Homeostatic proliferation of Des-TCR CD8 T cells during systemic inflammation does not break tolerance
| Donor T cells . | TLR ligand . | Incidence of tumor growth (P815.Kb.B7) . | Mean percent IFNγ-positive CD8 T cells ± SD . | |
|---|---|---|---|---|
| 1 | No cell transfer | CpG | 8 of 8* | |
| 2 | Tolerant Des-TCR CD8 | 9 of 9* and | ||
| 9 of 9† | 0.5 ± 0.4† | |||
| 3 | Tolerant Des-TCR CD8 | CpG | 8 of 8* and | |
| 8 of 9† | 0.6 ± 0.6† | |||
| 4 | Tolerant Des-TCR CD8 | 4× CpG | 5 of 5† | 1.1 ± 0.8† |
| 5 | Tolerant Des-TCR CD8 | Poly(I:C) | 10 of 10† | 0.7 ± 0.3† |
| 6 | Tolerant Des-TCR CD8 | LPS | 5 of 5† | 1.4 ± 0.4† |
| 7 | Tolerant Des-TCR CD8 | CpG/LPS | 5 of 5† | |
| 8 | Des-TCR.RAG2−/− (control mice) | 1 of 20 | 10.6 ± 7.8† | |
| Donor T cells . | TLR ligand . | Incidence of tumor growth (P815.Kb.B7) . | Mean percent IFNγ-positive CD8 T cells ± SD . | |
|---|---|---|---|---|
| 1 | No cell transfer | CpG | 8 of 8* | |
| 2 | Tolerant Des-TCR CD8 | 9 of 9* and | ||
| 9 of 9† | 0.5 ± 0.4† | |||
| 3 | Tolerant Des-TCR CD8 | CpG | 8 of 8* and | |
| 8 of 9† | 0.6 ± 0.6† | |||
| 4 | Tolerant Des-TCR CD8 | 4× CpG | 5 of 5† | 1.1 ± 0.8† |
| 5 | Tolerant Des-TCR CD8 | Poly(I:C) | 10 of 10† | 0.7 ± 0.3† |
| 6 | Tolerant Des-TCR CD8 | LPS | 5 of 5† | 1.4 ± 0.4† |
| 7 | Tolerant Des-TCR CD8 | CpG/LPS | 5 of 5† | |
| 8 | Des-TCR.RAG2−/− (control mice) | 1 of 20 | 10.6 ± 7.8† | |
As indicated, 120 μg of CpG-ODN was administered intraperitoneally 1 day before and 100 μg of poly(I:C) or 75 μg of LPS was administered intraperitoneally on the day of T-cell transfer. Group 4 received 120 μg of CpG on day −1 and 40 μg of CpG on days 2, 5, and 8. Group 7 received 40 μg of CpG on day −1 and 40 μg and LPS on day 0. Thirty thousand MACS-purified Des-TCR CD8 T cells (*) or
2 × 106 splenocytes (†) from thymectomized tolerant Des-TCR × 2.4KerIV-Kb.RAG2−/− donor mice were transferred intravenously into the respective RAG2−/− mice. After 3 days the mice were challenged subcutaneously with 2 × 105 P815.Kb.B7 tumor cells and tumor growth was followed. Upon large tumor burden the mice were sacrificed. Tumor-free mice were followed for up to 4 weeks. Des-TCR.RAG2−/− mice were used as a control for tumor rejection. For IFNγ staining, splenocytes were restimulated in vitro for 6 hours with anti-CD3 antibody (4 μg/mL) on day 18 after tumor injection. The pooled results were obtained in 3 independent experiments.