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                <title level="j">Journal of the American Society of Nephrology</title>
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                  <publisher>American Society of Nephrology</publisher>
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                  <biblScope unit="issue">8</biblScope>
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                  <date type="datePub">2015</date>
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                <term xml:lang="en">tolerance</term>
                <term xml:lang="en">pathology</term>
                <term xml:lang="en">kidney transplantation</term>
                <term xml:lang="en">immunology</term>
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              <p>The role of Foxp3 + regulatory T cells (Tregs) in operational tolerance remains elusive, as initial results revealed an increased frequency of this subset in tolerant patients but no functional differences compared with immunosuppressed recipients. In addition, recent studies of regulatory B cells strongly suggest that Tregs may not have a central role in kidney transplantation tolerance. However, recent investigations of the crucial role of Foxp3 demethylation in Treg function and the possibility of identifying distinct Foxp3 T cell subsets prompted us to more thoroughly characterize Tregs in operationally tolerant patients. Thus, we studied the level of demethylation of the Foxp3 Treg-specific demethylated region (TSDR) in circulating CD4 + T cells and analyzed Treg subset frequency in tolerant patients, healthy volunteers, patients with stable graft function under immunosuppression, and chronically rejecting recipients. We observed a higher proportion of CD4 + T cells with demethylated Foxp3 and a specific expansion of CD4 + CD45RA 2 Foxp3[hi] memory Tregs exclusively in tolerant patients. The memory Tregs of tolerant recipients exhibited increased Foxp3 TSDR demethylation, expressed higher levels of CD39 and glucocorticoid-induced TNF-related receptor, and harbored greater suppressive properties than memory Tregs from patients with stable graft function. Taken together, our data demonstrate that operationally tolerant patients mobilize an array of potentially suppressive cells, including not only regulatory B cells but also Tregs. Our results also indicate that tolerant patients have potent CD4 + CD45RA 2 Foxp3 hi memory Tregs with a specific Foxp3 TSDR demethylation pattern, which may contribute to the maintenance of graft tolerance.</p>
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