Production of Furin-Cleaved Papillomavirus Pseudovirions and Their Use for In Vitro Neutralization Assays of L1- or L2-Specific Antibodies.

Abstract

Immunization with Human Papillomavirus (HPV) L1 virus-like particles or L2 capsid protein elicits neutralizing antibodies that mediate protection. A high-throughput and sensitive in vitro neutralization assay is therefore valuable for prophylactic HPV vaccine studies. Over several hours during infection of the genital tract, virions take on a distinct intermediate conformation, including a required furin cleavage of L2 at its N-terminus. This intermediate is an important target for neutralization by L2-specific antibody, but it is very transiently exposed during in vitro infection of most cell lines resulting in insensitive measurement for L2, but not L1-specific neutralizing antibodies. To model this intermediate, we describe a protocol to generate furin-cleaved HPV pseudovirions (fc-PsV), which deliver an encapsidated reporter plasmid to facilitate infectivity measurements. We also describe a protocol for use of fc-PsV in a high-throughput in vitro neutralization assay for the sensitive measurement of both L1 and L2-specific neutralizing antibodies.

• Authors: Joshua W Wang, Ken Matsui, Yuanji Pan, Kihyuck Kwak, Shiwen Peng, Troy Kemp, Ligia Pinto, Richard B S Roden
• Journal: Current protocols in microbiology (Curr Protoc Microbiol) Vol. 38 Pg. 14B.5.1-26 (Aug 03 2015) ISSN: 1934-8533 [Electronic] United States
• PMID: 26237105 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2015 John Wiley & Sons, Inc.

Chemical References

• Antibodies, Neutralizing
• Antibodies, Viral
• Virosomes
• Furin

Topics

• Antibodies, Neutralizing (immunology)
• Antibodies, Viral (immunology)
• Furin (metabolism)
• High-Throughput Screening Assays
• Humans
• Neutralization Tests (methods)
• Papillomaviridae (genetics, immunology)
• Virosomes (genetics, immunology)