Journal: Frontiers in immunology

Article Title: Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.

doi: 10.3389/fimmu.2022.880262

Figure Lengend Snippet: FIGURE 3 | JHU083 inhibited T cells differentiation in vivo. Mice of Vehicle-AIH and JHU083-AIH group were intravenously injected with 8ml/kg ConA, and Vehicle- WT group was injected with normal saline of equal volume. JHU083-AIH group was gavaged with 0.3mg/kg JHU083 24h and 1h before ConA injection, and Vehicle- WT and Vehicle-AIH group were treated with equal vehicle at the same time points. (A) Flow cytometry was applied to analyze the Th1 cell percentage via IFNg expression, the Th2 cell percentage via IL4 expression, and the Th17 cell percentage via IL17A expression. (B) ELISA was applied to analyze serum IFNg (Th1 cells) and IL17 (Th17 cells) secretions. Flow cytometry was also applied to analyze the differentiation of CD8(+) T cells into cytotoxic T lymphocytes, evidenced by secreting IFNg (C) and Granzyme (D). Data were expressed as means ± SEM. *P < 0.05, **P <0.01, and ***P <0.001. ns, not significant.

Article Snippet: The antibodies used in the flow cytometry were as follows: CD4 (FITC, E-AB-F1097C), CD8a (PE, E-AB-F1104D), CD25 (PECy5, E-AB-F1102G), CD69 (PE-Cy7, E-AB-F1187H), IL4 (PE, E-AB-F1204UD), IL17A (APC, E-AB-F1272E) were purchased from Elabscience Biotechnology (Wuhan, China), antibodies IFNg (APC-Cy7, cat# 561479) was purchased from BD Biosciences (UK), and Granzyme B (PE-Cy7, cat# 372213) was purchased from Biolegend (San Diego, CA, USA).

Techniques: In Vivo, Injection, Saline, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in immunology

Article Title: Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.

doi: 10.3389/fimmu.2022.880262

Figure Lengend Snippet: FIGURE 5 | DON treatment suppressed T cells differentiation in vitro. Freshly separated spleen cells were stimulated with 1.5mg/ml ConA with or without treating 2.5mM DON, and cells were analyzed 24h after ConA stimulation. (A) The differentiation markers of CD4+ T cells (IFNg for Th1 cells, IL4 for Th2 cells and IL17 for Th17 cells) were detected using flowcytometry. (B) qRT-PCR was applied to analyze IFN-g, IL4 and IL17 mRNA levels. (C) ELISA was applied to analyze supernatant IFN-g and IL17 secretions. Flow cytometry was also applied to detect the production of activation markers of CTL, such as IFNg (D) and Granzyme B (E). Data were expressed as means ± SEM. **P <0.01 and ***P <0.001, ns, not significant.

Article Snippet: The antibodies used in the flow cytometry were as follows: CD4 (FITC, E-AB-F1097C), CD8a (PE, E-AB-F1104D), CD25 (PECy5, E-AB-F1102G), CD69 (PE-Cy7, E-AB-F1187H), IL4 (PE, E-AB-F1204UD), IL17A (APC, E-AB-F1272E) were purchased from Elabscience Biotechnology (Wuhan, China), antibodies IFNg (APC-Cy7, cat# 561479) was purchased from BD Biosciences (UK), and Granzyme B (PE-Cy7, cat# 372213) was purchased from Biolegend (San Diego, CA, USA).

Techniques: In Vitro, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Activation Assay

Journal: Annals of Dermatology

Article Title: Hyaluronan Oligosaccharides Improve Rosacea-Like Phenotype through Anti-Inflammatory and Epidermal Barrier-Improving Effects

doi: 10.5021/ad.2020.32.3.189

Figure Lengend Snippet: Treatment with hyaluronan oligosaccharides (oligo-HAs) reduced inflammatory cell infiltration, interleukin-17 (IL-17), and kallikrein 5 (KLK5) expressions. (A) Hematoxylin and eosin-stained tissue sections of LL-37 induced rosacea-like mice showed marked inflammatory cellular infiltration. Further treatments with oligo-HAs reduced cellular infiltration (H&E, ×100). (B) Majority of infiltrated cells were CD4+ T cells and infiltration of CD4+ T cells decreased in oligo-HAs treated group (CD4, ×100). (C) IL-17A expression was markedly increased in the epidermis of LL-37 induced rosacea-like mice. Oligo-HAs decreased the IL-17A expression regardless of the injection method (IL-17A, ×200). (D) Expression of KLK5 was increased in LL-37 induced rosacea-like mice (arrow head). Further treatment with oligo-HAs reduced KLK5 expression (KLK5, ×400). i.d.: intradermally, i.p.: intraperitoneally.

Article Snippet: To detect CD4, IL-17A, KLK5, filaggrin, and CD44 in histological specimens from mice, primary antibodies including a monoclonal mouse anti-CD4 antibody (BD Biosciences, Franklin Lakes, NJ, USA), monoclonal rat anti-mouse IL-17A antibody (Covalab, Villeurbanne, France), polyclonal rabbit anti-KLK5 antibody (Abcam, Cambridge, UK), polyclonal mouse anti-profilaggrin antibody (Covance, Princeton, NJ, USA), and monoclonal rat anti-human/mice CD44 antibody (eBioscience, San Diego, CA, USA) were used.

Techniques: Staining, Expressing, Injection