Baatrup G, Peterson I, Svehag S E, Brandslund I
J Immunol Methods. 1983 May 13;59(3):369-80. doi: 10.1016/0022-1759(83)90197-7.
A standardized radioassay for measuring the complement-mediated immune complex solubilizing capacity (CMSC) and the initial kinetics of the solubilization (IKS) reaction is described. The total complement (C)-mediated solubilizing capacity was determined after incubation of diluted serum and 125I-BSA-anti-BSA. Percentage C-mediated solubilization (CMS) was measured after centrifugation by determining the distribution of radioactivity. The dependency of CMSC upon factors such as serum dilution and buffer system used, amount of IC added to serum, serum storage conditions and centrifugation conditions was investigated in order to optimize the assay. The CVt of the standardized assay was 0.10-0.17 depending upon the CMSC level measured. Treatment which inactivates C factors (heating), interferes with C activation (EDTA) or activates and consumes C components (zymosan) markedly reduces the CMSC. Preliminary investigation of pathological sera showed that both IKS and CMSC were clearly reduced in SLE sera. By contrast, rheumatoid arthritis sera exhibited normal IKS and only marginal reduction in CMSC.
本文描述了一种用于测量补体介导的免疫复合物溶解能力(CMSC)和溶解反应初始动力学(IKS)的标准化放射分析方法。在稀释血清与¹²⁵I-牛血清白蛋白-抗牛血清白蛋白孵育后,测定总补体(C)介导的溶解能力。通过测定放射性分布,在离心后测量补体介导的溶解百分比(CMS)。为优化该分析方法,研究了CMSC对血清稀释度、所用缓冲系统、加入血清的免疫复合物量、血清储存条件和离心条件等因素的依赖性。根据所测CMSC水平,标准化分析方法的变异系数(CVt)为0.10 - 0.17。使补体因子失活(加热)、干扰补体激活(乙二胺四乙酸)或激活并消耗补体成分(酵母聚糖)的处理会显著降低CMSC。对病理血清的初步研究表明,系统性红斑狼疮(SLE)血清中的IKS和CMSC均明显降低。相比之下,类风湿性关节炎血清的IKS正常,CMSC仅略有降低。