Frequency of CD4⁺ TGFβRII⁺ T cells in tissues of tolerized C57BL/6 mice after single challenge with OVA plus CT.

A. Mononuclear cells were isolated from PPs, MLNs, iLP and spleens of C57BL/6 mice 7 days after 30 mg of OVA feeding (0) and at 0.5, 2 and 4 h after single challenge with 1 mg of OVA plus 10 µg of CT (OVA plus CT). Cells were then stained with FITC-conjugated anti-hTGFβRII, APC-labeled anti-CD4, and biotin-tagged anti-CD3 mAbs followed by PerCP-Cy™5.5-conjugated streptavidin. B. Cells were isolated as described above and stained with FITC-conjugated anti-hTGFβRII, APC-labeled anti-CD4, PE-labeled anti-hFoxp3, and biotin-tagged anti-CD3 mAbs followed by PerCP-Cy™5.5-conjugated streptavidin. Samples were subjected to flow cytometry analysis by FACSCalibur®. n = 5 mice per time point, *p<0.05 compared to sham-tolerized and OVA plus CT challenged controls.