人用重组溶瘤单纯疱疹病毒Ⅱ型(OH2)生物学特性的稳定性

Stability of biological properties of recombinant oncolytic herpes virus type Ⅱ(OH2) for human use

目的分析人用重组溶瘤单纯疱疹病毒Ⅱ型(oncolytic herpes simplex virus type 2,OH2)生物学特性的稳定性。方法将OH2在Vero细胞中连续传20代,测定不同时间点收获的不同代次(P4、P10、P20)病毒的滴度,并绘制病毒的生长曲线;ELISA夹心法检测表达的h GM-CSF的含量;TF-1细胞培养法检测表达的h GM-CSF的活性;CCK-8试剂盒检测OH2的体外癌细胞杀伤能力。结果 OH2在Vero细胞中连续传20代后,不同代次病毒(P4、P10、P20)的生长曲线模式、表达的h GM-CSF含量及活性、体外癌细胞的杀伤能力均无明显差异。结论 OH2生物学特性的稳定性良好,具有新生物药开发前景。

Objective To analyze the stability of biological properties of recombinant oncolytic herpes virus type 2（OH2）for human use.Methods OH2 was subcultured in Vero cells for 20 passages,and the titers of various passages（P4,P10 and P20）harvested at various time points were determined,based on which a growth curve was plotted.The content of expressed h GM-CSF was determined by sandwich ELISA,while the activity by TF-1 cell culture method.The in vitro killing effect of OH2 on tumor cells was determined by CCK-8 kit.Results After subculture in Vero cells for 20 passages,no significant differences were observed in the patterns of growth curves,contents and activities of h GM-CSF expressed and in vitro killing effect on tumor cells of OH2 of various passages.Conclusion OH2 showed stable biological properties and was of good prospect of development.