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. 2010 Mar 31;84(12):5958–5974. doi: 10.1128/JVI.00161-10

FIG. 3.

FIG. 3.

Isolation and infection of human primary CD4+ T cells. (A to D) Naïve CD4+ T cells were isolated from human whole blood from three donors (9.1, 9.2, and 9.3). Six days after isolation, the cells were analyzed by flow cytometry for expression of surface receptors CD4 (A), CD45RA (B), CD45RO (C), and CD27 (D). Histogram overlays include negative controls (shaded gray) and naïve CD4+ T cells (black outline). Fluorescence channels 6 (FL 6), 8, 4, and 9 are shown in panels A, B, C, and D, respectively. (E) Naïve CD4+ T cells were activated with CD3/CD28 antigen beads for 3 days after isolation to promote T-cell activation and expansion. At 7 days postisolation, cells from each donor were infected by one of seven different LGIT subtype variants at a low MOI. Cells were cultured in activating conditions until day 14, at which cells were transferred to minimal growth medium (1 ng/ml IL-7 and 10 U/ml IL-2).