Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to G ) PBMCs from the patients with irAE were cultured in the plate coated with anti-CD3 and anti-CD28 (10 μg/ml) in the presence of IgG1 isotype control or anti–human IL-6R (50 μg/ml), anti–human IL-12p40 (50 μg/ml), and anti–human IFNAR1 (50 μg/ml) for 3 days; n = 9. (A) Expression of CD38 and CD127 on CD8 + T cells. Right: Percentage of CD38 + CD127 − CD8 + T cells. (B and C) CD8 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (B) Expression of granzyme B and IFN-γ. Right: Percentage of granzyme B + IFN-γ + CD8 + T cells. (C) Expression of perforin and IFN-γ. Right: Percentage of perforin + IFN-γ + CD8 + T cells. [(D) and (E)] MFIs of GluCy5 (D), TMRM, and MTDR (E) in CD8 + T cells were presented. [(F) and (G)] CD4 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (F) Expression of IL-21 and IL-2. Right: Percentage of IL-21 + IL-2 + CD4 + T cells. (G) Expression of CD38 and CXCR5 on CD4 + T cells. Right: Percentage of CD38 + CXCR5 − CD4 + T cells. Data in graphs represent mean ± SEM. Significance was tested paired Student’s t test [(A) to (G)].

Article Snippet: PBMCs from patients with irAE were thawed and rested in a complete medium for 1 hour and then activated with plate-coated anti–human CD3 and anti–human CD28 (10 μg/ml) with IgG1 isotype control (Bio X Cell, catalog no. CP174) or a combination of anti–human IL-6R 50 μg/ml; Bio X Cell, catalog no. SIM0014), anti–human IL-12p40 (50 μg/ml; Bio X Cell, catalog no. SIM0020), and anti–human IFNAR1 (50 μg/ml; Bio X Cell, catalog no. SIM0022) for 3 days.

Techniques: Cell Culture, Control, Expressing

Journal: bioRxiv

Article Title: Type I interferon signaling promotes mucosal inflammation in murine models of colitis

doi: 10.64898/2026.03.02.709022

Figure Lengend Snippet: (A) Feature plots of IFN receptors, including IFN-I receptor ( IFNAR1/IFNAR2 ), IFN-II receptor ( IFNGR1/IFNGR2 ), IFN-III receptor ( IFNLR1/IL10RB ), in epithelial cells and CD14 + myeloid cells, from the scRNA-seq analysis of human colonic biopsies in . (B) Feature plots of IFN receptors, from the scRNA-seq analysis of DSS-induced colitis in . (C) Flow cytometry analysis of IFNAR1 + cells in mouse colon with or without DSS exposure, gated by immune cell marker CD45 and epithelial cell marker EpCAM. (D) Proportions of IFNAR1 + cells classified as immune cells (CD45 + /EpCAM - ) or epithelial cells (CD45 - /EpCAM + ) in mouse colon with or without DSS exposure. n=5 per group. (E-F) Flow cytometry analysis and quantification of median fluorescence intensity (MFI) of IFNAR1 expression in immune cells (CD45 + /EpCAM - ) and epithelial cells (CD45 - /EpCAM + ) from DSS-treated mouse colon.

Article Snippet: For pharmacologic inhibition of IFNAR1, mice were intraperitoneally injected with 0.5 mg anti-mouse IFNAR1 antibody (BioXcell, BE0241) or mouse IgG1 isotype control (BioXcell, BE0083) on day 2 and day 5 after DSS exposure.

Techniques: Flow Cytometry, Marker, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Type I interferon signaling promotes mucosal inflammation in murine models of colitis

doi: 10.64898/2026.03.02.709022

Figure Lengend Snippet: (A) Schematic illustrating how the serine-to-alanine substitution at position 535 of murine IFNAR1 protein affects phosphorylation and ubiquitination-dependent degradation. (B) qRT-PCR analysis of selected IFN-I signature genes in spleen and colon from WT and SA mice at baseline. Data were normalized to the mean of WT mice (set as 1). n=5-10 per group. (C) qRT-PCR analysis of selected IFN-I signature genes in bone marrow-derived macrophages (BMDMs) from WT and SA mice, at baseline (untreated) and after stimulation by interferon-β (IFNβ, 200 IU/mL x 8 hours) or lipopolysaccharides (LPS, 1 µg/mL x 8 hours). Data were normalized to the mean of WT BMDMs at baseline (set as 1). n=5-10 per group. (D-E) Mass cytometry (CyTOF) analysis of colon from WT (n=4) and SA (n=6) mice at baseline. (D) UMAP plots with cells colored by identity. (E) Proportions of immune cells (CD45 + /EpCAM - ), epithelial cells (CD45 - /EpCAM + ), and stromal cells (CD45 - /EpCAM - ). ns: not significant. (F) Heatmap showing the mean expression of target proteins in the antibody panel (Supplemental Table 4), with those having p <0.05 by SAM (Significance Analysis of Microarrays) for their median expression indicated by red asterisks.

Article Snippet: For pharmacologic inhibition of IFNAR1, mice were intraperitoneally injected with 0.5 mg anti-mouse IFNAR1 antibody (BioXcell, BE0241) or mouse IgG1 isotype control (BioXcell, BE0083) on day 2 and day 5 after DSS exposure.

Techniques: Phospho-proteomics, Ubiquitin Proteomics, Quantitative RT-PCR, Derivative Assay, Mass Cytometry, Expressing

Journal: bioRxiv

Article Title: Type I interferon signaling promotes mucosal inflammation in murine models of colitis

doi: 10.64898/2026.03.02.709022

Figure Lengend Snippet: Weight change (A and E) , clinical disease activity index (CDAI) (B and F) , colon length (C and G) , and qRT-PCR analysis of selected inflammatory cytokines and IFN-I signature genes (D and H) in colon in DSS-induced colitis (A-D) and piroxicam-accelerated enterocolitis (E-H) . (I-P) Schematics of experimental design (I and M) , clinical disease activity index (CDAI) (J and N) , colon length (K and O) , and qRT-PCR analysis of selected inflammatory cytokines and IFN-I signature genes (L and P) to assess the effect of tamoxifen-induced knockout of IFNAR1 (I-L) and administration of blocking anti-IFNAR1 antibody (M-P) on DSS-induced colitis.

Article Snippet: For pharmacologic inhibition of IFNAR1, mice were intraperitoneally injected with 0.5 mg anti-mouse IFNAR1 antibody (BioXcell, BE0241) or mouse IgG1 isotype control (BioXcell, BE0083) on day 2 and day 5 after DSS exposure.

Techniques: Activity Assay, Quantitative RT-PCR, Knock-Out, Blocking Assay