Journal: Journal of Translational Medicine
Article Title: Niclosamide suppresses the expansion of follicular helper T cells and alleviates disease severity in two murine models of lupus via STAT3
Figure Lengend Snippet: Niclosamide inhibits T FH -like cell differentiation and B cell IgG production in vitro. Naive CD4 + T cells were purified from the spleens of MRL/ lpr and C57BL/6 mice. For T FH -like cell differentiation, purified CD4 + T cells were activated with mouse T-activator CD3/CD28 Dynabeads, and treated with 20 ng/ml IL-6, 20 ng/ml IL-21, 10 μg/ml anti-IL-4, 10 μg/ml anti-IFN-γ, and 20 μg/ml anti-TGF-β for 4 days with or without niclosamide. a Left, T FH -like cells (CXCR5 + PD-1 + , gated on CD4 + ) isolated from MRL/ lpr mice were analyzed by flow cytometry. Right, the percentage of T FH -like cells is shown. b p-STAT3 and TCF-1 expressions in T FH -like cells isolated from MRL/ lpr mice were analyzed by western blot. c mRNA expression levels of Bcl-6 , CXCR5 , and Blimp-1 in T FH -like cells isolated from MRL/ lpr mice were measured by real-time PCR. d T FH -like cells and B cells isolated from C57BL/6 mice were co-cultured with or without T FH -like cells for 3 days, and then the concentrations of IgG in the supernatants were detected by ELISA. Data shown as mean ± SD. One-way ANOVA was performed. ** P
Article Snippet: For TFH -like cell differentiation, purified CD4+ T cells were seeded at 1 × 106 cells/well and were activated with mouse T-activator CD3/CD28 Dynabeads™ (Invitrogen), and treated with 20 ng/ml IL-6, 20 ng/ml IL-21, 10 μg/ml anti-IL-4, 10 μg/ml anti-IFN-γ, and 20 μg/ml anti-TGF-β (R & D Systems) for 4 days with or without niclosamide.
Techniques: Cell Differentiation, In Vitro, Purification, Mouse Assay, Isolation, Flow Cytometry, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay