Figure 3.

Impaired functions of thymocytes and splenocytes from M2-γc− mice. Thymocytes (A) or splenocytes (B) (1 × 10⁵) were cultured in triplicate in flat-bottomed 96-well plates, stimulated with RPMI, anti-CD3 antibody (145–2C11, 10 μg/ml), anti-CD3 antibody plus human IL-2 (1 nM, Takeda, Osaka), mouse IL-4 (10 ng/ml, Genzyme), mouse IL-7 (10 ng/ml), or Con A (5 μg/ml) plus human IL-2 (2 nM). Cells were pulsed with 0.5 μCi of [³H]thymidine (Amersham) for the last 16 h of the 88-h culture period, and the [³H]thymidine uptake levels were measured by using Top Count (Packard). Growth assays were performed in triplicate as described (32). The assay was performed several times, and the results were highly reproducible. (C) Cytokine production of splenic T cells from transgene-reconstituted γc null mice. Splenocytes (1 × 10⁶ cells/ml) were incubated in either media without or with anti-CD3 antibody (10 μg/ml) for 72 h (for IFNγ and IL-4) or 48 h (for IL-2). The supernatants were subsequently collected, and the levels of cytokines were determined by ELISA.