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. 2013 Dec 2;18(4):612–630. doi: 10.2478/s11658-013-0107-y

The expression of the eotaxins IL-6 and CXCL8 in human epithelial cells from various levels of the respiratory tract

Magdalena Paplińska-Goryca 1,✉, Patrycja Nejman-Gryz 1, Ryszarda Chazan 1, Hanna Grubek-Jaworska 1
PMCID: PMC6275597  PMID: 24297684

Abstract

Airway epithelium acts as multifunctional site of response in the respiratory tract. Epithelial activity plays an important part in the pathophysiology of obstructive lung disease. In this study, we compare normal human epithelial cells from various levels of the respiratory tract in terms of their reactivity to pro-allergic and pro-inflammatory stimulation. Normal human nasal, bronchial and small airway epithelial cells were stimulated with IL-4 and IL-13. The expressions of the eotaxins IL-6 and CXCL8 were evaluated at the mRNA and protein levels. The effects of pre-treatment with IFN-γ on the cell reactivity were measured, and the responses to TNF-α, LPS and IFN-γ were evaluated. All of the studied primary cells expressed CCL26, IL-6 and IL-8 after IL-4 or IL-13 stimulation. IFN-γ pre-treatment resulted in decreased CCL26 and increased IL-6 expression in the nasal and small airway cells, but this effect was not observed in the bronchial cells. IL-6 and CXCL8 were produced in varying degrees by all of the epithelial primary cells in cultures stimulated with TNF-α, LPS or IFN-γ. We showed that epithelial cells from the various levels of the respiratory tract act in a united way, responding in a similar manner to stimulation with IL-4 and IL-13, showing similar reactivity to TNF-α and LPS, and giving an almost unified response to IFN-γ pre-stimulation.

Electronic Supplementary Material

Supplementary material is available for this article at 10.2478/s11658-013-0107-y and is accessible for authorized users.

Key words: CXCL8, Eotaxin-3, Inflammation, Interferon gamma, Interleukin 6, Respiratory epithelium, United airways

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Abbreviations used

ATCC

American Type Culture Collection

BEGM

bronchial epithelial cell growth medium

COPD

chronic obstructive pulmonary disease

IFN-γ

interferon γ

LPS

lipopolysaccharide

TGF-β

transforming growth factor beta

TNF-α

tumor necrosis factor alpha

References

  • 1.Lloyd CM, Saglani S. Asthma and allergy: the emerging epithelium. Nat. Med. 2010;16:273–274. doi: 10.1038/nm0310-273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Knight DA, Holgate ST. The airway epithelium: structural and functional properties in health and disease. Respirology. 2003;8:432–446. doi: 10.1046/j.1440-1843.2003.00493.x. [DOI] [PubMed] [Google Scholar]
  • 3.Liu Y-J. Thymic stromal lymphopoietin: master switch for allergic inflammation. J. Exp. Med. 2006;203:269–273. doi: 10.1084/jem.20051745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Proud D, Leigh R. Epithelial cells and airway diseases. Immunol. Rev. 2011;242:186–204. doi: 10.1111/j.1600-065X.2011.01033.x. [DOI] [PubMed] [Google Scholar]
  • 5.Cao J, Ren G, Gong Y, Dong S, Yin Y, Zhang L. Bronchial epithelial cells release IL-6, CXCL1 and IL-8 upon mast cell interaction. Cytokine. 2011;56:823–831. doi: 10.1016/j.cyto.2011.09.016. [DOI] [PubMed] [Google Scholar]
  • 6.Danila E, Jurgauskiene L, Norkuniene J, Malickaite R. BAL fluid cells in newly diagnosed pulmonary sarcoidosis with different clinical activity. Ups. J. Med. Sci. 2009;114:26–31. doi: 10.1080/03009730802579729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Siva R, Green RH, Brightling CE, Shelley M, Hargadon B, McKenna S, Monteiro W, Berry M, Parker D, Wardlaw AJ, Pavord ID. Eosinophilic airway inflammation and exacerbations of COPD: a randomised controlled trial. Eur. Respir. J. 2007;29:906–913. doi: 10.1183/09031936.00146306. [DOI] [PubMed] [Google Scholar]
  • 8.Brightling CE, Symon FA, Birring SS, Bradding P, Pavord ID, Wardlaw AJ. TH2 cytokine expression in bronchoalveolar lavage fluid T lymphocytes and bronchial submucosa is a feature of asthma and eosinophilic bronchitis. J. Allergy Clin. Immunol. 2002;110:899–905. doi: 10.1067/mai.2002.129698. [DOI] [PubMed] [Google Scholar]
  • 9.Gordon SB, Read RC. Macrophage defences against respiratory tract infections. Br. Med. Bull. 2002;61:45–61. doi: 10.1093/bmb/61.1.45. [DOI] [PubMed] [Google Scholar]
  • 10.Douwes J, Gibson P, Pekkanen J, Pearce N. Non-eosinophilic asthma: importance and possible mechanisms. Thorax. 2002;57:643–648. doi: 10.1136/thorax.57.7.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Van Wetering S, Zuyderduyn S, Ninaber DK, van Sterkenburg MAJA, Rabe KF, Hiemstra S. Epithelial differentiation is a determinant in the production of eotaxin-2 and -3 by bronchial epithelial cells in response to IL-4 and IL-13. Mol. Immunol. 2007;44:803–811. doi: 10.1016/j.molimm.2006.04.008. [DOI] [PubMed] [Google Scholar]
  • 12.Tomkinson A, Duez C, Cieslewicz G, Pratt JC, Joetham A, Shanafelt MC, Gundel R, Gelfand EW. A murine IL-4 receptor antagonist that inhibits IL-4- and IL-13-induced responses prevents antigen-induced airway eosinophilia and airway hyperresponsiveness. J. Immunol. 2001;166:5792–5800. doi: 10.4049/jimmunol.166.9.5792. [DOI] [PubMed] [Google Scholar]
  • 13.Takizawa H, Ohtoshi T, Yamashita N, Oka T, Ito K. Interleukin 6-receptor expression on human bronchial epithelial cells: regulation by IL-1 and IL-6. Am. J. Physiol. 1996;270:346–352. doi: 10.1152/ajplung.1996.270.3.L346. [DOI] [PubMed] [Google Scholar]
  • 14.Cao J, Wong CK, Yin Y, Lam CWK. Activation of human bronchial epithelial cells by inflammatory cytokines IL-27 and TNF-alpha: implications for immunopathophysiology of airway inflammation. J. Cell. Physiol. 2010;223:788–797. doi: 10.1002/jcp.22094. [DOI] [PubMed] [Google Scholar]
  • 15.Ge Q, Moir LM, Black JL, Oliver BG, Burgess JK. TGFβ1 induces IL-6 and inhibits IL-8 release in human bronchial epithelial cells: the role of Smad2/3. J. Cell. Physiol. 2010;225:846–854. doi: 10.1002/jcp.22295. [DOI] [PubMed] [Google Scholar]
  • 16.Xie XH, Law HKW, Wang LJ, Li X, Yang XQ, Liu EM. Lipopolysaccharide induces IL-6 production in respiratory syncytial virusinfected airway epithelial cells through the toll-like receptor 4 signaling pathway. Pediatr. Res. 2009;65:156–162. doi: 10.1203/PDR.0b013e318191f5c6. [DOI] [PubMed] [Google Scholar]
  • 17.Xia C, Shichang Z, Tao L, Yong L, Yingjie W. Maintenance of rat hepatocytes under inflammation by coculture with human orbital fat-derived stem cells. Cell. Mol. Biol. Lett. 2012;17:182–195. doi: 10.2478/s11658-012-0004-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Denning GM, Wollenweber LA, Railsback MA, Cox CD, Stoll LL, Britigan BE. Pseudomonas pyocyanin increases interleukin-8 expression by human airway epithelial cells. Infect. Immun. 1998;66:5777–5784. doi: 10.1128/iai.66.12.5777-5784.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Nakanaga T, Nadel JA, Ueki IF, Koff JL, Shao MXG. Regulation of interleukin-8 via an airway epithelial signaling cascade. Am. J. Physiol. Lung Cell. Mol. Physiol. 2007;292:1289–1296. doi: 10.1152/ajplung.00356.2006. [DOI] [PubMed] [Google Scholar]
  • 20.Li J, Kartha S, Iasvovskaia S, Tan A, Bhat RK, Manaligod JM, Page K, Brasier AR, Hershenson MB. Regulation of human airway epithelial cell IL-8 expression by MAP kinases. Am. J. Physiol. Lung Cell. Mol. Physiol. 2002;283:690–699. doi: 10.1152/ajplung.00060.2002. [DOI] [PubMed] [Google Scholar]
  • 21.Compalati E, Ridolo E, Passalacqua G, Braido F, Villa E, Canonica GW. The link between allergic rhinitis and asthma: the united airways disease. Expert Rev. Clin. Immunol. 2010;6:413–423. doi: 10.1586/eci.10.15. [DOI] [PubMed] [Google Scholar]
  • 22.Togias A. Rhinitis and asthma: evidence for respiratory system integration. J. Allergy Clin. Immunol. 2003;111:1171–1183. doi: 10.1067/mai.2003.1592. [DOI] [PubMed] [Google Scholar]
  • 23.Feng CH, Miller MD, Simon RA. The united allergic airway: connections between allergic rhinitis, asthma, and chronic sinusitis. Am. J. Rhinol. Allergy. 2012;26:187–190. doi: 10.2500/ajra.2012.26.3762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Devalia JL, Sapsford RJ, Wells CW, Richman P, Davies RJ. Culture and comparison of human bronchial and nasal epithelial cells in vitro. Respir. Med. 1990;84:303–312. doi: 10.1016/S0954-6111(08)80058-3. [DOI] [PubMed] [Google Scholar]
  • 25.McDougall CM, Blaylock MG, Douglas JG, Brooker RJ, Helms PJ, Walsh GM. Nasal epithelial cells as surrogates for bronchial epithelial cells in airway inflammation studies. Am. J. Respir. Cell. Mol. Biol. 2008;39:560–568. doi: 10.1165/rcmb.2007-0325OC. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Comer DM, Elborn JS, Ennis M. Comparison of nasal and bronchial epithelial cells obtained from patients with COPD. PLoS ONE. 2012;7:e32924. doi: 10.1371/journal.pone.0032924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Thavagnanam S, Parker JC, McBrien ME, Skibinski G, Heaney LG, Shields MD. Effects of IL-13 on mucociliary differentiation of pediatric asthmatic bronchial epithelial cells. Pediatr. Res. 2011;69:95–100. doi: 10.1203/PDR.0b013e318204edb5. [DOI] [PubMed] [Google Scholar]
  • 28.Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25:402–408. doi: 10.1006/meth.2001.1262. [DOI] [PubMed] [Google Scholar]
  • 29.Pringle EJ, Richardson HB, Miller D, Cornish DS, Devereux GS, Walsh GM, Turner SW. Nasal and bronchial airway epithelial cell mediator release in children. Pediatr. Pulmonol. 2012;47:1215–1225. doi: 10.1002/ppul.22672. [DOI] [PubMed] [Google Scholar]
  • 30.Kobayashi I, Yamamoto S, Nishi N, Tsuji K, Imayoshi M, Inada S, Ichiamaru T, Hamasaki Y. Regulatory mechanisms of Th2 cytokineinduced eotaxin-3 production in bronchial epithelial cells: possible role of interleukin 4 receptor and nuclear factor-kappaB. Ann. Allergy Asthma Immunol. 2004;93:390–397. doi: 10.1016/S1081-1206(10)61399-3. [DOI] [PubMed] [Google Scholar]
  • 31.Komiya A, Nagase H, Yamada H, Sekiya T, Yamaguchi M, Sano Y, Hanai N, Furuya A, Ohta K, Matsushima K, Yoshie O, Yamamoto K, Hirai K. Concerted expression of eotaxin-1, eotaxin-2, and eotaxin-3 in human bronchial epithelial cells. Cell. Immunol. 2003;225:91–100. doi: 10.1016/j.cellimm.2003.10.001. [DOI] [PubMed] [Google Scholar]
  • 32.Faul JL, Tormey VJ, Leonard C, Burke CM, Farmer J, Horne SJ, Poulter LW. Lung immunopathology in cases of sudden asthma death. Eur. Respir. J. 1997;10:301–307. doi: 10.1183/09031936.97.10020301. [DOI] [PubMed] [Google Scholar]
  • 33.Carroll N, Cooke C, James A. The distribution of eosinophils and lymphocytes in the large and small airways of asthmatics. Eur. Respir. J. 1997;10:292–300. doi: 10.1183/09031936.97.10020292. [DOI] [PubMed] [Google Scholar]
  • 34.Lopez-Souza N, Favoreto S, Wong H, Ward T, Yagi S, Schnurr D, Finkbeiner WE, Dolganov GM, Widdicombe JH, Boushey HA, Avila PC. In vitro susceptibility to rhinovirus infection is greater for bronchial than for nasal airway epithelial cells in human subjects. J. Allergy Clin. Immunol. 2009;123:1384–1390.e2. doi: 10.1016/j.jaci.2009.03.010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Becker S, Koren HS, Henke DC. Interleukin-8 expression in normal nasal epithelium and its modulation by infection with respiratory syncytial virus and cytokines tumor necrosis factor, interleukin-1, and interleukin-6. Am. J. Respir. Cell Mol. Biol. 1993;8:20–27. doi: 10.1165/ajrcmb/8.1.20. [DOI] [PubMed] [Google Scholar]
  • 36.Adler KB, Fischer BM, Wright DT, Cohn LA, Becker S. Interactions between respiratory epithelial cells and cytokines: relationships to lung inflammation. Ann. N. Y. Acad. Sci. 1994;725:128–145. doi: 10.1111/j.1749-6632.1994.tb00275.x. [DOI] [PubMed] [Google Scholar]
  • 37.Villarete LH, Remick DG. Transcriptional and post-transcriptional regulation of interleukin-8. Am. J. Pathol. 1996;149:1685–1693. [PMC free article] [PubMed] [Google Scholar]
  • 38.Ma P, Cui X, Wang S, Zhang J, Nishanian EV, Wang W, Wesley RA, Danner RL. Nitric oxide post-transcriptionally up-regulates LPSinduced IL-8 expression through p38 MAPK activation. J. Leukoc. Biol. 2004;76:278–287. doi: 10.1189/jlb.1203653. [DOI] [PubMed] [Google Scholar]
  • 39.Yu Y, Chadee K. Prostaglandin E2 stimulates IL-8 gene expression in human colonic epithelial cells by a posttranscriptional mechanism. J. Immunol. 1998;161:3746–3752. [PubMed] [Google Scholar]
  • 40.Yu Y, Zeng H, Lyons S, Carlson A, Merlin D, Neish AS, Gewirtz AT. TLR5-mediated activation of p38 MAPK regulates epithelial IL-8 expression via posttranscriptional mechanism. Am. J. Physiol. Gastrointest. Liver. Physiol. 2003;285:282–290. doi: 10.1152/ajpgi.00503.2002. [DOI] [PubMed] [Google Scholar]
  • 41.Blume C, Swindle EJ, Dennison P, Jayasekera NP, Dudley S, Monk P, Behrendt H, Schmidt-Weber CB, Holgate ST, Howarth PH, Traidl-Hoffmann C, Davies DE. Barrier responses of human bronchial epithelial cells to grass pollen exposure. Eur. Respir. J. 2013;42:87–97. doi: 10.1183/09031936.00075612. [DOI] [PubMed] [Google Scholar]
  • 42.Van Wissen M, Snoek M, Smids B, Jansen HM, Lutter R. IFN-gamma amplifies IL-6 and IL-8 responses by airway epithelial-like cells via indoleamine 2,3-dioxygenase. J. Immunol. 2002;169:7039–7044. doi: 10.4049/jimmunol.169.12.7039. [DOI] [PubMed] [Google Scholar]
  • 43.Heller NM, Matsukura S, Georas SN, Boothby MR, Rothman PB, Stellato C, Schleimer RP. Interferon-gamma inhibits STAT6 signal transduction and gene expression in human airway epithelial cells. Am. J. Respir. Cell. Mol. Biol. 2004;31:573–582. doi: 10.1165/rcmb.2004-0195OC. [DOI] [PubMed] [Google Scholar]
  • 44.Kraft M, Djukanovic RM, Wilson SM, Holgate ST, Martin RJ. Alveolar tissue inflammation in asthma. Am. J. Respir. Crit. Care Med. 1996;154:1505–1510. doi: 10.1164/ajrccm.154.5.8912772. [DOI] [PubMed] [Google Scholar]

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