title: Robust HPV-18 production in organotypic cultures of primary human keratinocytes.
abstract: The productive program of the human papillomaviruses takes place in terminally differentiating squamous epithelia. In this chapter, we provide the protocols for robust production of HPV-18 in organotypic cultures of early passages of primary human keratinocytes. A critical step is the generation of genomic HPV plasmids in vivo by using Cre-loxP-mediated excisional recombination from a vector plasmid. We discuss the rationale for this approach. This system produces high yields of infectious virus and facilitates genetic analyses of HPV protein functions and their regulation in the context of recapitulated host tissue environment.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL, 35294-0005, USA.
authors.forename: Hsu-Kun
authors.initials: HK
authors.lastname: Wang
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Broker
authors.forename: Louise T
authors.initials: LT
authors.lastname: Chow
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics; D036022:Capsid Proteins / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D016679:Genome, Viral; D052161:Human papillomavirus 18 / Q000254:growth & development*; D006801:Humans; D015603:Keratinocytes / Q000821:virology*; D051379:Mice; D010957:Plasmids / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D014771:Virion / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D004279:DNA, Viral