International network for comparison of HIV neutralization assays: the NeutNet report II.Report as inadecuate

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Reference: Heyndrickx, L, Heath, A, Sheik-Khalil, E et al., (2012). International network for comparison of HIV neutralization assays: the NeutNet report II. PloS one, 7 (5), Article: e36438.Citable link to this page:


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

Abstract: BACKGROUND: Neutralizing antibodies provide markers for vaccine-induced protective immunity in many viral infections. By analogy, HIV-1 neutralizing antibodies induced by immunization may well predict vaccine effectiveness. Assessment of neutralizing antibodies is therefore of primary importance, but is hampered by the fact that we do not know which assay(s) can provide measures of protective immunity. An international collaboration (NeutNet) involving 18 different laboratories previously compared different assays using monoclonal antibodies (mAbs) and soluble CD4 (Phase I study). METHODS: In the present study (Phase II), polyclonal reagents were evaluated by 13 laboratories. Each laboratory evaluated nine plasmas against an 8 virus panel representing different genetic subtypes and phenotypes. TriMab, a mixture of three mAbs, was used as a positive control allowing comparison of the results with Phase I in a total of nine different assays. The assays used either uncloned virus produced in peripheral blood mononuclear cells (PBMCs) (Virus Infectivity Assays, VIA), or Env (gp160)-pseudotyped viruses (pseudoviruses, PSV) produced in HEK293T cells from molecular clones or from uncloned virus. Target cells included PBMC and genetically engineered cell lines in either single- or multiple-cycle infection format. Infection was quantified by using a range of assay read-outs including extra- or intra-cellular p24 antigen detection, luciferase, beta-galactosidase or green fluorescent protein (GFP) reporter gene expression. FINDINGS: Using TriMab, results of Phase I and Phase II were generally in agreement for six of the eight viruses tested and confirmed that the PSV assay is more sensitive than PBMC (p = 0.014). Comparisons with the polyclonal reagents showed that sensitivities were dependent on both virus and plasma. CONCLUSIONS: Here we further demonstrate clear differences in assay sensitivities that were dependent on both the neutralizing reagent and the virus. Consistent with the Phase I study, we recommend parallel use of PSV and VIA for vaccine evaluation.

Peer Review status:Peer reviewedPublication status:PublishedVersion:Publisher's version Funder: European Community   Funder: WHO/UNAIDS HIV Vaccine Initiative   Funder: Bill and Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery   Funder: Departamento de DST, Aids e Hepatites Virais, MS-Brasil   Notes:Copyright 2012 Heyndrickx 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: 2012

pages:Article: e36438Identifiers

Urn: uuid:f9d3f0b0-355b-4e4e-805e-a051d18f5a1f

Source identifier: 332748

Eissn: 1932-6203


Issn: 1932-6203 Item Description

Type: Journal article;

Language: eng

Version: Publisher's versionKeywords: Humans Hela Cells HIV Infections Biological Assay AIDS Vaccines HIV Antibodies Sensitivity and Specificity HIV-1 Antibodies, Neutralizing Female Male Tiny URL: pubs:332748


Author: Heyndrickx, L - - - Heath, A - - - Sheik-Khalil, E - - - Alcami, J - - - Bongertz, V - - - Jansson, M - - - Malnati, M - - - Mont



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