Fig 2: Dysregulated Immune and Inflammatory Responses in Mtb-Infected HO-1−/− Mice(A–M) Cytokine analysis of IL-1β (A), IL-2 (B), IL-3 (C), IL-5 (D), IL-6 (E), IL-10 (F), IL-17A (G), MIP-1α (H), MIP-1β (I), KC (J), GM-CSF (K), G-CSF (L), TGF-β (M) in the BALF of uninfected and Mtb-infected HO-1+/+ and HO-1−/− mice at 6, 12, and 18 weeks.(N and O) Percentage of neutrophils in the BALF (N) and lungs (O) of Mtb-infected HO-1+/+ and HO-1−/− mice at 6, 12, and 18 weeks post-infection.(P–S) Other monocytes and granulocytes in the lungs of Mtb-infected HO-1+/+ and HO-1−/− mice.(P and Q) Representative scatterplot for CD11b+Ly6C+F4/80+ monocytes (P) and percent differences (Q) at 6 weeks post-infection.(R and S) Representative scatterplot for CD11b+Ly6G+F4/80+Gr-1+ granulocytes (R) and percent differences at 12 and 18 weeks post-infection (S).n = 4 for each time point. Statistical testing was performed using the unpaired Student’s t test. Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Fig 3: The effect of altered gp130 signalling on MDSCs during intestinal inflammation.Wild type (WT), gp130757F/F, LysMcre/STAT3flox and gp130757F/F LysMcre/STAT3flox mice were treated with the acute (A–E) or chronic (F) DSS protocol to induce colitis and compared with untreated mice (H2O). (A) Cell populations from spleens (SPL), mesenteric lymph nodes (MLN), intraepithelial lymphocytes (IEL) and lamina propria mononuclear cells (LPMC) of wild type mice were characterized by flow cytometry for subpopulations of CD11b+Ly6ClowLy6G+CD49d− granulocytic MDSCs (G-MDSC) and CD11b+Ly6ChighLy6G−CD49d+ monocytic MDSCs (M-MDSC). (B) Colonic cell population from IELs and LPMCs of untreated WT mice (H2O) and DSS-treated WT and gp130757F/F mice were characterized by flow cytometry for subpopulations of CD11c+F4/80−Ly6G−Ly6C− dendritic cells (DC), CD11c−CD11b+Ly6G−F4/80+ monocytes (Mo), CD11b−F4/80+Ly6C−Ly6G− macrophages (MΦ) and CD11b+Ly6G+Ly6ClowF4/80−CD11c– granulocytes (Gr). (C) Colonic cell population from IELs and LPMCs of DSS-treated wild type (WT) and gp130757F/F mice were characterized by flow cytometry for subpopulations of CD11b+Ly6ClowLy6G+CD49d− granulocytic MDSCs (G-MDSC) and CD11b+Ly6ChighLy6G−CD49d+monocytic MDSCs (M-MDSC). (D) Arginase activity was determined in homogenized colon tissue of DSS-treated or untreated (H2O) wild type (WT) and gp130757F/F mice (Di) and also in polymorphonuclear Gr1highLy6G+ MDSCs isolated from spleens of wild type (WT) and gp130757F/F mice (Dii). (E) Granulocytic Gr1highLy6G+ MDSCs (G-MDSCs) were isolated from spleens of wild type (WT), gp130757F/F, LysMcre/STAT3flox and gp130757F/F LysMcre/STAT3flox mice and 1 × 106 cells were cultured for 24 hours. Cell culture supernatants were analysed by luminex array. (F) Cell populations from lamina propria mononuclear cells (LPMCs) were isolated from the colon of mice with chronic DSS-induced colitis and characterized by flow cytometry for MDSC subpopulations (CD11b+CD45+CD11c-/int, Gr1high/low). *indicates P < 0.05 and **indicates P < 0.01 (compared to untreated control or wild type); #indicates P < 0.05 (compared to LysMcre/STAT3flox or gp130757F/F mice). Bars represent the mean±SEM of at least three independent experiments (n = 5–12 per group).

Fig 4: RGS10 and Chronic Systemic Inflammation (CSI) synergize to regulate the frequency and functional dynamics of natural killer cells and antigen-presenting cells in peripheral circulation. Male and female mouse PBMCs were immunophenotyped via flow cytometry. A & F) Frequency of natural killer cell out of total CD45 + cells. B & G) Frequency of dendritic cells out of total CD45 + cells. C &H) Frequency of monocytes out of total CD45 + cells. D & I) Mean fluorescent intensity (MFI) of MHCII on Dendritic cells. E & J) MFI of MHCII on monocytes. L & O) MFI of LY6C (BB660) on monocytes. M & P) Frequency of LY6C-high monocytes out of total monocytes. N & Q) Frequency of LY6C-low monocytes out of total monocytes. Samples that are statistically different do not share the same letter. Significance for main effects is as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Fig 5: RGS10 and Chronic Systemic Inflammation (CSI) impair capacity for antigen presentation in brain-associated immune cells and induce myeloid cell activation. Male and female mouse CNS-associated immune cells were immunophenotyped via flow cytometry. A & E) Frequency of dendritic cells out of CD45 + cells. B & F) Frequency of natural killer cells out of CD45 + cells. I & L) Mean fluorescent intensity (MFI) of MHCII on dendritic cells. J & M) MFI of MHCII on monocytes. K & N) MFI of MHCII on microglia. O & R) Frequency of MHCII + dendritic cells out of total dendritic cells. P &S) Frequency MHCII + monocytes out of total monocytes. Q &T) Frequency MHCII + microglia out of total microglia. U & X) MFI of CD45 on microglia. V & Y) MFI of CD45 on monocytes. W & Z) MFI of LY6C on monocytes. Significance for main effects is as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001