Detection of neutralizing antibodies against human papillomaviruses (HPV) by inhibition of gene transfer mediated by HPV pseudovirions.
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.
Bousarghin
Latifa
L
Laboratoire de Virologie Moléculaire, INSERM EMIU 00-10, Tours, France.
Combita-Rojas
Alba-Lucia
AL
Touzé
Antoine
A
El Mehdaoui
Slimane
S
Sizaret
Pierre-Yves
PY
Bravo
Maria-Mercedes
MM
Coursaget
Pierre
P
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D022761:Gene Transfer, Horizontal; D006801:Humans; D007114:Immunization; D007223:Infant; D051379:Mice; D008854:Microscopy, Electron; D027383:Papillomaviridae / Q000235:genetics / Q000276:immunology*; D030361:Papillomavirus Infections / Q000276:immunology; D014771:Virion / Q000235:genetics*
D000914:Antibodies, Viral; D004279:DNA, Viral