Non-typeable Haemophilus influenzae (NTHi) was stored, grown and harvested as described [16, 26] . The cell pellet was washed and resuspended in 20 ml 0.9% sodium chloride solution. This suspension was passed three times through an EmulsiFlex C5 cell disruptor (Avestin) at greater then 10,000 psi, then diluted to 4-5 mg/ml in 0.9% sodium chloride solution by bicinchoninic assay (Pierce) and centrifuged at 15,0006g for 10 min. The supernatant was collected, the protein concentration was adjusted to 2.5 mg/ml, and the lysate was sterilized by passage through a 0.2 mm filter and frozen in 8 ml aliquots at 280uC. Treatment of mice with aerosolized lysate was performed as described [16] , delivering 8 mL of suspension during each 30 min exposure.