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. Author manuscript; available in PMC: 2009 Aug 15.
Published in final edited form as: J Immunol. 2008 Aug 15;181(4):2855–2868. doi: 10.4049/jimmunol.181.4.2855

Figure 1. CD28 costimulation is required to consistently obtain expanded cultures enriched for Foxp3 expression.

Figure 1

A. Analysis of CD25 expression in purified CD4 T cells (Left panel), CD25 (Middle Panel) and Foxp3 (Right Panel) expression after CD127 depletion and CD25 selection of CD4 T cells. This data is representative of the enrichment of all input Treg populations used in this study. B. Expression of CD64 and CD86 was analyzed on K64 (Left panel) and K64.86 (Right panel) aAPCs by flow cytometry. C. 200,000 enriched Tregs were stimulated with anti-CD3 Ab loaded K32 aAPCs, and anti-CD3 loaded K32.86 aAPCs cultured in the presence and absence of rapamycin (RAPA) for two weeks and population doubling rate was measured by cell counting. Each data point represents the average of four independent experiments (error bars represent standard deviation). D and E Analysis of Foxp3 (D) or CD27 and CD62-L (double positive) (E) expression on Tregs expanded with anti-CD3 Ab loaded K32 or K32.86 aAPCs in the presence of rapamycin. The panels on the left show a representative experiment and the box plot on the right is data compiled from 4 independent experiments.