Fig 1: IFNγ mediates STAT1-dependent CXCL9 production in cDC1 independent of the microbiota.a–f Splenocytes from 3-week-old mice of the indicated genotypes were cultured for 4 h with Brefeldin A and Monensin. XCR1+ cDC1 were then analyzed for cytokine production by flow cytometry. a CXCL9 and IL-12p40 staining profiles and b quantification in cDC1 from Clec9a+/creStat1fl/fl (n = 5) and Stat1fl/fl (n = 7) littermates. c Quantification of IL-12p40+ and CXCL9+ cDC1 from 3-week-old Clec9a+/creIfnar1fl/fl (n = 6) and Ifnar1fl/fl littermates or age-matched wild type controls (n = 4). d Quantification of IL-12p40+ and CXCL9+ cDC1 from 3-week-old and Ifngr1+/− (n = 6; for MFI n = 3) or Ifngr1−/− mice (n = 8; for MFI n = 4). e, f cDC1 from 3-week-old germ-free (GF) and age-matched specific pathogen-free (SPF) mice (n = 4) (e) or 3-week-old wildling and age-matched SPF mice (n = 4) (f) were analyzed as above and IL-12p40+ and CXCL9+ cDC1 were quantified. Each dot represents one biological replicate pooled from two independent experiments (a–d). Data in (e, f) are from one experiment. Horizontal bars represent mean, error bars represent SD. Statistical analysis was performed and p-values were determined using two-tailed Welch’s t-test. Only statistically significant comparisons are indicated. Source data are provided as a Source Data file.
Fig 2: Chow diet boosts CXCL9 production from splenic cDC1 and the effector differentiation of food antigen specific CD8+ T cells.a, b SPF mice were either conventionally weaned and separated from dams on day 18 (chow, n = 6) or weaning was delayed by placing mice on formula milk from day 16 while restricting access to chow (milk, n = 5). Mice were separated from dams at day 18 and milk diet was continued until day 28 after birth. a Frequency of IL-12p40+ and CXCL9+ cDC1. b Frequency of IFNγ-positive leukocytes, CD8+ T cells, NK and NKT cells. c, d Splenocytes from 3-week-old germ-free (GF) Myd88−/−TrifLPS2/LPS2 or age-matched GF wild type controls were analyzed (n = 5 from one experiment). c Frequency of IL-12p40+ and CXCL9+ cDC1. d Frequency of IFNγ+ leukocytes, CD8+ T cells, NK and NKT cells is shown. e–g Weaning was delayed in SPF mice as in (a). At 27 days old mice received 1 × 105 naïve OTI cells. On the next two consecutive days, mice were gavaged with OVA and three days after the last gavage, OTI T cells in spleens were analyzed. e Experimental scheme. f Number of OTI cells (chow (n = 10); milk (n = 9). g Frequency of IFNγ- and Granzyme B- producing OTI T cells in spleen. (GzmB: chow, n = 10; milk, n = 9), IFNγ (chow, n = 7; milk, n = 6). h–k Female mice were weaned and maintained on chow (n = 6) or switched to a standard purified (n = 6) or ketogenic purified (n = 7) diet at 6–7 weeks-old for three weeks. i Body weight at readout. j Spleen leukocyte counts, frequency of CD11c+MHCII+ cells and frequency of cDC1 within CD11c+MHCII+ cells. k Quantification of IL-12p40+ and CXCL9+ splenic cDC1. Dots represent biological replicates representative of two independent experiments (a, b) or pooled from two (g, h–k, f, IFNγ) or three (f, GzmB) independent experiments. Horizontal bars represent mean, error bars represent SD. Statistical analysis: two-tailed Welch’s t-test (a–d), linear mixed-effects models (two-tailed) (g), or one-way ANOVA with two-tailed Tukey’s multiple comparisons (i–k). p-values for statistically significant comparisons are shown. l Model of the diet-driven regulatory circuit. Source data are provided as a Source Data file.
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