Fig. 5

Adoptive transfer of Foxp3-transfected CD19⁺ B cells protects from CIA. Transfected B cells (1 × 10⁷) with Foxp3 vector were injected i.v. into CIA mice (each group n = 10) on days 7 and 28, which had been immunized with CII/CFA. a, b Evaluation of the severity of arthritis (a) by measurement of the increase in paw swelling and histologic examinations (b) of the joints of control mice or mice injected with transfected B cells. Representative photographs from each group are shown. Original magnifications, ×40, ×200. c Percentage of CD4⁺Foxp3⁺CTLA4⁺ Treg cells in the spleens, mesenteric lymph nodes (mLN) and draining lymph nodes (dLN) from control CIA or mice injected Foxp3-transfected CD19⁺ B cells. Cells were stained with CD4-PerCP cy5.5 mAb before permeabilization and stained using Foxp3-FITC and CTLA4-PE mAb. FACS analyses showed that Foxp3⁺CLTA4⁺ cells gated on CD4⁺ T cells. d, e Spleens from mice of each group were stained with anti-CD4 (green) and anti-IL-17 (red) (upper image) or anti-CD4 (red), anti-CD25 (blue), and anti-Foxp3 (green) (lower image) antibodies. Populations of CD4⁺CD25⁺Foxp3⁺ and CD4⁺IL-17⁺ T cells were analyzed using confocal laser microscopy and Flow cytometry. f Mock and Foxp3 transfected B cells (1 × 10⁷) were injected i.v. into CIA mice (each group n = 10) on days 7 and 28, which had been immunized with CII/CFA. Original magnification, ×400. Data are expressed as the means ± SD. *p < 0.05 compared with control CIA mice