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Journal of Clinical Microbiology, March 2002, p. 926-932, Vol. 40, No. 3
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.3.926-932.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Detection of Neutralizing Antibodies against Human Papillomaviruses (HPV) by Inhibition of Gene Transfer Mediated by HPV Pseudovirions

Laboratoire de Virologie Moléculaire, INSERM EMIU 00-10, and USC INRA, Faculté des Sciences Pharmaceutiques,¹ Laboratoire de Microscopie Electronique, Faculté de Médecine,² IFR Biologie Comparée des Transposons et des Virus, Tours, France, and Laboratorio de Immunologia, Instituto Nacional del Cancer, Bogotà, Colombia³

Received 8 June 2001/ Returned for modification 8 November 2001/ Accepted 21 December 2001

The goal of this study was to develop a human papillomavirus (HPV) neutralization assay using HPV pseudovirions generated in vitro. For this purpose, gene transfer efficiency of HPV virus-like particles (VLPs) was improved by using direct interaction between a reporter plasmid and the VLPs. Electron microscopic observation of the interaction between DNA molecules and VLPs revealed that VLPs always interact with a single DNA molecule and that VLPs bind to the end of linearized DNA molecules. An 100-fold improvement in the gene transfer was obtained by simple interaction between a linearized DNA molecule and VLPs. Moreover, direct interaction methods offer the possibility of transferring plasmids a size higher than that of the papillomavirus genome. The approach that we developed to generate HPV-16 and HPV-31 pseudovirions proved to be suitable for testing neutralizing antibodies in human sera both after immunization and after natural infection.

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