Table 1.
Spleen cells were isolated and cultured for 72 hours under the indicated conditions with the following concentrations: IL-2 (100 u/ml), TGFβ (10 ng/ml) and αCD3/αCD28 (1 μg/ml immobilized, 10 μg/ml soluble, respectively).
| Culture Conditions | C57BL/6 Foxp3EGFP CD8 |
C57BL/6 Foxp3EGFP CD4 |
Pmel Foxp3EGFP CD8 |
Pmel Foxp3EGFP CD4 |
OTII Foxp3EGFP CD4 |
|---|---|---|---|---|---|
| Media Only | 0.9 +/- 0.6 | 2.1 +/- 1.3 | 2.2 +/- 0.1 | 9.6 +/- 5.9 | 0.8 +/- 0.1 |
| IL-2 | 1.9 +/- 0.5 | 8.2 +/- 1.8 | 2.9 +/- 0.1 | 11.7 +/- 1.3 | 0.6 +/- 0.3 |
| TGF- β | 1.8 +/- 0.1 | 4.2 +/- 1.1 | 4.8 +/- 0.8 | 13.2 +/- 1.6 | 0 +/- 0.6 |
| IL-2/TGF-β | 2.0 +/- 1.3 | 20.4 +/- 1.4 | 1.4 +/- 0.8 | 20.7 +/- 3.4 | 0.8 +/- 0.5 |
| αCD3/αCD28 | 0.9 +/- 1.8 | 3.1 +/- 0.2 | 2.3 +/- 0.3 | 17.2 +/- 1.7 | 3.0 +/- 0.1 |
| αCD3/αCD28/IL-2 | 1.3 +/- 1.6 | 4.6 +/- 0.7 | 21.3 +/- 0.8 | 13.8 +/- 0.5 | 3.0 +/- 0.2 |
| αCD3/αCD28/TGF-β | 18.8 +/- 3.0 | 18.8 +/- 1.0 | 27.3 +/- 1.8 | 30.7 +/- 2.8 | 7.7 +/- 2.4 |
| αCD3/αCD28/IL-2/TGFβ | 25.1 +/- 0.4 | 34.4 +/- 1.1 | 27.9 +/- 0.2 | 46.2 +/- 1.1 | 8.4 +/- 0.4 |
Cells were then sorted for CD8, CD4, and EGFP expression. Results are a mean of triplicates +/- S.D. These experiments were repeated at least 3 times with comparable results.