Dexamethasone suppresses tumor-specific T cell proliferation and differentiation but not activity in antigen-experienced mice.

Healthy mice were administered activated pmel-1 T cells labeled with the congenic marker Thy1.1 on day -21 and vaccinated for the first time with a gp100 vaccine on day -19. From day -2 until day 6, the mice were administered 3 mg/kg dexamethasone daily. On day 0, they were administered the vaccine a second time. Spleens were harvested for flow cytometry on day 7. The cells were stained with Zombie Yellow, APC-conjugated anti-Thy1.1, APC/Cyanine7-conjugated anti-CD8, Pacific Blue-conjugated anti-CD3 mAbs, PE/Dazzle594-conjugated anti-B220, anti-F4/80, anti-NK1.1 and anti-Ly6G, followed by culture with 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl) amino]- 2-deoxy-D-glucose (2-NBDG, Peptide Institute, Osaka, Japan) at 80 μM for 30 min at 37°C in glucose-free RPMI-1640 containing 10% FBS.