Infection of HeLa cells by avian infectious bronchitis virus is dependent on cell status
Infection of HeLa cells by avian infectious bronchitis virus is dependent on cell status
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DOI：
10.1080/03079450701447291
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发表时间：
2007-01-01
期刊：
影响因子：
2.8
通讯作者：
Chen, H. C.
中科院分区：
文献类型：
--
作者：
Chen, H. Y.;Guo, A. Z.;Chen, H. C.
研究鸡传染性支气管炎病毒（IBV）在人细胞系中的适应性，有助于了解冠状病毒种间感染的可能机制。目前的研究解决了以前的报告中证实的HeLa人细胞系中IBV的复制不良。结果表明，IBV M41、H52、H120和Gray株能在HeLa细胞中增殖，并产生明显的细胞病变。新鲜分散的HeLa细胞中的病毒滴度比细胞单层中的病毒滴度高1000倍。胰蛋白酶不是病毒复制的决定因子，表明病毒在HeLa细胞中的复制受到限制是细胞内事件的结果，而不是与宿主细胞结合或融合的结果。这些IBV毒株在新鲜分散的HeLa细胞中复制到10（3.4 +/-0.2）/0.1 ml中值组织培养感染剂量的平均滴度，并在前12代中保持该滴度。然后在第13代中出现约10倍增加（10（4.20 +/- 0.19）/0.1 ml），并维持至第16代，之后在第17代中再次出现更大幅度的增加至10（6.6 +/- 0.3/）0.1 ml。该滴度保持至第24代，此时实验终止。扩增IBV M41 S1基因，并对第0、5和21代进行测序。在第21代，S1蛋白中只有一个氨基酸替换。HeLa细胞上唾液酸的存在有助于有效的病毒复制，而人氨肽酶N不参与感染。随着传代次数的增加，血凝素活性逐渐降低。这些结果表明，病毒的适应性可能是由宿主细胞的修饰，如受体糖基化和不同的受体利用，而不是病毒基因突变决定的。
To investigate the adaptation of avian infectious bronchitis virus (IBV) in a human cell line may be beneficial to understanding the potential mechanisms of coronavirus interspecies infection. The current study addressed the poor replication of IBV in the HeLa human cell line demonstrated in previous reports. We showed that IBV strains M41, H52, H120 and Gray could be propagated in HeLa cells with distinct cytopathic effect. The virus titre in freshly dispersed HeLa cells was 1000-fold higher than in cell monolayers. Trypsin was not the determinant for the viral replication, suggesting that the restriction of IBV replication in HeLa cells is the result of intracellular events rather than the binding to or fusion with host cells. These IBV strains replicated to an average titre of 10(3.4 +/- 0.2)/0.1 ml median tissue culture infectious doses in freshly dispersed HeLa cells and maintained this titre for the first 12 passages. Then an approximately 10-fold increase (10(4.20 +/- 0.19)/0.1 ml) occurred in passage 13, which was maintained to passage 16, after which there was another, bigger rise to 10(6.6 +/- 0.3/)0.1 ml in passage 17. This titre was maintained until passage 24 when the experiment was terminated. The IBV M41 S1 gene was amplified and sequenced for passages 0, 5 and 21. There was only one amino acid replacement in the S1 protein, in passage 21. The presence of sialic acid on HeLa cells contributed to efficient virus replication, while human aminopeptidase N was not involved in the infection. Haemagglutinin activity gradually reduced with increased passages. These results indicated that the virus adaptation would probably be determined by host cell modification such as receptor glycosylation and different receptor utilization instead of viral gene mutation.