Production of infectious human papillomavirus independently of viral replication and epithelial cell differentiation
Production of infectious human papillomavirus independently of viral replication and epithelial cell differentiation
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DOI：
10.1073/pnas.0504020102
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发表时间：
2005-06-28
影响因子：
11.1
通讯作者：
Ahlquist, P
中科院分区：
文献类型：
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作者：
Pyeon, D;Lambert, PF;Ahlquist, P
乳头瘤病毒是与良性和恶性上皮病变相关的小DNA病毒，包括> 95%的宫颈癌和约20%的头颈癌。由于乳头瘤病毒复制和病毒体产生与上皮细胞分化有关，因此感染性乳头瘤病毒病毒体产生已限于繁琐的器官型培养和小鼠异种移植。获得有用数量的野生型或突变型人乳头瘤病毒（HPV）病毒体的困难极大地限制了对乳头瘤病毒生物学的许多方面的研究。为了克服这些限制，我们开发了一种系统，将全长乳头瘤病毒基因组复制成感染性病毒粒子，独立于病毒DNA复制和上皮分化。这种基于瞬时转染的系统每个细胞培养皿产生的感染性病毒比劳动密集型培养多1,000倍。此外，我们表明，这种方法可以轻松生成含有野生型，突变型或嵌合型乳头瘤病毒基因组的感染性颗粒，克服了研究复制，宿主相互作用以及疫苗和药物开发的许多方面的障碍，这些障碍受到感染性病毒粒子可用性不足的限制。
Papillomavi ruses are small DNA viruses that are associated with benign and malignant epithelial lesions, including > 95 % of cervical cancers and approximate to 20 % of head and neck cancers. Because papillomavirus replication and virion production are tied to epithelial cell differentiation, infectious papillomavirus virion production has been limited to cumbersome organotypic cultures and mouse xenografts. Consequent difficulties in obtaining useful amounts of wild-type or mutant human papillomavirus (HPV) virions have greatly limited studies on many aspects of papillomavirus biology. To overcome these limitations, we developed a system to encapsidate the full-length papillomaviral genome into infectious virions, independently of viral DNA replication and epithelial differentiation. This transient-transfection-based system produces > 1,000 times more infectious virus per cell culture dish than the much more labor-intensive organotypic culture. Furthermore, we show that this method allows the facile generation of infectious particles containing wild-type, mutant, or chimeric papillomaviral genomes, overcoming barriers to studying many facets of replication, host interactions, and vaccine and drug development, which has been limited by the insufficient availability of infectious virions.