Proliferation of Thy1.1 cells following adoptive transfer
| Adoptively transferred cells* . | Lung T cells, × 106† . | Spleen T cells, × 106‡ . | ||||
|---|---|---|---|---|---|---|
| Day 1 . | Day 3 . | Day 5 . | Day 1 . | Day 3 . | Day 5 . | |
| CD4+ 5 × 106 | ||||||
| Gated on Thy1.1 | 0.02 | 1.4 | 22.7 | 0.03 | 0.06 | 0.15 |
| Gated on total lymphocytes | 2.0 | 6.3 | 33.6 | 9.0 | 3.0 | 3.5 |
| CD8+ 5 × 106 | ||||||
| Gated on Thy1.1 | 0.02 | 0.02 | 0.03 | 0.03 | 0.13 | 0.12 |
| Gated on total lymphocytes | 1.2 | 1.1 | 1.8 | 7.6 | 4.0 | 3.8 |
| CD4+ + CD8+ | ||||||
| Gated on Thy1.1 CD4, 2.5 × 106 each | 0.02 | 0.79 | 14.3 | 0.01 | 0.07 | 0.15 |
| Gated on Thy1.1 CD8 | 0.01 | 0.02 | 1.4 | 0.001 | 0.01 | 0.01 |
| Gated on total lymphocytes | 2.2 | 4.3 | 26.2 | 8.8 | 4.2 | 4.9 |
| Irradiated CD4+ 107 + CD8+ 2.5 × 106 | ||||||
| Gated on Thy1.1 CD4 | 0.14 | 0.12 | 0.05 | 0.05 | 0.014 | 0.004 |
| Gated on Thy1.1 CD8 | 0.06 | 0.04 | 0.34 | 0.00 | 0.011 | 0.027 |
| Gated on total lymphocytes | 3.5 | 2.6 | 4.8 | 8.4 | 3.8 | 3.0 |
| Adoptively transferred cells* . | Lung T cells, × 106† . | Spleen T cells, × 106‡ . | ||||
|---|---|---|---|---|---|---|
| Day 1 . | Day 3 . | Day 5 . | Day 1 . | Day 3 . | Day 5 . | |
| CD4+ 5 × 106 | ||||||
| Gated on Thy1.1 | 0.02 | 1.4 | 22.7 | 0.03 | 0.06 | 0.15 |
| Gated on total lymphocytes | 2.0 | 6.3 | 33.6 | 9.0 | 3.0 | 3.5 |
| CD8+ 5 × 106 | ||||||
| Gated on Thy1.1 | 0.02 | 0.02 | 0.03 | 0.03 | 0.13 | 0.12 |
| Gated on total lymphocytes | 1.2 | 1.1 | 1.8 | 7.6 | 4.0 | 3.8 |
| CD4+ + CD8+ | ||||||
| Gated on Thy1.1 CD4, 2.5 × 106 each | 0.02 | 0.79 | 14.3 | 0.01 | 0.07 | 0.15 |
| Gated on Thy1.1 CD8 | 0.01 | 0.02 | 1.4 | 0.001 | 0.01 | 0.01 |
| Gated on total lymphocytes | 2.2 | 4.3 | 26.2 | 8.8 | 4.2 | 4.9 |
| Irradiated CD4+ 107 + CD8+ 2.5 × 106 | ||||||
| Gated on Thy1.1 CD4 | 0.14 | 0.12 | 0.05 | 0.05 | 0.014 | 0.004 |
| Gated on Thy1.1 CD8 | 0.06 | 0.04 | 0.34 | 0.00 | 0.011 | 0.027 |
| Gated on total lymphocytes | 3.5 | 2.6 | 4.8 | 8.4 | 3.8 | 3.0 |
Thy1.1 TDLN CD62Llow, CD4+, or CD8+ subsets were activated for 2 cycles with anti-CD3/IL-2/IL-7 or anti-CD3/IL-7/IL-23 respectively, and were transferred on day 29 to hosts bearing 10-day lung metastases, which were conditioned with 5 Gy TBI before cell transfer.
Lungs were harvested on the indicated days and enzymatically digested, and single cell suspension were analyzed by FACS. The number of Thy1.1+ cells was determined by their percentage of the total number of CD4+ or CD8+ cells.
Spleens were harvested on the indicated days, mechanically disrupted, and depleted of red blood cells (RBCs). The number of Thy1.1+ cells was determined by their percentage of the total number of CD4+ or CD8+ cells.