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. Author manuscript; available in PMC: 2020 Mar 16.
Published in final edited form as: Nat Immunol. 2019 Sep 16;20(10):1335–1347. doi: 10.1038/s41590-019-0480-4

Figure 4: Ptpn2 deletion increases Tim-3+ cell differentiation and proliferation.

Figure 4:

(a) Quantification of frequencies of co-transferred control or Ptpn2-deleted CD8+ T cells day 4 post LCMV Clone 13 infection. Frequencies at day 4 were normalized to input frequencies at day 0. Representative of two independent experiments, n = 5 mice. (b) Representative flow cytometry plots of Slamf6 and Tim-3 expression on splenic CD8+ T cells day 4 post LCMV Clone 13 infection for co-transferred control and Ptpn2-deleted cells. Representative of two independent experiments, n = 5 mice. (c) Quantification of Slamf6+ Tim-3–, Slamf6+ Tim-3+, and Slamf6– Tim-3+ subsets in (b). Representative of two independent experiments, n = 5 mice. (d) Quantification of Granzyme B expression of cells as in (a). Representative of two independent experiments, n = 5 mice. (e) Schematic of in vitro conversion and CTV proliferation assays using co-transferred control or Ptpn2-deleted CD8+ T cells isolated at day 8 post LCMV Clone 13 infection. (f) Quantification of Slamf6+ Tim-3–, Slamf6+ Tim-3+, and Slamf6– Tim-3+ subsets following in vitro stimulation (αCD3/CD28 with IL-2 and IFN-α) of control or Ptpn2-deleted CD8+ T cells isolated at day 8 post LCMV Clone 13 infection. Representative of two independent experiments, n = 5 mice. (g-h) Quantification of frequency of divisions following in vitro stimulation (αCD3/CD28 with IL-2) of (g) CTV-labeled Slamf6+ or (h) CTV-labeled Tim-3+, control or Ptpn2-deleted CD8+ T cells isolated at day 8 post LCMV Clone 13 infection. Representative of two independent experiments, n = 3 mice (g), 5 mice (h). (i) Schematic of in vivo persistence assay of Tim-3+ cells. (j) Quantification of number of recovered Tim-3+ control or Ptpn2-deleted cells in the spleen day 6 post LCMV Clone 13 infection, following the transfer of these cells which were previously isolated at day 8 post LCMV Clone 13 infection. Representative of two pooled experiments, n = 8 mice. Bar graphs represent mean and error bars represent standard deviation. Statistical significance was assessed by two-sided Student’s paired t-test (a, c-d, f-h) or two-sided Student’s unpaired t-test (j) (ns p>.05, * p≤.05, ** p≤.01, *** p≤.001, **** p≤.0001). See also Supplementary Figure 4.