八联重组痘苗病毒的体内外毒性研究

Virulence of recombinant vaccinia virus Tiantan strain with eight exogenous genes combined in vitro and in vivo

为评价本实验室构建的八联重组痘苗病毒rVTT-C-J-8+的体内外毒性,用MTS方法和结晶紫染色方法检测rVTT-C-J-8+的体外细胞杀伤能力;通过鼻腔接毒后小鼠的体重变化检测rVTT-C-J-8+的体内毒性;通过颅内接毒后的小鼠存活率检测rVTT-C-J-8+的神经毒性;通过兔斑形成试验检测rVTTC-J-8+对家兔的致病能力。MTS检测结果表明,rVTT-C-J-8+对5种细胞的生长抑制作用明显低于VTT;结晶紫染色表明,rVTT-C-J-8+只有在BHK21和MDCK细胞具有较明显的细胞间扩散水平;小鼠体重变化和兔斑形成试验结果表明,rVTT-C-J-8+对小鼠体重变化影响较小,同时对兔背部皮肤损伤较小,体内毒力和致病力明显小于VTT;小鼠神经毒性的检测表明,rVTT-C-J-8+对小鼠几乎无神经毒性。结果证明,八联重组痘苗病毒rVTT-C-J-8+具有较低的体内外毒性,副作用较小,为今后多联多价疫苗的生产提供了参考依据。

To assess the virulence of recombinant vaccinia virus contained eight exogenous genes rVTT-C-J-8^＋,the cytotoxicity of rVTT-C-J-8^＋was assessed by MTS and crystal violet staining assay in vitro.The virulence of rVTT-C-J-8^＋was assessed by lesion experiment in rabbit,body weight change experiment and neurotoxicity experiment mice.MTS result showed that the inhibiting effect of rVTT-CJ-8^＋on BHK21,HeLa,PK-15,MDCK and Vero cells was significantly lower than that of VTT.Crystal violet staining result showed that rVTT-C-J-8^＋had replication capacity only in BHK21 and MDCK cells.Rabbit lesion experiment and mice body weight change experiment results showed that in vivo virulence of rVTT-C-J-8^＋was significantly lower than that of VTT.Mice neurotoxicity experiment result showed that rVTT-C-J-8^＋has almost no neurotoxicity in mice.In conclusion,rVTT-C-J-8^＋has low virulence and less side effects,and provided a reference on the multi-vaccine production.