Simple, rapid, quantitative, syncytium-forming microassay for the detection of human immunodeficiency virus neutralizing antibody. NLM AIDSLINE Important note: Information in this article was accurate in 1988. The state of the art may have changed since the publication date.

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Simple, rapid, quantitative, syncytium-forming microassay for the detection of human immunodeficiency virus neutralizing antibody.

AIDS Res Hum Retroviruses. 1987 Fall;3(3):283-302. Unique Identifier : AIDSLINE MED/88134684
Nara PL; Hatch WC; Dunlop NM; Robey WG; Arthur LO; Gonda MA; Fischinger PJ; Virus Control Unit, National Cancer Institute-Frederick Cancer; Research Facility, MD 21701.

Abstract: A simple, rapid, quantitative syncytium-forming microassay for the detection of human immunodeficiency virus (HIV-I) isolates is described. A virus-syncytial sensitive clone of CEM cells (CEM-SS) was identified and made adherent to flat bottom 96-well microtiter dishes. Following the addition of virus, these cells develop easily quantifiable, adherent syncytia on a background of confluent, normal CEM-SS monolayer in 4 to 6 days. One-hit kinetics for syncytia formation were obtained at various multiplicities of infection. Syncytia are associated with complete virion production and cytoplasmic localization of the p24 core protein (detected by immunofluorescence). Total infectious virus can be accurately determined in this assay; these results showed a close correlation with p24 and gp120 induction when microtiter well supernatants were passed to fresh cells and evaluated by competitive radioimmunoassay. Studies of p24 antigen induction at and beyond the end point of syncytia formation indicate that there are no detectable nonsyncytial variants in standard HIV-I stocks. Six divergent HIV-I isolates (HTLV-IIIB, -RFII, -MN, -RUTZ, -CC, and LAV-1), as well as HTLV-IIIB and LAV-1 reisolated from persistently infected chimpanzees, produce quantifiable syncytia which vary slightly in their developmental morphology. Accurate neutralization titers are readily obtained from easily constructed multiplicity curves derived from serial dilutions of test sera. Inherent within this system is a flexible method for studying various kinetics of antibody/virus interactions, as well as blocking and interference studies with any candidate antiviral compounds.
Keywords: Antibodies, Viral/*ANALYSIS Antigens, Viral/ANALYSIS *Cell Fusion Cell Line Cytopathogenic Effect, Viral HIV/*IMMUNOLOGY/PHYSIOLOGY Neutralization Tests Support, U.S. Gov't, P.H.S. Virus Replication JOURNAL ARTICLE

KWDantibodies,viral/KWDanalysisantigens,viral/analysisKWDcellfusioncelllinecytopathogeniceffect,viralhiv/KWDimmunology/physiologyneutralizationtestssupport,uKWDsKWDgov't,pKWDhKWDsKWDvirusreplicationjournalarticle
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M8860444

Copyright © 1988 - National Library of Medicine. Reproduced under license with the National Library of Medicine, Bethesda, MD.

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