Figure 5. Ability of Kifunensine treated CD25+CD4+ cells to migrate in vivo.

(a + b) CBA were pre-treated with an anti-CD4 mAb i.v. at days -28 and -27. At day -27, mice also received specific donor blood transfusion (DST). CD25⁺CD4⁺ Treg were purified from the spleen of these animals at day 0, and incubated for 30 min with either KIF or PBS. 5×10⁵ Treg were adoptively transferred into CBA-RAG^−/− animals along with 10⁵ CFSE labeled BM3 T cells. One day later mice received a B10 skin graft and lymphoid organs were harvested at day 10. Treg cell (a) numbers on a log scale and (b) percentage of total cells were analyzed by FACS. Data from n = 4 animals per group. Data is representative of 3 repeats. (5c) 10⁵ CFSE labeled BM3 T cells were adoptively transferred into CBA-RAG-/- animals. One day later mice received a B10 skin graft and axillary LNs were harvested at day 10. (5d) 10⁵ CD25⁻CD4⁺ T cells were adoptively transferred into CBA-RAG-/- animals. One day later mice received a B10 skin graft and peripheral lymphoid organs were harvested at day 10. The number of T cells was quantified by FACS. Data from n = 5 animals per group. Data are representative of 2 independent experiments. * p<0.01. Error bars represent the standard deviation.