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. 2012 Jan 17;109(5):1613–1618. doi: 10.1073/pnas.1121307109

Fig. 3.

Fig. 3.

Costimulation by CD28 or CD81 targets different T-cell subsets. (A–C) Isolated T cells were cultured in the presence of anti-CD3 anti-CD81 and anti-CD28 mAbs, as indicated. The cells were stimulated for 24 h, stained for expression of the surface molecules, CD4, CD45RO, CD62L and CD69 or CD25 and analyzed by flow cytometry. (A) CD4-gated cells were further analyzed for CD45RO and CD62L expression. Shown is gating on naïve (CD62L high CD45RO low), central memory (CD62L high CD45RO high), and effector memory (CD62L low CD45RO high) T cells. (B) Increased CD69 expression on gated naïve (black solid line) CD4 cells in response to costimulation by the anti-CD81 mAbs 5A6 and 1D6 (Middle). Costimulation by CD28 affected mostly CD69 expression on gated central (light gray line) and effector (black dashed line) memory CD4 cells (Lower Left). Dual costimulation increased CD69 expression on all subsets (Lower Right). (C) CD25 expression on gated naïve and memory CD4 cells was increased on naïve cells costimulated by CD81 (Left) and on memory cells (Right) costimulated by CD28. (D) Isolated T cells were labeled with CFSE, cocultured with the indicated antibodies for 5 d and analyzed by flow cytometry. Shown is CFSE dilution in relation to CD45RO expression on CD4-gated cells. Data are representative of four independent experiments.