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PMID: 7678308 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Antibodies to discontinuous or conformationally sensitive epitopes on the gp120 glycoprotein of human immunodeficiency virus type 1 are highly prevalent in sera of infected humans.

Journal of virology ·Vol. 67 ·No. 2 ·1993-02-00 ·Pages 863-75

Moore JP, Ho DD

Abstract

We have used an indirect-capture enzyme-linked immunosorbent assay to quantitate the reactivity of sera from human immunodeficiency virus type 1 (HIV-1)-infected humans with native recombinant gp120 (HIV-1 IIIB or SF-2) or with the gp120 molecule (IIIB or SF-2) denatured by being boiled in the presence of dithiothreitol with or without sodium dodecyl sulfate. Denaturation of IIIB gp120 reduced the titers of sera from randomly selected donors by at least 100-fold, suggesting that the majority of cross-reactive anti-gp120 antibodies present are directed against discontinuous or otherwise conformationally sensitive epitopes. When SF-2 gp120 was used, four of eight serum samples reacted significantly with the denatured protein, albeit with ca. 3- to 50-fold reductions in titer. Only those sera reacting with denatured SF-2 gp120 bound significantly to solid-phase-adsorbed SF-2 V3 loop peptide, and none bound to IIIB V3 loop peptide. Almost all antibody binding to reduced SF-2 gp120 was blocked by preincubation with the SF-2 V3 loop peptide, as was about 50% of the binding to native SF-2 gp120. When sera from a laboratory worker or a chimpanzee infected with IIIB were tested, the pattern of reactivity was reversed, i.e., there was significant binding to reduced IIIB gp120, but not to reduced SF-2 gp120. Binding of these sera to reduced IIIB gp120 was 1 to 10% that to native IIIB gp120 and was substantially decreased by preincubation with IIIB (but not SF-2) V3 loop peptide. To analyze which discontinuous or conformational epitopes were predominant in HIV-1-positive sera, we prebound monoclonal antibodies (MAbs) to IIIB gp120 and then added alkaline phosphatase-labelled HIV-1-positive sera. MAbs (such as 15e) that recognize discontinuous epitopes and compete directly with CD4 reduced HIV-1-positive sera binding by about 50%, whereas neutralizing MAbs to the C4, V2, and V3 domains of gp120 were either not inhibitory or only weakly so. Thus, antibodies to the discontinuous CD4-binding site on gp120 are prevalent in HIV-1-positive sera, antibodies to linear epitopes are less common, most of the antibodies to linear epitopes are directed against the V3 region, and most cross-reactive antibodies are directed against discontinuous epitopes, including regions involved in CD4 binding.

MeSH Terms
Amino Acid Sequence Antibodies, Monoclonal Binding, Competitive CD4 Antigens/immunology Cross Reactions Epitopes HIV Antibodies/blood,immunology HIV Envelope Protein gp120/immunology HIV Seropositivity/immunology HIV-1/immunology Humans Molecular Sequence Data New York City/epidemiology Peptide Fragments/immunology Protein Conformation Protein Denaturation/immunology Virion/immunology
Chemicals
Antibodies, Monoclonal CD4 Antigens Epitopes HIV Antibodies HIV Envelope Protein gp120 Peptide Fragments
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Moore J P
Aaron Diamond AIDS Research Center, New York University School of Medicine, New York, New York 10016.
Ho D D
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.67.2.863-875.1993
Published
1993-02-00
Pages
863-75
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237440
Subset
IM
Grants
NIAID NIH HHS · 1 P30 AI27742-04 · United States
External Links
PubMed source DOI: 10.1128/JVI.67.2.863-875.1993 PMC Full Text View journal details
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