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- Alpert, Michael D. (author)
A Novel Assay for Antibody-Dependent Cell-Mediated Cytotoxicity against HIV-1-or SIV-Infected Cells Reveals Incomplete Overlap with Antibodies Measured by Neutralization and Binding Assays
- Article/chapterEnglish2012
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- 2012
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- LIBRIS-ID:oai:lup.lub.lu.se:196efeee-2bf6-48c4-9d00-4f1d5a41c84e
- https://lup.lub.lu.se/record/3283606URI
- https://doi.org/10.1128/JVI.01650-12DOI
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- Language:English
- Summary in:English
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- The resistance of human immunodeficiency virus type 1 (HIV-1) to antibody-mediated immunity often prevents the detection of antibodies that neutralize primary isolates of HIV-1. However, conventional assays for antibody functions other than neutralization are suboptimal. Current methods for measuring the killing of virus-infected cells by antibody-dependent cell-mediated cytotoxicity (ADCC) are limited by the number of natural killer (NK) cells obtainable from individual donors, donor-to-donor variation, and the use of nonphysiological targets. We therefore developed an ADCC assay based on NK cell lines that express human or macaque CD16 and a CD4(+) T-cell line that expresses luciferase from a Tat-inducible promoter upon HIV-1 or simian immunodeficiency virus (SIV) infection. NK cells and virus-infected targets are mixed in the presence of serial plasma dilutions, and ADCC is measured as the dose-dependent loss of luciferase activity. Using this approach, ADCC titers were measured in plasma samples from HIV-infected human donors and SIV-infected macaques. For the same plasma samples paired with the same test viruses, this assay was approximately 2 orders of magnitude more sensitive than optimized assays for neutralizing antibodies-frequently allowing the measurement of ADCC in the absence of detectable neutralization. Although ADCC correlated with other measures of Env-specific antibodies, neutralizing and gp120 binding titers did not consistently predict ADCC activity. Hence, this assay affords a sensitive method for measuring antibodies capable of directing ADCC against HIV- or SIV-infected cells expressing native conformations of the viral envelope glycoprotein and reveals incomplete overlap of the antibodies that direct ADCC and those measured in neutralization and binding assays.
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- Heyer, Lisa N. (author)
- Williams, David E. J. (author)
- Harvey, Jackson D. (author)
- Greenough, Thomas (author)
- Allhorn, MariaLund University,Lunds universitet,Infektionsmedicin,Sektion III,Institutionen för kliniska vetenskaper, Lund,Medicinska fakulteten,Infection Medicine (BMC),Section III,Department of Clinical Sciences, Lund,Faculty of Medicine(Swepub:lu)medk-mal (author)
- Evans, David T. (author)
- InfektionsmedicinSektion III (creator_code:org_t)
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- In:Journal of Virology86:22, s. 12039-120521098-5514
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