MOG35-55/I-A^b dimer modulates the expression of CD3 and TCR as well as the mTGF-β1 and IL-10 suppressive cytokines in MOG35-55-specific T cells.

<p>(A–D) The 2D2 splenocytes were stimulated with MOG35-55 (25 µg/mL) <i>in vitro</i> in the presence of MOG35-55/I-A^b dimer (1.2 nM) or PBS. After 5 days culture, cells were harvest and stained with anti-CD4-FITC mAb, anti-CD3-PE-Cy7 mAb, anti-TCR-Vα3.2-APC mAb, anti-mTGF-β1-PE and/or anti-IL-10-PE mAb. (A–B) The MFIs of (A) PE-cy7-CD3 and (B) APC-TCR Vα3.2 on CD4⁺ cells were analyzed by flow cytometry and expressed as the mean ± SD from 6 mice in each group. (C-D) MOG35-55/I-A^b dimer intervention induced a population of CD3^lowTCR^lowCD4⁺ cells with high level of (C) mTGF-β1 and (D) IL-10 expression. The right panel represents the mean ± SD of MFI of PE-mTGF-β1 and PE-IL-10 on CD3^highTCR^highCD4⁺/CD3^lowTCR^lowCD4⁺ cells in dimer intervention group (n = 3). (E) Splenocytes were isolated from mice treated with PBS or MOG35-55/I-A^b dimer on day 18 post-immunization and stained with anti-CD3-APC, anti-CD4-FITC, anti-TGF-β1-PE, and/or anti-IL-10-PE Abs. CD4⁺ cells were gated for analysis by flow cytometry. (a–b) The mean percentage of CD3^lowCD4⁺ cells and the MFI of APC-CD3 on CD4⁺ cells from 6 mice in each group are represented as the mean ± SD. (c) The MFIs of PE-mTGF-β1 and PE-IL-10 on CD3^lowCD4⁺/CD3^highCD4⁺ cells in dimer treated-group are expressed as the mean ± SD from 6 mice respectively. *, <i>P<</i>0.05; **, <i>P<</i>0.01; ***, <i>P<</i>0.001 and n.s., nonsignificant.