Efficient Therapeutic Function and Mechanisms of Human Polyclonal CD8⁺CD103⁺Foxp3⁺ Regulatory T Cells on Collagen-Induced Arthritis in Mice

Figure 3

Survival and stability of induced human CD8⁺Tregs transfused into CIA mice. Briefly, human CD8⁺Tregs labeled with CFSE were transfused into established CIA mice on day 28 by i.v. After 72 hours of adoptive infusion, the mice were sacrificed, and the cells in their blood (BD), spleen cells (SC), lymph nodes (LN), and paws (foot, FT, minced, and digested) were harvested and labeled using PE-conjugated anti-human CD8. The cells were subsequently fixed, permeabilized, and labeled with anti-human APC-Foxp3 and evaluated by FACS. CD8⁺CFSE⁺ cells were considered surviving CD8⁺Tregs, and the percentage of Foxp3-positive cells in CFSE⁺CD8⁺ surviving cells was investigated to evaluate the stability of CD8⁺ Tregs in CIA. (a) Survival of human CD8⁺Tregs after transfusion into CIA mice for 72 hours. Representative flow data showed surviving human CD8⁺CFSE⁺Treg (above) and Foxp3 expression in surviving CD8⁺Tregs (below). (b) Quantitative analysis of surviving human CD8⁺Tregs in vivo after transfusion for 72 hours. (c) Quantitative analysis of Foxp3 expression in surviving human CD8⁺Tregs in vivo after transfusion for 72 hours. Each mean represents data from 5 mice in each group; the bars indicate the ; , , and .

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