Figure 3

RA synovial memory T cells are deficient in IL-4-producing cells after in vitro priming. CD4⁺ memory effector T cells were placed into priming cultures to generate effector cells. Effector T cells were generated by priming in the presence of anti-CD28 mAb and cytokines, as described in Materials and methods, or with additional immobilized anti-CD3 mAb, OKT3, at the indicated concentrations (T3^lo, 0.01 μg/ml; and T3^hi, 1 μg/ml). After priming, the cells were harvested and rested in cytokines before restimulation to detect intracellular cytokines as described in Materials and methods; 10⁴ cells were collected for each analysis. The data shown were derived from independent experiments; for normal donors (n = 4), RAPB (n = 4), RASF (n = 4), and RAST (n = 3), low-dose OKT3 was omitted in one experiment. The frequency of each subset of cytokine-producing cells is shown. The total number of cytokine-producing cells in each sample is shown in the upper right corner. ^*P < 0.05, versus freshly isolated samples shown in Fig. 2 (two-tailed Student's t-test).