Fig 2: MK2 target gene expression is enriched in murine inflammatory monocytes and neutrophils(A) Single cell UMAP clustering of CD45+ enriched cell populations from the small intestine of mice at homeostasis or 4 days post-infection of H. polygyrus. Raw data from (Biton et al., 2018) (GEO: GSE106510). Original cell annotations of the 12 cell types were used.(B) MK2 target genes are commonly detected and highly expressed in inflammatory monocytes and neutrophils.(C) UMAP display of MK2 Target gene score for all cells demonstrates high MK2 target gene signature in inflammatory monocytes and neutrophils.(D) Overlay of violin plot and box plot shows that MK2 target gene expression is significantly enriched in intestinal inflammatory monocytes and neutrophils (∗∗∗ p value < 2.2 × 10−16, Wilcoxon rank-sum test).(E) FACS-mediated quantification of myeloid cell recruitment to colonic (TCT) or (F) ileal (TNFΔARE) mucosa in mice after ATI-450 or vehicle treatment. Myeloid cells are defined as CD45 + CD11b+ cell. Neutrophils and Inflammatory Monocytes are defined within myeloid cells gate based on GR1 and Ly6C surface staining. Data are represented as mean ± SEM. ∗ p value < 0.05; ∗∗ p value < 0.01 (Student's t-test vehicle vs. ATI-450).See also Figures S3 andS4.

Fig 3: Glucose promotes Treg cell differentiation in vitro(A–D) CBir1 Tg CD4+ T cells were cultured with CBir1 peptide and glucose at different concentrations (n = 3/group).(A and B) Representative FACS plots and the frequency of IFNγ+ cells after 5 days of culture under Th1 polarization condition.(C and D) Representative FACS plots and frequency of IL-17+ cells after 5 days of culture under Th17 polarization condition.(E and F) Splenic CD4+ T cells were isolated from Foxp3-GFP mice and cultured under Treg conditions (n = 4/group). Representative FACS plots and frequency of Foxp3+ cells after 5 days of culture.(G and H) CD4+ T cells were activated with anti-CD3/CD28 mAbs under Treg conditions and treated with or without glucose (7000 μg/mL) for two days (n = 3/group). IL-10 (G) and TGFβ (H) production were measured by ELISA.(I and J) CD45.2 CBir1 Tg CD4+ T cells were cultured with APCs and CBir1 peptide in the presence (Glu) or absence (Ctr) of 7000 μg/mL glucose (n = 3/group) under Treg polarization conditions for 5 days. The cells were harvested and then co-cultured with CFSE-labeled CD45.1 naive CD4+ T cells at the ratio of 1:1 for 60h. (I) The CFSE dilution and (J) the CD45.1 cell proliferation were determined by flow cytometry. One representative of 3-5 independent experiments with similar results was shown. Data were expressed as mean ± SEM. Statistical significance was tested by two-tailed one-way ANOVA(B, D, F, J) or two-tailed unpaired Student’s t test (G, H). ns, not significance; ∗p < 0,05; ∗∗p < 0.01; ∗∗∗∗p < 0.0001. See also Figure S1.