mouse chil1  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Primer sequence.

Article Snippet: Recombinant mouse CHIL1 (2649-CH-050) was from R&D Systems (Minneapolis, MN).

Techniques: Sequencing

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Microarray analysis was performed in N-Neu and A-Neu. (A) Microarray heatmap data showing the expression level of transcript between N-Neu and A-Neu. The mRNA expressions were expressed as follows: Bright green, under expression; black, no change; bright red, overexpression. n=3. Row-based Z-score normalized. (B) GO enrichment analysis of differentially expressed genes in S1P activated neutrophils. (C) Venn diagram of gene expression overlap between GO terms including Regulation of Cell Communication, Cytokine Secretion, Inflammatory Response and Extracellular Space clusters. (D) Volcano plot of A-Neu gene expression compared with N-Neu. The threshold values were fold change≥|2| and P value ≤ 0.05. Red dots and blue dots represent up-regulated and down-regulated genes, respectively. Gray dots indicate genes with no statistically significant differences. The arrow indicated the location of Chil1. A-Neu, activated neutrophil; N-Neu, non-activated neutrophil; Il1b, Interleukin 1 beta; Chil1, Chitinase-like 1; Ccl3, Chemokine (C-C motif) ligand 3; Il1a, Interleukin 1 alpha; Tnf, tumor necrosis factor.

Article Snippet: Recombinant mouse CHIL1 (2649-CH-050) was from R&D Systems (Minneapolis, MN).

Techniques: Microarray, Expressing, Over Expression, Gene Expression

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Chil1 was positively correlated with Ly6g in the injured liver of mice and could be secreted by A-Neu. (A) The mRNA expression of Chil1 was examined by RT-qPCR in the injured liver of MCDHF-treated mice in indicated times (n=6). (B) S1P stimulated neutrophils for 2 hours, then PBS washed the cells, and continued to culture for 6 hours, the secretion of CHIL1 in cell supernatant was detected by ELISA, n=4. (C) Correlation analysis between Chil1 mRNA expression with neutrophil marker Ly6g , macrophage marker Adgre1 , hepatocyte marker Alb and myofibroblast marker Acta2 in the injured liver of MCDHF-treated mice. Nonparametric test (Kruskal-Wallis test) was used in (A) Student’s t test was used in (B) Pearson’s test was used in (C) *P < 0.05 vs. MCDHF treated group for 0 days or control. MCDHF, methionine-choline-deficient and high-fat diet; S1P, Sphingosine 1-phosphate; Chil1, Chitinase-like 1; Ly6G, lymphocyte antigen 6 complex, locus G; Adgre1, adhesion G protein-coupled receptor E1; Acta2, actin alpha 2; Alb, albumin.

Article Snippet: Recombinant mouse CHIL1 (2649-CH-050) was from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Marker, Control

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Recombinant CHIL1 inhibited the pro-inflammatory phenotype of macrophages. (A) Schematic representation of the experimental design: bone marrow-derived macrophages were stimulated with LPS 10 ng/mL for 3 hours, washed with PBS, and then treated with Vehicle (PBS) or rCHIL1 (100 ng/mL) for 24 hours respectively. (B) Macrophages were harvested and the mRNA expression levels of genes Ccl4 , Tnf and Nos2 were evaluated using RT-qPCR, presented relative to Gapdh , (control group, n=3; LPS group, n=6). The protein expressions of CCL4, TNF and NOS2 in liver were detected by CBA (C, D) and WB (E) n=4. The results were presented as mean ± SD. Two-way ANOVA was used. *P < 0.05. rCHIL1, Recombinant CHIL1; Ccl4, C-C chemokine motif ligand 4; Tnf, tumor necrosis factor; Nos2, nitric oxide synthase 2; LPS, lipopolysaccharide.

Article Snippet: Recombinant mouse CHIL1 (2649-CH-050) was from R&D Systems (Minneapolis, MN).

Techniques: Recombinant, Derivative Assay, Expressing, Quantitative RT-PCR, Control

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Immunodepletion of CHIL1 in A-Neu supernatant attenuated the inhibitory effect on pro-inflammatory macrophages. (A) Schematic representation of the experimental design: bone marrow-derived macrophages were stimulated with LPS 10 ng/mL for 3 hours, washed with PBS, and then cultured in the presence of A-Neu supernatant (SUP) or A-Neu supernatant immunodepleted with anti-CHIL1 (SUP-anti-CHIL1). Expressions of CCL4, TNF and NOS2 in indicated groups were detected by RT-qPCR ( B , n=6), CBA ( C, D , n=4) and WB ( E , n=3). The results were presented as mean ± SD. One-way ANOVA was used. *P < 0.05. A-Neu, activated neutrophil; N-Neu, non-activated neutrophil; SUP, activated neutrophil supernatant; SUP-anti-CHIL1, activated neutrophil supernatant immunodepletion CHIL1; Ccl4, C-C chemokine motif ligand 4; Tnf, tumor necrosis factor; Nos2, nitric oxide synthase 2; LPS, lipopolysaccharide.

Article Snippet: Recombinant mouse CHIL1 (2649-CH-050) was from R&D Systems (Minneapolis, MN).

Techniques: Immunodepletion, Derivative Assay, Cell Culture, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: The schematic diagram of the main content of this study. A-Neu secrete CHIL1 and reduce liver inflammation by inhibiting the pro-inflammatory macrophage response. A-Neu, activated neutrophil; N-Neu, non-activated neutrophil; Ccl4, C-C chemokine motif ligand 4; Tnf, tumor necrosis factor; Nos2, nitric oxide synthase 2; S1P, Sphingosine 1-phosphate; Chil1, Chitinase-like 1.

Article Snippet: Recombinant mouse CHIL1 (2649-CH-050) was from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Primer sequence.

Article Snippet: Protein G-conjugated magnetic beads were incubated with sheep polyclonal anti-CHIL1 antibody (AF2649, R&D Systems).

Techniques: Sequencing

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Microarray analysis was performed in N-Neu and A-Neu. (A) Microarray heatmap data showing the expression level of transcript between N-Neu and A-Neu. The mRNA expressions were expressed as follows: Bright green, under expression; black, no change; bright red, overexpression. n=3. Row-based Z-score normalized. (B) GO enrichment analysis of differentially expressed genes in S1P activated neutrophils. (C) Venn diagram of gene expression overlap between GO terms including Regulation of Cell Communication, Cytokine Secretion, Inflammatory Response and Extracellular Space clusters. (D) Volcano plot of A-Neu gene expression compared with N-Neu. The threshold values were fold change≥|2| and P value ≤ 0.05. Red dots and blue dots represent up-regulated and down-regulated genes, respectively. Gray dots indicate genes with no statistically significant differences. The arrow indicated the location of Chil1. A-Neu, activated neutrophil; N-Neu, non-activated neutrophil; Il1b, Interleukin 1 beta; Chil1, Chitinase-like 1; Ccl3, Chemokine (C-C motif) ligand 3; Il1a, Interleukin 1 alpha; Tnf, tumor necrosis factor.

Article Snippet: Protein G-conjugated magnetic beads were incubated with sheep polyclonal anti-CHIL1 antibody (AF2649, R&D Systems).

Techniques: Microarray, Expressing, Over Expression, Gene Expression

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Chil1 was positively correlated with Ly6g in the injured liver of mice and could be secreted by A-Neu. (A) The mRNA expression of Chil1 was examined by RT-qPCR in the injured liver of MCDHF-treated mice in indicated times (n=6). (B) S1P stimulated neutrophils for 2 hours, then PBS washed the cells, and continued to culture for 6 hours, the secretion of CHIL1 in cell supernatant was detected by ELISA, n=4. (C) Correlation analysis between Chil1 mRNA expression with neutrophil marker Ly6g , macrophage marker Adgre1 , hepatocyte marker Alb and myofibroblast marker Acta2 in the injured liver of MCDHF-treated mice. Nonparametric test (Kruskal-Wallis test) was used in (A) Student’s t test was used in (B) Pearson’s test was used in (C) *P < 0.05 vs. MCDHF treated group for 0 days or control. MCDHF, methionine-choline-deficient and high-fat diet; S1P, Sphingosine 1-phosphate; Chil1, Chitinase-like 1; Ly6G, lymphocyte antigen 6 complex, locus G; Adgre1, adhesion G protein-coupled receptor E1; Acta2, actin alpha 2; Alb, albumin.

Article Snippet: Protein G-conjugated magnetic beads were incubated with sheep polyclonal anti-CHIL1 antibody (AF2649, R&D Systems).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Marker, Control

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Recombinant CHIL1 inhibited the pro-inflammatory phenotype of macrophages. (A) Schematic representation of the experimental design: bone marrow-derived macrophages were stimulated with LPS 10 ng/mL for 3 hours, washed with PBS, and then treated with Vehicle (PBS) or rCHIL1 (100 ng/mL) for 24 hours respectively. (B) Macrophages were harvested and the mRNA expression levels of genes Ccl4 , Tnf and Nos2 were evaluated using RT-qPCR, presented relative to Gapdh , (control group, n=3; LPS group, n=6). The protein expressions of CCL4, TNF and NOS2 in liver were detected by CBA (C, D) and WB (E) n=4. The results were presented as mean ± SD. Two-way ANOVA was used. *P < 0.05. rCHIL1, Recombinant CHIL1; Ccl4, C-C chemokine motif ligand 4; Tnf, tumor necrosis factor; Nos2, nitric oxide synthase 2; LPS, lipopolysaccharide.

Article Snippet: Protein G-conjugated magnetic beads were incubated with sheep polyclonal anti-CHIL1 antibody (AF2649, R&D Systems).

Techniques: Recombinant, Derivative Assay, Expressing, Quantitative RT-PCR, Control

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: Immunodepletion of CHIL1 in A-Neu supernatant attenuated the inhibitory effect on pro-inflammatory macrophages. (A) Schematic representation of the experimental design: bone marrow-derived macrophages were stimulated with LPS 10 ng/mL for 3 hours, washed with PBS, and then cultured in the presence of A-Neu supernatant (SUP) or A-Neu supernatant immunodepleted with anti-CHIL1 (SUP-anti-CHIL1). Expressions of CCL4, TNF and NOS2 in indicated groups were detected by RT-qPCR ( B , n=6), CBA ( C, D , n=4) and WB ( E , n=3). The results were presented as mean ± SD. One-way ANOVA was used. *P < 0.05. A-Neu, activated neutrophil; N-Neu, non-activated neutrophil; SUP, activated neutrophil supernatant; SUP-anti-CHIL1, activated neutrophil supernatant immunodepletion CHIL1; Ccl4, C-C chemokine motif ligand 4; Tnf, tumor necrosis factor; Nos2, nitric oxide synthase 2; LPS, lipopolysaccharide.

Article Snippet: Protein G-conjugated magnetic beads were incubated with sheep polyclonal anti-CHIL1 antibody (AF2649, R&D Systems).

Techniques: Immunodepletion, Derivative Assay, Cell Culture, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Activated Neutrophils Secrete Chitinase-Like 1 and Attenuate Liver Inflammation by Inhibiting Pro-Inflammatory Macrophage Responses

doi: 10.3389/fimmu.2022.824385

Figure Lengend Snippet: The schematic diagram of the main content of this study. A-Neu secrete CHIL1 and reduce liver inflammation by inhibiting the pro-inflammatory macrophage response. A-Neu, activated neutrophil; N-Neu, non-activated neutrophil; Ccl4, C-C chemokine motif ligand 4; Tnf, tumor necrosis factor; Nos2, nitric oxide synthase 2; S1P, Sphingosine 1-phosphate; Chil1, Chitinase-like 1.

Article Snippet: Protein G-conjugated magnetic beads were incubated with sheep polyclonal anti-CHIL1 antibody (AF2649, R&D Systems).

Techniques: