Figure 2.
Detection of apoptosis. Fas-mediated apoptosis was analyzed by testing patient sample (A) in tandem with normal control (B). T cells were stimulated with PHA. After 14 to 21 days of culture, 106 T cells were incubated with media, anti-Fas monoclonal antibody, steroid, or C2-ceramide for 48 hours. (A) Results are an example of defective Fas-mediated apoptosis (consistent with diagnosis of ALPS) in 1 patient sample with percentages indicating proportion dead as determined by FACS analysis for 7-AAD. X-axis depicts percentage dead by 7-AAD and y-axis depicts number of cells counted. Gating strategy for dead cells is determined by comparing to control-stimulated T cells treated with ethanol and stained with 7-AAD.
