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Division of Pulmonary and Critical Care Medicine, Departments of 1 Anesthesiology and 2 Medicine, Division of Nephrology and Nephrology Research Training Center, Departments of 3 Genomics and Pathobiology and 4 Medicine, 6 Department of Physiology and Biophysics, School of Medicine, University of Alabama at Birmingham, Birmingham, Alabama 35294; and 5 Environmental Protection Agency, Research Triangle Park, North Carolina 27711-0001
We investigated putative mechanisms by
which human surfactant protein A (SP-A) effects killing of
Klebsiella pneumoniae by human alveolar macrophages (AMs)
isolated from bronchoalveolar lavagates of patients with transplanted
lungs. Coincubation of AMs with human SP-A (25 µg/ml) and
Klebsiella resulted in a 68% decrease in total colony
forming units by 120 min compared with AMs infected with
Klebsiella in the absence of SP-A, and this SP-A-mediated
effect was abolished by preincubation with
NG-monomethyl-L-arginine. Incubation
of transplant AMs with SP-A increased intracellular Ca2+
concentration ([Ca2+]i) by 70% and nitrite
and nitrate (NOx) production by 45% (from 0.24 ± 0.02 to 1.3 ± 0.21 nmol · 106
AMs
1 · h1). Preincubation with
1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester inhibited the increase in
[Ca2+]i and abrogated the SP-A-mediated
Klebsiella phagocytosis and killing. In contrast, incubation
of AMs from normal volunteers with SP-A decreased both
[Ca2+]i and NOx production and
did not result in killing of Klebsiella. Significant killing
of Klebsiella was also seen in a cell-free system by
sustained production of peroxynitrite (>1 µM/min) at pH 5 but not at
pH 7.4. These findings indicate that SP-A mediates pathogen killing by
AMs from transplant lungs by stimulating phagocytosis and production of
reactive oxygen-nitrogen intermediates.
innate immunity; collectins; peroxynitrite; lung transplant; phagolysosome
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