Extended Data Fig. 12: CTL transfer model during L. monocytogenes infection.
From: Caspase-7 activates ASM to repair gasdermin and perforin pores
a, Percentage of bacterial burden localized to hepatocytes at 3 dpi. Equal weight liver sections were removed and the hepatocyte fraction was graphed as a percentage of the total burden by weight. 3 mice per genotype. b-c, Mice were infected IV with 5x103 L. monocytogenes and IP treated with combinations of isotype antibody or NK depleting antibody (100 μg anti-NK1.1 PK136), with IL-18 (0.2 μg recombinant mouse IL-18) or isotype control as indicated. Mouse numbers (b) n = 5 each group; (c) WT PBS n = 6, WT IL-18 n = 6, Casp7–/– n = 7 each. d, Timeline for adoptive transfers. Donor mice were PBS treated (naïve) or vaccinated with 1x106 ∆actA L. monocytogenes (immune) in Fig. 5. e-f, Liver and splenic burdens 8 days post primary infection with 5x103 L. monocytogenes. n = 6 mice each. g, Flow gating strategy for flow experiments depicted in (h-i). h-i, Donor mice were NK depleted (> 99% eliminated) and their CTLs enriched by negative selection (> 72% purity). j, Mice were adoptively transferred with the indicated numbers of purified CTLs (naïve were given 1x107 as the maximum transferred number), infected with 5x104 L. monocytogenes and bacterial burdens were analysed in the liver and spleen at 3 dpi. 2 naïve mice, 3 per immunized group. k, Bulk splenocyte transfer (5x107, as previously described in23) with NK depletion was comparable to that of isolated CTLs. Naïve n = 3, immunized n = 4 mice. l, Mice were infected with 5x104 of L. monocytogenes and adoptively transferred with immunized WT CTLs as in d. Liver bacterial burdens were determined at 3 dpi. Mouse numbers: Casp7+/– (n = 2F), Casp7–/– (n = 3M+2F), and Casp6–/– (n = 5F+3M). Note full clearance in Casp7+/– mice may be due to gender disparities or simply to low numbers leading to stochasticity and poor sampling. m-n, Donor mice were PBS treated (naïve) or vaccinated with 1x106 ∆actA L. monocytogenes (immune). (m) Adoptive transfer data from Fig. 5b,c are shown on the same graph, as all the transfers were done in the same 2 pooled experiments. Numbers of mice (m) WT mice with Casp7–/– CTLs n = 7 each, naïve Prf1–/– CTLs n = 8, immune Prf1–/– CTLs n = 7; naïve knockout recipients n = 7 each, immune recipients Prf1–/– n = 8, Casp7–/– n = 7; (n) naïve WT n = 8, immune WT n = 10, Ifng–/– n = 9 each. Data are representative of two experiments (a, e-f, h) or pooled from two experiments (b-c, j–n). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (Two-sided Mann–Whitney U-test (a–c, k, l), or (j, m, n) one–way ANOVA with Tukey’s multiple comparison test). Bars indicate mean with standard deviation (a). Box plots show median, 25th–75th percentile, whiskers are minimum and maximum. Exact p and n values in Source Data EDF12
