recombinant murine il 22 protein  (R&D Systems)

Journal: Journal of Translational Medicine

Article Title: Low-dose 5-hydroxymethylfurfural mitigates radiation -induced intestinal toxicity via HIF2α-driven IL22/STAT3 signaling enhancement

doi: 10.1186/s12967-026-07757-3

Figure Lengend Snippet: Low-dose 5-HMF sensitizes intestinal crypts to IL-22‒STAT3 signaling and enhances organoid growth. ( A , B ) organoid formation assay. Crypts isolated from control or low-dose 5-HMF-treated mice were cultured with recombinant murine IL-22 (rmIL22; 50 ng/mL) for 3 days. Representative time-lapse images ( A ) and quantification of the organoid surface area (μm 2 ) from day 1 to day 3 after cultivation ( B ) are shown. ( C ) comparison of organoid growth stimulated by rmIL22 and CHIR99021 (3 μM), a Wnt/β-catenin agonist. Representative images of MTT-stained organoids. ( D ) MTT viability analysis (% positive, n = 6 wells/group). ( E , F ) immunoblot analysis ( E ) and quantification ( F ) of STAT3 phosphorylation (Tyr705) in organoids treated with or without rmIL22 (50 ng/mL, 48 h). The data are presented as the mean±SD. Statistical significance compared with the regular diet group. * p < 0.05, ** p < 0.01 and NS, no significant difference

Article Snippet: Recombinant murine IL-22 protein (rmIL22) and CHIR99021 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Tube Formation Assay, Isolation, Control, Cell Culture, Recombinant, Comparison, Staining, Western Blot, Phospho-proteomics

Journal: Journal of Translational Medicine

Article Title: Low-dose 5-hydroxymethylfurfural mitigates radiation -induced intestinal toxicity via HIF2α-driven IL22/STAT3 signaling enhancement

doi: 10.1186/s12967-026-07757-3

Figure Lengend Snippet: 5-HMF upregulates IL-22 receptor expression through HIF-dependent transcriptional activation. ( A , B ) expression of IL-22 receptor subunits in intestinal crypts. ( A ) mRNA and ( B ) protein levels of IL22R1 and IL10R2 in mice treated with vehicle, low-dose (50 mg/kg), or high-dose (100 mg/kg) 5-HMF. ( C ) Bioinformatics analysis of the IL22R1 promoter region. The schematic illustrates the positions of four putative hypoxia response elements (HREs; consensus core sequence) within the murine IL22R1 locus. Supporting evidence from publicly available human ChIP-seq data (GEO database) is shown below. The track labeled “Batch4_CHROM1_LOVO_EPAS1_RABBIT” visualizes HIF-2α (encoded by EPAS1) binding sites on chromosome 1 in the LOVO human colon epithelial cell line (experimental batch 4; rabbit antibody). The prominent peak (red box) indicates significant enrichment of HIF-2α binding within the upstream regulatory region of the human IL22R1 gene, supporting its status as a direct transcriptional target under HIF-mediated regulation. ( D ) mouse IL22R1 promoter scheme showing distal (−1525~-1449 bp) and proximal (+36 ~ +74 bp) HERs identified by sequence analysis. ( E ) dual-luciferase reporter assay in HEK-293 cells with an IL22R1 promoter-luciferase construct containing a wild-type sequence (−2000 wt) or mutations of distal or/and proximal HERs with transactivation by HIF-1, HIF-2, or GFP-control plasmids ( n = 3 transfections/group). Asterisks (*) indicate the comparison of HIF1 or HIF2 to their respective GFP controls. ** p < 0.01, * p < 0.001 and NS, no significant difference. The cross (†) indicates a comparison of the transactivation of the wild-type sequence to the sequences mutated by HIF1 (†) or by HIF2 (†). † p < 0.05. All the error bars represent the means±sds

Article Snippet: Recombinant murine IL-22 protein (rmIL22) and CHIR99021 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Activation Assay, Sequencing, ChIP-sequencing, Labeling, Binding Assay, Luciferase, Reporter Assay, Construct, Control, Transfection, Comparison

Journal: Journal of Translational Medicine

Article Title: Low-dose 5-hydroxymethylfurfural mitigates radiation -induced intestinal toxicity via HIF2α-driven IL22/STAT3 signaling enhancement

doi: 10.1186/s12967-026-07757-3

Figure Lengend Snippet: HIF-2α is required for 5-HMF-induced IL22R1 expression and the functional potentiation of IL-22 signaling. ( A , B ) HIF-2α inhibition abolishes 5-HMF-induced upregulation of IL22R1 at the protein level in intestinal crypts from mice treated with low-dose 5-HMF with or without the HIF-2α inhibitor PT-2385. ( C ) MTT-stained organoids were isolated from the mice in this study and cultured in medium in the presence or absence of rmIL22 and/or PT-2385 ( n = 3, 3 replicates/group). ( D ) quantification of the organoid surface area (μm 2 ×10 3 ) and MTT viability analysis (% positive, n = 6 wells/group). Statistical significance compared with the control group. * p < 0.05, ** p < 0.01, * p < 0.001 and NS, no significant difference. ( E ) schematic model summarizing the proposed mechanism by which low-dose 5-HMF mitigates radiation-induced intestinal injury. 5-HMF inhibits prolyl hydroxylase domain (PHD) enzyme activity, leading to the selective stabilization of HIF-2α in intestinal crypt cells. HIF-2α translocates to the nucleus and binds to hypoxia‒response elements (HREs) in the promoter of IL22R1, where it is upregulated. Increased IL22R1 surface expression enhances responsiveness to IL-22, leading to activation of the STAT3 signaling pathway. This promotes the proliferation and survival of intestinal stem cells (ISCs), facilitating epithelial regeneration and ultimately protecting against radiation-induced intestinal damage

Article Snippet: Recombinant murine IL-22 protein (rmIL22) and CHIR99021 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Functional Assay, Inhibition, Staining, Isolation, Cell Culture, Control, Activity Assay, Activation Assay

Journal: Cancer Discovery

Article Title: Aryl Hydrocarbon Receptor Ligands Drive Pancreatic Cancer Initiation and Progression through Protumorigenic T-cell Polarization

doi: 10.1158/2159-8290.CD-25-0377

Figure Lengend Snippet: Cigarette smoke and TCDD increase IL22 production in an AHR-dependent manner. A − C, In vitro naïve CD4 + T-cell polarization schema ( A ). CYP1A1 mRNA expression, IL22 mRNA expression, and IL22 protein concentration in culture supernatant after in vitro naïve CD4 + T-cell polarization with CSE ( B and C, n = 3/group), 6-formylindolo(3,2-b)carbazole (FICZ) and TCDD ( D, n = 2–3/group), and TCDD with AHR inhibitor CH-223191 ( E, n = 3/group). F, Experimental schema of naïve CD4 + T-cell polarization with human splenocytes. G, CYP1B1 and IL22 mRNA expression and IL22 protein concentration in the culture supernatant after naïve CD4 + T-cell polarization with TCDD and CH-223191 ( n = 3 technical replicates/group). H, Experimental schema, wherein the culture supernatant of polarized naïve CD4 + T cells was placed upon 7940b PDAC cells for 15 minutes, and protein was then isolated for Western blot. I, Western blot showing pSTAT3, total STAT3, and GAPDH expression among 7940b PDAC cells treated with unpolarized Th0 T-cell media, TCDD-polarized T-cell media, TCDD-polarized media with IL22-binding protein (IL22bp), TCDD alone, negative control, and IL22 alone. J, Schema of orthotopic tumor model with CSE or TCDD. K, IL22 mRNA expression in orthotopic tumors treated with CSE ( n = 3–5/group) or TCDD ( n = 4/group). Data represented as the mean ± SD unless otherwise noted (portion of the figure created with BioRender.com ).

Article Snippet: Controls included 1 nmol/L TCDD in complete DMEM (TCDD alone), negative control with DMEM media alone, and positive control with IL22 (20 ng/mL, R&D Systems, #582-ML-010) in complete DMEM.

Techniques: In Vitro, Expressing, Protein Concentration, Isolation, Western Blot, Binding Assay, Negative Control

Journal: Cancer Discovery

Article Title: Aryl Hydrocarbon Receptor Ligands Drive Pancreatic Cancer Initiation and Progression through Protumorigenic T-cell Polarization

doi: 10.1158/2159-8290.CD-25-0377

Figure Lengend Snippet: TCDD induced PDAC growth is mediated by IL22 signaling and Treg infiltration. A, Genetic makeup of the IL22-tdTomato reporter mouse (Catch-22) and experimental schema showing 3-week treatment course with TCDD. B and C, Representative mfIHC ( B ) and quantification of IL22 + immune cells ( C ) in the duodenum of Catch-22 mice treated with vehicle or TCDD for 3 weeks, n = 3 mice/group, n = 25–26 regions of interest/group. D, Genetic makeup of the IL22 eGFP reporter mouse, with quantification of the proportion of T cells, IL22 + T cells, ILCs, and IL22 + ILCs among pancreata of mice treated with 3 weeks of vehicle or TCDD, n = 3/group. E and F, Experimental schema, n = 6–9/group, and tumor weight and volume for treatment of WT mice with TCDD, IL22 −/− mice with TCDD, and IL22 −/− mice with vehicle for 19 days. G and H, Representative mfIHC ( G, scale bars, 50 μm, inlaid white arrows identifying CD3 + FOXP3 + Tregs), and quantification of T cell, Th cell, CD8 + T cell, and Treg infiltration via mfIHC ( H ), n = 4–5 mice/group, and quantification of 17 regions of interest/group. Phenotypes were identified as follows: T cells (CD3 + ), Th cells (CD3 + CD8 − FOXP3 − ), CD8 + T cells (CD3 + CD8 + ), and Tregs (CD3 + FOXP3 + CD8 − ). I, Quantification of T cells, CD4 + T cells, CD8 + T cells, and Tregs via flow cytometry of orthotopic tumors of WT mice treated with vehicle or TCDD, n = 3–4/group. J, Experimental design, n = 4–8/group, and tumor weight and volume of the orthotopic model of WT mice or FOXP3- DTR mice, allowing for inducible depletion of Tregs, treated with vehicle or TCDD for 16 days. Groups: WT treated with vehicle, WT treated with TCDD, and FOXP3- DTR treated with TCDD. K, Experimental schema and representative gating strategy on CD45 + CD3 + intratumoral T cells from orthotopic tumors of WT mice treated with vehicle or TCDD for 19 days. L, Quantification of IL22 + cells among CD4 + T cells, FOXP3 + cells among CD4 + T cells, and IL22 + cells among FOXP3 + Tregs. Data are represented as the mean ± SD unless otherwise noted. DAPI, 4′6-diamidino-2-phenylindole.

Article Snippet: Controls included 1 nmol/L TCDD in complete DMEM (TCDD alone), negative control with DMEM media alone, and positive control with IL22 (20 ng/mL, R&D Systems, #582-ML-010) in complete DMEM.

Techniques: Flow Cytometry

Journal: Cancer Discovery

Article Title: Aryl Hydrocarbon Receptor Ligands Drive Pancreatic Cancer Initiation and Progression through Protumorigenic T-cell Polarization

doi: 10.1158/2159-8290.CD-25-0377

Figure Lengend Snippet: TCDD-mediated IL22 production and Treg accumulation is dependent upon CD4 + T cell AHR signaling. A, CYP1A1 and CYP1B1 mRNA expression from CD4 + TILs isolated from orthotopic tumors of WT mice treated with vehicle or TCDD for 19 days. B, Genetic makeup of the CD4 Cre AHR mice with inducible CD4-specific deletion of AHR signaling. C, IL22 mRNA expression and protein concentration in the culture supernatant of naïve CD4 + T cells isolated from WT or CD4 Cre AHR mice and polarized under nonstimulating (Th0) conditions, with TCDD, or with TCDD and CH-223191, an AHR inhibitor. D − F, Experimental design ( D ), tumor weight and volume ( E ), and CYP1A1 and IL22 mRNA expression ( F ) from tumors of an orthotopic model utilizing WT mice and CD4-Cre AHR −/− mice treated with vehicle or TCDD, n = 4–7 mice/group. G and H, Representative mfIHC ( G, scale bars, 50 μm, inlaid white arrows identifying CD3 + CD8 + T cell), and quantification ( H ) of CD8 + T cell (CD3 + CD8 + FOXP3 − ) and Treg (CD3 + FOXP3 + CD8 − ) infiltration among orthotopic tumors of WT or CD4 Cre AHR mice treated with vehicle or TCDD, n = 17–39 regions of interest/group. I, Experimental design and ( J ) tumor weight and volume for WT mice treated with vehicle, AHR inhibitor CH-223191, TCDD, or TCDD and the AHR inhibitor for 19 days, n = 7–10/group. Data represented as the mean ± SD unless otherwise noted. 4′6-diamidino-2-phenylindole.

Article Snippet: Controls included 1 nmol/L TCDD in complete DMEM (TCDD alone), negative control with DMEM media alone, and positive control with IL22 (20 ng/mL, R&D Systems, #582-ML-010) in complete DMEM.

Techniques: Expressing, Isolation, Protein Concentration