Table 2.
Concentration and Time-dependent Inhibition of IL-2 Production by rGAL-1 upon Antigen Cell Stimulation of an Aq-restricted, CII-specific T Cell Hybridoma
| HCQ.6 cells (5 × 105/ml) | Spleen APC (5 × 106/ml) | CII (50 μg/ml) | rGAL-1 | TDG (100 mM) | IL-2 |
|---|---|---|---|---|---|
| μg/ml | pg/ml | ||||
| + | + | + | 611.00 ± 32.15 | ||
| + | + | + | 4 (18 h) | – | 25.90 ± 3.2 |
| + | + | + | 4 (4 h) | – | 29.95 ± 2.21 |
| + | + | + | 4 (2 h) | – | 56.52 ± 4.53 |
| + | + | + | 0.4 (18 h) | – | 48.77 ± 6.25 |
| + | + | + | 0.4 (4 h) | – | 97.13 ± 9.37 |
| + | + | + | 0.4 (2 h) | – | 382.59 ± 12.91 |
| + | + | + | 0.04 (18 h) | – | 362.10 ± 21.89 |
| + | + | + | 0.04 (4 h) | – | 399.60 ± 20.82 |
| + | + | + | 0.04 (2 h) | – | 400.02 ± 12.93 |
| + | + | + | 4 (18 h) | + | 119.22 ± 7.93 |
| + | + | + | 4 (4 h) | + | 203.57 ± 9.64 |
| + | − | + | – | 33.57 ± 1.96 | |
| + | + | – | – | 29.45 ± 2.53 |
HCQ.6 (5 × 105 cells/ml), an Aq-restricted, CII-specific T cell hybridoma, was stimulated with bovine CII (50 μg/ml) presented by splenocytes from naive DBA/1 mice (5 × 106 cells/ml) in the presence of rGAL-1 (0.04, 0.4, or 4 μg/ml), added for different time periods (2, 4, or 18 h) as indicated. Specificity was assayed by adding TDG (100 mM) to the culture. The anti–GAL-1 Ab (1:100) had no neutralizing activity (data not shown). Cells were cocultured for 24 h and IL-2 was measured in the supernatants by a capture ELISA. Anti-CD3–stimulated HCQ.6 cells reached maximal levels of IL-2 secretion (1,147 ± 165 pg/ml).