Investigating antibody neutralization of lyssaviruses using lentiviral pseudotypes: a cross-species comparison

Wright, E., Temperton, N.J., Marston, D.A., McElhinney, L.M., Fooks, A.R. and Weiss, R.A. 2008. Investigating antibody neutralization of lyssaviruses using lentiviral pseudotypes: a cross-species comparison. Journal of General Virology. 89 (9), pp. 2204-2213.

TitleInvestigating antibody neutralization of lyssaviruses using lentiviral pseudotypes: a cross-species comparison
AuthorsWright, E., Temperton, N.J., Marston, D.A., McElhinney, L.M., Fooks, A.R. and Weiss, R.A.
Abstract

Cross-neutralization between rabies virus (RABV) and two European bat lyssaviruses (EBLV-1 and -2) was analysed using lentiviral pseudotypes as antigen vectors. Glycoprotein (G-protein) cDNA from RABV challenge virus standard-11 (CVS-11) and EBLV-1 and -2 were cloned and co-expressed with human immunodeficiency virus (HIV) or murine leukemia virus (MLV) gag–pol and packageable green fluorescent protein (GFP) or luciferase reporter genes in human cells. The harvested lentiviral (HIV) vector infected over 40 % of baby hamster kidney (BHK) target cells, providing high-titre pseudotype stocks. Tests on blinded antibody-positive (n=15) and -negative (n=45) sera, predetermined by the fluorescent antibody virus neutralization (FAVN) test approved by the World Health Organization (WHO) and Office International des Epizooties (OIE), revealed that the CVS-11 pseudotype assay had 100 % concordance with FAVN and strongly correlated with neutralization titres (r2=0.89). Cross-neutralization tests using sera from RABV-vaccinated humans and animals on pseudotypes with CVS-11, EBLV-1 and EBLV-2 envelopes showed that the relative neutralization titres correlated broadly with the degree of G-protein diversity. Pseudotypes have three major advantages over live-virus neutralization tests: (i) they can be handled in low-biohazard-level laboratories; (ii) the use of reporter genes such as GFP or β-galactosidase will allow the assay to be undertaken at low cost in laboratories worldwide; (iii) each assay requires <10 μl serum. This robust microassay will improve our understanding of the protective humoral immunity that current rabies vaccines confer against emerging lyssaviruses, and will be applicable to surveillance studies, thus helping to control the spread of rabies.

JournalJournal of General Virology
Journal citation89 (9), pp. 2204-2213
ISSN0022-1317
YearSep 2008
PublisherSociety for General Microbiology
FileWright_et_al_2008_as_published.pdf
Digital Object Identifier (DOI)doi:10.1099/vir.0.2008/000349-0
Publication dates
PublishedSep 2008

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