Tobaly-Tapiero Joëlle, Bittoun Patricia, Saïb Ali
CNRS UPR9051, Hôpital Saint-Louis, Centre Hayem, Paris, France.
Methods Mol Biol. 2005;304:125-37. doi: 10.1385/1-59259-907-9:125.
The isolation of a retrovirus from peripheral blood lymphocytes/monocytes can be a difficult task, requiring the fulfillment of three essential parameters. First, this viral agent must infect such cells in vivo. Second, these circulating cells should harbor wild-type proviruses. Finally, the viral agent has to express, at least when these cells are cultured in vitro, the structural proteins necessary for the production of viral particles. Foamy viruses (FVs), also known as spumaviruses, are complex retroviruses whose genomic organization has been known since the cloning of the prototypic primate foamy virus type 1. These retroviruses infect most cell lines in culture, but circulating lymphocytes seem to represent their major reservoir in vivo. FV infection leads to the formation of multinucleated giant cells, resulting from the fusion of adjacent infected cells, which present multiple vacuoles giving the monolayer culture a foam aspect. These two features, combined with electron microscopy studies, have helped investigators in their attempt to isolate new FVs. These viruses were described and isolated from different animal species, mostly in nonhuman primates. Here we present the successive steps leading to the isolation of the equine foamy virus from peripheral blood lymphocytes of infected horses.
从外周血淋巴细胞/单核细胞中分离逆转录病毒可能是一项艰巨的任务，需要满足三个基本条件。首先，这种病毒因子必须在体内感染此类细胞。其次，这些循环细胞应携带野生型前病毒。最后，该病毒因子必须表达，至少在这些细胞体外培养时，表达产生病毒颗粒所需的结构蛋白。泡沫病毒（FV），也称为泡沫逆转录病毒，是复杂的逆转录病毒，自克隆原型灵长类泡沫病毒1型以来，其基因组结构已为人所知。这些逆转录病毒可感染培养中的大多数细胞系，但循环淋巴细胞似乎是它们在体内的主要储存库。FV感染导致多核巨细胞的形成，这是由相邻感染细胞融合所致，这些细胞呈现多个空泡，使单层培养物呈现泡沫状外观。这两个特征，再加上电子显微镜研究，有助于研究人员尝试分离新的FV。这些病毒已从不同动物物种中描述和分离出来，主要是在非人类灵长类动物中。在这里，我们展示了从感染马的外周血淋巴细胞中分离马泡沫病毒的连续步骤。
