(A) Human cytokine macroarray analysis shows NTHi-induced release of cytokines by middle ear epithelial cells. Cell culture medium was harvested at 48 hrs after NTHi lysate treatment and the released cytokines were detected using asolid phase multiplexed protein assay in a sandwich ELISA format. Cells were treated with NTHi lysate (NTHi) or vehicle (No Tx). (B) Real time quantitative PCR analysis shows regulation of β-defensin 2 (DEFB4) mRNA by cytokines and chemokines. Only IL-1α treatment results in an up-regulation of β-defensin 2 (DEFB4) mRNA in HMEEC and A549 cells, but not in Hela cells.