人源细胞激活试验法应用于中药注射剂过敏反应检查的研究

Study on the Application of in Vitro Human Cell Line Activation Test with Chinese Medicine Injection

关键词:  人源细胞激活试验  皮肤变态反应  替代方法

Key words:  human cell line activation test (h-CLAT)  skin sensitization  alternative methods

摘要:

目的 研究体外人源细胞激活试验法（human cell line activation test，h-CLAT）用于中药注射剂过敏反应检查的可行性，探讨中药注射剂的Ⅳ型变态反应体外检测的可行性。方法 参考OECD指导原则442E的h-CLAT法，应用流式细胞仪在类树突样的人源白血病细胞系THP-1上对4种中药注射剂进行体外过敏反应检查，根据THP-1细胞表面抗原CD86和CD54的相对荧光强度评价4种中药注射剂体外过敏反应；同时采用体内过敏反应对4种注射剂进行评价。结果 体外皮肤过敏试验表明，树突状细胞表面特异抗原CD86相对荧光强度值<150，CD54的相对荧光强度值<200，根据判定标准结果为阴性，与体内法结果一致。结论 h-CLAT法具有检验周期短、测试方法简便、评价客观等优点，是一种较好的检查中药注射剂过敏反应的替代方法。

Abstract:

OBJECTIVE To evaluate the possibility of allergic response test for Chinese medicine injections using human cell line activation test in vitro, and to explore the feasibility of in vitro test of type Ⅳ allergy reaction for traditional Chinese medicine injection. METHODS In terms of OECD Guideline 442E, 4 different Chinese medicine injections were tested both in human cell line activation test with human leukemia THP-1 cells in vitro and allergic response in vivo at the same time. RESULTS From the result of in vitro skin sensitization test, relative fluorescence intensity of specific surface antigen of dendritic cells CD86 was below 150 and CD54 below 200, which was negative according to the criteria. The results was the same as in vivo test. CONCLUSION In vitro test has its advantages of short cycle, relatively simple and objective, and is a good alternative method for allergic response test for Chinese medicine injections.