Augmentation of 5-lipoxygenase activity and expression during dengue serotype-2 infection

Wai Mun Loke, Angelia Yee Chow, Karen Lam Mok Sing, Chung-Yung J Lee, Barry Halliwell, Erle C H Lim, Amy M L Quek, Eng Eong Ooi, Raymond C S Seet, Wai Mun Loke, Angelia Yee Chow, Karen Lam Mok Sing, Chung-Yung J Lee, Barry Halliwell, Erle C H Lim, Amy M L Quek, Eng Eong Ooi, Raymond C S Seet

Abstract

Background: Leukotriene B4, a 5-lipoxygenase product of arachidonic acid with potent chemotactic effects on neutrophils, has not been assessed in dengue patients. In this study, plasma leukotriene B4 and serum high-sensitivity C-reactive protein levels were determined in adult patients during the febrile, convalescent and defervescent stages of dengue serotype-2 (DENV-2) infection, and compared with those of age-matched healthy and non-dengue febrile subjects. In vitro studies were performed to examine the effects of live and heat-inactivated DENV-2 on the activities and expression of 5-lipoxygenase in human neutrophils.

Results: Plasma leukotriene B4 was elevated during the febrile stages of dengue infection compared to levels during convalescence and in study controls. Plasma leukotriene B4 also correlated with serum high-sensitivity C-reactive protein in dengue patients (febrile, r = 0.91, p < 0.001; defervescence, r = 0.87, p < 0.001; convalescence, r = 0.87, p < 0.001). Exposure of human neutrophils to DENV-2 resulted in a significant rise in leukotriene B4; the extent of increase, however, did not differ between exposure to live and heat-inactivated DENV-2. Pre-incubation of either live or heat-inactivated DENV-2 resulted in reduced leukotriene B4 release by neutrophils, indicating that contact with dengue antigens (and not replication) triggers the neutrophil response. Production of leukotriene B4 was associated with an increase in 5-lipoxygenase expression in human neutrophils; addition of MK886 (a 5-lipoxygenase activating protein inhibitor) attenuated further increase in leukotriene B4 production.

Conclusion: These findings provide important clinical and mechanistic data on the involvement of 5-lipoxygenase and its metabolites in dengue infection. Further studies are needed to elucidate the therapeutic implications of these findings.

Figures

Figure 1
Figure 1
Production of leukotriene B4 by neutrophils incubated with or without DENV-2. (A) Production of leukotriene B4 (ng/106 cells) by freshly isolated peripheral neutrophils over 7-hour incubation in the presence and absence of live dengue viruses exposure (DENV-2), heat-inactivated dengue viruses exposure (hiDENV-2), and A23187 (A) treatment (N = 5). * p < 0.05 vs untreated cells using one-way ANOVA analysis of the area under the curves with Bonferroni adjustments. (B) Leukotriene B4 production (ng/106 cells) from freshly isolated peripheral neutrophils after 5 hours of incubation with or without live dengue viruses exposure (DENV-2), heat-inactivated dengue viruses exposure (hiDENV-2), MK886 (MK) and A23187 (A) treatment (N = 5). Bars with the same subscripts are not significantly different from each other using one-way ANOVA analysis with Bonferroni adjustments.
Figure 2
Figure 2
5-Lipoxygenase expression by neutrophils incubated with or without dengue viruses. The proteins were isolated from neutrophils and were electrophoretically transferred from the gel onto Hybond-C Extranitrocellulose membrane. The membrane was then cut, based on the size of proteins of interest, to separate the 5-lipoxygenase (79 kDa) and the control protein GAPDH (39 kDa). (A) Representative films from 5-lipoxygenase and GAPDH Western blots performed with untreated neutrophils and neutrophils incubated with live (DENV-2) or heat-inactivated dengue viruses (hiDENV-2). (B) Expression of 5-lipoxygenase protein by untreated neutrophils (×) and neutrophils exposed to live (DENV-2, □) or heat-inactivated dengue viruses (hiDENV-2, ■) (N = 3). * p < 0.05 vs untreated cells using one-way ANOVA analysis of the area under the curves with Bonferroni adjustments.
Figure 3
Figure 3
Production of leukotriene B4 by neutrophils incubated with or without anti-DENV-2 antibody. Production of leukotriene B4 (ng/106 cells) from freshly isolated peripheral neutrophils after 5-hour incubation in the presence and absence of anti-DENV-2 antibody (antiDENV), with live DENV-2 (DENV), heat-inactivated DENV-2 (hiDENV) and A23187 (A) treatment (N = 5). * p < 0.05 vs DENV + A using one-way ANOVA analysis with Bonferroni adjustments. # p < 0.05 vs hiDENV + A using one-way ANOVA analysis with Bonferroni adjustments.

References

    1. Bhatt S, Gething PW, Brady OJ, Messina JP, Farlow AW, Moyes CL, The global distribution and burden of dengue. Nature. 2013. doi:10.1038/nature12060.
    1. Ooi EE, Goh KT, Gubler DJ. Dengue prevention and 35 years of vector control in Singapore. Emerg Infect Dis. 2006;10:887–893.
    1. Seet RCS, Lee CY, Lim ECH, Quek AML, Yeo LL, Huang SH, Halliwell B. Oxidative damage in dengue fever. Free Rad Biol Med. 2009;10:375–380. doi: 10.1016/j.freeradbiomed.2009.04.035.
    1. Juffrie M, Van der Meer GM, Hack CE, Haasnoot K, Sutaryo A, Veerman AJP, Thijs LG. Inflammatory mediators in dengue virus infection in children: Interleukin-8 and its relationship to neutrophil degranulation. Infect Immun. 2000;10:702–707. doi: 10.1128/IAI.68.2.702-707.2000.
    1. Butthep P, Bunyaratvej A, Bhamarapravati N. Dengue virus and endothelial cell: a related phenomenon to thrombocytopenia and granulocytopenia in dengue hemorrhagic fever. Southeast Asian J Trop Med Public Health. 1993;10:246–249.
    1. Ganz T. Defensins: antimicrobial peptides of innate immunity. Nat Rev Immunol. 2003;10:710–720. doi: 10.1038/nri1180.
    1. Lehrer RI, Ganz T. Cathelicidins: a family of endogenous antimicrobial peptides. Curr Opin Hematol. 2002;10:18–22. doi: 10.1097/00062752-200201000-00004.
    1. Yokomizo T, Izumi T, Shimizu T. Leukotriene B4: metabolism and signal transduction. Arch Biochem Biophys. 2001;10:231–241. doi: 10.1006/abbi.2000.2168.
    1. Subbarao K, Jala VR, Mathis S, Suttles J, Zacharias W, Ahamed J, Ali H, Tseng MT, Haribabu B. Role of Leukotriene B4 receptors in the development of atherosclerosis: potential mechanisms. Arterioscler Thromb Vasc Biol. 2004;10:369–375.
    1. Flamand L, Tremblay MJ, Borgeat P. Leukotriene B4 triggers the in vitro and in vivo release of potent antimicrobial agents. J Immunol. 2007;10:8036–8045.
    1. Tager AM, Bromley SK, Medoff BD, Islam SA, Bercury SD, Friedrich EB, Carafone AD, Gerszten RE, Luster AD. Leukotriene B4 receptor BLT1 mediates early effector T cell recruitment. Nat Immunol. 2003;10:982–990. doi: 10.1038/ni970.
    1. Lotzer K, Funk KD, Habenich AJR. The 5-lipoxygenase pathway in arterial wall biology and atherosclerosis. Biochim Biophys Acta. 2005;10:30–37.
    1. Seet RC, Lee CY, Chan BP, Sharma VK, Teoh HL, Venketasubramanian N, Lim EC, Chong WL, Looi WF, Huang SH, Ong BK, Halliwell B. Oxidative damage in ischemic stroke revealed using multiple biomarkers. Stroke. 2011;10:2326–2329. doi: 10.1161/STROKEAHA.111.618835.
    1. Loke WM, Lam KMJ, Chong WL, Chew SE, Quek AML, Lim ECH, Seet RCS. Products of 5-lipoxygenase and myeloperoxidase activities are increased in young male cigarette smokers. Free Radic Res. 2012;10:1230–1237. doi: 10.3109/10715762.2012.701291.
    1. García G, González N, Pérez AB, Sierra B, Aguirre E, Rizo D, Izquierdo A, Sánchez L, Díaz D, Lezcay M. Long-term persistence of clinical symptoms in dengue-infected persons and its association with immunological disorders. Int J Infect Dis. 2011;10:e38–e43. doi: 10.1016/j.ijid.2010.09.008.
    1. Ananaba GA, Anderson LJ. Antibody enhancement of respiratory syncytial virus stimulation of leukotriene production by a macrophagelike cell line. J Virol. 1991;10:5052–5060.
    1. Medina JF, Barrios C, Funk CD, Larsson O, Haeggstrom J, Radmark O. Human fibroblasts show expression of the leukotriene-A4-hydrolase gene, which is increased after simian-virus-40 transformation. Euro J Biochem. 1990;10:27–31. doi: 10.1111/j.1432-1033.1990.tb19089.x.
    1. Gosselin J, Borgeat P. Epstein-Barr virus modulates 5-lipoxygenase product synthesis in human peripheral blood mononuclear cells. Blood. 1997;10:2122–2130.
    1. Qiu H, Straat K, Rahbar A, Wan M, Soderberg-Naucler C, Haeggstrom JZ. Human CMV infection induces 5-lipoxygenase expression and leukotriene B4 production in vascular smooth muscle cells. J Exp Med. 2008;10:19–24. doi: 10.1084/jem.20070201.
    1. Wahl LM, Corcoran ML, Pyle SW, Arthur LO, Harel-Bellan A, Farrar WL. Human immunodeficiency virus glycoprotein (GP120) induction of monocyte arachidonic acid metabolites and interleukin 1. Proc Natl Acad Sci USA. 1989;10:621–625. doi: 10.1073/pnas.86.2.621.
    1. Volovitz B, Faden H, Ogra PL. Release of leukotriene C4 in respiratory tract during acute viral infection. J Pediatr. 1988;10:218–222. doi: 10.1016/S0022-3476(88)80058-1.
    1. Widegren H, Andersson M, Borgeat P, Flamand L, Johnston S, Greiff L. LTB4 increases nasal neutrophil activity and conditions neutrophils to exert antiviral effects. Respir Med. 2011;10:997–1006. doi: 10.1016/j.rmed.2010.12.021.
    1. Bertin J, Barat C, Bélanger D, Tremblay MJ. Leukotrienes inhibit early stages of HIV-1 infection in monocyte-derived microglia-like cells. J Neuroinflammation. 2012;10:55–70. doi: 10.1186/1742-2094-9-55.
    1. Levine H, Kessler DA, Rappel W-J. Directional sensing in eukaryotic chemotaxis: A balanced inactivation model. Proc Natl Acad Sci. 2006;10:9761–9766. doi: 10.1073/pnas.0601302103.
    1. Luster AD, Tager AM. T-cell trafficking in asthma: lipid mediators grease the way. Nat Rev Immunol. 2004;10:711–724. doi: 10.1038/nri1438.
    1. Zhang YY, Walker JL, Huang A, Keaney JF, Clish CB, Serhan CN, Loscalzo J. Expression of 5-lipoxygenase in pulmonary artery endothelial cells. Biochem J. 2002;10:267–276. doi: 10.1042/0264-6021:3610267.
    1. Flamand L, Borgeat P, Lalonde R, Gosselin J. Release of anti-HIV mediators after administration of leukotriene B4 to humans. J Infect Dis. 2004;10:2001–2009. doi: 10.1086/386374.
    1. Rola-Pleszczynski M, Chavaillaz PA, Lemaire I. Stimulation of interleukin 2 and interferon gamma production by leukotriene B4 in human lymphocyte cultures. Prostaglandins Leukot Med. 1986;10:207–210. doi: 10.1016/0262-1746(86)90187-3.
    1. Rola-Pleszczynski M, Lemaire I. Leukotrienes augment interleukin 1 production by human monocytes. J Immunol. 1985;10:3958–3961.
    1. Henderson WR Jr. The role of leukotrienes in inflammation. Ann Intern Med. 1994;10:684–697. doi: 10.7326/0003-4819-121-9-199411010-00010.
    1. Libby P. Inflammation in atherosclerosis. Nature. 2002;10:868–874. doi: 10.1038/nature01323.
    1. Huang L, Zhao A, Wong F, Ayala JM, Struthers M, Ujjainwalla F, Wright SD, Springer MS, Evans J, Cui J. Leukotriene B4 strongly increases monocyte chemoattractant protein-1 in human monocytes. Arterioscler Thromb Vasc Biol. 2004;10:1783–1788. doi: 10.1161/01.ATV.0000140063.06341.09.
    1. Aiello RJ, Bourassa P-A, Lindsey S, Weng W, Freeman A, Showell HJ. Leukotriene B4 receptor antagonism reduces monocytic foam cells in mice. Arterioscler Thromb Vasc Biol. 2002;10:443–449. doi: 10.1161/hq0302.105593.
    1. Chen M, Lam BK, Kanaoka Y, Nigrovic PA, Audoly LP, Austen KF, Lee DM. Neutrophil-derived leukotriene B4 is required for inflammatory arthritis. J Exp Med. 2006;10:837–842. doi: 10.1084/jem.20052371.
    1. Wang D, DuBois N. Measurement of eicosanoids in cancer tissues. Methods Enzymol. 2007;10:27–50.
    1. Lee CYJ, Huang SH, Jenner AM, Halliwell B. Measurement of F2-isoprostanes, hydroxyeicosatetraenoic products, and oxysterols from a single plasma sample. Free Radical Biol Med. 2008;10:1314–1322. doi: 10.1016/j.freeradbiomed.2007.12.026.
    1. Loke WM, Proudfoot JM, Stewart S, McKinley AJ, Needs PW, Kroon PA, Hodgson JM, Croft KD. Metabolic transformation has a profound effect on anti-inflammatory activity of flavonoids such as quercetin: lack of association between antioxidant and lipoxygenase inhibitory activity. Biochem Pharmacol. 2008;10:1045–1053. doi: 10.1016/j.bcp.2007.11.002.
    1. Palmer R, Salmon J. Release of leukotriene B4 from human neutrophils and its relationship to degranulation induced by N-formyl-methionyl-leucyl-phenylalanine, serum-treated zymosan and the ionophore A23187. Immunology. 1983;10:65–73.
    1. Rouzer CA, Ford-Hutchinson AW, Morton HE, Gillard JW. MK886, a potent and specific leukotriene biosynthesis inhibitor blocks and reverses the membrane association of 5-lipoxygenase in ionophore-challenged leukocytes. J Biol Chem. 1990;10:1436–1442.

Source: PubMed

Sponsorer og samarbeidspartnere

Medisinsk tilstand

Legemiddelintervensjoner

3
Abonnere