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: 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 IFNγ expression, the Th2 cell percentage via IL4 expression, and the Th17 cell percentage via IL17A expression. (B) ELISA was applied to analyze serum IFNγ (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 IFNγ (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 (PE-Cy5, 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 IFNγ (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: DON treatment suppressed T cells differentiation in vitro . Freshly separated spleen cells were stimulated with 1.5mg/ml ConA with or without treating 2.5μM DON, and cells were analyzed 24h after ConA stimulation. (A) The differentiation markers of CD4+ T cells (IFNγ for Th1 cells, IL4 for Th2 cells and IL17 for Th17 cells) were detected using flowcytometry. (B) qRT-PCR was applied to analyze IFN-γ, IL4 and IL17 mRNA levels. (C) ELISA was applied to analyze supernatant IFN-γ and IL17 secretions. Flow cytometry was also applied to detect the production of activation markers of CTL, such as IFNγ (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 (PE-Cy5, 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 IFNγ (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: Technology and Health Care

Article Title: Prediction and identification of epitopes in the Echinococcus multilocularis thrombospondin 3 antigen

doi: 10.3233/THC-212983

Figure Lengend Snippet: The enzyme-linked immunosorbent assay test on the cytokines of mouse spleen lymphocytes immunized with thrombospondin 3. A: The interleukin-2 test; B: the interferon-y test; C: the tumor necrosis factor- β test; D: the interleukin-4 test; E: the interleukin-17 test. P * ⁣ * * < 0.001.

Article Snippet: PE-labeled anti-mouse IL-4 flow antibody , BD, USA.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Technology and Health Care

Article Title: Prediction and identification of epitopes in the Echinococcus multilocularis thrombospondin 3 antigen

doi: 10.3233/THC-212983

Figure Lengend Snippet: The enzyme-linked immune absorbent spot test of the cytokines of mouse spleen lymphocytes immunized with thrombospondin 3. A: The interleukin-2 test; B: the interferon- γ test; C: the tumor necrosis factor- β test; D: the interleukin-4 test. P * ⁣ * * < 0.001.

Article Snippet: PE-labeled anti-mouse IL-4 flow antibody , BD, USA.

Techniques: Spot Test

Journal: Technology and Health Care

Article Title: Prediction and identification of epitopes in the Echinococcus multilocularis thrombospondin 3 antigen

doi: 10.3233/THC-212983

Figure Lengend Snippet: Experimental reagents

Article Snippet: PE-labeled anti-mouse IL-4 flow antibody , BD, USA.

Techniques: Cell Culture, Labeling, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot