Skip to main content
. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: Am J Transplant. 2017 Jun 14;17(12):3098–3113. doi: 10.1111/ajt.14338

Figure 1. In vitro expansion of naïve CD4 T-cells in the presence of Low Trp+Kyn induces suppressive function.

Figure 1

Figure 1

Figure 1

tTreg (CD4+25++127−) and naïve CD4+ T-cells (CD4+ 25–45RA+) were purified from peripheral blood using magnetic beads, stimulated with a cell line expressing CD86 and CD64 (KT64/86) to which anti-CD3 mAb is bound, and cultured for 14 days as described. A) Representative example and average fold expansion for the indicated cell types on day 14 of culture. B) Representative example and summary of CD127 and Foxp3 expression in tTregs and naive T-cells cultured ± Low Trp+Kyn. C) Representative example and summary of relative Foxp3 expression (MFI) in tTregs and naive T-cells cultured ± Low Trp+Kyn. D) Representative example and summary of CD25 and Foxp3 expression in tTreg and naive T-cells cultured ± Low Trp+Kyn. E) Representative example and average CD25 expression (± SEM) on CD4-gated cells. F) Suppressive function of cultured cells was determined using a CFSE-based proliferation assay in which expanded tTreg or naive T-cells were incubated with CFSE-loaded allogeneic PBMC at doses from 1:4 to 1:16 (expanded cell:PBMC) and stimulated with anti-CD3 beads for 4 days.