Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells
Effects of signaling molecule inhibitors on the cell surface expression of ICAM-1 and induction of cytokines/chemokines from coculture of BEAS-2B cells and basophils/eosinophils in the presence of LIGHT (100 ng/mL). Cocultures of (a, b, c, d) basophils or (e, f) eosinophils (3 × 105 cells) and confluent BEAS-2B cells (1 × 105 cells) were pretreated with BAY11-7082 (1 μM), SP600125 (3 μM), SB203580 (7.5 μM), U0126 (10 μM), or LY294002 (10 μM) for 1 h, followed by incubation with or without LIGHT (100 μg/mL) in the presence of inhibitors for a further 24 h. Cell surface expression of ICAM-1 on 5,000 cells was analyzed by flow cytometry as MFI. Results are expressed as mean plus SEM of three independent experiments with three blood samples in bar charts. Release of cytokines/chemokines and MMP-9 in culture supernatant was measured by Milliplex assay. Results are expressed as mean plus SEM. DMSO (0.1%) was used as the vehicle control. *, ** when compared with vehicle or negative control.
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