Journal: Cell Reports Medicine

Article Title: mRNAs encoding IL-12 and a decoy-resistant variant of IL-18 synergize to engineer T cells for efficacious intratumoral adoptive immunotherapy

doi: 10.1016/j.xcrm.2023.100978

Figure Lengend Snippet:

Article Snippet: Anti-mouse affinity purified IL-22 , R and D Systems , Polyclonal; Cat# AF582, RRID: AB_355457.

Techniques: In Vivo, Activation Assay, Affinity Purification, Virus, Recombinant, Staining, Membrane, Plasmid Preparation, SYBR Green Assay, Cell Isolation, Enzyme-linked Immunosorbent Assay, Transgenic Assay, Control, Negative Control, Retroviral, Software

Journal: American Journal of Translational Research

Article Title: IL-22/IL-22R1 axis is involved in myocardial injury of a mouse cecal ligation and puncture model

doi:

Figure Lengend Snippet: The specific primer pairs for genes used in this study

Article Snippet: After incubated with 5% BSA in PBS for 30 min, sections were incubated with primary antibodies (1:500 diluted rabbit anti-mouse IL-22 polyclonal antibody (ab18499, Abcam, USA); 1:250 diluted rat anti-mouse IL-22R1 polyclonal antibodies (MAB42941, R&D Systems Co., USA); 1:1000 diluted rabbit anti-mouse IL-22RA2 (IL-22BP, ab203211, Abcam, USA)) or 2% BSA in PBS as negative control at 4°C overnight.

Techniques:

Journal: American Journal of Translational Research

Article Title: IL-22/IL-22R1 axis is involved in myocardial injury of a mouse cecal ligation and puncture model

doi:

Figure Lengend Snippet: The mRNA expression of IL-22, IL-22R1 and IL-22BP in heart tissues of both groups determined by real-time PCR. Heart tissues harvested from the mice in both groups at the four time points and stored in liquid nitrogen were subjected to the total RNA extraction using Trizol reagent and then used for cDNA synthesis and the subsequent real-time PCR (n = 6 for each time point). Data are shown as box plot. IL-22 mRNA level was sharply reduced by surgery at the first 8 h and then gradually increased to normal level at 72 h in sham group (n = 6). However, it kept at low level till 72 h in the CLP group. Compared with the mRNA expression of IL-22, the mRNA expression of IL-22R1 and IL-22BP had a completely reverse change trend.

Article Snippet: After incubated with 5% BSA in PBS for 30 min, sections were incubated with primary antibodies (1:500 diluted rabbit anti-mouse IL-22 polyclonal antibody (ab18499, Abcam, USA); 1:250 diluted rat anti-mouse IL-22R1 polyclonal antibodies (MAB42941, R&D Systems Co., USA); 1:1000 diluted rabbit anti-mouse IL-22RA2 (IL-22BP, ab203211, Abcam, USA)) or 2% BSA in PBS as negative control at 4°C overnight.

Techniques: Expressing, Real-time Polymerase Chain Reaction, RNA Extraction, cDNA Synthesis

Journal: American Journal of Translational Research

Article Title: IL-22/IL-22R1 axis is involved in myocardial injury of a mouse cecal ligation and puncture model

doi:

Figure Lengend Snippet: Protein expression of IL-22, IL-22R1 and IL-22BP in heart tissues in both groups determined by IHC (PV9001/9004 method, 400 ×). The 4 μm sections cut from paraffin-embedded heart tissues were subjected to IHC assay with primary antibodies IL-22, IL-22R1 and IL-22BP, respectively. After that, the sections were incubated with the corresponding secondary IgG (PV9001/PV9004 kit) and then developed by DAB kit. The immunoreactivity of the cells were semi-quantitatively transferred into scores and statistically analyzed at the right column. Data are shown as means ± S.E. (n = 6).

Article Snippet: After incubated with 5% BSA in PBS for 30 min, sections were incubated with primary antibodies (1:500 diluted rabbit anti-mouse IL-22 polyclonal antibody (ab18499, Abcam, USA); 1:250 diluted rat anti-mouse IL-22R1 polyclonal antibodies (MAB42941, R&D Systems Co., USA); 1:1000 diluted rabbit anti-mouse IL-22RA2 (IL-22BP, ab203211, Abcam, USA)) or 2% BSA in PBS as negative control at 4°C overnight.

Techniques: Expressing, Incubation

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: IL-22 is involved in liver regeneration after hepatectomy

doi: 10.1152/ajpgi.00075.2009

Figure Lengend Snippet: Expression of hepatic IL-22 receptor-α (IL-22Rα) mRNA after partial hepatectomy. Total RNA was extracted from liver tissues and RT-PCR was used to measure IL-22Rα mRNA expression. mRNA expression in sham-operated control mice was set as 100%. A: significant increases in IL-22Rα mRNA are seen at 12, 24, and 48 h posthepatectomy, with peak levels seen at 12 h posthepatectomy (*P < 0.01 vs. sham, **P < 0.05 vs. sham). Levels return to baseline at 72 h postoperatively. Results are expressed as means ± SE; n = 5 for each group. B: a representative gel.

Article Snippet: Recombinant IL-22 protein, anti-IL-22 antibody, Quantikine mouse IL-6 ELISA kit, and mouse HGF Duoset ELISA Development kit were all obtained from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: IL-22 is involved in liver regeneration after hepatectomy

doi: 10.1152/ajpgi.00075.2009

Figure Lengend Snippet: Serum IL-22 levels after partial hepatectomy. Mice underwent sham laparotomy or partial hepatectomy and were euthanized at 1, 3, 6, 12, 24, 48, or 72 h after 70% hepatectomy, serum was collected, and IL-22 levels were measured by ELISA. Increases in serum IL-22 were seen at 6, 12, 24, and 48 h and reached statistical significance at 12 h (*P < 0.05 vs. sham). These data correlate with the increases seen in hepatic IL-22R mRNA levels.

Article Snippet: Recombinant IL-22 protein, anti-IL-22 antibody, Quantikine mouse IL-6 ELISA kit, and mouse HGF Duoset ELISA Development kit were all obtained from R&D Systems (Minneapolis, MN).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: IL-22 is involved in liver regeneration after hepatectomy

doi: 10.1152/ajpgi.00075.2009

Figure Lengend Snippet: Bromodeoxyuridine (BrdU) staining in mice undergoing 70% hepatectomy and treatment with anti-IL-22 antibody or control IgG antibody. A: mice were treated with IL-22 antibody or control IgG 30 min before partial hepatectomy. Hepatocyte proliferation was measured at 36 h (*P < 0.005), 48 h (*P < 0.0001), and 72 h (*P < 0.05) posthepatectomy. Hepatocyte proliferation was significantly decreased in animals treated with IL-22 antibody compared with animals treated with control IgG (36 h: *P < 0.005; 48 h: *P < 0.0001; 72 h: *P < 0.05). Graphs illustrate means ± SE; 4 animals were used per group and 5 low-power fields (LPF) were observed per mouse. B: comparative photographs of BrdU staining in mice treated with control IgG (a, c, e, g) and mice treated with IL-22 antibody (b, d, f, h) are presented. All pictures were taken at ×200 magnification.

Article Snippet: Recombinant IL-22 protein, anti-IL-22 antibody, Quantikine mouse IL-6 ELISA kit, and mouse HGF Duoset ELISA Development kit were all obtained from R&D Systems (Minneapolis, MN).

Techniques: BrdU Staining

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: IL-22 is involved in liver regeneration after hepatectomy

doi: 10.1152/ajpgi.00075.2009

Figure Lengend Snippet: Western blot analysis for signal transducer and activator of transcription-3 (stat-3) activation after partial hepatectomy and treatment with saline or exogenous IL-22. Representative Western blots are illustrated. GAPDH levels are also shown to demonstrate equal loading of the gels. A: significant increase in phosphorylated stat-3 (p-stat-3) levels in mice treated with exogenous IL-22 at baseline and 30 min after hepatectomy, compared with mice treated with saline alone. B: representative densitometry, as a semiquantitative measure of the p-stat-3 levels.

Article Snippet: Recombinant IL-22 protein, anti-IL-22 antibody, Quantikine mouse IL-6 ELISA kit, and mouse HGF Duoset ELISA Development kit were all obtained from R&D Systems (Minneapolis, MN).

Techniques: Western Blot, Activation Assay, Saline

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: IL-22 is involved in liver regeneration after hepatectomy

doi: 10.1152/ajpgi.00075.2009

Figure Lengend Snippet: Serum and hepatic IL-6 protein levels after partial hepatectomy and treated with recombinant IL-22 (rIL-22) or IL-22 antibody. Administration of rIL-22 to mice before partial hepatectomy increased both hepatic and serum levels of IL-6 compared with animals treated with vehicle; these effects were statistically significant at 1 h posthepatectomy (*P < 0.05). Treatment with anti-IL-22 antibody had no significant effects on serum or hepatic IL-6 levels. A: serum IL-6 levels. B: hepatic IL-6 levels. hep, hepatectomy; AG, Antigen; CAB, control antibody.

Article Snippet: Recombinant IL-22 protein, anti-IL-22 antibody, Quantikine mouse IL-6 ELISA kit, and mouse HGF Duoset ELISA Development kit were all obtained from R&D Systems (Minneapolis, MN).

Techniques: Recombinant

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: IL-22 is involved in liver regeneration after hepatectomy

doi: 10.1152/ajpgi.00075.2009

Figure Lengend Snippet: Western blot analysis for TGF-α after partial hepatectomy and treatment with anti-IL-22 antibodies, control IgG, rIL-22, or saline. Administration of IL-22-neutralizing antibody to mice before partial hepatectomy decreased hepatic TGF-α levels compared with animals treated with control IgG, particularly at later time points. Administration of rIL-22 had minimal effects on TGF-α levels. Representative Western blots are illustrated; GAPDH levels are also shown to demonstrate equal loading of the gels.

Article Snippet: Recombinant IL-22 protein, anti-IL-22 antibody, Quantikine mouse IL-6 ELISA kit, and mouse HGF Duoset ELISA Development kit were all obtained from R&D Systems (Minneapolis, MN).

Techniques: Western Blot, Saline

Journal: Science immunology

Article Title: Oral epithelial IL-22/STAT3 signaling licenses IL-17-mediated immunity to oral mucosal candidiasis

doi: 10.1126/sciimmunol.aba0570

Figure Lengend Snippet: The indicated mice were subjected to OPC. A. Fungal burdens were assessed on day 5 p.i. Bars show geometric mean ± SD. Data were pooled from 2 independent experiments. B. BM from indicated donors was transferred into irradiated recipients. After 6–9 weeks, mice were subjected to OPC and fungal burdens assessed on day 5 p.i. Data were pooled from 2 experiments. C. Frozen sections from WT tongues were co-stained with DAPI and Abs against K13, K14 or IL-22RA1. Suprabasal and basal epithelial layers are indicated. Images are representative of a minimum of 3 sections. Size bar = 200 μm. D. Top: Fungal burdens were assessed on day 5 p.i. Data are pooled from 3 independent experiments. Bottom: IF staining of tongues from the indicated mice were co-stained with DAPI and α-IL-22RA1 Abs. Size bar = 200 μm. E. Top: All mice except Il22−/− were administered tamoxifen for 5 days prior to OPC, and fungal burden assessed on day 5 p.i. Bars show geometric mean ± SD. Bottom: Frozen sections from tongues from the indicated mice were co-stained with DAPI and α-IL-22RA1. Size bar = 200 μm. Data were pooled from 3 independent experiments and analyzed by ANOVA with Mann-Whitney U test.

Article Snippet: Abs: IL-22RA1 (R&D Systems, clone MAB42941), Keratin 13 and Keratin 14 (Abcam, EPR3671 and {"type":"entrez-protein","attrs":{"text":"EPR17350","term_id":"523383453","term_text":"EPR17350"}} EPR17350 ; Invitrogen LL002), Ki67 and anti-p-STAT3 (Tyr705) (Cell Signaling, #9145), IL-17RA (Amgen, clone M751).

Techniques: Irradiation, Staining, MANN-WHITNEY

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model

doi: 10.1016/j.jcmgh.2018.06.008

Figure Lengend Snippet: Il22ra1 is expressed heterogeneously throughout the crypt. ( A ) The Il22ra1 gene expression profile characterized in FACS-isolated total epithelium (CD326+), absorptive/goblet differentiated cells (Sox9-EGFP neg ), TA progenitor cells (Sox9-EGFP sublow ), ISCs (Sox9-EGFP low ), enteroendocrine/tuft cells (Sox9-EGFP high ), and Paneth cells (Sox9-EGFP high , CD24 high ). Technical replicate n = 3; biological N = 3 mice. Significance was calculated by 1-way analysis of variance with Tukey multiple comparisons; bars that are not connected by the same letter are statistically significant ( P < .05). ( B ) Left : t-SNE analysis of single-cell RNA sequence analysis of mouse small intestinal epithelium. Each color represents a different population defined in the original analysis based on lineage-specific transcriptomic signatures. Right : Table depicts the number of cells in each lineage category expressing IL22ra1. ( C ) Cells from the t-SNE profiles in the graph in panel B are highlighted specifically for the expression of IL22ra1 levels in all epithelial cells. Darker shades of grey represent higher expression levels. Pink circles represent no expression. ( D ) The same analysis in panel C except only ISCs are shown. ( E ) The same analysis in panel C except only TA progenitors are shown. ( F ) Representative immunohistochemistry of IL22RA1 (red) and cell nuclei (blue) in a mouse ileal crypt. ( G ) FACS analysis of fixed cell populations described in panel A stained for IL22RA1. Technical replicate n = 3; biological N = 3 mice. Bars represent parts of whole. EC, enterocyte; EE, enteroendocrine; EEC, enteroendocrine; Max, maximum; Min, minimum.

Article Snippet: Primary antibodies and dilutions used were as follows: KI67 (rabbit, 1:100, M7249; Dako), LYZ (goat, 1:500, sc-12091; Santa Cruz, Dallas, TX), CD326 epithelial cell adhesion molecule (EPCAM)/CD326 (rat, 1:500, H8201, clone G8.8; Biolegend, San Diego, CA), IL22RA1 (rat, 1:50, FAB42941P; R&D Systems), OLFM4 (rabbit, 1:250, 39141; Cell Signaling Technology).

Techniques: Expressing, Isolation, Sequencing, Immunohistochemistry, Staining