Kawana K, Yoshikawa H, Taketani Y, Yoshiike K, Kanda T
Division of Molecular Genetics, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan.
J Virol. 1998 Dec;72(12):10298-300. doi: 10.1128/JVI.72.12.10298-10300.1998.
Lack of permissive and productive cell cultures for the human papillomaviruses (HPVs) has hindered the study of virus-neutralizing antibodies and infection. We developed a cell-free system generating infectious HPV16 pseudovirions. HPV16 L1/L2 capsids, which had been self-assembled in insect cells (Sf9) expressing virion proteins L1 and L2, were disassembled with 2-mercaptoethanol (2-ME), a reducing agent, and reassembled by removal of 2-ME in the presence of a beta-galactosidase expression plasmid. Plasmid DNA purified together with the reassembled capsids was resistant to DNase I digestion. The reassembled pseudovirions mediated DNA transfer to COS-1 cells, as monitored by induced beta-galactosidase activity. Transfer was inhibited by anti-HPV16 L1 antiserum but not by antisera against L1s of HPV6 and HPV18. Construction in vitro of HPV pseudovirions containing marker plasmids would be potentially useful in developing methods to assay virus-neutralizing antibodies and to transfer exogenous genes to HPV-susceptible cells.
缺乏用于人乳头瘤病毒（HPV）的允许性和生产性细胞培养物阻碍了对病毒中和抗体及感染的研究。我们开发了一种无细胞系统来产生感染性HPV16假病毒颗粒。在表达病毒体蛋白L1和L2的昆虫细胞（Sf9）中自组装的HPV16 L1/L2衣壳，用还原剂2-巯基乙醇（2-ME）进行拆解，并在β-半乳糖苷酶表达质粒存在的情况下通过去除2-ME进行重新组装。与重新组装的衣壳一起纯化的质粒DNA对DNase I消化具有抗性。如通过诱导的β-半乳糖苷酶活性监测的那样，重新组装的假病毒颗粒介导了DNA向COS-1细胞的转移。转移受到抗HPV16 L1抗血清的抑制，但不受针对HPV6和HPV18的L1的抗血清的抑制。体外构建含有标记质粒的HPV假病毒颗粒在开发检测病毒中和抗体及将外源基因转移至HPV易感细胞的方法方面可能会很有用。
