Fig 2: ST2 suppression in the peritoneal lavage and lung during Heligmosomoides polygyrus infection.Mice were infected with 200 H. polygyrus L3 larvae, and at day seven post-infection peritoneal lavage (PL) and lung cells were stained for flow cytometry of ST2 on peritoneal lavage KLRG1+CD25+lineage–CD45+ ILC2s (A, B), lung ICOS+lineage–CD45+ ILC2s (C, D), or peritoneal lavage SSChilineage–CD45+ cells (E, F). Geometric mean fluorescence intensity of ST2 is shown in A, C and E, representative ST2 histograms on gated cells is shown in B, D and F. Data from a single experiment with five mice per group. Error bars show SEM.

Fig 3: Itga1-/- HSV-gB/Kb CD8+ T Cells Are Activated, Enter into the Skin, and Clear Replicating HSVWT and Itga1-/- mice were infected with HSV-KOS.(A and B) WT and Itga1-/- HSV-gB/Kb CD8+ T cells differentiate comparably following HSV infection. (A) Proportion of HSV-gB/Kb CD8+ T cells expressing CD44, KLRG1, and CD62L in spleen, DLN, and skin 7 days post-infection. Median, IQR. (B) Left panel: frequency of HSV-gB/Kb CD8+ T cells expressing granzyme B+ in spleen and skin 7 days post-infection. Right panel: granzyme B geometric mean fluorescence intensity gated on CD44hi HSV-gB/Kb CD8+ granzyme B+ T cells. Median, IQR.(C) Left panel: frequency of IFN-?+ among total CD8+ T cells after in vitro re-stimulation of day 9 skin cells or splenocytes with gB(498–505) peptide. Right panel: IFN-? geometric mean fluorescence intensity gated on CD44hi HSV-gB/Kb CD8+ IFN-?+ T cells. Median, IQR.(D) HSV-gB/Kb CD8+ T cells enter comparably into HSV-infected skin of WT and Itga1-/- mice. Numbers of HSV-gB/Kb T cells recovered from the spleen, DLN, and skin of WT versus Itga1-/- mice 7 days post-infection. Median, IQR.(E) Both WT and Itga1-/- mice clear replicating HSV. Skin HSV titers 4, 7, 9, and 14 days post-infection. Median, IQR.(F) HSV-gB/Kb CD8+ T cells migrate into the epidermis of HSV-infected skin of WT and Itga1-/- mice. Numbers of HSV-gB/Kb CD8+ T cells recovered from the epidermis and dermis of WT versus Itga1-/- mice 14 days post-infection. Median, IQR.Data are compiled from three experiments (A, B left panel, and E) or from two experiments (C left panel, D) or are representative of two experiments (B, C right panel). No significant differences between groups were observed using Mann-Whitney tests.

Fig 4: Cardiolipin Is Necessary for In Vivo and In Vitro CD8+ T Cell Responses(A) Schematic of the cardiolipin synthesis pathway.(B) PTPMT1 expression in CD8+ T cells from PTPMT1 WT (Ptpmt1flox/floxCD4-Cre-/-) and PTPMT1-?T (Ptpmt1flox/floxCD4-Cre+/-) mice.(C) Cardiolipin content in freshly isolated Ptpmt1 WT and Ptpmt1-/- CD8+ T cells.(D) Percentage of CD8+ T cells in spleens from PTPMT1 WT and PTPMT1-?T mice. Dots represent individual mice.(E) PTPMT1 WT and PTPMT1-?T mice infected i.v. with 1 × 106 colony-forming units (CFU) LmOVA ?Acta and challenged i.v. with 5 × 107 CFU LmOVA 21 days after primary infection. Blood analyzed at indicated time.(F) Percentage of CD8+ T cells in PTPMT1 WT and PTPMT1-?T mice infected as in (E). Dots represent individual mice.(G) Percentage of CD8+ Tet+ T cells in PTPMT1 WT and PTPMT1-?T mice infected as in (E). Dots represent individual mice.(H) Bacterial burden shown as CFU per µg of liver of PTPMT1 WT and PTPMT1-?T mice 7 days post-LmOVA ?Acta challenge. Dots represent individual mice.(I) TN (CD62LhiCD44lo), TE (CD62LloCD44hi), and TM (CD62LhiCD44hi) CD8+ T cells from PTPMT1 WT and PTPMT1-?T mice 7 days after primary infection. Dots represent individual mice.(J) EEC, SLEC, MPEC, and DPEC CD8+ T cells from PTPMT1 WT and PTPMT1-?T mice 7 days after primary infection according to KLRG1 and CD127 expression. Dots represent individual mice.(K) OCR/ECAR ratio of WT and Ptpmt1-/- CD8+ T cells analyzed 7 days after secondary infection. Dots represent individual mice.(L) Ptpmt1 WT and Ptpmt1-/- CD8+ T cells stimulated with PMA/Iono for 4 h. Also shown is the percentage of CD8+ T cells producing IFN-? and IFN-? MFI of cytokine-producing cells.(M) Cytoplasmic calcium levels at baseline and after aCD3 crosslinking via IgG, tapsigargin (TG), and ionomycin (iono) in Ptpmt1 WT and Ptpmt1-/- CD8+ T cells. Representative of 3 experiments.(N) CD5, CD44, CD25, and CD69 expression in Ptpmt1 WT and Ptpmt1-/- CD8+ T cells 24 h after activation with aCD3/aCD28 + IL-2.(O) Survival of Ptpmt1 WT and Ptpmt1-/- CD8+ T cells 24 h after activation.(P) Cell cycle analysis assessed by FxCycle + Ki67 staining of Ptpmt1 WT and Ptpmt1-/- CD8+ T cells 24 h and 48 h after activation with aCD3/aCD28 + IL-2.(Q) Active proliferation assessed by Ki67 expression in Ptpmt1 WT and Ptpmt1-/- CD8+ T cells 24 h and 48 h after activation with aCD3/aCD28 + IL-2.(R) Cell proliferation assessed by CTV dilution of Ptpmt1 WT and Ptpmt1-/- CD8+ T cells 72 h after activation with aCD3/aCD28 + IL-2. Histogram representative of 4 independent experiments.Data shown as mean ± SEM of =3 independent experiments unless indicated. Statistical comparisons for 2 groups calculated by unpaired two-tailed Student’s t test, *p < 0.05; **p < 0.01; ***p < 0.001.See also Figure S2.