Kilgallon W, Amlot P L, Williams B D
Clin Exp Immunol. 1983 Aug;53(2):308-18.
Immune complexes (IC) present in sera from patients with Hodgkin's disease (HD) were isolated using three different affinity columns: C1q-degalan, anti-C1q sepharose and conglutinin (K)-degalan. The isolated IC were analysed by immunoprecipitation, SDS-PAGE and sucrose density gradients and compared with IC similarly isolated from patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and in vitro prepared BSA-anti-BSA complexes. Isolated material from each disease, and BSA-anti-BSA complexes contained proteins compatible with true immune complexes--IgM, IgG, C1q and C3 breakdown components. Albumin, fibronectin and CRP, whose affinity for IgM, C1q and C3 are known, were co-isolated along with IC material. The size of isolated IC in HD ranged from 8-40S on sucrose density gradients. Despite the operational difference in detecting and isolating HD complexes via the C1q ligand (C1q-degalan or anti-C1q column) and C3bi (K-degalan), material purified by both methods showed remarkable similarity on SDS-PAGE and immunoprecipitation analysis. Although IC isolated from different diseases showed disparate banding patterns on SDS-PAGE this was attributed to a variation in the relative concentrations of constituent proteins--IgM, IgG and C3 breakdown products. IgM, IgG and C3 bind loosely, and non-specifically, to macromolecular aggregates formed around immune complexes. Using the anti-C1q column, most of this material could be eluted using 0.02M EDTA. Least protein, yet the most specific for antigen and antibody was eluted at pH 3.0.
使用三种不同的亲和柱（C1q - 脱半乳糖、抗C1q琼脂糖和胶固素（K） - 脱半乳糖）从霍奇金病（HD）患者血清中分离免疫复合物（IC）。通过免疫沉淀、SDS - PAGE和蔗糖密度梯度对分离出的IC进行分析，并与从类风湿性关节炎（RA）、系统性红斑狼疮（SLE）患者中类似分离出的IC以及体外制备的牛血清白蛋白 - 抗牛血清白蛋白复合物进行比较。每种疾病分离出的物质以及牛血清白蛋白 - 抗牛血清白蛋白复合物都含有与真正免疫复合物相容的蛋白质——IgM、IgG、C1q和C3降解成分。已知对IgM、C1q和C3有亲和力的白蛋白、纤连蛋白和CRP与IC物质一起被共分离出来。在蔗糖密度梯度上，HD中分离出的IC大小范围为8 - 40S。尽管通过C1q配体（C1q - 脱半乳糖或抗C1q柱）和C3bi（K - 脱半乳糖）检测和分离HD复合物存在操作差异，但通过两种方法纯化的物质在SDS - PAGE和免疫沉淀分析中显示出显著的相似性。虽然从不同疾病中分离出的IC在SDS - PAGE上显示出不同的条带模式，但这归因于组成蛋白质——IgM、IgG和C3降解产物的相对浓度变化。IgM、IgG和C3与免疫复合物周围形成的大分子聚集体松散且非特异性地结合。使用抗C1q柱，大部分这种物质可以用0.02M EDTA洗脱。在pH值为3.0时洗脱的蛋白质最少，但对抗抗原和抗体最具特异性。
