NDLI: Production of infectious bovine papillomavirus from cloned viral DNA by using an organotypic raft/xenograft technique

Content Provider	PubMed Central
Author	Mcbride, Alison A. Dlugosz, Andrzej Baker, Carl C.
Copyright Year	2000
Abstract	Bovine papillomavirus type 1 (BPV-1) induces fibropapillomas in its natural host and can transform fibroblasts in culture. The viral genome is maintained as an episome within fibroblasts, which has allowed extensive genetic analyses of the viral functions required for DNA replication, gene expression, and transformation. Much less is known about BPV-1 gene expression and replication in bovine epithelial cells because the study of the complete viral life cycle requires an experimental system capable of generating a fully differentiated stratified bovine epithelium. Using a combination of organotypic raft cultures and xenografts on nude mice, we have developed a system in which BPV-1 can replicate and produce infectious viral particles. Organotypic cultures were established with bovine keratinocytes plated on a collagen raft containing BPV-1-transformed fibroblasts. These keratinocytes were infected with virus particles isolated from a bovine wart or were transfected with cloned BPV-1 DNA. Several days after the rafts were lifted to the air interface, they were grafted on nude mice. After 6–8 weeks, large xenografts were produced that exhibited a hyperplastic and hyperkeratotic epithelium overlying a large dermal fibroma. These lesions were strikingly similar to a fibropapilloma caused by BPV-1 in the natural host. Amplified viral DNA and capsid antigens were detected in the suprabasal cells of the epithelium. Moreover, infectious virus particles could be isolated from these lesions and quantitated by a focus formation assay on mouse cells in culture. Interestingly, analysis of grafts produced with infected and uninfected fibroblasts indicated that the fibroma component was not required for productive infection or morphological changes characteristic of papillomavirus-infected epithelium. This system will be a powerful tool for the genetic analysis of the roles of the viral gene products in the complete viral life cycle.
Starting Page	5534
File Format	PDF
ISSN	10916490
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	10
Volume Number	97
Language	English
Publisher	The National Academy of Sciences
Publisher Date	2000-05-09
Access Restriction	Open
Rights Holder	The National Academy of Sciences
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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Production of infectious bovine papillomavirus from cloned viral DNA by using an organotypic raft/xenograft technique.

Mouse cells transformed by bovine papillomavirus contain only extrachromosomal viral DNA sequences.

Mouse cells transformed by bovine papillomavirus contain only extrachromosomal viral DNA sequences.

Bovine papillomavirus contains multiple transforming genes.

DNA bending is induced in an enhancer by the DNA-binding domain of the bovine papillomavirus E2 protein.

DNA bending is induced in an enhancer by the DNA-binding domain of the bovine papillomavirus E2 protein.

The E2 'gene' of bovine papillomavirus encodes an enhancer-binding protein.

Identification of a second transforming region in bovine papillomavirus DNA.

Infectious RNA derived by transcription from cloned cDNA copies of the genomic RNA of an insect virus.

Production of infectious bovine papillomavirus from cloned viral DNA by using an organotypic raft/xenograft technique

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Production of infectious bovine papillomavirus from cloned viral DNA by using an organotypic raft/xenograft technique.

Mouse cells transformed by bovine papillomavirus contain only extrachromosomal viral DNA sequences.

Mouse cells transformed by bovine papillomavirus contain only extrachromosomal viral DNA sequences.

Bovine papillomavirus contains multiple transforming genes.

DNA bending is induced in an enhancer by the DNA-binding domain of the bovine papillomavirus E2 protein.

DNA bending is induced in an enhancer by the DNA-binding domain of the bovine papillomavirus E2 protein.

The E2 'gene' of bovine papillomavirus encodes an enhancer-binding protein.

Identification of a second transforming region in bovine papillomavirus DNA.

Infectious RNA derived by transcription from cloned cDNA copies of the genomic RNA of an insect virus.

Production of infectious bovine papillomavirus from cloned viral DNA by using an organotypic raft/xenograft technique