Alloantigen induced Foxp3+CD25+CD4+ Tregs were differentiated from naïve,
polyclonal BALB/c CD4+ T cells using the combination of B6 DCs, IL-2, and
TGF-β1. The suppressors tested were (S) were either alloantigen induced
Tregs (A) or freshly isolated nTregs (B) and
were mixed in various ratios with 105 CD4+CD25−
cells from BALB/c [responders(R)], and stimulated with irradiated spleen cells
from B6 mice (A and B). (C) AlloTregs generated from BALB/c
CD4+ T cells with B6 DCs, IL-2, and TGF-β1 were mixed with
105 CD4+CD25− cells from CBA responder T
cells in various ratios, and stimulated with irradiated SJL spleen cells.
Proliferation was assessed by [3H]thymidine uptake. Data from each of
three consecutive experiments are shown, and expressed as mean (±SE) of
three replicates within each experiment. P values calculated using the
Kruskal-Wallis test.