Figure 5

NTHi supernatant does not cause cell lysis, but it does result in the preferential detachment of ciliated cells. A) Media supernatants were collected from bovine bronchial cell cultures treated with 50% NTHi supernatants. Supernatants were assayed for lactate dehydrogenase (LDH) activity. As a positive control, cells were intentionally lysed using sonication. The cells that were exposed to 100% NTHi for 1–48 hours showed no elevation in LDH. This indicates that NTHi does not cause cytotoxicity (*p < 0.05 + control vs. NTHi) (n = 6) B) BBECs were exposed to 50% NTHi supernatant for 1–48 hours. The supernatant from these cultures was pelleted to collect any ciliated cells that had detached. Pellets were vacuum transferred and blotted for 13S dynein. Detached ciliated cells were detected at 18–48 hr NTHi supernatant exposure. Positive control is purified bovine tracheal axonemes (0.1 μg) (n = 3).