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Molecular evolution of hepatitis A virus in a human diploid cell line

World Journal of Gastroenterology 2007年第34期13卷 4630-4635页

作者： Cai-Hua Tang Jiang-Sen Mao Shao-Ai Chai Yong Chen Fang-Cheng Zhuang Institute of Infectious Diseases the First Affiliated Hospital College of Medical Science Zhejiang UniversityHangzhou 310003 Zhejiang Province China Zhejiang Academy of Medical Science Hangzhou 310013 Zh-ejiang'Province China

AIM： To investigate the hoLspoLs, direction, and the time course of evolution of hepatitis A virus in the process of consecutive cell culture passage in human KMB17 diploid cells.METHODS： Wild type hepatitis A virus H2w was serially propagated in KMB17 cells until passage 30, and the fulllength genomes of H2w and its six chosen progenies were determined by directly sequencing RT-PCR products amplified from viral genomic RNA. Alignment comparison of sequences from H2w with its six progenies and phylogenetic analysis of the whole VP1 region from H2w, progenies of H2w, and other cell culture adapted hepatitis A virus were then carried out to obtain data on the molecular evolution of hepatitis A virus in the process of consecutive passage in KMB17 cells.RESULTS： Most of the mutations occurred by passage 5 and several hotspots related to adaptation of the virus during cell growth were observed. After that stage, few additional mutations occurred through the remaining duration of passage in KMB17 cells except for mutation in the virulence determinants, which occurred in the vicinity of passage 15. The phylogenetic analysis of the whole VP1 region suggested that the progenies of H2w evolved closely to other cell culture adapted hepatitis A virus, i.e. MBB, L-A-l, other than its progenitor H2w.CONCLUSION： Hepatitis A virus served as a useful model for studying molecular evolution of viruses in a given environment. The information obtained in this study may provide assistance in cultivating the next generation of a seed virus for live hepatitis A vaccine production.

关键词： Molecular evolution Virus evolution phylogenyanalysis Virulence gene Hepat’s A virus

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