T cells          (No tumors)     (pg/ml) T+HEp2-vec (pg/ml) T+HEp2-IκB(S32AS36A) (pg/ml) 
day T cells -/+IL2   - IFN-g + IFN-g - IFN-g + IFN-g
15 - 0.0±0.0 1.5±0.7 5 ± 2.8 11 ± 0.0 7.5 ± 4.9
  + 4 ±0.0 16.5±0.7 8 ± 1.4 185 ± 10 104.5±18
             
19 - 9 ±0.0 6 ± 4.2 3.5 ± 2.1 16 ± 0.0 5±0.0
  + 51.5 ±25 14.5 ± 9.2 10 ± 0.0 341.5 ± 20 231±42
             
23 - 0.0 ±0.0 0.0 ±0.0 0.0 ±0.0 23 ± 0.0 7.5±0.7
  + 9.5 ±0.7 13.5 ± 4.9 26.5 ± 2.1 1243 ± 25 465.5±16
Purified CD8+ T cells were treated with and without IL-2 (500 u/ml) overnight before their co-culture with and without IFN-γ (200 u/ml) treated vector alone and IκB(S32AS36A) transfected HEp2 cells (E:T ratio 1:1). After two weekly stimulations with freshly supplemented IL2, the supernatants were removed from the CD8+ T /HEp2 cell cocultures at the indicated days in the table and assayed for released GM-CSF by a specific and sensitive ELISA. IFN-γ treated HEp2 cell transfectants were washed three times before they were added to IL-2 treated CD8+ T cells. No secretion of GM-CSF from vector-alone or IκB(S32AS36A) transfected HEp2 cells in the absence of CD8+ T cells could be observed (data not shown). The p value for the difference between secretion of GM-CSF in the co-cultures of CD8+ T cells with control HEp2-vec and IκB(S32AS36A) transfected HEp2 cells are less than 0.05. 1 of 3 representative experiments is shown in this table.
Table 2: Kinetics of increased GM-CSF release when CD8+ T cells were co-cultured with untreated and IFN-γ treated HEp2-IκB(S32AS36A) cells.