Theofilopoulos A N, Wilson C B, Dixon F J
J Clin Invest. 1976 Jan;57(1):169-82. doi: 10.1172/JCI108257.
A sensitivie and simple procedure for the detection and quantitation of soluble complement (C)- fixing immune complexes in sera of patients with various disease states has been developed by utilizing C receptors on Raji cells. These cells lack membrane-bound immunoglobulin but have receptors for IgG Fc, C3b, C3d, and possibly with other C proteins. Uptake experiments showed that both aggregated human gamma globulin (AHG) and 7S IgG bound to receptors for IgG Fc; however, AHG reacted with C bound to cells only via receptors for C and this binding was much more efficient than via IgG Fc receptors. AHG was used as an in vitro model of human immune complexes and its uptake by Raji cells was quantitated by 125I-radiolabeled antihuman IgG. The limit of sensitivity of this test was 6 mug AHG/ml serum. The ability of Raji cells to detect AHG in serum depended on the amount of radioactive antibody used and the size of aggregates. The presence of an excess of C somewhat inhibited binding of AHG containing C to Raji cells. The efficient binding of AHG by receptors for C on Raji cells was used for the detection and quantitation of immune complexes in human sera. Raji cells were incubated with sera to be tested and then reacted with excess radiolabeled antihuman IgG; the amount of radioactivity bound to the washed cells was determined and referred to a standard curve of radioactive antibody uptake by cells previously incubated with increasing amounts of AHG in serum. Thereby immune complexes were detected and quantitated in serum hepatitis, systemic lupus erythematosus, vasculitis, subacute sclerosing panencephalitis, dengue hemorrhagic fever, and malignancies.
通过利用Raji细胞上的补体(C)受体,已开发出一种灵敏且简单的程序,用于检测和定量各种疾病状态患者血清中的可溶性补体结合免疫复合物。这些细胞缺乏膜结合免疫球蛋白,但具有IgG Fc、C3b、C3d以及可能其他C蛋白的受体。摄取实验表明,聚合人γ球蛋白(AHG)和7S IgG均与IgG Fc受体结合;然而,AHG仅通过C受体与细胞结合的C发生反应,且这种结合比通过IgG Fc受体更有效。AHG被用作人类免疫复合物的体外模型,其被Raji细胞摄取的情况通过125I放射性标记的抗人IgG进行定量。该检测的灵敏度极限为6μg AHG/ml血清。Raji细胞检测血清中AHG的能力取决于所用放射性抗体的量和聚集体的大小。过量C的存在会在一定程度上抑制含C的AHG与Raji细胞的结合。Raji细胞上C受体对AHG的有效结合被用于检测和定量人血清中的免疫复合物。将Raji细胞与待测血清孵育,然后与过量的放射性标记抗人IgG反应;测定与洗涤后的细胞结合的放射性量,并参照先前用血清中逐渐增加量的AHG孵育的细胞对放射性抗体摄取的标准曲线。从而在血清性肝炎、系统性红斑狼疮、血管炎、亚急性硬化性全脑炎、登革出血热和恶性肿瘤中检测和定量免疫复合物。