Detection of circulating p24 antigen-positive CD4+ cells during HIV infection by flow cytometry

Human Immunodeficiency Virus Type 1 Neutralization Measured by Flow Cytometric Quantitation of Single-Round Infection of Primary Human T Cells

“…The variability inherent in neutralization assays that quantify infection indirectly by measurement of secreted p24-Ag led us to develop an assay that directly enumerates the first-round infection of individual lymphocytes. Numerous prior studies have demonstrated the flow cytometric detection of infected lymphocytes in culture and in HIV-1-infected patients by intracellular staining with an anti-p24 antibody (14,15,31,32,44,67,73). Folghera et al used flow cytometric detection of intracellular p24 to measure neutralization of HIV-IIIB in H9 target cells (22).…”

Applications of Flow Cytometry to Clinical Microbiology

“…This method proved to be more sensitive and accurate for quantitative studies and faster than other methods of HIV-1 detection, such as the reverse transcriptase (RT) assay or determination of syncytium formation. Detection of HIV antigens on peripheral blood mononuclear cells by FCM is a useful method for monitoring HIV replication in vivo by monitoring the number of circulating CD4 ϩ cells positive for p24 (63,131,235), p24 and nef (213), or p18 and p24 (107). In all these works, the percentage of cells expressing these antigens was statistically correlated with the clinical status of the patient.…”

γδ T lymphocyte responses to HIV

“…5% paraformaldehyde in PBS, washed and permeabilized with 0 . 3% Tween 20 in PBS [36] and incubated with either a supernate from mouse hybridoma 31-90-25 (reactive with HIV p24) or MK-D6 hybridoma supernate (utilized as a negative control for human cells) for 1 h at 48C. The cells were washed twice, incubated with FITC-conjugated goat anti-mouse antibody for 30 min at 48C, washed twice, and the percentage of HIV p24-positive cells (based on the analysis of 5000-10 000 cells) was determined by flow cytometry.…”