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: Frontiers in Immunology

Article Title: Treatment of Surgical Brain Injury by Immune Tolerance Induced by Peripheral Intravenous Injection of Biotargeting Nanoparticles Loaded With Brain Antigens

doi: 10.3389/fimmu.2019.00743

Figure Lengend Snippet: (A) Neurological function scoring histogram. (B) Histogram of peripheral blood CD4 + T/CD8 + T cell ratios. (C) Histogram of the proinflammatory cytokine IL-2, IL-4, and TGF-β1.*compared with Group A, P < 0.05; # compared with Group B, P < 0.05. (D) Iba-1 expression scoring of microglial cells measured at 21 d after SBI. *compared with Group A, P < 0.05; # compared with Group B, P < 0.05. The brown-positive cells are activated microglial cells, the yellow arrows indicate the somata of the activated microglia, and the red arrows indicate the processes of the activated microglia (x40). The black arrows indicate positive results (the brown cells) (x20). The scale bars are 20 μm. (E) GFAP expression scoring of astrocytes measured at 21 d after SBI. *compared with Group A, P < 0.05, **compared with Group A, P < 0.01; # icompared with Group B, P < 0.05. The scale bars are 50 μm.

Article Snippet: According to the ELISA kit protocol (Boster Biological Technology, Wuhan, China) plus the TMB color drops, then proinflammatory factor IL-2, anti-inflammatory factor IL-4 and inflammation suppression factor TGF-β1 concentrations were measured at 1, 7, 14, and 21 d postoperatively by using the enzyme reader at 450 nm.

Techniques: Expressing

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