补体活化的体外评价模型建立及应用

Establishment of in-vitro model of complement activation and its application

补体激活相关的假性过敏反应(complement activation-related pseudo-allergic reactions,CARPA)占全部免疫介导的直接过敏反应的77%。由于CARPA临床普遍性,以及该反应与药物理化性质的相关性,补体活性检测被建议作为药物免疫毒性和药物一致性的评价指标之一。但是,由于补体活性的体内检测较为复杂、不同个体之间以及人与动物之间存在免疫差异,使得有必要建立一个标准、快捷的模型,用于评价药物对补体活性的影响。本研究使用人混合血清作为标准反应血清,通过加入热聚集Ig G和酵母多糖激活血清得到标准阳性对照,将SC5b-9、C5a、C4d和Bb作为检测指标并确定评价标准,建立了药物对补体活性影响的体外评价模型。应用该评价模型,以抗真菌类药物两性霉素B的不同处方和注射用伏立康唑为研究对象,对这些药物的免疫毒性以及仿制与市售制剂的一致性进行了初步评价。

Complement activation-related pseudo-allergic reactions （CARPA） may represent 77% of all immune-mediated immediate hypersensitivity reactions. Because of the universality of the CARPA response and correlation between it and drug properties, complement activity tests are recommended as one of the tests for immunotoxicity and bioequivalence of drugs. However, in-vivo tests of complement activation are complicated, and the immunological differences between different individuals and between human and animal, making it very necessary to establish a standard and sample evaluation model for testing the effects of drugs on complement activity. In this study, the standard reaction serum was prepared by pooling sera collected from 40 healthy blood donors; a standard positive control was prepared by incubation with a heat-agglutinated IgG and zymosan A; SC5b-9, C5a, C4d and Bb were chosen as the test targets and evaluation criteria of the results was defined, all of these constituted the in-vitro model. By using this in-vitro model, the immunological toxicity of the different prescription of antifungal drug amphotericin B, and voriconazole for injection, and the bioequivalence of amphotericin B liposome formulations were studied.