Skip to main content
NCBI home page
Thorax logoLink to Thorax
. 1996 Dec;51(12):1223–1228. doi: 10.1136/thx.51.12.1223

Eicosanoid mediator expression in mononuclear and polymorphonuclear cells in normal subjects and patients with atopic asthma and cystic fibrosis.

L M Kuitert 1, R Newton 1, N C Barnes 1, I M Adcock 1, P J Barnes 1
PMCID: PMC472767  PMID: 8994519

Abstract

BACKGROUND: Eicosanoids such as leukotrienes, prostaglandins, lipoxins, and 15-hydroperoxyeicosatetraenoic acid (15-HETE) cause bronchoconstriction, increased microvascular permeability, mucus secretion, and polymorph chemotaxis. These pro-inflammatory effects are important in diseases such as asthma and cystic fibrosis where the levels of mediators are increased both in the stable and acute state. A study was conducted to examine the expression of the mRNA for the enzymes of the eicosanoid pathways (5-lipoxygenase (5-LO), 5-lipoxygenase activating protein (FLAP), cyclo-oxygenases 1 and 2 (COX-1, COX-2), and 15-lipoxygenase (15-LO)) in normal subjects and in patients with stable atopic asthma and stable cystic fibrosis. METHODS: Reverse transcription polymerase chain reaction (RT-PCR) was used to examine the expression of total RNA for 5-LO, FLAP, COX-1, COX-2, and 15-LO in peripheral blood polymorphonuclear cells and mononuclear cells from the three subjects groups. RESULTS: The expression of mRNA for 5-LO and FLAP was similar in normal subjects and in patients with asthma and cystic fibrosis. COX-1 was increased in both cell types in asthmatic patients. COX-2 and 15-LO were increased in polymorphs of patients with atopic asthma but not in mononuclear cells. COX-2 and 15-LO were undetectable in either cell type in patients with cystic fibrosis whereas COX-1 levels in polymorphs were similar to those in patients with asthma. CONCLUSIONS: The increased leukotriene production in asthma and cystic fibrosis is not explained by an increase in transcription of 5-LO and FLAP. Transcription of 15-LO and COX-2 is increased in atopic asthma. Transcription of COX-1 is increased in both atopic asthma and cystic fibrosis.

Full text

PDF
1223

Images in this article

1224Image
on p.1224

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ali A., Ford-Hutchinson A. W., Nicholson D. W. Activation of protein kinase C down-regulates leukotriene C4 synthase activity and attenuates cysteinyl leukotriene production in an eosinophilic substrain of HL-60 cells. J Immunol. 1994 Jul 15;153(2):776–788. [PubMed] [Google Scholar]
  2. Arm J. P., Lee T. H. Sulphidopeptide leukotrienes in asthma. Clin Sci (Lond) 1993 May;84(5):501–510. doi: 10.1042/cs0840501. [DOI] [PubMed] [Google Scholar]
  3. Badr K. F. 15-Lipoxygenase products as leukotriene antagonists: therapeutic potential in glomerulonephritis. Kidney Int Suppl. 1992 Oct;38:S101–S108. [PubMed] [Google Scholar]
  4. Bennett C. F., Chiang M. Y., Monia B. P., Crooke S. T. Regulation of 5-lipoxygenase and 5-lipoxygenase-activating protein expression in HL-60 cells. Biochem J. 1993 Jan 1;289(Pt 1):33–39. doi: 10.1042/bj2890033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bigby T. D., Holtzman M. J. Enhanced 5-lipoxygenase activity in lung macrophages compared to monocytes from normal subjects. J Immunol. 1987 Mar 1;138(5):1546–1550. [PubMed] [Google Scholar]
  6. Bonfield T. L., Konstan M. W., Burfeind P., Panuska J. R., Hilliard J. B., Berger M. Normal bronchial epithelial cells constitutively produce the anti-inflammatory cytokine interleukin-10, which is downregulated in cystic fibrosis. Am J Respir Cell Mol Biol. 1995 Sep;13(3):257–261. doi: 10.1165/ajrcmb.13.3.7544594. [DOI] [PubMed] [Google Scholar]
  7. Christie P. E., Spur B. W., Lee T. H. The effects of lipoxin A4 on airway responses in asthmatic subjects. Am Rev Respir Dis. 1992 Jun;145(6):1281–1284. doi: 10.1164/ajrccm/145.6.1281. [DOI] [PubMed] [Google Scholar]
  8. Coffey M. J., Wilcoxen S. E., Peters-Golden M. Increases in 5-lipoxygenase activating protein expression account for enhanced capacity for 5-lipoxygenase metabolism that accompanies differentiation of peripheral blood monocytes into alveolar macrophages. Am J Respir Cell Mol Biol. 1994 Aug;11(2):153–158. doi: 10.1165/ajrcmb.11.2.8049076. [DOI] [PubMed] [Google Scholar]
  9. Conrad D. J., Kuhn H., Mulkins M., Highland E., Sigal E. Specific inflammatory cytokines regulate the expression of human monocyte 15-lipoxygenase. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):217–221. doi: 10.1073/pnas.89.1.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fischer D. B., Christman J. W., Badr K. F. Fifteen-S-hydroxyeicosatetraenoic acid (15-S-HETE) specifically antagonizes the chemotactic action and glomerular synthesis of leukotriene B4 in the rat. Kidney Int. 1992 May;41(5):1155–1160. doi: 10.1038/ki.1992.176. [DOI] [PubMed] [Google Scholar]
  11. Gough N. M. Rapid and quantitative preparation of cytoplasmic RNA from small numbers of cells. Anal Biochem. 1988 Aug 15;173(1):93–95. doi: 10.1016/0003-2697(88)90164-9. [DOI] [PubMed] [Google Scholar]
  12. Hempel S. L., Monick M. M., Hunninghake G. W. Lipopolysaccharide induces prostaglandin H synthase-2 protein and mRNA in human alveolar macrophages and blood monocytes. J Clin Invest. 1994 Jan;93(1):391–396. doi: 10.1172/JCI116971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Israel E., Rubin P., Kemp J. P., Grossman J., Pierson W., Siegel S. C., Tinkelman D., Murray J. J., Busse W., Segal A. T. The effect of inhibition of 5-lipoxygenase by zileuton in mild-to-moderate asthma. Ann Intern Med. 1993 Dec 1;119(11):1059–1066. doi: 10.7326/0003-4819-119-11-199312010-00001. [DOI] [PubMed] [Google Scholar]
  14. Kargman S., Rouzer C. A. Studies on the regulation, biosynthesis, and activation of 5-lipoxygenase in differentiated HL60 cells. J Biol Chem. 1989 Aug 5;264(22):13313–13320. [PubMed] [Google Scholar]
  15. Lee T. H., Crea A. E., Gant V., Spur B. W., Marron B. E., Nicolaou K. C., Reardon E., Brezinski M., Serhan C. N. Identification of lipoxin A4 and its relationship to the sulfidopeptide leukotrienes C4, D4, and E4 in the bronchoalveolar lavage fluids obtained from patients with selected pulmonary diseases. Am Rev Respir Dis. 1990 Jun;141(6):1453–1458. doi: 10.1164/ajrccm/141.6.1453. [DOI] [PubMed] [Google Scholar]
  16. Lewis R. A., Austen K. F., Soberman R. J. Leukotrienes and other products of the 5-lipoxygenase pathway. Biochemistry and relation to pathobiology in human diseases. N Engl J Med. 1990 Sep 6;323(10):645–655. doi: 10.1056/NEJM199009063231006. [DOI] [PubMed] [Google Scholar]
  17. Lin A. H., Bienkowski M. J., Gorman R. R. Regulation of prostaglandin H synthase mRNA levels and prostaglandin biosynthesis by platelet-derived growth factor. J Biol Chem. 1989 Oct 15;264(29):17379–17383. [PubMed] [Google Scholar]
  18. Malaviya R., Malaviya R., Jakschik B. A. Reversible translocation of 5-lipoxygenase in mast cells upon IgE/antigen stimulation. J Biol Chem. 1993 Mar 5;268(7):4939–4944. [PubMed] [Google Scholar]
  19. Miadonna A., Tedeschi A., Brasca C., Folco G., Sala A., Murphy R. C. Mediator release after endobronchial antigen challenge in patients with respiratory allergy. J Allergy Clin Immunol. 1990 May;85(5):906–913. doi: 10.1016/0091-6749(90)90076-g. [DOI] [PubMed] [Google Scholar]
  20. Mitchell J. A., Larkin S., Williams T. J. Cyclooxygenase-2: regulation and relevance in inflammation. Biochem Pharmacol. 1995 Nov 9;50(10):1535–1542. doi: 10.1016/0006-2952(95)00212-x. [DOI] [PubMed] [Google Scholar]
  21. Nadel J. A., Conrad D. J., Ueki I. F., Schuster A., Sigal E. Immunocytochemical localization of arachidonate 15-lipoxygenase in erythrocytes, leukocytes, and airway cells. J Clin Invest. 1991 Apr;87(4):1139–1145. doi: 10.1172/JCI115110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nassar G. M., Morrow J. D., Roberts L. J., 2nd, Lakkis F. G., Badr K. F. Induction of 15-lipoxygenase by interleukin-13 in human blood monocytes. J Biol Chem. 1994 Nov 4;269(44):27631–27634. [PubMed] [Google Scholar]
  23. O'Banion M. K., Winn V. D., Young D. A. cDNA cloning and functional activity of a glucocorticoid-regulated inflammatory cyclooxygenase. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4888–4892. doi: 10.1073/pnas.89.11.4888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pueringer R. J., Bahns C. C., Hunninghake G. W. Alveolar macrophages have greater amounts of the enzyme 5-lipoxygenase than do monocytes. J Appl Physiol (1985) 1992 Aug;73(2):781–786. doi: 10.1152/jappl.1992.73.2.781. [DOI] [PubMed] [Google Scholar]
  25. Sampson A. P., Green C. P., Spencer D. A., Piper P. J., Price J. F. Leukotrienes in the blood and urine of children with acute asthma. Ann N Y Acad Sci. 1991;629:437–439. doi: 10.1111/j.1749-6632.1991.tb38011.x. [DOI] [PubMed] [Google Scholar]
  26. Sampson A. P., Spencer D. A., Green C. P., Piper P. J., Price J. F. Leukotrienes in the sputum and urine of cystic fibrosis children. Br J Clin Pharmacol. 1990 Dec;30(6):861–869. doi: 10.1111/j.1365-2125.1990.tb05452.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sampson A. P., Thomas R. U., Costello J. F., Piper P. J. Enhanced leukotriene synthesis in leukocytes of atopic and asthmatic subjects. Br J Clin Pharmacol. 1992 Apr;33(4):423–430. doi: 10.1111/j.1365-2125.1992.tb04062.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Serhan C. N. Lipoxin biosynthesis and its impact in inflammatory and vascular events. Biochim Biophys Acta. 1994 Apr 14;1212(1):1–25. doi: 10.1016/0005-2760(94)90185-6. [DOI] [PubMed] [Google Scholar]
  29. Shannon V. R., Chanez P., Bousquet J., Holtzman M. J. Histochemical evidence for induction of arachidonate 15-lipoxygenase in airway disease. Am Rev Respir Dis. 1993 Apr;147(4):1024–1028. doi: 10.1164/ajrccm/147.4.1024. [DOI] [PubMed] [Google Scholar]
  30. Sigal E., Conrad D. J. Human 15-lipoxygenase: a potential effector molecule for interleukin-4. Adv Prostaglandin Thromboxane Leukot Res. 1994;22:309–316. [PubMed] [Google Scholar]
  31. Zakrzewski J. T., Barnes N. C., Piper P. J., Costello J. F. Detection of sputum eicosanoids in cystic fibrosis and in normal saliva by bioassay and radioimmunoassay. Br J Clin Pharmacol. 1987 Jan;23(1):19–27. doi: 10.1111/j.1365-2125.1987.tb03004.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Thorax are provided here courtesy of BMJ Publishing Group

RESOURCES