Our findings outlined in these studies support the possibility that local intragraft expression of IP-10 facilitates the migration of expanded Tregs into the graft. Consistent with our observations, CXCR3+ cells isolated from inflamed livers were found to have KU-57788 mw suppressive function 40, 41. Also, FOXP3+ T cells have been observed within renal allografts

in association with rejection 50. These findings as well as others 16, 17, 51 strongly suggest that alloactivated Tregs migrate into allografts where they have the potential to suppress the local inflammatory response. Our observations are suggestive that CXCR3 faciltates the peripheral migration of Tregs into allografts and that this subset has the potential to suppress ongoing rejection. It is well established that mTOR inhibitors augment the expansion of Tregs 47, 48 and promote tolerance induction in vivo 48, 52, 53. We find that the mTOR inhibitor rapamycin also permits the expansion of CXCR3hi Tregs in vitro, and we found higher numbers of circulating FOXP3+CXCR3+ Tregs in transplant recipients treated with mTOR inhibitors versus those treated with calcineurin inhibitors as part of their maintenance immunosuppressive therapy. Our studies involved small numbers of patients, but they are suggestive that the use of

mTOR-inhibitor therapy may enable the expansion of CXCR3+ Tregs in vivo, and may have an impact on long-term graft survival. SB203580 chemical structure Further evaluation of this observation in a larger cohort of patients may identify if expansion of this subset, for instance in association with the use of mTOR inhibitors, may serve as a biomarker and/or predict long-term graft survival. In summary, although CXCR3 is classically reported to be expressed on T effector cells, these new findings demonstrate that it is also expressed on populations of immunoregulatory T cells. Our findings explain the variable effects of CXCR3 blockade

in allograft SDHB rejection 32, 42, in as much as it was not previously known that CXCR3 may mediate the local trafficking of Tregs. Thus, an important implication of our observations is that the activation and expansion of CXCR3-expressing Tregs in vivo will facilitate the compartmentalization of T-cell regulatory subsets within allografts. Mouse anti-human CD4-FITC (RPA-T4), anti-human CD4-PE (RPA-T4), anti-human CD4-PECy7 (RPA-T4), anti-human CD39-FITC (A1), anti-human CCR7-PE (3D12), anti-human CCR5-FITC (HEK/1/85a) and anti-human FOXP3-FITC (206D) were obtained from Biolegend (San Diego, CA). Mouse anti-human FOXP3-APC (3G3), mouse anti-human CD62L-APC (DREG-56) and mouse anti-human CCR4-FITC were purchased from Miltenyi Biotec (Auburn, CA), eBioscience (San Diego, CA) and R&D Systems (Minneapolis, MN) respectively.