRF protein was offered to PNH-E, it became firmly attached to the cell. Approximately 1,000 molecules of HRF per cell reduced the characteristic susceptibility of these cells to reactive lysis by C5b-9 to nearly normal levels. The results suggest that HRF, which is known to control the action of C8 and C9 on normal human E membranes, is deficient in PNH, as well as acetylcholinesterase and DAF.
通过用抗人衰变加速因子（DAF）和豚鼠血清裂解未受影响的正常红细胞，富集了两名阵发性夜间血红蛋白尿（PNH）患者的受影响红细胞。用抗同源限制因子（HRF）抗血清通过SDS-PAGE和免疫印迹法溶解并检查未裂解的、缺乏DAF的细胞（PNH-E）的膜。在正常对照中很容易检测到65kD补体调节蛋白，但在PNH-E膜的两个样本中完全缺乏。功能研究同样表明PNH-E缺乏HRF活性。当将放射性标记的分离HRF蛋白提供给PNH-E时，它牢固地附着在细胞上。每个细胞约1000个HRF分子将这些细胞对C5b-9反应性裂解的特征敏感性降低到接近正常水平。结果表明，已知控制C8和C9对正常人红细胞膜作用的HRF在PNH中缺乏，乙酰胆碱酯酶和DAF也是如此。
