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. 2017 Feb 28;91(6):e02296-16. doi: 10.1128/JVI.02296-16

FIG 4.

FIG 4

Ultrasensitive p24 quantification allows short-term detection of HIV-1 reactivation in cells from HIV-infected individuals on cART. CD4+ T cells from HIV-infected individuals on cART were stimulated with PHA and IL-2 to induce HIV-1 reactivation, stained with anti-Gag antibodies at the indicated time points, and analyzed by flow cytometry. Dead cells were excluded based on forward scatter (FSC) and side scatter (SSC) profiles. p24 in culture supernatants was assessed by the ultrasensitive assay. (A) One representative example (participant 6) of HIV-1 reactivation detected by flow cytometry. (B) Comparison of quantification of Gag p24 levels (pg/ml; blue) by the ultrasensitive p24 assay and frequency of Gag p24+ cells measured by flow cytometry (black) to reveal viral reactivation in cells from six HIV-infected individuals displaying detectable reactivation by flow cytometry. (C) CD4+ T cells from up to 29 cART-treated individuals (red) and 4 HIV-negative controls (blue) were activated and analyzed by ultrasensitive p24 quantification after 1, 3, or 7 days. Each dot represents cells isolated from one individual. Black bars indicate medians.