differential responses of human regulatory t cells (treg) and effector t cells to rapamycin微分响应人类的调节性t细胞(treg)和效应t细胞雷帕霉素.pdfVIP

Differential Responses of Human Regulatory T Cells (Treg) and Effector T Cells to Rapamycin Laura Strauss, Malgorzata Czystowska, Marta Szajnik, Magis Mandapathil, Theresa L. Whiteside* Department of Pathology, University of Pittsburgh School of Medicine and University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania, United States of America Abstract Background: The immunosuppressive drug rapamycin (RAPA) promotes the expansion of CD4+ CD25highFoxp3+ regulatory T cells via mechanisms that remain unknown. Here, we studied expansion, IL-2R-c chain signaling, survival pathways and resistance to apoptosis in human Treg responding to RAPA. + + + neg Methodology/Principal Findings: CD4 CD25 and CD4 CD25 T cells were isolated from PBMC of normal controls (n = 21) using AutoMACS. These T cell subsets were cultured in the presence of anti-CD3/CD28 antibodies and 1000 IU/mL IL-2 for 3 to 6 weeks. RAPA (1–100 nM) was added to half of the cultures. After harvest, the cell phenotype, signaling via the PI3K/ mTOR and STAT pathways, expression of survival proteins and Annexin V binding were determined and compared to values obtained with freshly-separated CD4+ high + neg CD25 and CD4 CD25 T cells. Suppressor function was tested in co-cultures with autologous CFSE-labeled CD4+ neg + neg CD25 or CD8 CD25 T-cell responders. The frequency and suppressor activity of Treg were increased after culture of CD4+CD25+ T cells in the presence of 1–100 nM RAPA (p ,0.001). RAPA-expanded Treg were + high + + neg largely CD4 CD25