Govorkova E A, Murti G, Meignier B, de Taisne C, Webster R G
Department of Virology & Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
J Virol. 1996 Aug;70(8):5519-24. doi: 10.1128/JVI.70.8.5519-5524.1996.
The preparation of live, attenuated human influenza virus vaccines and of large quantities of inactivated vaccines after the emergence or reemergence of a pandemic influenza virus will require an alternative host cell system, because embryonated chicken eggs will likely be insufficient and suboptimal. Preliminary studies indicated that an African green monkey kidney cell line (Vero) is a suitable system for the primary isolation and cultivation of influenza A viruses (E. A. Govorkova, N. V. Kaverin, L. V. Gubareva, B. Meignier, and R. G. Webster, J. Infect. Dis. 172:250-253, 1995). We now demonstrate for the first time that Vero cells are suitable for isolation and productive replication of influenza B viruses and determine the biological and genetic properties of both influenza A and B viruses in Vero cells; additionally, we characterize the receptors on Vero cells compared with those on Madin-Darby canine kidney (MDCK) cells. Sequence analysis indicated that the hemagglutinin of Vero cell-derived influenza B viruses was identical to that of MDCK-grown counterparts but differed from that of egg-grown viruses at amino acid positions 196 to 198. Fluorescence-activated cell sorting analysis showed that although Vero cells possess predominantly alpha2,3 galactose-linked sialic acid, they are fully susceptible to infection with either human influenza A or B viruses. Moreover, all virus-specific polypeptides were synthesized in the same proportions in Vero cells as in MDCK cells. Electron microscopic and immunofluorescence studies confirmed that infected Vero cells undergo the same morphological changes as do other polarized epithelia] cells. Taken together, these results indicate that Vero cell lines could serve as an alternative host system for the cultivation of influenza A and B viruses, providing adequate quantities of either virus to meet the vaccine requirements imposed by an emerging pandemic.
在大流行性流感病毒出现或再次出现后,制备减毒活流感病毒疫苗和大量灭活疫苗将需要一种替代宿主细胞系统,因为鸡胚可能数量不足且并非最佳选择。初步研究表明,非洲绿猴肾细胞系(Vero)是甲型流感病毒初次分离和培养的合适系统(E. A. Govorkova、N. V. Kaverin、L. V. Gubareva、B. Meignier和R. G. Webster,《传染病杂志》172:250 - 253,1995年)。我们现在首次证明Vero细胞适用于乙型流感病毒的分离和有效复制,并确定甲型和乙型流感病毒在Vero细胞中的生物学和遗传学特性;此外,我们将Vero细胞上的受体与犬肾传代细胞(MDCK)上的受体进行了比较。序列分析表明,Vero细胞来源的乙型流感病毒的血凝素与MDCK细胞培养的对应病毒相同,但在氨基酸位置196至198处与鸡胚培养的病毒不同。荧光激活细胞分选分析表明,虽然Vero细胞主要含有α2,3连接的半乳糖唾液酸,但它们对甲型或乙型人流感病毒的感染完全敏感。此外,所有病毒特异性多肽在Vero细胞中的合成比例与在MDCK细胞中的相同。电子显微镜和免疫荧光研究证实,受感染的Vero细胞与其他极化上皮细胞一样会发生相同的形态变化。综上所述,这些结果表明Vero细胞系可作为甲型和乙型流感病毒培养的替代宿主系统,提供足够数量的任何一种病毒以满足新出现的大流行所带来的疫苗需求。