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目的制备hIL-18和TK双基因修饰的人脑胶质瘤活疫苗,检测HSV-TK/GCV系统对疫苗的体外调控作用。方法采用表达HSV-TK基因的慢病毒液感染U87-hIL18细胞,经G418和BSD抗性基因筛选获得U87-hIL18-TK单克隆细胞株,扩增传代获得双基因修饰疫苗,Elisa检测疫苗上清IL-18浓度,RT-PCR检测疫苗TK基因表达,MTT法检测更昔洛韦(GCV)对体外培养疫苗的杀伤效应。结果 TK基因成功转导入U87-hIL18细胞中,获得稳定表达IL-18、TK基因的U87-hIL18-TK胶质瘤活疫苗,体外杀伤效应显示10μg/mlGCV对活疫苗的杀伤率达90.3%。结论成功构建免疫基因和自杀基因共同修饰的胶质瘤活细胞疫苗,其在体内的安全性及免疫效能有待进一步研究。
Objective To prepare hIL-18 and TK double gene modified human glioma live vaccine to detect HSV-TK / GCV system in vitro regulation of the vaccine. Methods U87-hIL18 cells were infected with lentivirus expressing HSV-TK gene. U87-hIL18-TK monoclonal cell line was screened by G418 and BSD resistance genes. The double-gene modified vaccine was amplified and passaged. IL-18 concentration, TK gene expression was detected by RT-PCR and MTT assay was used to detect the killing effect of ganciclovir (GCV) on in vitro vaccine. Results The TK gene was successfully transfected into U87-hIL18 cells. The live U87-hIL18-TK glioma vaccine stably expressing IL-18 and TK genes was obtained. The killing effect of 10μg / ml GCV on live vaccine was 90.3%. Conclusion The successful construction of a glioma live cell vaccine co-modified with both immunogenic and suicide genes remains to be further studied in vivo for its safety and immunological efficacy.