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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。
In order to evaluate the in vitro and in vivo cytotoxicity of rVTT-CJ-8 +, an eight-recombinant recombinant vaccinia virus constructed in our laboratory, the cell killing ability of rVTT-CJ-8 + in vitro was detected by MTS method and crystal violet staining method. The in vivo toxicity of rVTT-CJ-8 + was detected by the change of body weight of rats. The neurotoxicity of rVTT-CJ-8 + was detected by the survival rate of mice after intracranial exposure. The rVTTC-J-8 + Rabbit’s pathogenicity. MTS assay showed that the inhibitory effect of rVTT-CJ-8 + on the growth of 5 cell lines was significantly lower than that of VTT. Crystal violet staining showed that rVTT-CJ-8 + had more obvious intercellular diffusion in BHK21 and MDCK cells. The body weight changes and rabbit plaque formation test results showed that rVTT-CJ-8 + had little effect on the weight changes of the mice and at the same time had less damage to the skin on the back of the rabbits. The virulence and virulence of the mice were significantly less than that of the VTT. Toxicity tests showed that rVTT-CJ-8 + had almost no neurotoxicity in mice. The results showed that the recombinant vaccinia virus rVTT-C-J-8 + had lower toxicity in vivo and less side effects, which provided a reference for the future production of polyvalent vaccine.