International network for comparison of HIV neutralization assays

The neutnet report II

Leo Heyndrickx, Alan Heath, Enas Sheik-Khalil, Jose Alcami, Vera Bongertz, Marianne Jansson, Mauro Malnati, David Montefiori, Christiane Moog, Lynn Morris, Saladin Osmanov, Victoria Polonis, Meghna Ramaswamy, Quentin Sattentau, Monica Tolazzi, Hanneke Schuitemaker, Betty Willems, Terri Wrin, Eva Maria Fenyö, Gabriella Scarlatti

Research output: Contribution to journalArticle

24 Citations (Scopus)

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.

Original languageEnglish
Article numbere36438
JournalPLoS One
Volume7
Issue number5
DOIs
Publication statusPublished - May 9 2012

Fingerprint

neutralization tests
Viruses
Assays
HIV
Pseudoviridae
viruses
assays
mononuclear leukocytes
Neutralizing Antibodies
neutralizing antibodies
Blood Cells
Blood
vaccines
Immunity
monoclonal antibodies
Marker Vaccines
pathogenicity
Vaccines
immunity
Monoclonal Antibodies

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Heyndrickx, L., Heath, A., Sheik-Khalil, E., Alcami, J., Bongertz, V., Jansson, M., ... Scarlatti, G. (2012). International network for comparison of HIV neutralization assays: The neutnet report II. PLoS One, 7(5), [e36438]. https://doi.org/10.1371/journal.pone.0036438

International network for comparison of HIV neutralization assays : The neutnet report II. / Heyndrickx, Leo; Heath, Alan; Sheik-Khalil, Enas; Alcami, Jose; Bongertz, Vera; Jansson, Marianne; Malnati, Mauro; Montefiori, David; Moog, Christiane; Morris, Lynn; Osmanov, Saladin; Polonis, Victoria; Ramaswamy, Meghna; Sattentau, Quentin; Tolazzi, Monica; Schuitemaker, Hanneke; Willems, Betty; Wrin, Terri; Fenyö, Eva Maria; Scarlatti, Gabriella.

In: PLoS One, Vol. 7, No. 5, e36438, 09.05.2012.

Research output: Contribution to journalArticle

Heyndrickx, L, Heath, A, Sheik-Khalil, E, Alcami, J, Bongertz, V, Jansson, M, Malnati, M, Montefiori, D, Moog, C, Morris, L, Osmanov, S, Polonis, V, Ramaswamy, M, Sattentau, Q, Tolazzi, M, Schuitemaker, H, Willems, B, Wrin, T, Fenyö, EM & Scarlatti, G 2012, 'International network for comparison of HIV neutralization assays: The neutnet report II', PLoS One, vol. 7, no. 5, e36438. https://doi.org/10.1371/journal.pone.0036438
Heyndrickx L, Heath A, Sheik-Khalil E, Alcami J, Bongertz V, Jansson M et al. International network for comparison of HIV neutralization assays: The neutnet report II. PLoS One. 2012 May 9;7(5). e36438. https://doi.org/10.1371/journal.pone.0036438
Heyndrickx, Leo ; Heath, Alan ; Sheik-Khalil, Enas ; Alcami, Jose ; Bongertz, Vera ; Jansson, Marianne ; Malnati, Mauro ; Montefiori, David ; Moog, Christiane ; Morris, Lynn ; Osmanov, Saladin ; Polonis, Victoria ; Ramaswamy, Meghna ; Sattentau, Quentin ; Tolazzi, Monica ; Schuitemaker, Hanneke ; Willems, Betty ; Wrin, Terri ; Fenyö, Eva Maria ; Scarlatti, Gabriella. / International network for comparison of HIV neutralization assays : The neutnet report II. In: PLoS One. 2012 ; Vol. 7, No. 5.
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AU - Morris, Lynn

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