Effects of human polymorphonuclear leukocyte elastase upon surfactant proteins in vitro.

Recent evidence has suggested that elastase is released by polymorphonuclear leukocytes (PMN) recruited from the pulmonary microcirculation into the alveoli during acute lung injury. This study was undertaken to test the hypothesis that elastase from PMN (PMN elastase) damages or degrades one or more of the surfactant proteins (SP-A, SP-B and SP-C) of the lung, and thereby alters its function. We attempted to use amounts of PMN elastase and quantities of surfactant that would be plausible in the lungs of patients with ARDS. Surfactant from normal dog lungs (2 mg phospholipid, 200 micrograms protein), and purified SP-A (20 micrograms), SP-B (10 micrograms) and SP-C (10 micrograms) from the surfactant (identified by SDS-PAGE and N-terminal amino acid sequences) were incubated for 4-8 h at 37 degrees C with various amounts (0.25-1.0 U) of human PMN elastase purified by affinity chromatography. SDS-PAGE and amino acid composition analysis of the surfactant as well as of the purified SP-A, SP-B, and SP-C showed that degradation of these proteins progressed with incubation time and with the amount of PMN elastase, and was accompanied by decreases in isopycnic density (g/cm3) and surface adsorption, and increase of surface tension of the surfactant. No effects were observed with heat inactivated PMN elastase (95 degrees C, 30 min) or with PMN elastase in the presence of human alpha-1 protease inhibitor (2 micrograms/microgram elastase). Phospholipid compositions of the surfactant after exposure to PMN elastase were not significantly different from those of the controls, suggesting that SP-A, SP-B, and SP-C play a major role in altering the surfactant properties. SP-A was also degraded by elastase and trypsin from pancreas whereas SP-B and SP-C remained intact, providing a natural surfactant without SP-A. Surface adsorption rate of the SP-A deficient surfactant was lower than that of the control, but was much higher than that of the surfactant with completely degraded SP-A, SP-B, and SP-C, suggesting that hydrophobic SP-B and SP-C are the essential components in enhancing adsorption. We conclude that proteolytic degradation of SP-A, SP-B, and SP-C causes the decrease of surfactant isopycnic density, and is responsible for retarding adsorption resulting in surfactant dysfunction.