Figure 5.

Induction of the iT_reg from naïve CD4⁺CD45RA⁺ T cells (Th₀) by allogeneic N-CMFs depends on MHC class II. T cells were cultured without or with allogeneic CMFs at a ratio 1:10 for 7 days in 24 well plates in presence/absence of anti-MHC class II mAb mix (anti- HLA-DR, -DQ and -DP cocktail, clones L243 and IVA12) or isotype mix control. (A) The RNA from the N-CMF primed T cells was analyzed for FoxP3 mRNA expression by using real time RT-PCR. The mRNA level for FoxP3 was normalized to 18S. Data represent mean of mRNA Δ fold increase ± SEfrom duplicates in three experiments (n=6) *P<0.01. (B) The surface CD25 and intracellular FoxP3 expression in the N-CMF primed T cells was analyzed using flow cytometry. Harvested T cells were stained with anti-CD4 FITC, anti-CD25PE-Cy7, and anti-FoxP3-APC mAbs. The appropriate isotype controls were included in the experiments. A representative experiment is shown (n=5). *p<0.01.