Selective inhibition of IL-4 secretion in CD4⁺ PBT treated with ASA.

(A) IL-4, IL-13, IFN-γ, and IL-2 secretion in PBT (purified to ≥ 97% CD3⁺CD4⁺) treated with a therapeutic concentration of ASA (10⁻³ M) or the corresponding amount of DMSO carrier (0.1%) 15 minutes before 20-hour stimulation with A23187 (0.5 μg/mL) and PMA (25 ng/mL). Shown is the mean ± SEM percentage of IL-4 (615.7 ± 70.5 pg/mL), IL-13 (253.2 ± 110.5 pg/mL), IFN-γ (1.1 ± 0.1 ng/mL), and IL-2 (158.2 ± 67.5 U/mL) secretion from DMSO-treated controls in 3 independent experiments done in duplicate. The asterisk indicates P < .05 (Wilcoxon test) relative to controls. (B) Purified PBT (≥ 97% CD3⁺CD4⁺) were treated with increasing concentrations of ASA (10⁻⁴-3 × 10⁻³ M) or corresponding amounts of DMSO before stimulation with A23187 (0.5 μg/mL) and PMA (25 ng/mL). The mean ± SEM percentage of control IL-4 (●, 595.1 ± 28.9 pg/mL) and IL-2 (○, 364.4 ± 39.7 U/mL) secretion in 4 independent experiments done in duplicate is shown. The asterisk indicates P < .05 (Wilcoxon test) relative to DMSO-treated controls. (C) IL-4 concentrations in the supernatants of enriched PBT preparations (33%-54% CD3⁺CD4⁺) treated with 10⁻³ M ASA (▪) or 0.1% DMSO (■) before stimulation with anti-CD3 ([αCD3]; 3 μg/mL) and anti-CD28 ([αCD28]; 5 μg/mL) monoclonal antibodies or with A23187 (0.5 μg/mL) and PMA (25 ng/mL). The mean ± SEM result from 3 independent experiments done in duplicate is shown. The asterisk indicates P < .05 (Wilcoxon test) relative to DMSO-treated controls. (D) IL-4 concentrations in the supernatants of enriched PBT primed with PHA (5 μg/mL; control) in the presence or absence of IL-4 (50 ng/mL), then treated with ASA (▪, 10⁻³ M) or DMSO (■, 0.1%) 15 minutes before restimulation with A23187 (0.5 μg/mL) and PMA (25 ng/mL). The mean ± SEM result from 3 independent experiments done in duplicate is shown. The asterisk indicates P < .05 (Wilcoxon test) relative to DMSO-treated controls.