International Network for Comparison of HIV Neutralization Assays: The NeutNet Report

被引：106

作者：

Fenyo, Eva Maria ^{[1
]}

Heath, Alan ^{[6
]}

Dispinseri, Stefania ^{[4
]}

Holmes, Harvey ^{[6
]}

Lusso, Paolo ^{[8
]}

Zolla-Pazner, Susan ^{[18
]}

Donners, Helen ^{[5
]}

Heyndrickx, Leo ^{[5
]}

Alcami, Jose ^{[2
]}

Bongertz, Vera ^{[3
]}

Jassoy, Christian ^{[7
]}

Malnati, Mauro ^{[8
]}

Montefiori, David ^{[9
]}

Moog, Christiane ^{[10
]}

Morris, Lynn ^{[11
]}

Osmanov, Saladin ^{[12
]}

Polonis, Victoria ^{[13
]}

Sattentau, Quentin ^{[14
]}

Schuitemaker, Hanneke ^{[15
]}

Sutthent, Ruengpung ^{[16
]}

Wrin, Terri ^{[17
]}

Scarlatti, Gabriella ^{[4
]}

机构：

[1] Lund Univ, Dept Microbiol Dermatol & Infect, Lund, Sweden

[2] Inst Salud Carlos III, Unidad Immunopatol SIDA, Madrid, Spain

[3] Fdn Oswaldo Crusz, Lab AIDS & Mol Immunol, Rio De Janeiro, Brazil

[4] Ist Sci San Raffaele, Viral Evolut & Transmission Unit, Milan, Italy

[5] Inst Trop Med, Dept Microbiol, Virol Unit, Antwerp, Belgium

[6] Natl Inst Biol Standards & Control, Potters Bar, Herts, England

[7] Univ Leipzig, Inst Virol, Leipzig, Germany

[8] Ist Sci San Raffaele, Unit Human Virol, Milan, Italy

[9] Duke Univ, Med Ctr, Dept Surg, Durham, NC 27706 USA

[10] Univ Louis Pasteur, F-67070 Strasbourg, France

[11] Natl Inst Communicable Dis, Johannesburg, South Africa

[12] WHO, IVR HVI, Geneva, Switzerland

[13] Henry Jackson Fdn Advancement Military Med, Dept Vaccine Res, Rockville, MD USA

[14] Univ Oxford, Sir William Dunn Sch Pathol, Oxford OX1 2JD, England

[15] Univ Amsterdam, Academ Med Ctr, CINIMA, Dept Expt Immunol, Landsteiner Lab, NL-1012 WX Amsterdam, Netherlands

[16] Mahidol Univ, Siriraj Hosp, Natl HIV Repository & Bioinformat Ctr, Fac Microbiol, Bangkok, Thailand

[17] Monogram Biosci Inc, San Francisco, CA USA

[18] NYU, Sch Med, New York, NY USA

来源：

PLOS ONE | 2009年 / 4卷 / 02期

基金：

瑞典研究理事会;

关键词：

D O I：

10.1371/journal.pone.0004505

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：12