利用VSV△G＊伪型病毒建立安全、快速的西尼罗病毒抗体检测方法的尝试

目的为探讨利用水泡性口炎病毒（vesicular stomatitis virus，VSV）假病毒粒子系统包装西尼罗病毒（West Nile virus，WNV）的E蛋白可行性，构建3种E蛋白基因的真核表达质粒。以期用于WNV感染的流行病学调查。方法将表达WNN囊膜蛋白E的质粒体外转染293T细胞，然后感染VSV△G＊G，并对E蛋白表达质粒进行改造，分别在基因序列前加入信号肽，在终止密码子之前加入VSV△G蛋白的细胞质尾序列。结果包装出的VSV△G＊-WNVE滴度没有明显升高。提示WNV E蛋白与VSV的囊膜匹配性不佳，VSV△G＊-WNVE作为病毒中和抗原代替野毒进行抗体筛查还需要进一步提高转染和包装效率。结论VSV作为高效、安全的新型载体、活病毒疫苗载体和肿瘤基因治疗载体以及高危病毒的研究工具具有巨大的研究价值和应用潜力。

Generating vesicular stomatitis virus pseudotypes bearing the West Nile virus envelope glycoprotein for rapid and safe neutralization test

Objective In order to discuss she potential applicability of VSV pseudotype particles coated by WNVE used in the study of WNV infection. Methods 293T cells were transfected with plasmids for the expression of envelope glycoproteirus E of WNV and were then infected with VSV△G ＊ G. Pseudotype VSV with the WNV （VSV△G ＊ -WNVE） envelope glycoproteins were harvested from the culture supematant. Results The number of infectious units （IU） of the pseudotype VSVs ranged from 10^3 to 10^4/ml. To increase the IU of VSV△G ＊ -WNVE, a glycoprotein cytoplasmic domain of VSV was inserted before TAA of WNV E gene. The IU of the pseudotype VSVs still ranged from 10^3 to 10^4/ml. Conclusions These results show that we need another hard work till VSV△G ＊ -WNVE can be used as a rapid, specific, and safe neutralization test for detecting WNV-neutralizing antibodies as an effective substitute for the use of native hantaviruses.