氯胺-T氧化C5、体外组装补体膜攻击复合物及其鉴定  

Assembling and identification of membrane attack complex after C5 activation in vitro

作　　者：周镜然[1] 朱锡华[1] 白云[1] 姜曼[1] 黎万玲[1] 

机构地区：[1]第三军医大学基础医学部全军免疫学研究所,重庆400038

出　　处：《第三军医大学学报》2002年第10期1229-1230,共2页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 397890 10 ) ;国家重点基础研究发展规划资助项目 ("973"项目 ) (G19990 54 2 0 0 )

摘　　要：目的　利用氧化的方法在无酶解反应发生的情况下获得活化形式的C5 ,进而实现以纯化的补体终末成分体外组装补体膜攻击复合物 (MAC)。方法　以蛋氨酸氧化剂氯氨 T与纯化的C5相互作用后 ,加入C6组装具溶血活性的C5 6复合物。以此复合物与NHS或纯化的后续终末补体成分组装MAC ,检测复合物的溶血活性。结果　反应性溶破实验证实 ,氯胺 T氧化C5与C6组装形成功能C5 6复合物 ,后者可与NHS或纯化的补体终末成分C7～C9体外组装具溶血活性的MAC ,并且所形成的C5 6复合物在 2 4h达到最大的溶血活性 ,48h仍保持稳定。结论　氯胺 T在不裂解C5的条件下产生了C5b样活性分子 ,该分子能与C6组装形成稳定的活性复合物 ,并可与其它补体终末成分组装产生活性MAC。Objective To form hemolytica membrane attack complex(MAC)with purified human terminal components of complement after activation of C5 with chloramine T(Cl T), a methionine oxidizing agent Methods Purified human C5 was incubated with Cl T in barbital buffer C6 was added to form a stable C56 complex 10 min later After incubation with NHS or purified C7, C8 and C9, MAC formation and lysis of non sensitized guinea pig red cells (reactive lysis) were assayed Results Cl T treated C5 acquired a binding site for C6 to form C56; After it incubated with C6 and subsequent addition of NHS or C7, C8 and C9, a membrane attack complex was formed which lysed non sensitized guinea pig red cells The hemolytic activity was in a dose dependent fashion Conclusion Cl T oxidating C5 leads to form C5b like functional activity molecules which can bind to C6 and form hemolytic MAC

关 键 词：C5 补体膜攻击复合物 补体终末成分 氯氨-T 

分 类 号：R392[医药卫生—免疫学]