P04-18. Comparison of HIV neutralization assays for use in vaccine research and clinical trials, phase II: results from the NeutNet working group

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P04-18. Comparison of HIV neutralization assays for use in vaccine research and clinical trials, phase II: results from the NeutNet working group
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  BioMed   Central Page 1 of 2 (page number not for citation purposes) Retrovirology  Open Access Poster presentation P04-18. Comparison of HIV neutralization assays for use in vaccine research and clinical trials, phase II: results from the NeutNet  working group GScarlatti* 1 , JAlcami 2 , VBongertz 3 , EFenyö 4 , AHeath 5 , LHeyndrickx  6 , HHolmes 5 , MJansson 4 , LLopalco 1 , MMalnati 1 , DMontefiori 7 , CMoog  8 , LMorris 9 , SOsmanov  10 , VPolonis 11 , MRamaswamy  5 , QSattentau 12 , HSchuitemaker  13  and TWrin 14  Address: 1 Division of Immunology, Transplant and Infectious Diseases, DIBIT-San Raffaele Scientific Institute, Milan, Italy, 2 Centro Nacional de Microbiologia, Instituto de Salude Carlos III,, Madrid, Spain, 3 Department of Immunology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil, 4 OrganizationDepartment of Laboratory Medicine, Lund University, Lund, Sweden, 5 National Institute for Biological Standards and Control, London, UK, 6 Institute of Tropical Medicine, Microbiology Department, Antwerp, Belgium, 7 Duke University, Laboratory for AIDS Vaccine Research and Development, Durham, NC, USA, 8 University Louis Pasteur, Strasbourg, France, 9 National Institute for Communicable Diseases, Johannesburg, South Africa, 10  WHO-UNAIDS HIV Vaccine Initiative, Geneva, Switzerland, 11 Henry Jackson Foundation for the Advancement of Military Medicine, Rockville, MD, USA, 12 University of Oxford, Oxford, UK, 13  Academic Medical Center, Amsterdam, Netherlands and 14 MonogramBiosciences, Inc., South San Francisco, CA, USA * Corresponding author Background In vitro assessment of HIV neutralization for pathogenesisor vaccine efficacy studies is a complex task, attributableto several confounding variables surrounding virus, anti-bodies and host cells employed. NeutNet, a collaborationinvolving 18 independent laboratories from 12 countries,showed during the first phase clear differences in neutral-ization assay sensitivity that were dependent on both theantibody (TriMab, 4E10, sCD4) and the virus used http:/ /www.PlosOne.org . The second phase of NeutNet focusedon testing polyclonal reagents against a panel of viruses with 17 different assays. Methods Each laboratory evaluated TriMab, 8 HIV-positive and oneseronegative sera at a given range of dilutions against 8 viruses representing different subtypes and phenotypes with 17 different assays. Assays utilized uncloned virussupernatant (virus infectivity assays-VIA) or Env-pseudo-typed viruses (PSV assays). Target cells included PBMCsand engineered cell lines in a single- or multiple-cycleinfection format. A range of read-outs were used, whichincluded intra- and extra-cellular p24 detection, and luci-ferase or beta-galactosidase reporter gene expression. Results Neutralization with TriMab showed variation for bothPSV and VI assays when comparing results of phase I andII. Negative serum gave sporadic neutralization in bothtypes of assays only when the inhibitory concentration(IC) 50 was considered. IC50 showed more variation thanIC 75 and 90 for PBMC-based VIA. PSV assays were in gen-eral not more sensitive than VIA. Variation was dependent on both sera and viruses used. Specific assay to assay com-parison showed impact of the target cell used. Conclusion In agreement with our phase I observations, we hereobserved that also for polyclonal agents, the assay condi-tions seem to influence the outcome of HIV-1 neutraliza- from AIDS Vaccine 2009Paris, France. 19–22 October 2009Published: 22 October 2009 Retrovirology   2009, 6 (Suppl 3):P46doi:10.1186/1742-4690-6-S3-P46 <supplement> <title> <p>AIDSVaccine 2009</p> </title><editor>AnnaLauraRoss</editor> <note>Meetingabstracts – Asingle PDF containingallabstracts inthis Supplement is available <ahref="http://www.biomedcentral.com/content/files/pdf/1742-4690-6-S3-full.pdf">here</a>.</note> <url>http://www.biomedcentral.com/content/pdf/1471-2105-10-S12-info.pdf</url></supplement> This abstract is available from: http://www.retrovirology.com/content/6/S3/P46© 2009 Scarlatti et al; licensee BioMed Central Ltd.  Publish with BioMed Central  and every scientist can read your work free of charge "BioMed Central will be the most significant development for disseminating the results of biomedical research in our lifetime." Sir Paul Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical communitypeer reviewed and published immediately upon acceptancecited in PubMed and archived on PubMed Central yours — you keep the copyright Submit your manuscript here: http://www.biomedcentral.com/info/publishing_adv.asp BioMed central Retrovirology   2009, 6 (Suppl 3):P46http://www.retrovirology.com/content/6/S3/P46Page 2 of 2 (page number not for citation purposes) tion in vitro . Since protective neutralizing immunity in vivo is not yet defined, no single assay can be recommended toachieve optimal information on the neutralization poten-tial of a serum or agent.
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