Detection of
CD4+CD25+Foxp3+Tregs and Th cells in spleens and MLNs by flow cytometry
Spleen and MLN lymphocyte single cell suspensions were prepared, and
1×106 cells were added to each tube. The cells were
resuspended in PBS and incubated for 30 min in the dark with the
following antibody combinations: FITC-rat-anti-mouse CD4,
PE-rat-anti-mouse Foxp3 and APC-rat-anti-mouse CD25; FITC-rat-anti-mouse
IL-17 and PE-Rat-anti-mouse IL-12; FITC-rat-anti-mouse IL-10 and
PE-rat-anti-mouse IFN-γ ; or PE-rat-anti-mouse IL-4 (all antibodies were
obtained from BD, USA). The cells were washed and resuspended again for
flow cytometry analysis. All data were obtained from three separate
experiments.