A flow cytometric method for measuring neutralization of HIV-1 subtype B and E primary isolates.

摘要: Background Clinical trials testing candidate human immunodeficiency virus type 1 (HIV-1) vaccines have required the use of HIV neutralization assays to detect responses specific geographic subtypes HIV-1. The variability in results seen with current p24 assay endpoints prompted us assess utility flow cytometry for monitoring HIV-1 primary isolates. Methods A modified was performed using CD8-depleted peripheral blood mononuclear cells (PBMC). were fixed, permeabilized, stained a directly conjugated monoclonal antibody, and analyzed by cytometry. subtype B′ E isolates tested pooled sera or plasma from infected patients. Results Primary isolate cultures (without neutralizing antibody) showed 18% 42% p24+ cells, depending on virus. Less than 0.2% detected uninfected cultures. Subtype-specific viruses observed serum pools; ranged 0% 99% reduction cells. Conclusions Flow cytometric detection intracellular can be used as an endpoint isolates. This enumerative method has advantage identifying subsets at early culture timepoint. It also provides alternative quantitative assays. Cytometry 40:141–150, 2000. © 2000 Wiley-Liss, Inc.