Th (T-helper) 2 and Th1 clonal cell lines treated with AG-490 show preferential inhibition of interleukin (IL) 4 protein but not after anti-CD3 stimulation.

(A) Th2 clone D10 or Th1 cells (clone 29) were treated with AG-490 or vehicle alone at 37°C for 16 hours. Enzyme-linked immunosorbent analysis (ELISA) results for each cytokine (indicated on ordinate) were plotted on the abscissa as the percentage of inhibition of the mean (± SE; n = 6) from ELISA values as follows: IL-4–dimethyl sulfoxide (DMSO), 497 ± 91 pg/mL, and AG-490, below detection (< 60 pg/mL); IL-5–DMSO, 34 984 ± 4422 pg/mL, and AG-490, 34 585 ± 8504 pg/mL; IL-10–DMSO, 54 689 ± 1554 pg/mL, and AG-490, 38 038 ± 3518 pg/mL; and IL-13–DMSO, 53 907 ± 6398 pg/mL, and AG-490, 66 439 ± 1995 pg/mL. Effects of AG-490 on clone 29 Th1 cytokine values (n = 6) were as follows: interferon γ–DMSO, 1062 ± 96 pg/mL, and AG-490, 1011 ± 18 pg/mL; tumor necrosis factor (TNF) α–DMSO, 1523 ± 108 pg/mL, and AG-490, 1378 ± 96 pg/mL; TNF-β–DMSO 375 ± 23 pg/mL, and AG-490, 307 ± 21 pg/mL; and IL-2–DMSO 13 ± 1 pg/mL, and AG-490, 12.8 ± 1 pg/mL. (B) AG-490 did not block production of IL-4 by D10 and IL-2 by clone 29 after stimulation with anti-CD3. D10 and clone 29 cells were washed, stimulated by coated anti-CD3 antibodies, and treated in the presence or absence of 50 μmol/L AG-490 for 16 hours.