GCs do not interfere with early TCR signaling, but inhibit AICD in vivo. DC27.1 T cells (A) and primary murine day 5 T cells (B) were (re-) stimulated by anti-CD3 antibody in the presence or absence of 10^-7 M Dex for the indicated periods of time. CD95L protein expression on the cell surface was assayed by FACS analysis. (C) DC27.1 T cells were stimulated for 5 hours by anti-CD3 antibody, washed, and subsequently cultured for another 3 hours. For the last 1, 2, or 3 hours of this treatment 10^-7 M Dex was added (lanes 2, 3, and 4, respectively). The mRNA expression of CD95L and β-actin was detected by RT-PCR. Of 3 experiments, 1 representative is shown. (D) C57BL/6 mice (8 weeks old) were pretreated for 24 hours with or without Dex-phosphate (300 μg/L) in the drinking water and subsequently injected intravenously with 5 μg anti-CD3 (+) or isotype control (-) antibody. Then, 20 hours later splenocytes were stained with anti-TCRβ and anti-B220 antibodies and analyzed by FACS (combined results of 5 independent experiments; each diamond represents one mouse; horizontal bars represent the mean; n = 8; t-value for CD3 vs CD3/Dex, P < .005).