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Reference: Fenyo, EM, Heath, A, Dispinseri, S et al., (2009). International network for comparison of HIV neutralization assays: the NeutNet report. PloS one, 4 (2), e4505.Citable link to this page:


International network for comparison of HIV neutralization assays: the NeutNet report.

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.

Peer Review status:Peer reviewedPublication status:PublishedVersion:Publisher's version Funder: European Community   Funder: WHO/UNAIDS HIV Vaccine Initiative   Funder: The Bill and Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery   Funder: Departamento de DST, Aids e Hepatites Virais, MS-Brasil   Funder: EUROPRISE Network of Excellence of the EU   Funder: Instituto de Salud Carlos III   Funder: FIPSE Foundation   Funder: Fund for Scientific Research in Belgium   Funder: Istituto Superiore di Sanita -VI Programma Nazionale   Funder: Swedish Research Council   Funder: Swedish International Development Cooperation Agency/Department for Research Cooperation   Funder: International AIDS Vaccine Initiative (IAVI) Neutralizing Antibody Consortium (NAC)   Funder: National Institutes of Health   Funder: New York University Center for AIDS Research   Notes:Copyright 2009 Fenyo et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Bibliographic Details

Publisher: Public Library of Science

Publisher Website:

Journal: PloS onesee more from them

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Issue Date: 2009


Urn: uuid:f486d928-34e4-4478-8b0b-77b3aa5f1ce2

Source identifier: 18368

Eissn: 1932-6203


Issn: 1932-6203 Item Description

Type: Journal article;

Language: eng

Version: Publisher's versionKeywords: Humans HIV Infections Neutralization Tests Indicators and Reagents International Cooperation Tiny URL: pubs:18368


Autor: Fenyo, EM - - - Heath, A - - - Dispinseri, S - - - Holmes, H - - - Lusso, P - - - Zolla-Pazner, S - - - Donners, H - - - Heyndric



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