Fig 5.

Latency in resting CD4⁺ T cells cocultured with EC. (A) Resting CD4⁺ T cells with or without EC coculture were infected, and on day 9 postinfection, GFP⁻ cells were sorted and stimulated with PMA-ionomycin or cultured alone for 2 days. Raltegravir was added to prevent de novo viral integration. GFP expression in resting T cells from a EC⁻ coculture are shown, either unstimulated or stimulated with PMA-I. (B) Experiment similar to that in panel A. GFP expressions were compared in unstimulated cultures versus PMA-I-stimulated cultures. Samples were taken in duplicate (ACT) or quadruplicate (all others) depending on the number of cells available. (C) Resting CD4⁺ T cells were isolated from four suppressed patients and cultured alone, with CD3/28 activation, or with EC cocultures in limiting dilutions. Frequencies of latently infected cells (IUPM) were determined by Poisson statistics. (D) Latently infected long-lived resting T cells (B2T) were cultured alone (B2T alone), with activation (CD3/28 or PMA), or with EC cocultures for 6 days. Samples were taken in quadruplicates, and means ± standard errors are plotted in panels B and C, representative of three and two experiments, respectively. *, P < 0.05, Student t test.