Figure 6.

Protective effects of protein transduction domain (PTD)-mouse forkhead box protein 3 (mFoxP3) were associated with regulating the balance between T helper type 17 (Th170 and regulatory T cells (T_regs). Lymphocytes were obtained from DBA/1 mice treated with PTD-mFoxP3, mFoxP3 or MTX after CII immunization, and analysed by flow cytometry (FCM) for CD4⁺ FoxP3⁺ T cells or CD4⁺ interleukin (IL)-17A⁺ cells after in-vitro restimulation with phorbol myristate acetate (PMA)/ionomycin in the presence of GolgiStop for 4 h. (a) Representative dot-plots of T_reg cells from draining lymph nodes of collagen-induced arthritis (CIA) mouse. For staining of T_reg cells, anti-CD4-fluorescein isothiocyanate (FITC) and anti-CD25-phycoerythrin (PE) surface-stained cells were fixed and permeabilized, and then stained with anti-FoxP3-allophycocyanin (APC). (b) Frequency of T_reg cells subset in the CD4⁺ T lymphocytes of each group. Data are expressed as the mean ± standard deviation (s.d.), n = 5, *P < 0·05 versus phosphate-buffered saline (PBS)-treated group. (c) Representative dot-plots of Th17 cells from draining lymph nodes of CIA mouse isolated after treated for 4 weeks. For staining of Th17 cells, anti-CD4-FITC surface-stained cells were fixed and permeabilized, and then stained with anti-IL-17A-APC. (d) Frequency of Th17 cells subset in the CD4⁺ T lymphocytes of each group. Data are expressed as the mean ± s.d., n = 5, **P < 0·01 versus PBS-treated group. (e) The ratio of T_reg-like/Th17 cells in the CD4⁺ T lymphocytes of each group. Data are expressed as the mean ± s.d., n = 5, **P < 0·01; ***P < 0·001 versus PBS-treated group.