Mahalingam R, Wellish M, White T, Soike K, Cohrs R, Kleinschmidt-DeMasters B K, Gilden D H
Department of Neurology, University of Colorado Health Sciences Center, Denver 80262, USA.
J Neurovirol. 1998 Aug;4(4):438-44. doi: 10.3109/13550289809114543.
Clinical, pathologic, immunologic and virologic features of simian varicella virus (SVV) infection in primates closely resemble varicella-zoster virus (VZV) infection in humans. Such similarities provide a rationale to analyze SVV infection in primates as a model of varicella pathogenesis and latency. Thus, we constructed an SVV-expressing green fluorescent protein (SVV-GFP) by inserting the GFP gene into the unique short segment of the virus genome by homologous recombination. Analysis of recombinant viral DNA and the expressed proteins of plaque-purified SVV-GFP confirmed the location of the GFP insert and that the recombinant SVV expressed the 27 kDa GFP. Infection of monkey kidney cells in tissue culture with SVV-GFP revealed bright green fluorescence associated with the characteristic focal cytopathic effect produced by SVV infection. Microscopic examination of lung from a 3-month-old African green monkey 10 days after infection with SVV-GFP revealed bright green fluorescence in areas of acute necrotizing pneumonitis. SVV-GFP allows ready identification of cells infected with SVV both in vitro and in vivo, and will be useful for further analysis of varicella pathogenesis and latency in experimentally infected animals--studies not possible in humans.
灵长类动物中猿猴水痘病毒(SVV)感染的临床、病理、免疫和病毒学特征与人类水痘-带状疱疹病毒(VZV)感染极为相似。这些相似之处为将灵长类动物中的SVV感染作为水痘发病机制和潜伏模型进行分析提供了理论依据。因此,我们通过同源重组将绿色荧光蛋白(GFP)基因插入病毒基因组的独特短片段中,构建了一种表达SVV的绿色荧光蛋白(SVV-GFP)。对噬斑纯化的SVV-GFP的重组病毒DNA和表达蛋白进行分析,证实了GFP插入片段的位置,并且重组SVV表达了27 kDa的GFP。用SVV-GFP在组织培养中感染猴肾细胞,发现明亮的绿色荧光与SVV感染产生的特征性局灶性细胞病变效应相关。对一只3个月大的非洲绿猴在感染SVV-GFP 10天后的肺组织进行显微镜检查,发现在急性坏死性肺炎区域有明亮的绿色荧光。SVV-GFP能够在体外和体内轻松识别感染SVV的细胞,并且将有助于进一步分析实验感染动物中水痘的发病机制和潜伏期——这是在人类中无法进行的研究。