Abstract
Abstract The development of Foxp3+ CD4+ regulatory T (Treg) cells in the thymus is important for the preservation of self-tolerance and the prevention of spontaneous autoimmunity. It has been suggested that the CD4+CD8+ double positive (DP) stage is the earliest that Foxp3+ expression can be observed. We hypothesized that the antigen specificity of Foxp3+ DP cells may differ from Foxp3+ CD4+CD8- (CD4SP) cells due to the different antigenic repertoires presented by cortical thymic epithelial cells versus medullary APCs. However, purification of Foxp3+ DP cells proved difficult as many of those events on flow cytometry represented doublets of Foxp3+ CD4SP cells and Foxp3- DP cells. Our data therefore suggest that the frequency of Foxp3+ cells that can be found at the DP stage is around 1%, which is much smaller than previously reported. Rather, our current data suggests that thymic Treg cell development occurs primarily at the HSAhi CD4SP stage involving a two-step process involving both TCR-instructive and TCR-independent steps.
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