Laboratory of Zoonoses, School of Veterinary Medicine Kitasato University, Japan.
J Virol Methods. 2010 Oct;169(1):232-8. doi: 10.1016/j.jviromet.2010.07.023. Epub 2010 Jul 30.
Although various cultured cells are used for propagating influenza A viruses, the types of cells which can support replication of and plaque production by low pathogenic avian influenza (LPAI) viruses without supplementary trypsin are limited. In this study, the infectivity and growth kinetics of as well as plaque production by LPAI viruses in Caco-2 cells were investigated. The suitability of this cell line for virus isolation was examined and compared with virus isolation in embryonated chicken eggs. Generation of Caco-2 mediated viral variants, if any, was assessed phenotypically and genotypically. It was found that Caco-2 cells can readily support continued replication of LPAI viruses without supplementary trypsin. Viruses replicate to high titer compared to embryonated chicken eggs, and more efficiently than in MDCK cells, without trypsin. Also, LPAI viruses produced plaques in Caco-2 cells. However, these cells were found to be less sensitive than embryonated chicken eggs for virus isolation. Notably, no phenotypic and genotypic changes of the viruses were observed during viral passages (at least up to 10th passage) in Caco-2 cells. These findings indicate that Caco-2 cells may provide an appropriate substrate for studying and cultivating AIVs.
尽管各种培养细胞被用于繁殖甲型流感病毒,但能够在没有补充胰蛋白酶的情况下支持低致病性禽流感(LPAI)病毒复制和产生蚀斑的细胞类型是有限的。在这项研究中,研究了 LPAI 病毒在 Caco-2 细胞中的感染性和生长动力学以及产生蚀斑的情况。检查了该细胞系用于病毒分离的适用性,并与鸡胚中的病毒分离进行了比较。如果有任何 Caco-2 介导的病毒变异体产生,则对其表型和基因型进行了评估。结果发现,Caco-2 细胞无需补充胰蛋白酶即可轻易支持 LPAI 病毒的持续复制。与鸡胚相比,病毒的复制达到了更高的滴度,并且在没有胰蛋白酶的情况下比 MDCK 细胞更有效。此外,LPAI 病毒在 Caco-2 细胞中产生蚀斑。然而,与鸡胚相比,这些细胞用于病毒分离的敏感性较低。值得注意的是,在 Caco-2 细胞中(至少在第 10 次传代)进行病毒传代时,未观察到病毒的表型和基因型变化。这些发现表明,Caco-2 细胞可能为研究和培养 AIV 提供合适的基质。