建立DBT细胞模型比较药物对MHV-A59病毒的抗毒作用

目的:利用病毒学方法建立对小鼠肝炎病毒(MHVA59)易感的星形胶质瘤细胞(DBT)模型,在细胞水平上建立抗冠状病毒药物的筛选的实验方法。方法:通过测定细胞病变效应(CPE)的半定量法及噬斑定量法测定组织培养感染量(TCID50);以PEG沉淀法部分纯化病毒,利用免疫学方法免疫动物制备抗MHVA59的多克隆抗体;以血清学方法进行中和实验及酶联免疫方法定量测定病毒抗体的滴度及特异性;以100倍的TCID50病毒量感染细胞同时加不同稀释度的药物,在同一条件下,动态观察不同时间的细胞的CPE变化。以能保护CPE<50%为阳性。并比较药物的最低抗毒浓度。结果:制备的抗体对MHV病毒具有特异性的翻转作用;合成新药Cinanserin和Pkumdlcvs1003对MHV所致的细胞感染具有不同程度的保护作用。结论:建立的MHVA59敏感的DBT细胞模型可以用于抗冠状病毒药物的筛选。

Comparison of drugs effect on MHV-A59 virus by established DBT cell model

AIM: To establish sensitive DBT cell model to MHV-A59 by virology method, and develop selective method of anti virus drugs in cell level. [WTHZ]METHODS: Test 50% percent tissue culture infective dose (TCID_ 50) by cytopathetic effect (CPE) and plaque methods. Preparation of antibody against virus by partially purified virus immunized animal. Selective anti virus drug adding 100 fold of TICD50 virus dose infected DBT cell as positive results by protesting lower 50% CPE, and tested drugs lowest virus inhibit concentration. RESULTS: Prepared antibody has specificity reverse virus action, and new synthesized drugs had various degree protective actions to virus infective. CONCLUSION: The DBT cell model sensitive to MHV-A59 can select anti coronaviruses drug.