Table 1.
Cytotoxicity and cytokine production by CTL on coincubation with various cell lines1
| Tumour cells | Histology | MHC class I isotype control | mAb | Lysis (%) | IFN-γ (pg/ml) | TNF-α (pg/ml) | IL-4 (pg/ml) |
|---|---|---|---|---|---|---|---|
| C15 | Colon carcinoma | + | 48.7 ± 4.1 | 2000 ± 40* | 450 ± 30* | <20 | |
| Anti-CD4 | 40.0 ± 3.9 | 1260 ± 30 | 285 ± 15 | <20 | |||
| Anti-CD8 | 9.8 ± 1.8 | 360 ± 10 | 180 ± 15 | <20 | |||
| Anti-I-Ab | 38.9 ± 4.2 | 1190 ± 25 | 270 ± 15 | <0 | |||
| Anti-H-2Kb/H-2Db | 10.9 ± 2.1 | 425 ± 15 | 165 ± 15 | <0 | |||
| MC-38 | Colon carcinoma | + | 10.8 ± 2.4 | 1105 ± 25 | 180 ± 15 | <0 | |
| EL4 | Thymoma | + | 4.3 ± 2.0 | 1075 ± 20 | 135 ± 15 | <0 | |
| RMA | Lymphoma | + | 10.9 ± 2.6 | 1030 ± 25 | 150 ± 15 | <0 | |
| B16-F10 | Melanoma | – | 7.5 ± 2.1 | 990 ± 20 | 135 ± 15 | <0 | |
| SCC VII | Squamous cell carcinoma | + | 10.6 ± 3.0 | 885 ± 20 | 135 ± 15 | <0 | |
| AG104A | Fibrosarcoma | + | 2.3 ± 1.6 | 955 ± 20 | 150 ± 15 | <0 | |
| TSA | Mammary carcinoma | + | 5.2 ± 2.0 | 825 ± 15 | 165 ± 15 | <0 | |
| YAC-1 | Lymphoma | ND | 8.8 ± 2.8 | 1010 ± 20 | 150 ± 15 | <0 | |
| – | 1090 ± 25 | 165 ± 15 | <0 |
Cytotoxicity against various cell lines was measured in a 6 hr 51Cr release assay at an E : T ratio of 25 : 1. CTL were generated from 3H1-pulsed DC immunized mice as described. CTL (2 × 105/well) were incubated with tumour cells (2 × 104/well) or media alone in 96-well tissue culture plates at 37° for 6 hr. Cell-free supernatants were harvested and analysed for the production of IFN-γ, TNF-α, and IL-4 by standard ELISA kits.17 For inhibition experiments, blocking mAb was added to the culture at a concentration of 10 µg/ml. Surface MHC class I antigen expression was analysed by flow cytometry. The representative data of three independent experiments is shown.
P < 0.002 with respect to other culture conditions for IFN-γ and TNF-α production.
ND, not determined.