Fig 2: LPSlow-macrophage injections influence the chemokine network in cGvHD mice. (A, B) Flow cytometric analysis of splenic CD4+ and CD8+ T-cells CXCR3 and CCR5 expression at day 40. (C, D) Flow cytometric analysis of dermal CD4+ and CD8+ T-cells CCR4 and CCR10 expression at day 40. Gating strategy is represented in Supplementary Figure S4. Data represent the mean fluorescence intensity of the chemokine receptors expression on T cells obtained from at least n = 7 biologically independent mice. The ANOVA test with Bonferroni correction was used to detect significant differences between the groups. NS, non-significant; *p = 0.05; **p = 0.01; ***p = 0.001. (E, F) RT-qPCR assessment of ccl17 and ccl27 mRNA levels in the skin (E) and cxcl2 and ccl2 mRNA levels in the liver (F) of cGvHD mice at day 40 of the experiment. Results were expressed as fold increase versus the syngeneic group, derived from at least n = 7 biologically independent mice. Unpaired T-test was used to detect the significance between the groups. NS, non-significant; *p = 0.05; **p = 0.01; ***p = 0.001.

Fig 3: Development of coinhibitory–receptor rich Tr1 cells in the spleen during experimental malaria caused by infection of triple reporter (Il10gfp × Ifngyfp × Foxp3rfp) C57BL/6 mice with PbA.Splenic CD4+T cells were identified by flow cytometry at day 5 after infection (A). Clustering of cells based on expression of IFN-γ, IL-10, FoxP3, CD49b, LAG3, PD1, TIGIT, TIM3, CCR2, and CCR5 (B) and individual molecule expression by tSNE plot (C) was performed. The MFI of staining for all 20 cell clusters identified was presented as a heat map, along with the relative frequency of each cluster (D). Selected clusters were then overlayed on all CD4+T cells and the expression of IL-10 and IFN-γ, LAG3 and CD49b, or PD1 and TIGIT is shown (E). n = 5 individual mice (B–E). Arcsinh scaled MFI values used to generate heat maps are shown in Supplemental Table 5.