Journal: Cell Reports Medicine

Article Title: TYK2 inhibition enhances Treg differentiation and function while preventing Th1 and Th17 differentiation

doi: 10.1016/j.xcrm.2025.102303

Figure Lengend Snippet: TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region (TSDR) after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: TSDR Pyrosequencing primer: AGAAATTTGTGGGGTGGG , Boardman et al. , N/A.

Techniques: Inhibition, Isolation, Cell Culture, Expressing, Control, Methylation, Concentration Assay