Extended Data Fig. 6: IFN-γ mediated changes to myeloid PD-L1 expression.

Cohorts of 8 week-old male C57BL/6 mice (n = 10/group) were treated with an isotype antibody or IFNγ blocking antibody and half of each group (n = 5) were infected with 20 cysts of ME49 intraperitoneally (IP). Splenocytes and Peritoneal exudate cells (PEC) were isolated 72 hours later and analyzed via high-parameter flow cytometry. (a) Comparative histograms evaluating 72 hour timepoint changes in the MFI of PD-L1 expression amongst splenocytes between experimental groups within leukocyte subsets: neutrophils (CD3⁻, B220⁻, CD19⁻, NK1.1⁻, Ly6G⁺, Ly6C⁺, CD11b⁺), cDC1s (CD3⁻, B220⁻, CD19⁻, NK1.1⁻, Ly6G⁻, CD64⁻, CD11c⁺, MHC-II⁺, XCR1⁺), cDC2s (CD3⁻, B220⁻, CD19⁻, NK1.1⁻, Ly6G⁻, CD64⁻, CD11c⁺, MHC-II⁺, SIRPα⁺), monocytes (CD3⁻, B220⁻, CD19⁻, NK1.1⁻, Ly6G⁻, CD64⁺, CD11b⁺, MHC-II⁺, Ly6C⁺), macrophages (CD3⁻, B220⁻, CD19⁻, NK1.1⁻, Ly6G⁻, CD64⁺, CD11b⁺, MHC⁻II⁺, Ly6C⁻) and Treg cells (B220⁻, CD19⁻, Ly6G⁻, NK1.1⁻, CD3⁺, CD4⁺, Foxp3⁺) (n = 5/group, 2-way ANOVA with Tukey’s multiple comparisons test, * = p = 0.0239, ** = p < 0.01, **** = p < 0.0001, 2 experimental replicates). (b) Cohorts of 8 week-old female STAT1^flox mice without any cre expressing alleles (n = 5), or STAT1^flox mice crossed onto either the CD11c^cre (n = 4) or LysM^cre (n = 5) background were infected with 20 cysts of ME49 IP. Splenocytes and PEC were harvested on day 7 of infection and analyzed via flow-cytometry. (b) Histogram comparisons of PD-L1 MFI changes in splenic monocytes and macrophages following conditional deletion of STAT1 (2-way ANOVA with Tukey’s multiple comparisons test, * = p = 0.0475, **** = p < 0.0001, 2 experimental replicates). All data presented are means + /- SEM and show individual data points.

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