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

Wright, Edward and Temperton, Nigel J. and Marston, Denise A. and McElhinney, Lorraine M. and Fooks, Anthony R. and Weiss, Robin A. (2008) Investigating antibody neutralization of lyssaviruses using lentiviral pseudotypes: A cross-species comparison. Journal of General Virology, 89 (9). pp. 2204-2213. ISSN 0022-1317. (Full text available)

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http://dx.doi.org/10.1099/vir.0.2008/000349-0

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) gagpol 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=515) and -negative (n=545) 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. © 2008 Crown Copyright.

Item Type: Article
Uncontrolled keywords: beta galactosidase, complementary DNA, glycoprotein, green fluorescent protein, lentivirus vector, luciferase, neutralizing antibody, G protein, vesicular stomatitis virus, membrane protein, rabies vaccine, virus antibody, virus envelope protein, virus protein, article, cell strain BHK, controlled study, correlation analysis, cross reaction, enzyme assay, human, human cell, Human immunodeficiency virus, humoral immunity, molecular cloning, Murine leukemia virus, nonhuman, nucleotide sequence, priority journal, Rabies virus, reporter gene, species comparison, virus immunity, virus neutralization, animal, bat, biosynthesis, cat, cell line, comparative study, dog, genetic transfection, genetics, hamster, immunology, Lentivirinae, mouse, serodiagnosis, species difference, virology, Animalia, Cricetinae, Human immunodeficiency virus, Murine leukemia virus, Rabies virus, Animals, Antibodies, Viral, Cats, Cell Line, Chiroptera, Cricetinae, Dogs, Humans, Lentivirus, Lyssavirus, Membrane Glycoproteins, Mice, Neutralization Tests, Rabies Vaccines, Rabies virus, Species Specificity, Transfection, Viral Envelope Proteins, Viral Proteins
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Faculties > Science Technology and Medical Studies > Medway School of Pharmacy
Depositing User: Nigel Temperton
Date Deposited: 02 Aug 2012 10:35
Last Modified: 10 Jun 2014 08:08
Resource URI: http://kar.kent.ac.uk/id/eprint/29995 (The current URI for this page, for reference purposes)
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