In vitro generation and type-specific neutralization of a human papillomavirus type 16 virion pseudotype

Richard S Roden, Heather L. Greenstone, Reinhard Kirnbauer, Frank P. Booy, Joel Jessie, Douglas R. Lowy, John T. Schiller

Research output: Contribution to journalArticle

Abstract

We report a system for generating infectious papillomaviruses in vitro that facilitates the analysis of papillomavirus assembly, infectivity, and serologic relatedness. Cultured hamster BPHE-1 cells harboring autonomously replicating bovine papillomavirus type 1 (BPV1) genomes were infected with recombinant Semliki Forest viruses that express the structural proteins of BPV1. When plated on C127 cells, extracts from cells expressing L1 and L2 together induced numerous transformed foci that could be specifically prevented by BPV neutralizing antibodies, demonstrating that BPV infection was responsible for the focal transformation. Extracts from BPHE-1 cells expressing L1 or L2 separately were not infectious. Although Semliki Forest virus-expressed L1 self-assembled into virus-like particles (VLPs), viral DNA was detected in particles only when L2 was coexpressed with L1, indicating that genome encapsidation requires L2. Expression of human papillomavirus type 16 (HPV16) L1 and L2 together in BPHE-1 cells also yielded infectious virus. These pseudotyped virions were neutralized by antiserum to HPV16 VLPs derived from European (114/K) or African (Z-1194) HPV16 variants but not by antisera to BPV VLPs, to a poorly assembling mutant HPV16 L1 protein, or to VLPs of closely related genital HPV types. Extracts from BPHE-1 cells coexpressing BPV L1 and HPV16 L2 or HPV16 L1 and BPV L2 were not infectious. We conclude that (i) mouse C127 cells express the cell surface receptor for HPV16 and are able to uncoat HPV16 capsids; (ii) if a papillomavirus DNA packaging signal exists, then it is conserved between the BPV and HPV16 genomes; (iii) functional L1-L2 interaction exhibits type specificity; and (iv) protection by HPV virus-like particle vaccines is likely to be type specific.

Original languageEnglish (US)
Pages (from-to)5875-5883
Number of pages9
JournalJournal of Virology
Volume70
Issue number9
StatePublished - Sep 1996
Externally publishedYes

Fingerprint

Human papillomavirus 16
Papillomaviridae
virion
neutralization
Virion
virus-like particles
Bovine papillomavirus 1
Semliki forest virus
Genome
Semliki Forest virus
cells
Immune Sera
Virus-Like Particle Vaccines
DNA Packaging
Viral Structural Proteins
Papillomavirus Vaccines
genome
antiserum
In Vitro Techniques
Capsid

ASJC Scopus subject areas

  • Immunology

Cite this

Roden, R. S., Greenstone, H. L., Kirnbauer, R., Booy, F. P., Jessie, J., Lowy, D. R., & Schiller, J. T. (1996). In vitro generation and type-specific neutralization of a human papillomavirus type 16 virion pseudotype. Journal of Virology, 70(9), 5875-5883.

In vitro generation and type-specific neutralization of a human papillomavirus type 16 virion pseudotype. / Roden, Richard S; Greenstone, Heather L.; Kirnbauer, Reinhard; Booy, Frank P.; Jessie, Joel; Lowy, Douglas R.; Schiller, John T.

In: Journal of Virology, Vol. 70, No. 9, 09.1996, p. 5875-5883.

Research output: Contribution to journalArticle

Roden, RS, Greenstone, HL, Kirnbauer, R, Booy, FP, Jessie, J, Lowy, DR & Schiller, JT 1996, 'In vitro generation and type-specific neutralization of a human papillomavirus type 16 virion pseudotype', Journal of Virology, vol. 70, no. 9, pp. 5875-5883.
Roden RS, Greenstone HL, Kirnbauer R, Booy FP, Jessie J, Lowy DR et al. In vitro generation and type-specific neutralization of a human papillomavirus type 16 virion pseudotype. Journal of Virology. 1996 Sep;70(9):5875-5883.
Roden, Richard S ; Greenstone, Heather L. ; Kirnbauer, Reinhard ; Booy, Frank P. ; Jessie, Joel ; Lowy, Douglas R. ; Schiller, John T. / In vitro generation and type-specific neutralization of a human papillomavirus type 16 virion pseudotype. In: Journal of Virology. 1996 ; Vol. 70, No. 9. pp. 5875-5883.
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