Design, Development, and Interpretation of HIV Neutralization Assays

Whalley, Alice; Nguyen, Mai; Morrow, W. J. W.

doi:10.1089/vim.1991.4.201

Cited by 3 publications

(1 citation statement)

References 52 publications

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“…Neutralization assay. A quantitative neutralization assay with HIV-1 laboratory strains was performed as previously described (16,35). Monolayers of CEM-SS target cells were cultured with different HIV strains in the presence or absence of different neutralizing Abs, and 3 to 5 days later, numbers of syncytium-forming units were determined.…”

Section: Methodsmentioning

confidence: 99%

Analysis of the cross-reactive anti-gp120 antibody population in human immunodeficiency virus-infected asymptomatic individuals

Hariharan

Nara

Caralli

et al. 1993

J Virol

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This study was undertaken to analyze the specificity and neutralizing properties of cross-reactive anti-gpl20 antibodies (Abs) in the sera of two human immunodeficiency virus (HIV)-infected asymptomatic individuals. Two panels of murine monoclonal anti-idiotype Abs (anti-id MAbs) were established against cross-reactive polyclonal anti-gpl20 Abs purified from HIV+ sera by sequential affinity chromatography using gpl20sand gpl2OMB-Sepharose columns. These panels of anti-id MAbs were then used to affinity purify idiotype-positive (Id') anti-gpl20 Abs from HIV+ sera. The recovery of each of these Id' Abs by purification indicated that several idiotypically distinct cross-reactive anti-gpl20 Abs are present in sera over a wide range of concentrations. Immunological and biological studies showed that although all of the Id' Abs were reactive

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Section: Methodsmentioning

confidence: 99%

Analysis of the cross-reactive anti-gp120 antibody population in human immunodeficiency virus-infected asymptomatic individuals

Hariharan

Nara

Caralli

et al. 1993

J Virol

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Anti-idiotype monoclonal antibody elicits broadly neutralizing anti-gp120 antibodies in monkeys.

Kang

Nara²,

Chamat³

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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Murine monoclonal antibodies (mAbs) were raised against human, polyclonal, anti-gpl20 antibodies (Abl) and were selected for binding to broadly neutralizing antigpl20 antibodies in sera positive for human immunodeficiency virus (HIV). One anti-idiotype mAb (Ab2), 3C9, was found to be specific for human anti-gpl20 antibodies directed against an epitope around the conserved CD4 attachment site of gpl20. The 3C9 reactive human anti-gpl20 antibodies (3C9' Ab) neutralized MN, huB, RF, and four primary isolates of HIV type 1 (HIV-1). Cynomolgus monkeys were immunized with 3C9 in adjuvant to test whether this anti-idiotype mAb could induce neutralizing anti-gpl20 antibodies. The results show that purified anti-anti-idiotype antibodies (Ab3) from 3C9 immune sera bind to an epitope around the CD4 attachment site of gpl2OSF and gpl2OmII. Furthermore, purified gpl20-specific Ab3 neutralize MN, fIB, and RF isolates. These results demonstrate that primates immunized with an antiidiotype mAb produce broadly neutralizing anti-HIV-1 antibodies. Since this anti-idiotype mAb was selected by identifying a clonotypic marker, its biological activity can be explained as the result of clonotypic B-cell stimulation.The immunodominant neutralizing site against human immunodeficiency virus type 1 (HIV-1) resides in the third hypervariable region (V3) of gpl20. The hypervariability of the amino acid composition outside and within this region suggests that HIV may be able to escape the effects of V3-specific neutralizing antibodies by mutation (1). The second major neutralizing epitope of gpl20 is the CD4 attachment site. A recent report has demonstrated the conformational dependence of this region (2). Furthermore, our study has shown that several epitopes reside around the CD4 attachment site (3). Antibodies specific for these epitopes generally exhibit broad neutralizing activities against multiple strains of HIV-1 (2-5). Thus, the CD4 attachment site of gpl20 is an attractive target for immunotherapy as the problems of antigenic variation may be minimized. However, the natural antibody response to this region is weaker, at least in early infection (1, 2), and the neutralizing activities of the CD4 site-specific antibodies are less potent than that of V3-specific antibodies (6).In the present study, we explored the use of anti-idiotype antibodies, to induce CD4 site-directed, broadly neutralizing antibodies in nonhuman primates. The reason for using this approach was to overcome the limiting immunogenic potential of the CD4 epitopes. We believed that the inherent antigenically imposed restriction associated with virus-based vaccines could be overcome by using antibodies directed against idiotopes on B-cell receptors with specificity for the CD4 attachment site of gpl20. Targeting such B cells with specific anti-idiotype (or anti-clonotype) antibodies may stimulate the synthesis of specific antibodies. In this regard, we utilized broadly neutralizing human polyclonal anti-gpl20 antibodies as template idiotype antibody (Abl) to...

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Recommendations for the Design and Use of Standard Virus Panels To Assess Neutralizing Antibody Responses Elicited by Candidate Human Immunodeficiency Virus Type 1 Vaccines

Mascola

DʼSouza

Gilbert

et al. 2005

J Virol

230

233

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Design, Development, and Interpretation of HIV Neutralization Assays

Cited by 3 publications

References 52 publications

Analysis of the cross-reactive anti-gp120 antibody population in human immunodeficiency virus-infected asymptomatic individuals

Analysis of the cross-reactive anti-gp120 antibody population in human immunodeficiency virus-infected asymptomatic individuals

Anti-idiotype monoclonal antibody elicits broadly neutralizing anti-gp120 antibodies in monkeys.

Recommendations for the Design and Use of Standard Virus Panels To Assess Neutralizing Antibody Responses Elicited by Candidate Human Immunodeficiency Virus Type 1 Vaccines

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