Percentage of inhibition of LAG-3–induced up-regulation of CD40, CD80, CD83, and CD86 expression and IL-12 p40 and TNF-α secretion
. | SB203580 (p38MAPKi) 40 μM . | PD98059 (p42/44ERKi) 50 μM . | LY294002 (PI3Ki) 25 μM . | Herbimycin A (TKi) 5 μM . | Piceatannol (p72SYKi) 100 μM . |
|---|---|---|---|---|---|
| CD40, % | ND | ND | 62 | ND | 116 |
| CD80, % | 65 | 26 | 101 | 84 | 90 |
| CD83, % | 77 | 50 | 100 | 66 | 65 |
| CD86, % | 22 | 28 | 65 | 46 | 60 |
| IL-12 p40, % | 86 | 76 | 86 | 56 | 83 |
| TNF-α, % | 52 | 85 | 88 | 76 | 82 |
. | SB203580 (p38MAPKi) 40 μM . | PD98059 (p42/44ERKi) 50 μM . | LY294002 (PI3Ki) 25 μM . | Herbimycin A (TKi) 5 μM . | Piceatannol (p72SYKi) 100 μM . |
|---|---|---|---|---|---|
| CD40, % | ND | ND | 62 | ND | 116 |
| CD80, % | 65 | 26 | 101 | 84 | 90 |
| CD83, % | 77 | 50 | 100 | 66 | 65 |
| CD86, % | 22 | 28 | 65 | 46 | 60 |
| IL-12 p40, % | 86 | 76 | 86 | 56 | 83 |
| TNF-α, % | 52 | 85 | 88 | 76 | 82 |
Peripheral blood monocytes were incubated with GM-CSF and IL-4 for 7 days. Immature DCs were then either treated with DMSO alone (negative control, not shown) or preincubated with the indicated inhibitors for 2 hours and then exposed to 10 μg/mL LAG-3Ig for 24 hours. Data are shown as mean of percentages of at least 3 independent experiments. Surface expression of the indicated markers was assessed by FACS analysis. Mean cellular fluorescence values were used to calculate the percentage of inhibition of LAG-3—induced up-regulation of CD40, CD80, CD83, and CD86. Cell culture supernatants were collected, and cytokine concentration was measured by ELISA. Concentration values were used to calculate the percentage of inhibition of LAG-3—induced up-regulation. Following LAG-3Ig addition, concentrations rose from 7 ± 2 and 0.4 ± 0.1 ng/mL to 61 ± 10 and 3.9 ± 0.1 ng/mL for IL-12 p40 and TNF-α, respectively. ND indicates not determined.