FIG. 2.
Reactivation of HIV-1 from latently infected cultured TCM cells. (A) Fourteen CD4+ purified samples were treated with IL-2 alone or IL-2+αCD3/αCD28 for 48 h. ICp24 was analyzed by flow cytometry. Significance was calculated using a two-tailed paired t-test analysis (p values provided). Asterisk indicates data corresponding to dot plot figures in (B). (B) Representative dot plots of IL-2 and IL-2+αCD3/αCD28-stimulated UL fractions. (C) Four CD4+ purified samples were treated with IL-2 alone or IL-2+αCD3/αCD28 for 48 h. CA HIV-1 RNA copies were measured by quantitative polymerase chain reaction (qPCR) in triplicate samples. Normalization of cell-associated HIV-1 RNA to a cellular RNA would not be feasible for the comparison of HIV-1 transcripts produced from quiescent cells to those generated from cells treated with a strong cell activation stimulus. We, therefore, report HIV-1 RNA values normalized to input cell number. Mean values are plotted and error bars denote standard deviations. (D) UL fractions were stimulated with 330 nM SAHA, 10 μg/ml PAM3CSK4, 100 nM bryostatin-1, 100 nM ingenol 3,20-dibenzoate and ICp24 and viability from Donor 5-7 (E) were measured using flow cytometry. Significance was calculated using a two-tailed paired t-test analysis (p values provided).