Lung surfactant protein A (SP-A) activates a phosphoinositide/calcium signaling pathway in alveolar macrophages.
Lung surfactant protein A (SP-A), the main protein component of lung surfactant which lines the alveoli, strongly enhances serum-independent phagocytosis of bacteria by rat alveolar macrophages. We tested if the effect of SP-A is due to interaction with the macrophages or to opsonization of the bacteria. In phagocytosis assays with fluorescein isothiocyanate labeled bacteria, SP-A had no opsonic effect on Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, but enhanced phagocytosis by acting only on the macrophages. We characterized this activation mechanism. With single cell measurements of fura-2 loaded cells we demonstrate that SP-A raises the intracellular free calcium ion concentration 6 to 8 seconds after addition. This calcium mobilization is dose-dependent in that increased SP-A concentrations lead to a higher percentage of responding cells. Additionally, SP-A leads to a dose-dependent and transient generation of inositol 1,4.5-trisphosphate. Release of intracellular stored calcium by SP-A is a prerequisite for its stimulatory effect on phagocytosis, since SP-A-induced enhancement of phagocytosis can be impaired by prior addition of thapsigargin, a Ca(2+)-ATPase inhibitor that leads to depletion of intracellular calcium stores. We conclude that SP-A activates a phosphoinositide/calcium signaling pathway in alveolar macrophages leading to enhanced serum-independent phagocytosis of bacteria.
Ohmer-Schröck
D
D
University Konstanz, Faculty of Biology, Germany.
Schlatterer
C
C
Plattner
H
H
Schlepper-Schäfer
J
J
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D008262:Macrophage Activation; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000378:metabolism; D009895:Opsonin Proteins / Q000494:pharmacology; D010587:Phagocytosis / Q000187:drug effects; D010716:Phosphatidylinositols / Q000378:metabolism*; D011510:Proteolipids / Q000494:pharmacology*; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects*; D013211:Staphylococcus aureus / Q000187:drug effects
D009895:Opsonin Proteins; D010716:Phosphatidylinositols; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D011994:Recombinant Proteins; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium