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Title: International network for comparison of HIV neutralization assays: The NeutNet report
Authors: Eva Maria Fenyö
Alan Heath
Stefania Dispinseri
Harvey Holmes
Paolo Lusso
Susan Zolla-Pazner
Helen Donners
Leo Heyndrickx
Jose Alcami
Vera Bongertz
Christian Jassoy
Mauro Malnati
David Montefiori
Christiane Moog
Lynn Morris
Saladin Osmanov
Victoria Polonis
Quentin Sattentau
Hanneke Schuitemaker
Ruengpung Sutthent
Terri Wrin
Gabriella Scarlatti
Instituto de Salud Carlos III
Fundação Oswaldo
IRCCS San Raffaele Scientific Institute
Prins Leopold Instituut voor Tropische Geneeskunde
Lunds Universitet
National Institute for Biological Standards and Control
Universitat Leipzig
Duke University School of Medicine
Universite de Strasbourg
National Institute for Communicable Diseases
Organisation Mondiale de la Sante
Sir William Dunn School of Pathology
University of Amsterdam
Mahidol University
Monogram Biosciences
NYU School of Medicine
National Institute of Allergy and Infectious Diseases
Keywords: Agricultural and Biological Sciences;Biochemistry, Genetics and Molecular Biology;Medicine
Issue Date: 20-Feb-2009
Citation: PLoS ONE. Vol.4, No.2 (2009)
Abstract: Background: Neutralizing antibody assessments play a central role in human immunodeficiency virus type-1 (HIV-1) vaccine development but it is unclear which assay, or combination of assays, will provide reliable measures of correlates of protection. To address this, an international collaboration (NeutNet) involving 18 independent participants was organized to compare different assays. Methods: Each laboratory evaluated four neutralizing reagents (TriMab, 447-52D, 4E10, sCD4) at a given range of concentrations against a panel of 11 viruses representing a wide range of genetic subtypes and phenotypes. A total of 16 different assays were compared. The assays utilized either uncloned virus produced in peripheral blood mononuclear cells (PBMCs) (virus infectivity assays, VI assays), or their Env-pseudotyped (gp160) derivatives produced in 293T cells (PSV assays) from molecular clones or uncloned virus. Target cells included PBMC and genetically-engineered cell lines in either a single-or multiple-cycle infection format. Infection was quantified by using a range of assay read-outs that included extracellular or intracellular p24 antigen detection, RNA quantification and luciferase and beta-galactosidase reporter gene expression. Findings: PSV assays were generally more sensitive than VI assays, but there were important differences according to the virus and inhibitor used. For example, for TriMab, the mean IC50 was always lower in PSV than in VI assays. However, with 4E10 or sCD4 some viruses were neutralized with a lower IC50 in VI assays than in the PSV assays. Inter-laboratory concordance was slightly better for PSV than for VI assays with some viruses, but for other viruses agreement between laboratories was limited and depended on both the virus and the neutralizing reagent. Conclusions: The NeutNet project demonstrated clear differences in assay sensitivity that were dependent on both the neutralizing reagent and the virus. No single assay was capable of detecting the entire spectrum of neutralizing activities. Since it is not known which in vitro assay correlates with in vivo protection, a range of neutralization assays is recommended for vaccine evaluation. © 2009 Fenyö et al.
ISSN: 19326203
Appears in Collections:Scopus 2006-2010

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