Biryukov Jennifer, Cruz Linda, Ryndock Eric J, Meyers Craig
Department of Microbiology and Immunology, College of Medicine, The Pennsylvania State University, Mailbox H107, 500 University Drive, Hershey, PA, 17033, USA.
Methods Mol Biol. 2015;1249:317-31. doi: 10.1007/978-1-4939-2013-6_24.
In a natural infection, human papillomavirus (HPV) replicates in a stratified and differentiated epithelium. We have developed an in vitro organotypic raft culture system that allows researchers to study HPV in its natural environment. Not only does this system reproduce the differentiation-dependent replication cycle of HPV, but it also allows for the production of high titers of native HPV virions. Currently, much of the HPV research has been done utilizing synthetic particles produced in transfection systems. However, by production of native virions, this research can now be continued using native particles. This chapter presents methods for producing, titering, and qualitating, via infectivity assay, native virus produced from organotypic raft culture.
在自然感染中，人乳头瘤病毒（HPV）在分层且分化的上皮细胞中复制。我们开发了一种体外器官型筏式培养系统，使研究人员能够在其自然环境中研究HPV。该系统不仅重现了HPV依赖分化的复制周期，还能产生高滴度的天然HPV病毒粒子。目前，许多HPV研究是利用转染系统产生的合成颗粒进行的。然而，通过生产天然病毒粒子，现在可以继续使用天然颗粒开展这项研究。本章介绍了通过感染性测定法，对从器官型筏式培养物中产生的天然病毒进行生产、滴定和定性的方法。
