Synthesis of leukotriene B4, and prostanoids by human alveolar macrophages: analysis by gas chromatography/ mass spectrometry |
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Authors: | J. MacDermot C.R. Kelsey K.A. Waddell R. Richmond R.K. Knight P.J. Cole C.T. Dollery D.N. Landon I.A. Blair |
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Affiliation: | 1. Department of Clinical Pharmacology, Royal Postgraduate Medical School, London W12 OHS, UK |
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Abstract: | Human alveolar macrophages, obtained during diagnostic bronchoscoy, were maintained in monolayer culture. Challenge of these cells (>95% purity) with 1.2 mg/ml zymosan A particles (opsonized with human serum) was followed by a rapid release of leukotriene B4 into the medium, 7.28 ± 5.99 ng/mg cell protein at 2 h mean ± S.D4, n = 4). Leukotriene B4 was identified and measured by a novel technique employing capillary column gas chromatography coupled to negative ion chemical ionization mass spectrometry. The release of thromboxane B2, prostaglandins D2, E2, F2α and the lysosomal enzyme N-acetyl-β-D- glucosaminidase was also measured. Thromboxane B2 was the most abundant metabolite of arachidonic acid released into the culture medium (65.2 ± 14.8 ng/mg cell protein 2 h after the addition of zymosanA, n = 4), and the synthesis of thromboxane B2 was inhibited by >90% in 1 μM Na flurbiprofen. Inhibition of cyclooxygenase activity was accompanied by a 2-fold increase in leukotriene B4 synthesis. |
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