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. Author manuscript; available in PMC: 2017 Oct 1.

Published in final edited form as: Am J Transplant. 2016 Jul 11;16(10):2994–3006. doi: 10.1111/ajt.13883

(A) Percent of proliferation and Ki67 expression by CD4⁺ naïve and Tmem subsets in response to allostimulation in the presence of rapamycin ± TGFβ1/Fc. (B) Similarly, CD8⁺ naïve and Tmem subsets were also evaluated for percent of proliferation and Ki67 expression. CFSE-labeled PBMC were cultured for 4 – 5 days with T cell-depleted allogeneic PBMC in the absence or presence of rapamycin ± TGFβ1/Fc. At the end of the culture, T cell subsets were assessed by flow cytometry based on CD45RA and CD28 expression: naïve (Tn)- CD45RA⁺CD28⁺, central memory (Tcm)- CD45RA⁻CD28⁺, effector memory (Tem)- CD45RA⁻CD28⁻, and terminally-differentiated effector memory (Temra)-CD45RA⁺CD28⁻. All results are representative data of three independent experiments using different responders and stimulator-responder pairs. PBMC, peripheral blood mononuclear cell.

(A) Percent of proliferation and Ki67 expression by CD4⁺ naïve and Tmem subsets in response to allostimulation in the presence of rapamycin ± TGFβ1/Fc. (B) Similarly, CD8⁺ naïve and Tmem subsets were also evaluated for percent of proliferation and Ki67 expression. CFSE-labeled PBMC were cultured for 4 – 5 days with T cell-depleted allogeneic PBMC in the absence or presence of rapamycin ± TGFβ1/Fc. At the end of the culture, T cell subsets were assessed by flow cytometry based on CD45RA and CD28 expression: naïve (Tn)- CD45RA⁺CD28⁺, central memory (Tcm)- CD45RA⁻CD28⁺, effector memory (Tem)- CD45RA⁻CD28⁻, and terminally-differentiated effector memory (Temra)-CD45RA⁺CD28⁻. All results are representative data of three independent experiments using different responders and stimulator-responder pairs. PBMC, peripheral blood mononuclear cell.