Figure 5

Effects of galectin (Gal)-9 on subpopulations of spleen T cells. Spleen cells were obtained from PBS- or Gal-9-treated cecal ligation and puncture (CLP) mice at 24 hours after a single intravenous injection. Flow cytometric analysis was performed to clarify which types of T cells are expanded by Gal-9 in the spleens of CLP mice. (A) The frequencies of CD4 and CD8 T cells. Spleen cells were stained with CD3, CD4, CD8 and Tim-3, and fluorescence-activated cell sorting (FACS) analysis was performed; n = 5 for each group (NS, not significant). (B) The frequency of Tim-3+ CD4 and CD8 T cells. Spleen cells were stained with CD3, CD4, CD8 and Tim-3, and FACS analysis was performed; n = 5 for each group (NS, not significant; ^***P <0.001). (C) The frequencies of natural killer T (NKT) cells and γδT cells. Spleen cells were stained with CD3, GL-3 and NK1.1, and FACS analysis was performed; n = 5 for each group (NS, not significant; ^**P <0.001). (D) Expansion of Tim-3+ NKT cells but not γδT cells. Spleen cells were stained with CD3, GL-3, NK1.1 and Tim-3, and FACS analysis was performed; n = 5 for each group (NS, not significant; ^***P <0.001). (E) Gal-9 treatment resulted in an increase of intracellular IL-17+ NKT cells. Spleen cells were stained with CD3, NK1.1 and intracellular IL-17, and FACS analysis was performed; n = 5 for each group (^*P <0.05).