Fig. 7.

Reduced survival of pathogenic AQP4-specific Th17 cells WT mice is associated with apoptosis. (A) GFP⁺ T cells were examined for viability and apoptosis by staining with Annexin V and 7-AAD followed by flow cytometry (28, 29)). Viable cells are negative for Annexin V and 7-AAD. Apoptotic cells are Annexin V positive, which are further distinguished by early stage (7-AAD negative) and late stage (7-AAD positive). A representative flow staining pattern is shown. The total apoptotic donor populations for CD4⁺GFP⁺ AQP4-specific and MOG-specific T cells are shown (magenta box) with the percentages indicated above. Percentages of late-stage and total apoptotic donor GFP⁺AQP4-specific T cells were higher than donor GFP⁺MOG-specific T cells in recipient WT mice at day 9 and day 21. (B) Apoptotic GFP⁺CD4⁺ T cells from recipient mice were examined in the spleen and CNS, and counting beads were used for quantification. The data shown represent a composite from three experiments, n = 6 per condition (mean ± SEM). Statistical analysis for comparison of surviving numbers of GFP⁺ donor cells was performed using a t test with the Holm–Sidak correction for multiple comparisons (**P < 0.01 and ***P < 0.001).