202397.fig.005a
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202397.fig.005b
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Figure 5: Caspase and GrB may be involved in different types of Th cell apoptotic mechanisms that are induced by different S. japonicum antigens. (a) Splenocytes from SWA-immunized mice were restimulated in vitro with SWA or PBS in the presence or absence of the GrB inhibitor Z-AAD-CMK. (b) Splenocytes from SEA immunized mice were restimulated in vitro with SEA or PBS in the presence or absence of the caspase inhibitor Z-VAD-FMK. After 36 hours, restimulated C D 4 + T cells in (a) and (b) were purified by MACS and stained with anti-IFN- 𝛾 -PE or anti-IL-4-PE mAbs. Apoptotic cells were labeled with an Apo-Direct TUNEL assay kit and then detected by FACS. The percentage of apoptotic Th1 or Th2 cells among the purified C D 4 + T cells was derived from the number of IFN- 𝛾 + or IL- 4 + cells and gated on the TUNEL positive population. Data are expressed as the mean ± SD of 18 mice from three independent experiments. 𝑃 < . 0 5 ; 𝑃 < . 0 1 . Left panels: One representative experiment of flow cytometric analysis with the average percentage of Th or apoptotic cells shown in the FACS data. Right panels: The statistical analysis of 18 mice from three independent experiments.