Suppression of Cytokine Production of Anergic Tyrosinase-Specific CD8+ T Cells After Restimulation
| First Coculture . | Cytokine Production After Second Coculture (pg/mL) . | ||||
|---|---|---|---|---|---|
| IL-2 . | IFN-γ . | IL-4 . | IL-10 . | TGF-β . | |
| Tyrosinase-specific CD8+ T cells and | |||||
| DC | 987 ± 23 | 3,765 ± 124 | ND | ND | ND |
| DC (IL-10) | 1,102 ± 76 | 4,087 ± 218 | ND | ND | ND |
| DC + tyrosinase peptide | 1,154 ± 205 | 3,867 ± 134 | ND | ND | ND |
| DC (IL-10) + tyrosinase peptide | 105 ± 20 | 258 ± 87 | ND | ND | ND |
| DC + MART-1 | 1,287 ± 156 | 3,841 ± 434 | ND | ND | ND |
| DC (IL-10) + MART-1 | 1,087 ± 187 | 4,187 ± 128 | ND | ND | ND |
| DC [HLA.A1+] + tyrosinase peptide | 1,376 ± 231 | 4,283 ± 349 | ND | ND | ND |
| DC (IL-10) [HLA.A1+] + tyrosinase peptide | 1,498 ± 327 | 3,762 ± 297 | ND | ND | ND |
| First Coculture . | Cytokine Production After Second Coculture (pg/mL) . | ||||
|---|---|---|---|---|---|
| IL-2 . | IFN-γ . | IL-4 . | IL-10 . | TGF-β . | |
| Tyrosinase-specific CD8+ T cells and | |||||
| DC | 987 ± 23 | 3,765 ± 124 | ND | ND | ND |
| DC (IL-10) | 1,102 ± 76 | 4,087 ± 218 | ND | ND | ND |
| DC + tyrosinase peptide | 1,154 ± 205 | 3,867 ± 134 | ND | ND | ND |
| DC (IL-10) + tyrosinase peptide | 105 ± 20 | 258 ± 87 | ND | ND | ND |
| DC + MART-1 | 1,287 ± 156 | 3,841 ± 434 | ND | ND | ND |
| DC (IL-10) + MART-1 | 1,087 ± 187 | 4,187 ± 128 | ND | ND | ND |
| DC [HLA.A1+] + tyrosinase peptide | 1,376 ± 231 | 4,283 ± 349 | ND | ND | ND |
| DC (IL-10) [HLA.A1+] + tyrosinase peptide | 1,498 ± 327 | 3,762 ± 297 | ND | ND | ND |
Tyrosinase-specific CD8+ T cells were cocultured with untreated or IL-10–treated DC in combination with the specific tyrosinase peptide, with an unspecific peptide (MART-1), or without antigen during the first coculture. Additional control experiments were performed using HLA-mismatched (HLA.A1+) untreated and IL-10–treated DC and the specific tyrosinase peptide. After 2 days, the T cells were rescued, cultured for 2 days in the presence of low levels of IL-2 (2 U/mL), and restimulated with untreated DC and the specific tyrosinase peptide. Subsequently, the supernatants of the T-cell culture were harvested and assessed for IL-2, IL-4, IL-10, VTGF-β, and IFN-γ content by ELISA.
Abbreviation: ND, not detectable.