CD4⁺CD25⁺ regulatory T cells suppress allograft rejection mediated by memory CD8⁺ T cells via a CD30-dependent mechanism
J. Clin. Invest. Zhenhua Dai, et al. 113:310 doi:10.1172/JCI19727 [Go to this article.]

Figure 2
Characterization of CD4⁺CD25⁺ Treg cells. (a) Freshly isolated spleen cells from naive or DST-treated WT mice were stained for surface markers CD4, CD25, CD62L, CD45RB, CTLA4, and CD30 and analyzed in histograms after gating on the CD4⁺CD25⁺ T cell population. The dotted lines represent isotype control Ab, and the solid gray and black lines represent naive and DST-induced Treg cells, respectively. One representative experiment of three is shown. (b) Median fluorescence intensity of CD30 on DST Treg cells is higher than on naive Treg cells (*P < 0.05, ANOVA). (c) Freshly isolated spleen cells were also stained for intracellular cytokine expression of IL-2 and IL-10 after gating on CD4⁺CD25⁺ T cells. (d) Treg cells suppress anti-CD3–induced proliferation of CD4⁺CD25^– T cells. Purified CD4⁺CD25⁺ Treg cells (3 × 10⁴) were cultured with or without naive CD4⁺CD25^– cells (6 × 10⁴) in the presence of soluble anti-CD3 (1.0 μg/ml) and APCs (6 × 10⁴) from BALB/c spleen cells for 72 hours. 50 U/ml of IL-2 was added to Treg cell culture alone to test whether the anergic status of Treg cells can be reversed. Results are presented as the mean of triplicate cultures.