Journal: Immunity

Article Title: Anti-commensal IgG Drives Intestinal Inflammation and Type 17 Immunity in Ulcerative Colitis

doi: 10.1016/j.immuni.2019.02.006

Figure Lengend Snippet: MNP FcγR A:I Ratio Modulates Intestinal Type 17 Immunity (A) Type-17-associated cytokine production by day-14 LPMCs stimulated with Ova and Ova-IC for 16 h (n = 5–9). Paired samples represent cells isolated from the same mouse. Data are pooled from 2 independent experiments. (B) qPCR of type 17 cytokines in WT and Fcgr2b −/− whole colonic tissue after cDSS versus controls (n = 4–9 per group). Data are normalized to uninflamed healthy colon. Medians are indicated. Data are representative of 2 independent experiments. (C) Flow-cytometry plots of colonic IL-17A-expressing T cell subsets in co-housed sex-matched WT and Fcgr2b −/− mice at day 21 after aDSS (n = 6–8 per group) versus controls (day 0) (n = 3–5 per group). (D) Quantification of absolute numbers of colonic IL-17A-producing T cells shown in (C). Medians are indicated. Data are representative of 3 independent experiments. (E) Quantification of absolute numbers of colonic IL-17A-producing T cell subsets in co-housed WT and Fcgr2b −/− mice at day 15 after aDSS and weekly treatment with anti-IL-1R1 IgG-blocking antibody or control IgG (n = 5–7 per group). Medians are indicated. Data are representative of two independent experiments. (F) Colonic IL-17A-expressing T cell subsets in M-TG and N-TG littermate controls at day 21 after aDSS (n = 5 or 6 per group) versus controls (n = 5 or 6 per group). Data are representative of two independent experiments. (G) Quantification of absolute cell counts of colonic IL-17A-producing T cell subsets as shown in (F). Medians are indicated. (H) Weight loss of Fcgr2b −/− mice treated with control or anti-IL-1R1 IgG antibodies after aDSS treatment (n = 5–7 per group). Mean ± SEM are indicated. Data are representative of two independent experiments. (I) Colonic neutrophil infiltration in WT and Fcgr2b −/− mice treated as in (H) (n = 5–7 per group). Medians are indicated. p values were calculated with a ratio paired t test (A), the nonparametric Mann-Whitney U test (B–G and I), or a two-way ANOVA (H). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. See also Figure S6 .

Article Snippet: InVivoMab anti-mouse IL-1R1 antibody , BioXCell , Cat#BE0256; RRID: AB_2661843.

Techniques: Isolation, Flow Cytometry, Expressing, Blocking Assay, Control, MANN-WHITNEY

Journal: Immunity

Article Title: Anti-commensal IgG Drives Intestinal Inflammation and Type 17 Immunity in Ulcerative Colitis

doi: 10.1016/j.immuni.2019.02.006

Figure Lengend Snippet:

Article Snippet: InVivoMab anti-mouse IL-1R1 antibody , BioXCell , Cat#BE0256; RRID: AB_2661843.

Techniques: Virus, Control, Recombinant, Staining, SYBR Green Assay, Protease Inhibitor, Protein Extraction, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Chromatography, Library Quantification, SNP Genotyping Assay, Gene Expression, Microarray, Infection, Software, Transfection, Transferring

Journal: International Journal of Molecular Sciences

Article Title: Myeloid Cell-Derived IL-1 Signaling Damps Neuregulin-1 from Fibroblasts to Suppress Colitis-Induced Early Repair of the Intestinal Epithelium

doi: 10.3390/ijms25084469

Figure Lengend Snippet: Neutrophils and macrophages exhibit dynamic changes during DSS-induced colitis. ( A – G ): Mice were treated with DSS in drinking water for 6 days (D6) followed by recovery with normal water. D18 and D12 represent recovery periods of 12 and 6 days after DSS treatment, respectively. D0 represents control mice with no DSS exposure (n = 6/group for all). ( A ) Representative images of colons from each group. ( B ) Colon length was measured after euthanasia. ( C ) Histological score of colon tissue sections. ( D ) Flow cytometry analysis of neutrophils (CD11b + Ly6G + ) and macrophages (CD11b − Ly6G + ). ( E ) Immunofluorescence staining of PDGFRα (red), IL-1R1 (green), and DAPI (blue) in colon tissues (n = 6/group). The scale bar represents 50 μm. ( F ) Integrated fluorescence density of PDGFRα and IL-1R1 (n = 6/group). ( G ) Relative mRNA expression of Pdgfra , Il1r1 , and Nrg1 in mouse colon tissues was measured by qPCR (n = 6/group). Data in ( A – G ) are representative of two independent experiments. Data in ( B – D , F , G ) are shown as the mean ± SD, followed by one-way ANOVA with Bonferroni’s multiple-comparisons test. (ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Antibodies against mouse PDGFRα (Cat# bs-0231R-AF594), IL-1R1 (Cat# bs-20698R-FITC), and neuregulin-1 (Cat# bs-1817R-Cy5) were purchased from Bioss (Beijing, China).

Techniques: Flow Cytometry, Immunofluorescence, Staining, Fluorescence, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Myeloid Cell-Derived IL-1 Signaling Damps Neuregulin-1 from Fibroblasts to Suppress Colitis-Induced Early Repair of the Intestinal Epithelium

doi: 10.3390/ijms25084469

Figure Lengend Snippet: Neuregulin-1 is co-expressed with IL-1R1 in fibroblasts and is suppressed by IL-1 signaling. ( A , B ) Mice were treated with 3% DSS for six days, and colonic cells were harvested to examine the proportions of neuregulin-1 + IL-R1 + , PDGFRα + IL-1R1 +, and neuregulin-1 + PDGFRα + population in whole colonic fibroblast ( A ) and neuregulin-1 + population in PDGFRα + IL-1R1 + fibroblast ( B ) by flow cytometry (n = 4/group). ( C ) Correlation analysis between normalized expression levels of Pdgfra , Il1r1 , and Nrg1 in mouse colon tissues (n = 18). Spearman’s correlation coefficient (r) is shown. ( D ) Mice were treated with 3% DSS for six days, and colonic CD45-EpCAM-CD31-fibroblasts were sorted and treated with recombinant mouse IL-1β for 24 h. Relative mRNA expression of Il1r1 and Nrg1 in these fibroblasts was measured by qPCR (n = 3/group). ( E ) Lung fibroblasts were obtained from normal C57BL/6J mice and were stimulated with LPS for 24 h followed by IL-1β addition for another 24 h. Relative mRNA expression of Il1r1 and Nrg1 in lung fibroblasts was measured by qPCR (n = 3 replicates/group). Data in ( A – E ) are representative of two independent experiments. Data in ( A , B , D , E ) are shown as the mean ± SD, followed by one-way ANOVA with Bonferroni’s multiple-comparisons test. (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: Antibodies against mouse PDGFRα (Cat# bs-0231R-AF594), IL-1R1 (Cat# bs-20698R-FITC), and neuregulin-1 (Cat# bs-1817R-Cy5) were purchased from Bioss (Beijing, China).

Techniques: Flow Cytometry, Expressing, Recombinant

Journal: International Journal of Molecular Sciences

Article Title: Myeloid Cell-Derived IL-1 Signaling Damps Neuregulin-1 from Fibroblasts to Suppress Colitis-Induced Early Repair of the Intestinal Epithelium

doi: 10.3390/ijms25084469

Figure Lengend Snippet: NRG1-IL-1R1 co-expressing intestinal fibroblasts also exist in IBD patients ( A ) Transcriptional levels of IL1B , IL1R1 , and NRG1 genes expressed in colon tissues from non-IBD control, patients with colonic CD, ileal CD, and UC diseases were determined by analyzing a public bulk RNA-seq dataset (GSE117993, Not IBD n = 55, cCD n = 31, iCD n = 60, UC n = 43). Data are shown as the mean ± SD, followed by the Kruskal–Wallis test with Dunn’s multiple-comparisons test. ( B ) Correlation analysis between expression levels of IL1R1 and NRG1 in colon tissues of all individuals from GSE117993 (n = 189). Spearman’s correlation coefficient (r) is shown. ( C ) Correlation analysis between expression levels of PDGFRA , IL1R1 , and NRG1 in colon tissues of IBD patients from the GSE128682 dataset (n = 44). Pearson’s correlation coefficient (r) is shown. ( D ) UMAP projection of 27,456 single cells from human healthy and UC cohorts, split by disease state and colored by cluster. HC, health control; UC, ulcerative colitis. ( E ) UMAP projection of 1643 fibroblasts, colored by cluster. ( F ) Feature plot showing the expression of NRG1 and IL1R1 , split by disease state. ( G ) UMAP projection depicting the binary expression pattern of IL1R1 and NRG1 in the identified fibroblast population, with colors indicating cells expressing only IL1R1 (blue), only NRG1 (pink), both genes (green), or neither (gray). ( H ) Correlation curves between NRG1 and IL1R1 in NRG1 + IL1R1 + fibroblasts, with each dot representing one cell (n = 175). p value from Spearman correlation test. ( I ) Bar plots showing the enriched items and differential pathways of GO analysis based on highly variable markers of NRG1 + IL1R1 + fibroblasts. ( J ) Bubble plot showing significant interactions between fibroblasts and epithelial cell clusters by ligand–receptor pairs of cytokines and growth factors. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Antibodies against mouse PDGFRα (Cat# bs-0231R-AF594), IL-1R1 (Cat# bs-20698R-FITC), and neuregulin-1 (Cat# bs-1817R-Cy5) were purchased from Bioss (Beijing, China).

Techniques: Expressing, RNA Sequencing Assay

Journal: Clinical and Translational Medicine

Article Title: Hepatic interleukin‐1 receptor type 1 signalling regulates insulin sensitivity in the early phases of nonalcoholic fatty liver disease

doi: 10.1002/ctm2.1048

Figure Lengend Snippet: Body weight gain and systemic metabolic alterations in Il1r1 Hep−/– and WT mice fed a high‐fat, high‐carbohydrate diet for 12 weeks. Eight to 10‐week‐old, male Il1r1 Hep−/– mice and WT littermates were metabolically challenged with a high‐fat, high‐carbohydrate diet (HFD) or received a corresponding control diet (CD) for 12 weeks. (A) Body weight curve, (B) % change in body weight, and (C) mean caloric intake during the entire experimental period. (D) Serum samples were drawn from fasted mice to measure the levels of adiponectin, total cholesterol, triglycerides, NEFA, glucose, insulin, ALT, CRP, and TPO after 12 weeks of dietary feeding and to determine HOMA‐IR and Adipo‐IR indices. Data in (A–C) represent mean of n = 7 Il1r1 Hep−/– CD, n = 10 WT CD, n = 15 Il1r1 Hep−/– HFD, and n = 15 WT HFD mice ± SEM. Data in (D) represent mean of n = 4 Il1r1 Hep−/– CD, n = 3 WT CD, n = 7 Il1r1 Hep−/– HFD and n = 6 WT HFD mice ± SEM (adiponectin), n = 6 Il1r1 Hep−/– CD, n = 8 WT CD, n = 11 Il1r1 Hep−/– HFD and n = 11 WT HFD mice ± SEM (total cholesterol, triglycerides), n = 5 Il1r1 Hep−/– CD, n = 5 WT CD, n = 6 Il1r1 Hep−/– HFD and n = 9 WT HFD mice ± SEM (NEFA, Adipo‐IR), n = 7 Il1r1 Hep−/– CD, n = 10 WT CD, n = 15 Il1r1 Hep−/– HFD and n = 15 WT HFD mice ± SEM (glucose, insulin, HOMA‐IR, and ALT), resp. n = 4 Il1r1 Hep−/– CD, n = 6 WT CD, n = 11 Il1r1 Hep−/– HFD and n = 9 WT HFD mice ± SEM (CRP, TPO). * p < .05 for Il1r1 Hep−/– versus WT and $ p < .05, $$ p < .01, $$$ p < .001 for CD versus HFD using two‐way method of ANOVA (A–D) and Kruskal‐Wallis H test (D [Insulin, HOMA‐IR, ALT, and TPO]) following the Bonferroni multiple comparison tests and Mann‐Whitney tests with a Bonferroni correction, respectively.

Article Snippet: Formalin‐fixed and paraffin‐embedded human liver samples of patients with simple hepatic steatosis (NAFLD activity score [NAS] < 3) and noncirrhotic nonalcoholic steatohepatitis (NASH, NAS > 5) were stained for IL‐1R1 (antibody from Novus Biologicals [Littleton, CO, USA] or Sigma‐Aldrich [St. Louis, MI, USA]) according to the manufacturer's protocols.

Techniques: Metabolic Labelling, Control, Comparison, MANN-WHITNEY

Journal: Clinical and Translational Medicine

Article Title: Hepatic interleukin‐1 receptor type 1 signalling regulates insulin sensitivity in the early phases of nonalcoholic fatty liver disease

doi: 10.1002/ctm2.1048

Figure Lengend Snippet: Effects of HFD feeding on the development of NAFL in Il1r1 Hep−/– and WT mice. (A) Absolute liver weight, liver‐to‐body‐weight ratio, (B) representative liver histology by H&E staining (scale bar: 100 μM) and SAF score, (C) biochemically determined liver triglyceride and (D) protein carbonyl and MDA content of Il1r1 Hep−/– and WT mice fed with the diets for 12 weeks. Data in A represent mean of n = 7 Il1r1 Hep−/– CD, n = 10 WT CD, n = 15 Il1r1 Hep−/– HFD and n = 15 WT HFD mice ± SEM, in B and C n = 6 Il1r1 Hep−/– CD, n = 8 WT CD, n = 11 Il1r1 Hep−/– HFD and n = 11 WT HFD mice ± SEM, and in D n = 4 Il1r1 Hep−/– CD, n = 4 WT CD, n = 8 Il1r1 Hep−/– HFD and n = 8 WT HFD mice ± SEM. $ p < .05, $$ p < .01, $$$ p < .001 for CD versus HFD using two‐way method of ANOVA (A and C) and Kruskal‐Wallis H test (B and D) following the Bonferroni multiple comparison tests and Mann Whitney tests with a Bonferroni correction, respectively. There was no statistical difference for Il1r1 Hep−/– versus WT with respect to these parameters

Article Snippet: Formalin‐fixed and paraffin‐embedded human liver samples of patients with simple hepatic steatosis (NAFLD activity score [NAS] < 3) and noncirrhotic nonalcoholic steatohepatitis (NASH, NAS > 5) were stained for IL‐1R1 (antibody from Novus Biologicals [Littleton, CO, USA] or Sigma‐Aldrich [St. Louis, MI, USA]) according to the manufacturer's protocols.

Techniques: Staining, Comparison, MANN-WHITNEY

Journal: Clinical and Translational Medicine

Article Title: Hepatic interleukin‐1 receptor type 1 signalling regulates insulin sensitivity in the early phases of nonalcoholic fatty liver disease

doi: 10.1002/ctm2.1048

Figure Lengend Snippet: HFD‐induced changes in hepatic lipid metabolism and mitochondrial function of Il1r1 Hep−/– and WT mice. Expression analysis of PPAR‐γ (A and B), PGC‐1α (C and D), NRF‐1, and TFAM (E), HO‐1 (F and G), and FXR‐α (H and I) in liver whole tissue lysates from Il1r1 Hep−/– and WT mice following 12‐week feeding of the HFD or the CD by qRT‐PCR (A, C, E, F, and H) and immunoblotting (B, D, G, and I). Data in A, C, E, F, and H represent mean of n = 4 Il1r1 Hep−/– CD, n = 4 WT CD, n = 7 Il1r1 Hep−/– HFD, and n = 7 WT HFD mice ± SEM. In B, D, G, and I representative immunoblots with densitometric analysis are shown. * p < .05, ** p < .01, *** p < .001 for Il1r1 Hep−/– versus WT and $ p < .05 for CD versus HFD using two‐way method of ANOVA following the Bonferroni multiple comparison tests (A, C, E, F, and H) and unpaired, two‐tailed Student's t ‐test (B, D, G, and I)

Article Snippet: Formalin‐fixed and paraffin‐embedded human liver samples of patients with simple hepatic steatosis (NAFLD activity score [NAS] < 3) and noncirrhotic nonalcoholic steatohepatitis (NASH, NAS > 5) were stained for IL‐1R1 (antibody from Novus Biologicals [Littleton, CO, USA] or Sigma‐Aldrich [St. Louis, MI, USA]) according to the manufacturer's protocols.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Comparison, Two Tailed Test

Journal: Clinical and Translational Medicine

Article Title: Hepatic interleukin‐1 receptor type 1 signalling regulates insulin sensitivity in the early phases of nonalcoholic fatty liver disease

doi: 10.1002/ctm2.1048

Figure Lengend Snippet: Insulin signalling and glucose metabolism in HFD‐fed Il1r1 Hep−/– and WT mice after 12 weeks. (A) I.p. glucose tolerance testing in Il1r1 Hep−/– and WT mice following 11‐week feeding of the HFD or the CD, (B) the area under the curve (AUC, 0–120 min), calculated using the trapezoidal rule, in all experimental groups, and (C) AUC of first (0–15 min), second (15–30 min), third (30–60 min), and fourth phase (60–120 min) in HFD‐fed Il1r1 Hep−/– mice vs. WT littermates. Molecules involved in hepatic insulin signalling were assessed by immunoblotting of (D) insulin receptor, (E) phospho‐IRS‐1 (Ser307) and total IRS‐1, (F) phospho‐AKT (Ser473) and total AKT following insulin injection, and (G) qRT‐PCR analysis of genes encoding IRS‐1, IRS‐2, and the gluconeogenic enzymes PEPCK, FBP1, G6PC, and PC in liver whole tissue lysates prepared from mice fed with the HFD or the CD for 12 weeks. Data in A–C represent mean of n = 6 Il1r1 Hep−/– CD, n = 8 WT CD, n = 11 Il1r1 Hep−/– HFD, and n = 11 WT HFD mice ± SEM. In D–F representative immunoblots with densitometric analysis are shown. Data in G represent mean of n = 4 Il1r1 Hep−/– CD, n = 4 WT CD, n = 7 Il1r1 Hep−/– HFD and n = 7 WT HFD mice ± SEM. * p < .05, ** p < .01, *** p < .001 for Il1r1 Hep−/– versus WT and $ p < 0.05, $$ p < 0.01, $$$ p < .001 for CD versus HFD using two‐way method of ANOVA following the Bonferroni multiple comparison tests (B and G) and unpaired, two‐tailed Student's t ‐test (C–F)

Article Snippet: Formalin‐fixed and paraffin‐embedded human liver samples of patients with simple hepatic steatosis (NAFLD activity score [NAS] < 3) and noncirrhotic nonalcoholic steatohepatitis (NASH, NAS > 5) were stained for IL‐1R1 (antibody from Novus Biologicals [Littleton, CO, USA] or Sigma‐Aldrich [St. Louis, MI, USA]) according to the manufacturer's protocols.

Techniques: Western Blot, Injection, Quantitative RT-PCR, Comparison, Two Tailed Test

Journal: Clinical and Translational Medicine

Article Title: Hepatic interleukin‐1 receptor type 1 signalling regulates insulin sensitivity in the early phases of nonalcoholic fatty liver disease

doi: 10.1002/ctm2.1048

Figure Lengend Snippet: HFD‐induced alterations of MAPK and NF‐κB p65 signalling in the liver of Il1r1 Hep−/– and WT mice. Immunoblotting of (A) phospho‐JNK1/2 (Thr183/Tyr185), (B) phospho‐ERK1/2 (Thr202/Tyr204), (C) phosho‐p38 MAPK (Thr180/Tyr182), (D) phospho‐NF‐κB p65 (Ser536) and total (A) JNK1/2, (B) ERK1/2, (C) p38 MAPK, and (D) NF‐κB p65 protein in liver whole tissue lysates of the different experimental groups after 12 weeks of dietary feeding. In A‐D representative immunoblots with densitometric analysis are shown. * p < .05, *** p < .001 for Il1r1 Hep−/– versus WT using unpaired, two‐tailed Student's t ‐test (A and B). There was no statistically significant difference between Il1r1 Hep−/– and WT mice on HFD with respect of the parameters in C and D

Article Snippet: Formalin‐fixed and paraffin‐embedded human liver samples of patients with simple hepatic steatosis (NAFLD activity score [NAS] < 3) and noncirrhotic nonalcoholic steatohepatitis (NASH, NAS > 5) were stained for IL‐1R1 (antibody from Novus Biologicals [Littleton, CO, USA] or Sigma‐Aldrich [St. Louis, MI, USA]) according to the manufacturer's protocols.

Techniques: Western Blot, Two Tailed Test

Journal: Clinical and Translational Medicine

Article Title: Hepatic interleukin‐1 receptor type 1 signalling regulates insulin sensitivity in the early phases of nonalcoholic fatty liver disease

doi: 10.1002/ctm2.1048

Figure Lengend Snippet: Insulin resistance is accompanied by adipose tissue inflammation in HFD‐fed mice. Inflammation including macrophage recruitment into the obese adipose tissue was assessed by qRT‐PCR‐based gene expression analyses of (A) cytokines, chemokines, (B) the macrophage‐marker F4/80, CCR2, (C) serum levels of CCL2, (D) relative hepatic mRNA expression of leptin, and (E) release of IL‐6 from ex vivo‐cultured adipocytes. Data in A, B, D represent mean of n = 5 Il1r1 Hep−/– CD, n = 4 WT CD, n = 8 Il1r1 Hep−/– HFD and n = 7 WT HFD mice ± SEM, in C n = 4 Il1r1 Hep−/– CD, n = 4 WT CD, n = 6 Il1r1 Hep−/– HFD and n = 6 WT HFD mice ± SEM, and in E mean of duplicate cultures of n = 3 Il1r1 Hep−/– CD, n = 3 WT CD, n = 8 Il1r1 Hep−/– HFD and n = 4 WT HFD mice ± SEM. * p < .05 for Il1r1 Hep−/– versus WT and $ p < .05, $$ p < .01, $$$ p < .001 for CD versus HFD using two‐way method of ANOVA following the Bonferroni multiple comparison tests (A–E)

Article Snippet: Formalin‐fixed and paraffin‐embedded human liver samples of patients with simple hepatic steatosis (NAFLD activity score [NAS] < 3) and noncirrhotic nonalcoholic steatohepatitis (NASH, NAS > 5) were stained for IL‐1R1 (antibody from Novus Biologicals [Littleton, CO, USA] or Sigma‐Aldrich [St. Louis, MI, USA]) according to the manufacturer's protocols.

Techniques: Quantitative RT-PCR, Gene Expression, Marker, Expressing, Ex Vivo, Cell Culture, Comparison