Figure 3.

Sulfatide treatment of Th cells did not induce cell death. A–C, Splenocytes were stimulated with plate-bound anti-CD3 and with the concentrations indicated of sulfatides and percentages of intracellular active form of caspase 3 (A), annexin V-positive 7AAD-negative (B), or annexin V-negative 7AAD-positive Th cells (C) were measured by flow cytometry after a 24 or 72 h period in culture. Results are either representative (A) or mean ± SEM (B, C) from four independent experiments. Data for gated Th cells are presented. D, Sorted splenic Th cells were stimulated with plate-bound anti-CD3 and with indicated concentrations of sulfatides and Z-VAD-FMK for 3 d, and proliferative responses were measured. Results represent mean ± SEM of the percentage of the responses from two separate experiments compared with cultures without sulfatides and without Z-VAD-FMK.