Table 2.
Adoptive transfer of CD4 T cells populations into B16-OVA tumor-bearing mice.
| Percent Foxp3EGFP Positive Cells Received From Spleen and Tumor | ||||||
|---|---|---|---|---|---|---|
| T Cell Subsets Adoptively Transferred |
Spleen day 5 |
Tumor day 5 |
Spleen day 10 |
Tumor day 10 |
Spleen day 15 |
Tumor day 15 |
| Bone marrow control (0%) |
0.0, 0.0 | 0.0, 0.0 | 0.0, 0.0 | |||
| C57BL/6 CD4 (0%) |
0.0, 0.0 | 0.3, 0.5 | 0.0,0.0 | 0.0, 0.0 | ||
| C57BL/6 nTreg CD4 (100%) |
1.4, 1.9 | 11.7, 15.2 | 14.8, 20.8 | |||
| C57BL/6 iTreg CD4 (26%) |
0.4, 0.3 | 4.5, 3.8 | 0.4, 2.0 | |||
| OTII CD4 (0%) |
0.0, 0.1 | 0.0, 0.0 | 0.0, 0.0 | 0.0, 0.0 | ||
| OTII iTreg CD4 (10.4%) |
0.2, 0.0 | 0.0, 0.0 | 0.9, 1.4 | 0.1, 0.1 | 0.3, 0.4 | |
|
SPLEEN day 7 |
TUMOR day 7 |
SPLEEN day 14 |
TUMOR day 14 |
SPLEEN day 25 |
TUMOR day 25 |
|
| Pmel CD8 (<1%) |
0.0, 0.0 | 0.1, 0.1 | 0.0, 0.0 | 0.1, 0.1 | 0.0, 0.0 | 0.0,0.0 |
C57BL/6 (CD45.2) mice bearing (five-day) B16-OVA tumors were irradiated with 900 cGy. The following day, all mice received bone marrow, from syngenic CD45.1 mice. Several different populations of cells were adoptively transferred into B16-OVA tumor-bearing mice. These cells were activated for 72 hours with αCD3/αCD28/IL-2, and in some cases flow-purified to yield a homogenous population of Foxp3EGFP-positive or negative cells. These cell populations included: (1) 105 C57BL/6 CD4 T cells (0% Foxp3EGFP); (2) 105 C57BL/6 CD4 nTreg (100% Foxp3EGFP); (3) 105 C57BL/6 CD4 TGFβ-induced iTreg (flow purified Foxp3EGFP-negative CD4 T cells, activated in the presence of TGFβ for 72 hours to yield ~26% Foxp3EGFP iTreg); (4) 105 OTII CD4 T cells (0% Foxp3EGFP); (5) 105 OTII CD4 iTreg (10.4% Foxp3EGFP); and 106 Pmel CD8 (<1% Foxp3EGFP). When sufficient numbers of TIL could be isolated for flow (at least 103), these were also surface phenotyped.