痘苗病毒对马属动物细胞的感染性研究

目的研究痘苗病毒天坛株是否能够感染马属动物细胞,并诱导免疫马产生免疫反应。方法首先采用重组绿色荧光蛋白(GFP)的痘苗病毒WR株感染驴胎皮肤细胞(FDD)后,通过流式细胞仪定期检测GFP表达水平(病毒感染率),探索最佳感染时间,进而用重组HIV-1 Gag蛋白的痘苗病毒天坛株感染FDD,通过免疫印记实验确认Gag蛋白在细胞内表达,并且进行胞内染色,通过流式细胞仪监测病毒的复制进程;使用痘苗病毒天坛株空载体接种实验马并监测其诱导的免疫反应。结果痘苗病毒感染FDD使产生病变效应,在其内表达外源蛋白,最佳时间为48h;使实验马产生针对病毒的结合抗体。结论痘苗病毒对FDD易感,痘苗病毒天坛株能使实验马感染,提示痘苗病毒重组EIAV抗原的载体疫苗可用于马体接种免疫。

Study on the infectivity of vaccinia virus in equine cells

To study whether the vaccinia virus Tian Tan strain(VTT) can infect equine cells and produce immune response in horses,recombinant vaccinia virus WR strain expressing green fluorescence protein(GFP) was used to infect fetal dermal donkey cells(FDD) and the virus replication was tested regularly by flow cytometry.The optional infection time was observed.HIV-1 Gag protein-expressing recombinant VTT(rVTT) was further used to infect FDD cells.Infection was confirmed by western blotting and intracellular protein staining.Finally,horses were vaccinated with VTT to inspect the immune response induced.Both strains of vaccinia virus could infect FDD cells and inducs cytopathic effects.Optimal infection time of FDD cells was 48h,at which point,binding antibodies could be detected in the horses sera.These results study suggest that the vector vaccine with recombinant eqaine infcetion anemia virus(EIAT) antigen of vaccinaa virus may be useful for vaccination to horses.