cf interleukin 17b il 17b r d systems  (R&D Systems)

Journal: Frontiers in immunology

Article Title: Interleukin 17 B regulates colonic myeloid cell infiltration in a mouse model of DSS-induced colitis.

doi: 10.3389/fimmu.2023.1055256

Figure Lengend Snippet: FIGURE 1 IL17B expression was increased in the colon tissue of IBD patients. (A) IL17B expression in different cell types isolated from endoscopic biopsies of patients with IBD in a RNAseq dataset GSE166924. n=17 (B) Quantification of IL17B expression of colon uninflamed tissues (n=66) and inflamed tissues (n=63) from UC patients with a microarray data set GDS3268. ***P < 0.001 determined by student t test. (C) IL17B expression in paired colon inflamed tissues and non-inflamed tissues from 23 IBD patients was detected by real-time PCR. Data represent mean ± SD, *P < 0.05; determined by paired student t test.

Article Snippet: Recombinant IL17B administration Il17b-/- mice andWTmice were induced colitis by feeding 2.5% or 3% DSS water respectively, and intraperitoneally (I.P.) injected with 0.5 mg of mouse IL17B (R&D Systems, Minneapolis, MN, USA) or PBS.

Techniques: Expressing, Isolation, Microarray, Real-time Polymerase Chain Reaction

Journal: Frontiers in immunology

Article Title: Interleukin 17 B regulates colonic myeloid cell infiltration in a mouse model of DSS-induced colitis.

doi: 10.3389/fimmu.2023.1055256

Figure Lengend Snippet: FIGURE 2 IL17B deficiency results in exacerbated DSS-induced colitis. WT and Il17b-/- mice were fed with 2.5% DSS water for 6 days. n = 5 - 6 mice/group. (A) Body weights were measured daily and body weight loss relative to initial weight were calculated. Data are presented as mean ± SD. **P < 0.01, ***P < 0.001, evaluated by 2way ANOVA analysis with Sidak`s multiple comparisons test. (B) Colons were removed at day 8 and the colon length was measured. (C) Representative H&E staining of colon tissues. Scale bar: 200 mm. (D) Quantification of histological score from colonic sections in (C, E) The supernatants of colon tissue homogenates were assayed for cytokine by ELISA. For (B, C), data are mean ± SD and are representative of at least 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, evaluated by unpaired t test.

Article Snippet: Recombinant IL17B administration Il17b-/- mice andWTmice were induced colitis by feeding 2.5% or 3% DSS water respectively, and intraperitoneally (I.P.) injected with 0.5 mg of mouse IL17B (R&D Systems, Minneapolis, MN, USA) or PBS.

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in immunology

Article Title: Interleukin 17 B regulates colonic myeloid cell infiltration in a mouse model of DSS-induced colitis.

doi: 10.3389/fimmu.2023.1055256

Figure Lengend Snippet: FIGURE 3 Rescue of IL17B alleviates the severity of colitis. Il17b-/- mice were induced colitis by feeding 2.5% DSS water and intraperitoneally (I.P.) injected with 0.5 mg of mouse IL17B (n = 4) or PBS (n=3). The treatment was performed 1 hour before the initial DSS administration and repeated every 48 hours for 4 times. (A) Body weight loss was measured daily. ***P < 0.001, ****P < 0.0001, evaluated by 2 way ANOVA analysis with Sidak`s multiple comparisons test. (B) The colon lengths were measured on day 8. (C) Representative H&E staining of colon tissues. Scale bar: 200 mm. (D) Quantification of histological score from colonic sections in (C, E) The supernatants of colon tissue homogenates were assayed for cytokine by ELISA. Data are mean ± SD. *P < 0.05, **P < 0.01, ****P < 0.0001, evaluated by unpaired t test.

Article Snippet: Recombinant IL17B administration Il17b-/- mice andWTmice were induced colitis by feeding 2.5% or 3% DSS water respectively, and intraperitoneally (I.P.) injected with 0.5 mg of mouse IL17B (R&D Systems, Minneapolis, MN, USA) or PBS.

Techniques: Injection, Staining, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in immunology

Article Title: Interleukin 17 B regulates colonic myeloid cell infiltration in a mouse model of DSS-induced colitis.

doi: 10.3389/fimmu.2023.1055256

Figure Lengend Snippet: FIGURE 4 Single cell transcriptome analysis of colon lamina propria immune cells in colitis in WT and Il17b-/- mice. Two WT and two Il17b-/- mice were fed with 2.5% DSS water to induce colitis. Lamina propria CD45+ immune cells were sorted by magnetic activated cell sorting and processed for scRNAseq. (A) UMAP plot shows the clustering of 12725 immune cells, colored by cell subset and cell types. (B) UMAP plots displaying marker genes expression for cell cluster. (C) Heatmap showing specific marker genes for major cell types. (D) UMAPs plots showing a comparison of the CD45+ cell clusters distribution in WT and Il17b-/- colitic mice. (E) Percentage of each cell cluster in CD45+ cells in WT and Il17b-/- colitic mice.

Article Snippet: Recombinant IL17B administration Il17b-/- mice andWTmice were induced colitis by feeding 2.5% or 3% DSS water respectively, and intraperitoneally (I.P.) injected with 0.5 mg of mouse IL17B (R&D Systems, Minneapolis, MN, USA) or PBS.

Techniques: FACS, Marker, Expressing, Comparison

Journal: Frontiers in immunology

Article Title: Interleukin 17 B regulates colonic myeloid cell infiltration in a mouse model of DSS-induced colitis.

doi: 10.3389/fimmu.2023.1055256

Figure Lengend Snippet: FIGURE 5 ScRNAseq analysis of myeloid cell response. (A) UMAP plot shows myeloid cell clusters. (B) The specific cell markers of neutrophils were projected on UMAP. (C) Gene enrichments of upregulated DEGs in cluster 1 and cluster 6. Top 20 of terms were shown. (D) UMAPs plots showing the difference of cell clusters distribution in WT and Il17b-/- colitic mice. (E) Percentage of cell cluster 1 and 6 in WT and Il17b-/- colitic mice. (F–H) WT and Il17b-/- mice (n = 4/group) were fed with 2.5% DSS water for 6 days. (F) The infiltration of CD11B+ S100A9+ in colon lamina propria was detected by flow cytometry on day 8 (right panel). The CD11B+ S100A9+ cells express Ly6G, confirming neutrophils phenotype (left panel). (G) The percentages of CD11B+ S100A9+ neutrophils in CD11B+ cells were compared between WT and Il17b-/- colitis mice. Bar graph showing the statistic of the CD11B+ S100A9+ cells in colitis mice. (H) The inflammatory genes expression in colonic lamina propria CD45+ cells was detected by real-time PCR. For bar graphs, data are expressed as mean ± SD. *P < 0.05; **P < 0.01, determined by unpaired t test. The experiment was performed 3 times.

Article Snippet: Recombinant IL17B administration Il17b-/- mice andWTmice were induced colitis by feeding 2.5% or 3% DSS water respectively, and intraperitoneally (I.P.) injected with 0.5 mg of mouse IL17B (R&D Systems, Minneapolis, MN, USA) or PBS.

Techniques: Cytometry, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in immunology

Article Title: Interleukin 17 B regulates colonic myeloid cell infiltration in a mouse model of DSS-induced colitis.

doi: 10.3389/fimmu.2023.1055256

Figure Lengend Snippet: FIGURE 6 Increased inflammatory response in colonic macrophages in Il17b-/- colitic mice. (A) The heatmap shows the upregulated signature genes in colonic macrophages. (B) UMAP plots displaying marker genes expression for cell clusters. (C) Percentage of cell clusters of colonic macrophagse in WT and Il17b-/- colitic mice. (D) Gene enrichment of upregulated DEGs in cluster 3 of colonic macrophages. (E) The mRNA expression of some DEGs in colon lamina propria CD45+ cells of Il17b-/- and WT colitis mice were detected by real-time PCR. n = 7 for WT group and 6 for Il17b-/- mice. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01, ****P < 0.0001, determined by unpaired t test.

Article Snippet: Recombinant IL17B administration Il17b-/- mice andWTmice were induced colitis by feeding 2.5% or 3% DSS water respectively, and intraperitoneally (I.P.) injected with 0.5 mg of mouse IL17B (R&D Systems, Minneapolis, MN, USA) or PBS.

Techniques: Marker, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in immunology

Article Title: Interleukin 17 B regulates colonic myeloid cell infiltration in a mouse model of DSS-induced colitis.

doi: 10.3389/fimmu.2023.1055256

Figure Lengend Snippet: FIGURE 7 Treatment of IL17B reduces LPS-induced inflammation. (A) Il17b-/- mice were induced colitis and treated with IL17B as shown in figure 3. The infiltration of CD11B+ S100A9+ neutrophils in colon lamina propria was detected by flow cytometry. Bar graph showing the statistic of the CD11B+ S100A9+ cells in colitis mice. (B) BMDM was treated with LPS and IL17B for 2 hours (n = 3). The cytokines and chemokines expression of cells were detected by real-time PCR. (C, D) Mice were I.P. treated with LPS (n = 4) and IL17B (n = 3) for 2 hours and the inflammatory responses in serum (C) and colon tissue (D) were analyzed by ELISA. Data are mean ± SD. Statistics in (B) was done by ordinary one-way ANOVA with Turkey`s multiple comparisons test. Statistics in (C) and (D) was done with unpaired t test. *P < 0.05; **P < 0.01. (E) Volcano plot showing the DEGs of BMDM treated with IL17B detected by bulk RNAseq (triplicate samples for each treatment). (F) Gene enrichment of upregulated DEGs of IL17B-treated BMDM. (G) Gene enrichment of downregulated DEGs of IL17B-treated BMDM.

Article Snippet: Recombinant IL17B administration Il17b-/- mice andWTmice were induced colitis by feeding 2.5% or 3% DSS water respectively, and intraperitoneally (I.P.) injected with 0.5 mg of mouse IL17B (R&D Systems, Minneapolis, MN, USA) or PBS.

Techniques: Cytometry, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay