Figure 6.

Pharmacological inhibition of PLD1 modulates the Th17/Treg balance in murine splenocytes. (A) Effect of the PLD1 inhibitor on the population of CD4⁺IL-17⁺ cells in the spleens of the indicated mice. (B) Effect of the PLD1 inhibitor on the population of CD4⁺Foxp3⁺ cells in the spleen of mice with CIA. (C,D) Effect of the PLD1 inhibitor on in vitro differentiation of Treg (C) and Th17 (D) cells. Naïve CD4⁺ T cells isolated from mice spleens were cultured under condition of Treg cells or Th17 cell differentiation. Representative flow cytometry profile and quantification for CD25⁺Foxp3⁺ Treg and CD25⁺IL-17⁺ Th17 cell populations. (E) Freshly isolated naïve CD4⁺ T cells (naïve T cells), labeled with CFSE were cocultured in the presence of anti-mouse CD3ε and anti-mouse CD28 for 72 h with induced Treg cells (iTreg) generated in the presence or absence of the PLD1 inhibitor. Proliferation of naïve CD4⁺ T cells was analyzed by flow cytometry for CFSE dilution. (F,G) Effect of the PLD1 inhibitor on the secretion (F) and expression (G) of IL-17 during differentiation of Th17 cells as analyzed by ELISA and q-PCR, respectively. n = 6 per group. * p < 0.05. Results are representative of at least six independent experiments, and shown as mean ± standard error of the mean (SEM).