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. 1997 Dec;52(12):1024–1029. doi: 10.1136/thx.52.12.1024

Enhancement of leukotriene B4 release in stimulated asthmatic neutrophils by platelet activating factor

K Shindo, K Koide, M Fukumura
PMCID: PMC1758460  PMID: 9516893

Abstract

BACKGROUND: The role of platelet activating factor (PAF) in asthma remains controversial. The priming effect of PAF on leukotriene B4 (LTB4) release, 5-lipoxygenase activity, and intracellular calcium levels in asthmatic neutrophils was examined. METHODS: LTB4 and other lipoxygenase metabolites in neutrophils obtained from 17 asthmatic patients and 15 control subjects were measured by reverse phase-high performance liquid chromatography (RP-HPLC). Intracellular calcium levels were monitored using the fluorescent probe fura-2. RESULTS: The mean (SD) basal LTB4, release from neutrophils was not significantly different between the two groups (0.05 (0.01) vs 0.03 (0.02) ng/10(6) cells); however, when stimulated with calcium ionophore A23187 (2.5 microM), neutrophils from asthma patients released more LTB4 than cells from control subjects (15.7 (1.2) vs 9.9 (1.6) ng/10(6) cells). Although PAF alone did not alter LTB4 release, it enhanced the response to subsequent A23187 stimulation. This effect was observed following treatment for five minutes with PAF at concentrations > 1.0 microM. The maximal effect was seen with 5.0 microM PAF + 2.5 microM A23187 (62.7 (2.2) vs 18.6 (2.3) ng/10(6) cells). Pretreatment with PAF also increased 5-lipoxygenase activity and intracellular calcium levels in neutrophils from asthmatic patients to a greater extent than in those from non-asthmatic patients. CONCLUSIONS: These findings indicate that, in neutrophils from asthmatic patients, PAF enhances LTB4 release and increases 5-lipoxygenase activity and intracellular calcium to a greater extent than in neutrophils from non-asthmatic patients. 




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Selected References

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  1. Beaubien B. B., Tippins J. R., Morris H. R. Platelet-activating factor stimulation of peptidoleukotriene release: inhibition by vasoactive polypeptide. Biochem Biophys Res Commun. 1984 Nov 30;125(1):105–108. doi: 10.1016/s0006-291x(84)80340-x. [DOI] [PubMed] [Google Scholar]
  2. Benveniste J., Henson P. M., Cochrane C. G. Leukocyte-dependent histamine release from rabbit platelets. The role of IgE, basophils, and a platelet-activating factor. J Exp Med. 1972 Dec 1;136(6):1356–1377. doi: 10.1084/jem.136.6.1356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  5. Chao W., Olson M. S. Platelet-activating factor: receptors and signal transduction. Biochem J. 1993 Jun 15;292(Pt 3):617–629. doi: 10.1042/bj2920617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chung K. F. Platelet-activating factor in inflammation and pulmonary disorders. Clin Sci (Lond) 1992 Aug;83(2):127–138. doi: 10.1042/cs0830127. [DOI] [PubMed] [Google Scholar]
  7. Koh Y. Y., Dupuis R., Pollice M., Albertine K. H., Fish J. E., Peters S. P. Neutrophils recruited to the lungs of humans by segmental antigen challenge display a reduced chemotactic response to leukotriene B4. Am J Respir Cell Mol Biol. 1993 May;8(5):493–499. doi: 10.1165/ajrcmb/8.5.493. [DOI] [PubMed] [Google Scholar]
  8. Kuitert L. M., Angus R. M., Barnes N. C., Barnes P. J., Bone M. F., Chung K. F., Fairfax A. J., Higenbotham T. W., O'Connor B. J., Piotrowska B. Effect of a novel potent platelet-activating factor antagonist, modipafant, in clinical asthma. Am J Respir Crit Care Med. 1995 May;151(5):1331–1335. doi: 10.1164/ajrccm.151.5.7735582. [DOI] [PubMed] [Google Scholar]
  9. Lin A. H., Morton D. R., Gorman R. R. Acetyl glyceryl ether phosphorylcholine stimulates leukotriene B4 synthesis in human polymorphonuclear leukocytes. J Clin Invest. 1982 Nov;70(5):1058–1065. doi: 10.1172/JCI110693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Meltzer S., Goldberg B., Lad P., Easton J. Superoxide generation and its modulation by adenosine in the neutrophils of subjects with asthma. J Allergy Clin Immunol. 1989 May;83(5):960–966. doi: 10.1016/0091-6749(89)90112-7. [DOI] [PubMed] [Google Scholar]
  11. Miwa M., Miyake T., Yamanaka T., Sugatani J., Suzuki Y., Sakata S., Araki Y., Matsumoto M. Characterization of serum platelet-activating factor (PAF) acetylhydrolase. Correlation between deficiency of serum PAF acetylhydrolase and respiratory symptoms in asthmatic children. J Clin Invest. 1988 Dec;82(6):1983–1991. doi: 10.1172/JCI113818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Munafo D. A., Shindo K., Baker J. R., Bigby T. D. Leukotriene A4 hydrolase in human bronchoalveolar lavage fluid. J Clin Invest. 1994 Mar;93(3):1042–1050. doi: 10.1172/JCI117053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Naccache P. H., Faucher N., Caon A. C., McColl S. R. Propionic acid-induced calcium mobilization in human neutrophils. J Cell Physiol. 1988 Jul;136(1):118–124. doi: 10.1002/jcp.1041360115. [DOI] [PubMed] [Google Scholar]
  14. Nadel J. A. Genetics reveals importance of platelet activating factor in asthma and possibly other inflammatory states. J Clin Invest. 1996 Jun 15;97(12):2689–2690. doi: 10.1172/JCI118720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Neijens H. J., Raatgeep R. E., Degenhart H. J., Duiverman E. J., Kerrebijn K. F. Altered leukocyte response in relation to the basic abnormality in children with asthma and bronchial hyperresponsiveness. Am Rev Respir Dis. 1984 Nov;130(5):744–747. doi: 10.1164/arrd.1984.130.5.744. [DOI] [PubMed] [Google Scholar]
  16. Postma D. S., Renkema T. E., Noordhoek J. A., Faber H., Sluiter H. J., Kauffman H. Association between nonspecific bronchial hyperreactivity and superoxide anion production by polymorphonuclear leukocytes in chronic air-flow obstruction. Am Rev Respir Dis. 1988 Jan;137(1):57–61. doi: 10.1164/ajrccm/137.1.57. [DOI] [PubMed] [Google Scholar]
  17. Pozzan T., Arslan P., Tsien R. Y., Rink T. J. Anti-immunoglobulin, cytoplasmic free calcium, and capping in B lymphocytes. J Cell Biol. 1982 Aug;94(2):335–340. doi: 10.1083/jcb.94.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rouzer C. A., Kargman S. Translocation of 5-lipoxygenase to the membrane in human leukocytes challenged with ionophore A23187. J Biol Chem. 1988 Aug 5;263(22):10980–10988. [PubMed] [Google Scholar]
  19. Scappaticci E., Libertucci D., Bottomicca F., Da Col R., Silvestro L., Tetta C., Camussi G. Platelet-activating factor in bronchoalveolar lavage from patients with sarcoidosis. Am Rev Respir Dis. 1992 Aug;146(2):433–438. doi: 10.1164/ajrccm/146.2.433. [DOI] [PubMed] [Google Scholar]
  20. Shindo K., Koide K., Hirai Y., Sumitomo M., Fukumura M. Priming effect of platelet activating factor on leukotriene C4 from stimulated eosinophils of asthmatic patients. Thorax. 1996 Feb;51(2):155–158. doi: 10.1136/thx.51.2.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stafforini D. M., Satoh K., Atkinson D. L., Tjoelker L. W., Eberhardt C., Yoshida H., Imaizumi T., Takamatsu S., Zimmerman G. A., McIntyre T. M. Platelet-activating factor acetylhydrolase deficiency. A missense mutation near the active site of an anti-inflammatory phospholipase. J Clin Invest. 1996 Jun 15;97(12):2784–2791. doi: 10.1172/JCI118733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stenton S. C., Court E. N., Kingston W. P., Goadby P., Kelly C. A., Duddridge M., Ward C., Hendrick D. J., Walters E. H. Platelet-activating factor in bronchoalveolar lavage fluid from asthmatic subjects. Eur Respir J. 1990 Apr;3(4):408–413. [PubMed] [Google Scholar]
  23. Tool A. T., Verhoeven A. J., Roos D., Koenderman L. Platelet-activating factor (PAF) acts as an intercellular messenger in the changes of cytosolic free Ca2+ in human neutrophils induced by opsonized particles. FEBS Lett. 1989 Dec 18;259(1):209–212. doi: 10.1016/0014-5793(89)81530-3. [DOI] [PubMed] [Google Scholar]
  24. Tsien R. Y. A non-disruptive technique for loading calcium buffers and indicators into cells. Nature. 1981 Apr 9;290(5806):527–528. doi: 10.1038/290527a0. [DOI] [PubMed] [Google Scholar]
  25. Turner C. R., Breslow R., Conklyn M. J., Andresen C. J., Patterson D. K., Lopez-Anaya A., Owens B., Lee P., Watson J. W., Showell H. J. In vitro and in vivo effects of leukotriene B4 antagonism in a primate model of asthma. J Clin Invest. 1996 Jan 15;97(2):381–387. doi: 10.1172/JCI118426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. White J. R., Naccache P. H., Molski T. F., Borgeat P., Sha'afi R. I. Direct demonstration of increased intracellular concentration of free calcium in rabbit and human neutrophils following stimulation by chemotactic factor. Biochem Biophys Res Commun. 1983 May 31;113(1):44–50. doi: 10.1016/0006-291x(83)90429-1. [DOI] [PubMed] [Google Scholar]

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