Publication: A flow cytometric method for measuring neutralization of HIV-1 subtype B and E primary isolates
Issued Date
2000-06-01
Resource Type
ISSN
01964763
Other identifier(s)
2-s2.0-0034212296
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Mahidol University
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SCOPUS
Bibliographic Citation
Cytometry. Vol.40, No.2 (2000), 141-150
Suggested Citation
Janice M. Darden, Victoria R. Polonis, Mark S. DeSouza, Somsak Chantakulkij, Arthur E. Brown, Deborah L. Birx, Kovit Pattanapanyasat A flow cytometric method for measuring neutralization of HIV-1 subtype B and E primary isolates. Cytometry. Vol.40, No.2 (2000), 141-150. doi:10.1002/(SICI)1097-0320(20000601)40:2<141::AID-CYTO8>3.0.CO;2-F Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/25871
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Title
A flow cytometric method for measuring neutralization of HIV-1 subtype B and E primary isolates
Abstract
Background: Clinical trials testing candidate human immunodeficiency virus type 1 (HIV-1) vaccines have required the use of HIV neutralization assays to detect responses to specific geographic subtypes of HIV-1. The variability in results seen with current p24 neutralization assay endpoints prompted us to assess the utility of flow cytometry for monitoring the neutralization of HIV-1 primary isolates. Methods: A modified neutralization assay was performed using CD8-depleted peripheral blood mononuclear cells (PBMC). The cells were fixed, permeabilized, stained with a directly conjugated HIV-1 p24 monoclonal antibody, and analyzed by flow cytometry. HIV-1 subtype B' and E primary isolates were tested using pooled sera or plasma from subtype B' or E infected patients. Results: Primary isolate cultures (without neutralizing antibody) showed from 18% to 42% p24+ cells, depending on the virus. Less than 0.2% p24+ cells were detected in uninfected cultures. Subtype-specific neutralization of viruses was observed using plasma or serum pools; neutralization ranged from 0% to 99% reduction of infected cells. Conclusions: Flow cytometric detection of intracellular HIV-1 p24 can be used as an endpoint assay to assess neutralization of HIV-1 subtypes B' and E primary isolates. This enumerative method has the advantage of identifying intracellular p24 in specific subsets at an early culture timepoint. It also provides an alternative quantitative end-point for HIV neutralization assays. (C) 2000 Wiley-Liss, Inc.