| | siRNA | IL-2 [pg/ml] | IFN-γ [pg/ml] | Proliferation [CPM] |
|---|
αCD3
| Ctrl | 1417 ± 144 | 965 ± 105 | 4106 ± 433 |
| | Lck | 443 ± 93 | 357 ± 50 | 540 ± 220 |
| | PLC-γ | 195 ± 28 | 146 ± 26 | 236 ± 19 |
| | ZAP70 | 400 ± 60 | 467 ± 149 | 568 ± 69 |
PMA + Ionomycin
| Ctrl | 1980 ± 2 | 5067 ± 346 | 19919 ± 787 |
| | Lck | 2167 ± 0 | 4158 ± 135 | 16640 ± 954 |
| | PLC-γ | 1676 ± 28 | 2939 ± 88 | 11285 ± 658 |
| | ZAP70 | 1991 ± 2 | 4603 ± 277 | 15987 ± 1906 |
- T cells were transfected with 0.9 μM Lck-, PLC-γ- and ZAP70-specific siRNA pool, ZAP70-specific stealth™ siRNA 1 and with control siRNA pool (Ctrl). 72 h after nucleofection, functionality of transfected cells was tested by restimulation with anti-mouse CD3 antibody or PMA and ionomycin in vitro. IL-2 and IFN-γ production was determined in the supernatant 24 h after restimulation by ELISA. Means of duplicate values are shown. Proliferation of T cells was detected by 3H-thymidine incorporation 96 h after restimulation. Values of 3H-thymidine incorporation are shown in means of triplicates.
