Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: HCC cells promote MC migration and sustain MC proliferation. (A, B) Timer3.0 database analysis of SCF expression in tumors and correlation analysis between SCF and MCs; (C) ELISA detection of SCF levels in tumor tissues and adjacent tissues; (D) Correlation analysis between TAMC density and SCF concentration in HCC tumor tissues; (E): Flow cytometry identification of expanded hMCs at week 4; (F, G) Transwell assay for hMC migration capacity; (H, I) Flow cytometry for hMC proliferation; (J, K) Flow cytometry for hMC cell cycle. Data are presented as mean ± SD. Figure (B) used independent samples t ‐tests ( n = 59). (D) Spearman 's rank correlation analysis ( n = 59); (G/I/K), one‐way ANOVA with Tukey's post‐hoc multiple comparison test ( N = 3). * indicates p < 0.05; *** indicates p < 0.001.

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Migration, Analysis, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Flow Cytometry, Transwell Assay, Comparison, X-Ray Photoelectron Spectroscopy

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: TAMCs recruit Tregs into the tumor microenvironment by secreting chemokines. (A) ELISA detection of CCL1, CCL5, CCL17, CCL22, and CCL28 secretion by TAMCs (after subtracting CCL1, CCL5, CCL17, CCL22, and CCL28 levels in TCCS); (B) ELISA detection of CCL17 levels in tumor tissues and adjacent tissues; (C) Correlation analysis between TAMCs and CCL17 concentration; D: Correlation analysis between Tregs and CCL17 concentration; (E, F) Transwell migration assay to assess Treg migration capacity. Data are presented as mean ± SD. (A, B), independent samples t ‐tests ( N = 3); (C, D) Spearman 's rank correlation analysis ( n = 59); (F), one‐way ANOVA and Tukey's multiple comparison test ( N = 3). * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Analysis, Concentration Assay, Transwell Migration Assay, Migration, Comparison

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: The TAMCs‐Tregs axis inhibits CD8 + T cell function. (A, B) Flow cytometry analysis of Treg‐mediated suppression of CD8 + T cell proliferation; (C–F) ELISA for Granzyme B, IFN‐γ, TNF‐α, and Perforin in supernatants from CD8 + T cell proliferation assays. Data are presented as mean ± SD. One‐way ANOVA was performed, followed by Tukey's multiple comparison test for post‐hoc analysis ( N = 3). * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Cell Function Assay, Flow Cytometry, Analysis, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: Combination therapy of anti‐PD‐1 antibody and CCR4 antagonists reshapes the tumor immune microenvironment in HCC. (A–F) Flow cytometry analysis of immune cell proportions (MCs, Tregs, CD8 + T cells) in tumor tissue; G–K: ELISA detection of Granzyme B, IFN‐γ, TNF‐α, Perforin, and CCL17 levels. Data are presented as mean ± SD. One‐way ANOVA was performed, followed by Tukey's multiple comparison test for post‐hoc analysis ( n = 6). * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Flow Cytometry, Analysis, Immune Cell, Tissue, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: Validation of the synergistic antitumor efficacy of the triple‐drug therapy in an in situ Hepa1‐6 HCC model. (A) Representative images of in situ tumors; (B): Tumor volume in each group; (C) Tumor weight in each group; (D) Average ratio of liver weight to body weight; (E, F): Ki67 immunohistochemical staining and quantitative analysis; (G–L) Flow cytometry detection of the proportions of various immune cells (mast cells, Tregs, CD8 + T cells); (M) ELISA detection of Granzyme B, IFN‐γ, TNF‐α, and perforin levels. Data are presented as mean ± SD. One‐way ANOVA was performed, followed by Tukey's multiple comparison test for post‐hoc analysis ( n = 6).

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Biomarker Discovery, In Situ, Immunohistochemical staining, Staining, Analysis, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: HCC cells promote MC migration and sustain MC proliferation. (A, B) Timer3.0 database analysis of SCF expression in tumors and correlation analysis between SCF and MCs; (C) ELISA detection of SCF levels in tumor tissues and adjacent tissues; (D) Correlation analysis between TAMC density and SCF concentration in HCC tumor tissues; (E): Flow cytometry identification of expanded hMCs at week 4; (F, G) Transwell assay for hMC migration capacity; (H, I) Flow cytometry for hMC proliferation; (J, K) Flow cytometry for hMC cell cycle. Data are presented as mean ± SD. Figure (B) used independent samples t ‐tests ( n = 59). (D) Spearman 's rank correlation analysis ( n = 59); (G/I/K), one‐way ANOVA with Tukey's post‐hoc multiple comparison test ( N = 3). * indicates p < 0.05; *** indicates p < 0.001.

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Migration, Analysis, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Flow Cytometry, Transwell Assay, Comparison, X-Ray Photoelectron Spectroscopy

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: TAMCs recruit Tregs into the tumor microenvironment by secreting chemokines. (A) ELISA detection of CCL1, CCL5, CCL17, CCL22, and CCL28 secretion by TAMCs (after subtracting CCL1, CCL5, CCL17, CCL22, and CCL28 levels in TCCS); (B) ELISA detection of CCL17 levels in tumor tissues and adjacent tissues; (C) Correlation analysis between TAMCs and CCL17 concentration; D: Correlation analysis between Tregs and CCL17 concentration; (E, F) Transwell migration assay to assess Treg migration capacity. Data are presented as mean ± SD. (A, B), independent samples t ‐tests ( N = 3); (C, D) Spearman 's rank correlation analysis ( n = 59); (F), one‐way ANOVA and Tukey's multiple comparison test ( N = 3). * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Analysis, Concentration Assay, Transwell Migration Assay, Migration, Comparison

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: The TAMCs‐Tregs axis inhibits CD8 + T cell function. (A, B) Flow cytometry analysis of Treg‐mediated suppression of CD8 + T cell proliferation; (C–F) ELISA for Granzyme B, IFN‐γ, TNF‐α, and Perforin in supernatants from CD8 + T cell proliferation assays. Data are presented as mean ± SD. One‐way ANOVA was performed, followed by Tukey's multiple comparison test for post‐hoc analysis ( N = 3). * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Cell Function Assay, Flow Cytometry, Analysis, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: Combination therapy of anti‐PD‐1 antibody and CCR4 antagonists reshapes the tumor immune microenvironment in HCC. (A–F) Flow cytometry analysis of immune cell proportions (MCs, Tregs, CD8 + T cells) in tumor tissue; G–K: ELISA detection of Granzyme B, IFN‐γ, TNF‐α, Perforin, and CCL17 levels. Data are presented as mean ± SD. One‐way ANOVA was performed, followed by Tukey's multiple comparison test for post‐hoc analysis ( n = 6). * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Flow Cytometry, Analysis, Immune Cell, Tissue, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Tumor‐Associated Mast Cells Drive Immunosuppression and Resistance to Anti‐PD‐1 Therapy in Hepatocellular Carcinoma Through CCL17/CCR4‐mediated Regulatory T‐Cell Recruitment

doi: 10.1002/jbt.71127

Figure Lengend Snippet: Validation of the synergistic antitumor efficacy of the triple‐drug therapy in an in situ Hepa1‐6 HCC model. (A) Representative images of in situ tumors; (B): Tumor volume in each group; (C) Tumor weight in each group; (D) Average ratio of liver weight to body weight; (E, F): Ki67 immunohistochemical staining and quantitative analysis; (G–L) Flow cytometry detection of the proportions of various immune cells (mast cells, Tregs, CD8 + T cells); (M) ELISA detection of Granzyme B, IFN‐γ, TNF‐α, and perforin levels. Data are presented as mean ± SD. One‐way ANOVA was performed, followed by Tukey's multiple comparison test for post‐hoc analysis ( n = 6).

Article Snippet: The following kits were utilized: human SCF ELISA Kit (E‐EL‐H1299, Elabscience), human CCL1 ELISA kit (ab314600, Abcam), human CCL5 ELISA kit (ab174446, Abcam), human CCL17 ELISA kit (ab183366, Abcam), Human CCL22 ELISA Kit (ab242931, Abcam), Human CCL28 ELISA Kit (ab99988, Abcam), Human Granzyme B ELISA Kit (E‐EL‐H1617, Elabscience), Human Interferon‐γ (IFN‐γ) ELISA Kit (E‐EL‐H0108, Elabscience), Human TNF‐α ELISA Kit (E‐EL‐H0109, Elabscience), Human Perforin 1 ELISA Kit (E‐EL‐H1123, Elabscience), Mouse Granzyme B ELISA Kit (E‐EL‐M0594, Elabscience), Mouse IFN‐γ ELISA Kit (E‐EL‐M0048, Elabscience), Mouse TNF‐α ELISA Kit (E‐EL‐M3063, Elabscience), and Mouse Perforin ELISA Kit (E‐EL‐M0890, Elabscience).

Techniques: Biomarker Discovery, In Situ, Immunohistochemical staining, Staining, Analysis, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: KU treatment mitigates EAE and reduces Th1 and Th17 cell proportions in mice. (A) Schematic timeline of EAE induction and KU administration (100 mg kg −1 , intragastric). Mice were immunized with MOG 35‐55 peptide, and KU or vehicle was administered from 11 to 21 dpi. (B) Line graph of clinical scores in the control (Ctrl), vehicle‐treated EAE, and KU‐treated EAE groups ( n = 8 per group). (C) Disease incidence curves of EAE mice. (D) Cumulative clinical scores and peak score in each group. (E) Representative HE staining and LFB staining of spinal cord from the Ctrl, vehicle‐treated EAE, and KU‐treated EAE groups. (F) Inflammation and demyelination scores based on HE and LFB staining ( n = 6 per group). (G) The proportion of CD3 + CD4 + T cells in the CNS and spleen (SP) ( n = 3–4 per group). (H) Absolute counts of CD3 + and CD4 + T cells in the CNS of each group. (I) The proportion of CD4 + IFNγ + (Th1) cells in the CNS and SP. (J) The proportion of CD4 + IL‐17A + (Th17) cells in the CNS and SP. (K) Relative mRNA expression of T‐bet and RORγt in the spinal cord measured by qPCR. (L) ELISA detection of serum IFNγ and IL‐17A concentrations in each group. Data are presented as mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. vehicle‐treated EAE.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Control, H&E Stain, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: KU modulates gut microbiota to ameliorate EAE. (A) Alpha diversity (Shannon index) of gut microbiota in the Ctrl, vehicle‐treated EAE, and KU‐treated EAE groups ( n = 8 per group). Pair‐wise comparisons were performed using Wilcoxon rank‐sum test. (B) Principal coordinate analysis (PCoA) of gut microbiota based on Bray‐Curtis distance (pseudo‐ F = 30.12, R 2 = 0.741, p < 0.001); each dot represents an individual sample. (C) Stacked bar plot showing the relative abundance of gut microbiota at the family level in the Ctrl, vehicle‐treated EAE, and KU‐treated EAE groups. (D) Schematic timeline of ABX treatment and FMT and EAE induction: EAE mice received FMT from vehicle‐treated or KU‐treated donor mice (100 mg kg −1 , intragastric). (E) Clinical scores of FMT‐Vehicle and FMT‐KU groups ( n = 9 per group). (F) Disease incidence curves of EAE mice. (G) Statistical analysis of cumulative clinical scores and peak score. (H) Representative HE staining and LFB staining of spinal cord tissues from FMT‐Vehicle and FMT‐KU mice. (I) Inflammation score and demyelination score in the spinal cord ( n = 4 per group). (J) Flow cytometric plots of CD4 + IFNγ + (Th1) and CD4 + IL‐17A + (Th17) cells in the CNS and SP. (K) Quantification of CD4 + IFNγ + and CD4 + IL‐17A + cell percentages in the CNS and SP ( n = 5 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. EAE mice that received FMT from vehicle‐treated donors.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Analysis, H&E Stain, Staining

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: KU ameliorates EAE in an A. muciniphila ‐dependent manner. (A) Heatmap showing the relative abundance of the top 10 gut microbial species in the Ctrl, vehicle‐treated EAE, and KU‐treated groups ( n = 8 per group). (B) Linear discriminant analysis effect size (LEfSe) analysis was performed to screen differentially abundant microbial taxa across the three groups. Taxa with an LDA score > 3 were defined as potential microbial biomarkers with statistical significance. (C) Schematic timeline of ABX pretreatment, KU administration, A. muciniphila supplementation, and EAE induction. (D) Clinical scores of EAE, EAE+ABX, EAE+ABX+KU, and EAE+ABX+KU+ A. muciniphila ( n = 8 per group). (E) Cumulative clinical scores and peak score in each group. (F) Representative HE staining and LFB staining of the spinal cord. (G) Spinal cord inflammation score and demyelination score ( n = 4 per group). (H) Flow cytometric plots of CD4 + IFNγ + (Th1) cells in the CNS and SP of each group. (I) Quantification of CD4 + IFNγ + cell percentages in the CNS and SP ( n = 4 per group). (J) Flow cytometric plots of CD4 + IL‐17A + (Th17) cells in the CNS and SP of each group. (K) Quantification of CD4 + IL‐17A + cell percentages in the CNS and SP ( n = 4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, p > 0.05.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Analysis, H&E Stain, Staining

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: IAAld alleviates EAE and reduces Th1 and Th17 cell proportions. (A) Schematic timeline of EAE induction and IAAld administration ( n = 8 per group). (B) Clinical scores of vehicle‐treated EAE, EAE+IAAld 40 mg/kg/day (IAAld 40), and EAE+IAAld 80 mg/kg/day (IAAld 80) groups. (C) Disease incidence curves of EAE mice with different doses of IAAld treatment. (D) Quantification of cumulative clinical scores and peak score in each group. (E) Representative HE staining and LFB staining of spinal cord tissues from the vehicle‐treated EAE, IAAld 40, and IAAld 80 groups. (F) Spinal cord inflammation score and demyelination score ( n = 3 per group). (G) Flow cytometric plots of CD4 + IFNγ + (Th1) and CD4 + IL‐17A + (Th17) cells in the CNS and SP of each group. (H) Quantification of CD4 + IFNγ + and CD4 + IL‐17A + cell percentages in the CNS and SP ( n = 4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: H&E Stain, Staining

Journal: Advanced Science

Article Title: Gut Microbiota‐Derived Indole‐3‐Acetaldehyde Contributes to the Disease‐Alleviating Effects of Kurarinone in Experimental Autoimmune Encephalomyelitis

doi: 10.1002/advs.78358

Figure Lengend Snippet: IAAld treatment mitigates EAE in an AhR‐dependent manner. (A) Relative mRNA expression levels of Ahr, Cyp1a1, and Cyp1b1 in the spinal cord of EAE mice treated with vehicle, IAAld 40, or IAAld 80 groups. (B) Schematic timeline depicting the experimental protocol for EAE induction, followed by administration of IAAld (80 mg/kg/day) and AhR inhibitor (AhRi; SR1, 50 mg/kg/day). (C) Clinical scores for EAE mice treated with vehicle, IAAld, and IAAld+AhRi groups ( n = 6 per group). (D) Disease incidence curves for EAE mice over time. (E) Quantification of cumulative clinical scores and peak score for each group. (F) Representative HE and LFB staining of spinal cord tissues for assessment of inflammation and demyelination, respectively. (G) Quantification of spinal cord inflammation and demyelination scores ( n = 3 per group). (H) Representative flow cytometry plots indicating the percentages of CD4 + IFNγ + (Th1) and CD4 + IL‐17A + (Th17) cells in the CNS and SP of each group. (I) Quantification of CD4 + IFNγ + and CD4 + IL‐17A + cell percentages in the CNS and SP across groups ( n = 3–4 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The mouse serum was detected using the IFNγ and IL‐17A ELISA kits (Elabscience, E‐EL‐M0047, E‐EL‐M0048) according to the kit instructions.

Techniques: Expressing, Staining, Flow Cytometry