Journal: Frontiers in Immunology

Article Title: Protective effects of YCHD on the autoimmune hepatitis mice model induced by Ad-CYP2D6 through modulating the Th1/Treg ratio and intestinal flora

doi: 10.3389/fimmu.2024.1488125

Figure Lengend Snippet: Effect of YCHD on Th1 and Treg cell differentiation. Lymphocytes isolated from the spleen and hepatic lymph nodes of the C57BL/6 mice were stimulated with or without PMA (50ng/ml), ionomycin (1μg/ml), and BFA (10μg/ml) for 5h at 37°. (A, B) The gating strategy of Th1 ang Treg cells isolated from the spleen defined as CD3 + CD4 + IFN-γ + cells and CD3 + CD4 + CD25 + FOXP3 + cells and the representative dot plots from each group are shown. (C) Frequency Th1 and Treg cells in the spleen of mice (n=8) in each group were analyzed by flow cytometry. (D, E) The Th1 and Treg population in the hepatic hilar lymph nodes was analyzed by staining the cell surface CD4 antigen and IFN-γ antigen, as well as by staining of cell surface CD4 antigen, CD25 antigen, and intracellular Foxp3, in the CD3 + T cells population. (F) Frequency Th1 and Treg cells in the hepatic hilar lymph nodes of mice (n=8) in each group were also analyzed by flow cytometry. *P<0.05, **P<0.01, compared with the control group. ##P<0.01, compared with the untreated group.

Article Snippet: Cells were blocked with 50μl blocking buffer (2% goat serum, 2% fetal bovine serum, and 96% PBS) for 30 minutes at 4 °, 5μl PerCP Anti-Mouse CD3 Antibody (E-AB-F1013F, Elabscience) and 0.5μl FITC anti-mouse CD4 (B374032, BioLegend) were added for surface staining (incubated at 4°C for 40 minutes).

Techniques: Cell Differentiation, Isolation, Flow Cytometry, Staining, Control