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Electronic cigarette inhalation alters innate immunity and airway cytokines while increasing the virulence of colonizing bacteria

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Abstract

Electronic (e)-cigarette use is rapidly rising, with 20 % of Americans ages 25–44 now using these drug delivery devices. E-cigarette users expose their airways, cells of host defense, and colonizing bacteria to e-cigarette vapor (EV). Here, we report that exposure of human epithelial cells at the air–liquid interface to fresh EV (vaped from an e-cigarette device) resulted in dose-dependent cell death. After exposure to EV, cells of host defense—epithelial cells, alveolar macrophages, and neutrophils—had reduced antimicrobial activity against Staphylococcus aureus (SA). Mouse inhalation of EV for 1 h daily for 4 weeks led to alterations in inflammatory markers within the airways and elevation of an acute phase reactant in serum. Upon exposure to e-cigarette vapor extract (EVE), airway colonizer SA had increased biofilm formation, adherence and invasion of epithelial cells, resistance to human antimicrobial peptide LL-37, and up-regulation of virulence genes. EVE-exposed SA were more virulent in a mouse model of pneumonia. These data suggest that e-cigarettes may be toxic to airway cells, suppress host defenses, and promote inflammation over time, while also promoting virulence of colonizing bacteria.

Key message

  • Acute exposure to e-cigarette vapor (EV) is cytotoxic to airway cells in vitro.

  • Acute exposure to EV decreases macrophage and neutrophil antimicrobial function.

  • Inhalation of EV alters immunomodulating cytokines in the airways of mice.

  • Inhalation of EV leads to increased markers of inflammation in BAL and serum.

  • Staphylococcus aureus become more virulent when exposed to EV.

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References

  1. King BA (2013) Notes from the field: electronic cigarette use among middle and high school students—United States, 2011-2012 Morbidity and Mortality Weekly Report (MMWR) Centers for Disease Control and Prevention (CDC), Atlanta, pp. 729–730.

  2. Czoli CD, Hammond D, White CM (2014) Electronic cigarettes in Canada: prevalence of use and perceptions among youth and young adults. Can J Public Health Revue Canadienne de Sante Publique 105:e97–e102

    PubMed  Google Scholar 

  3. Kinnunen JM, Ollila H, El-Amin SE, Pere LA, Lindfors PL, Rimpela AH (2014) Awareness and determinants of electronic cigarette use among Finnish adolescents in 2013: a population-based study. Tob Control. doi:10.1136/tobaccocontrol-2013-051512

    PubMed  PubMed Central  Google Scholar 

  4. Lee S, Grana RA, Glantz SA (2014) Electronic cigarette use among Korean adolescents: a cross-sectional study of market penetration, dual use, and relationship to quit attempts and former smoking. J Adolesc Health: Off Publ Soc Adolesc Med 54:684–690

    Article  Google Scholar 

  5. Bostean G, Trinidad DR, McCarthy WJ (2015) E-cigarette use among never-smoking California students. American journal of public health: e1-e3. DOI 10.2105/AJPH.2015.302899

  6. Sutfin EL, McCoy TP, Morrell HE, Hoeppner BB, Wolfson M (2013) Electronic cigarette use by college students. Drug Alcohol Depend 131:214–221

    Article  PubMed  PubMed Central  Google Scholar 

  7. Regan AK, Promoff G, Dube SR, Arrazola R (2013) Electronic nicotine delivery systems: adult use and awareness of the 'e-cigarette' in the USA. Tob Control 22:19–23

    Article  PubMed  Google Scholar 

  8. Brown J, West R, Beard E, Michie S, Shahab L, McNeill A (2014) Prevalence and characteristics of e-cigarette users in Great Britain: findings from a general population survey of smokers. Addict Behav 39:1120–1125

    Article  PubMed  PubMed Central  Google Scholar 

  9. Dawkins L, Turner J, Roberts A, Soar K (2013) 'Vaping' profiles and preferences: an online survey of electronic cigarette users. Addiction 108:1115–1125

    Article  PubMed  Google Scholar 

  10. Bauguess AJ (2014) How to make your own e-liquid (e-juice). ecigvape.com.

  11. Flouris AD, Chorti MS, Poulianiti KP, Jamurtas AZ, Kostikas K, Tzatzarakis MN, Wallace Hayes A, Tsatsakis AM, Koutedakis Y (2013) Acute impact of active and passive electronic cigarette smoking on serum cotinine and lung function. Inhal Toxicol 25:91–101

    Article  CAS  PubMed  Google Scholar 

  12. Liu GY (2009) Molecular pathogenesis of Staphylococcus aureus infection. Pediatr Res 65:71R–77R

    Article  PubMed  PubMed Central  Google Scholar 

  13. McEachern EK, Hwang JH, Sladewski KM, Nicatia S, Dewitz C, Mathew DP, Nizet V, Alexander LE (2015) Analysis of the effects of cigarette smoke on staphylococcal virulence phenotypes. Infect Immun 83:2443–2452

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Crotty Alexander LE, Shin S, Hwang JH (2015) Inflammatory diseases of the lung induced by conventional cigarette smoke: a review. Chest. doi:10.1378/chest.15-0409

    Google Scholar 

  15. Foster KA, Oster CG, Mayer MM, Avery ML, Audus KL (1998) Characterization of the A549 cell line as a type II pulmonary epithelial cell model for drug metabolism. Exp Cell Res 243:359–366

    Article  CAS  PubMed  Google Scholar 

  16. Grando SA (2008) Basic and clinical aspects of non-neuronal acetylcholine: biological and clinical significance of non-canonical ligands of epithelial nicotinic acetylcholine receptors. J Pharmacol Sci 106:174–179

    Article  CAS  PubMed  Google Scholar 

  17. Thunnissen FB (2009) Acetylcholine receptor pathway and lung cancer. J Thorac Oncol: Off Publ Int Assoc Study Lung Cancer 4:943–946

    Article  Google Scholar 

  18. Bermudez Y, Benavente CA, Meyer RG, Coyle WR, Jacobson MK, Jacobson EL (2011) Nicotinic acid receptor abnormalities in human skin cancer: implications for a role in epidermal differentiation. PLoS ONE 6, e20487. doi:10.1371/journal.pone.0020487

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Reynolds PR, Allison CH, Willnauer CP (2010) TTF-1 regulates alpha5 nicotinic acetylcholine receptor (nAChR) subunits in proximal and distal lung epithelium. Respir Res 11:175

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Ma X, Jia Y, Zu S, Li R, Zhao Y, Xiao D, Dang N, Wang Y (2014) alpha5 Nicotinic acetylcholine receptor mediates nicotine-induced HIF-1alpha and VEGF expression in non-small cell lung cancer. Toxicol Appl Pharmacol 278:172–179

    Article  CAS  PubMed  Google Scholar 

  21. Comer DM, Elborn JS, Ennis M (2014) Inflammatory and cytotoxic effects of acrolein, nicotine, acetylaldehyde and cigarette smoke extract on human nasal epithelial cells. BMC Pulm Med 14:32

    Article  PubMed  PubMed Central  Google Scholar 

  22. Streck E, Jorres RA, Huber RM, Bergner A (2010) Effects of cigarette smoke extract and nicotine on bronchial tone and acetylcholine-induced airway contraction in mouse lung slices. J Investig Allergol Clin Immunol 20:324–330

    CAS  PubMed  Google Scholar 

  23. Askarian F, Sangvik M, Hanssen AM, Snipen L, Sollid JU, Johannessen M (2014) Staphylococcus aureus nasal isolates from healthy individuals cause highly variable host cell responses in vitro: the Tromso Staph and Skin Study. Pathog Dis 70:158–166

    Article  CAS  PubMed  Google Scholar 

  24. Pegtel DM, Subramanian A, Sheen TS, Tsai CH, Golub TR, Thorley-Lawson DA (2005) Epstein-Barr-virus-encoded LMP2A induces primary epithelial cell migration and invasion: possible role in nasopharyngeal carcinoma metastasis. J Virol 79:15430–15442

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Condliffe AM, Kitchen E, Chilvers ER (1998) Neutrophil priming: pathophysiological consequences and underlying mechanisms. Clin Sci 94:461–471

    Article  CAS  PubMed  Google Scholar 

  26. Bermudez EA, Rifai N, Buring J, Manson JE, Ridker PM (2002) Interrelationships among circulating interleukin-6, C-reactive protein, and traditional cardiovascular risk factors in women. Arterioscler Thromb Vasc Biol 22:1668–1673

    Article  CAS  PubMed  Google Scholar 

  27. Rohde LE, Hennekens CH, Ridker PM (1999) Survey of C-reactive protein and cardiovascular risk factors in apparently healthy men. Am J Cardiol 84:1018–1022

    Article  CAS  PubMed  Google Scholar 

  28. Magnusson KE (1982) Hydrophobic interaction—a mechanism of bacterial binding. Scand J Infect Dis Suppl 33:32–36

    CAS  PubMed  Google Scholar 

  29. Dahlback B, Hermansson M, Kjelleberg S, Norkrans B (1981) The hydrophobicity of bacteria—an important factor in their initial adhesion at the air-water interface. Arch Microbiol 128:267–270

    Article  CAS  PubMed  Google Scholar 

  30. Yang ES, Tan J, Eells S, Rieg G, Tagudar G, Miller LG (2010) Body site colonization in patients with community-associated methicillin-resistant Staphylococcus aureus and other types of S. aureus skin infections. Clin Microbiol Infect: Off Publ Eur Soc Clin Microbiol Infect Dis 16:425–431

    Article  CAS  Google Scholar 

  31. Wang G (2008) Structures of human host defense cathelicidin LL-37 and its smallest antimicrobial peptide KR-12 in lipid micelles. J Biol Chem 283:32637–32643

    Article  CAS  PubMed  Google Scholar 

  32. Neville F, Cahuzac M, Konovalov O, Ishitsuka Y, Lee KY, Kuzmenko I, Kale GM, Gidalevitz D (2006) Lipid headgroup discrimination by antimicrobial peptide LL-37: insight into mechanism of action. Biophys J 90:1275–1287

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Neville F, Gidalevitz D, Kale G, Nelson A (2007) Electrochemical screening of anti-microbial peptide LL-37 interaction with phospholipids. Bioelectrochemistry 70:205–213

    Article  CAS  PubMed  Google Scholar 

  34. Park EJ, Lee GH, Yoon C, Jeong U, Kim Y, Cho MH, Kim DW (2015) Biodistribution and toxicity of spherical aluminum oxide nanoparticles. J Appl Toxicol: JAT. doi:10.1002/jat.3233

    Google Scholar 

  35. Viard I, Wehrli P, Bullani R, Schneider P, Holler N, Salomon D, Hunziker T, Saurat JH, Tschopp J, French LE (1998) Inhibition of toxic epidermal necrolysis by blockade of CD95 with human intravenous immunoglobulin. Science 282:490–493

    Article  CAS  PubMed  Google Scholar 

  36. Herzog E, Casey A, Lyng FM, Chambers G, Byrne HJ, Davoren M (2007) A new approach to the toxicity testing of carbon-based nanomaterials—the clonogenic assay. Toxicol Lett 174:49–60

    Article  CAS  PubMed  Google Scholar 

  37. Farsalinos KE, Romagna G, Allifranchini E, Ripamonti E, Bocchietto E, Todeschi S, Tsiapras D, Kyrzopoulos S, Voudris V (2013) Comparison of the cytotoxic potential of cigarette smoke and electronic cigarette vapour extract on cultured myocardial cells. Int J Environ Res Public Health 10(10):5146–5162

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Schweitzer KS, Chen SX, Law S, Van Demark M, Poirier C, Justice MJ, Hubbard WC, Kim ES, Lai X, Wang M et al (2015) Endothelial disruptive proinflammatory effects of nicotine and e-cigarette vapor exposures. Am J Physiol Lung Cell Mol Physiol 309(2):L175–L187

    Article  PubMed  Google Scholar 

  39. Savill J, Dransfield I, Gregory C, Haslett C (2002) A blast from the past: clearance of apoptotic cells regulates immune responses. Nat Rev Immunol 2:965–975

    Article  CAS  PubMed  Google Scholar 

  40. Kaczmarek A, Vandenabeele P, Krysko DV (2013) Necroptosis: the release of damage-associated molecular patterns and its physiological relevance. Immunity 38:209–223

    Article  CAS  PubMed  Google Scholar 

  41. El Mezayen R, El Gazzar M, Seeds MC, McCall CE, Dreskin SC, Nicolls MR (2007) Endogenous signals released from necrotic cells augment inflammatory responses to bacterial endotoxin. Immunol Lett 111:36–44

    Article  PubMed  PubMed Central  Google Scholar 

  42. Radsak MP, Taube C, Haselmayer P, Tenzer S, Salih HR, Wiewrodt R, Buhl R, Schild H (2007) Soluble triggering receptor expressed on myeloid cells 1 is released in patients with stable chronic obstructive pulmonary disease. Clin Dev Immunol 2007:52040

    Article  PubMed  PubMed Central  Google Scholar 

  43. Papanikolaou IC, Boki KA, Giamarellos-Bourboulis EJ, Kotsaki A, Kagouridis K, Karagiannidis N, Polychronopoulos VS (2015) Innate immunity alterations in idiopathic interstitial pneumonias and rheumatoid arthritis-associated interstitial lung diseases. Immunol Lett 163:179–186

    Article  CAS  PubMed  Google Scholar 

  44. Molad Y, Ofer-Shiber S, Pokroy-Shapira E, Oren S, Shay-Aharoni H, Babai I (2015) Soluble triggering receptor expressed on myeloid cells-1 is a biomarker of anti-CCP-positive, early rheumatoid arthritis. Eur J Clin Investig 45:557–564

    Article  CAS  Google Scholar 

  45. Sussan TE, Gajghate S, Thimmulappa RK, Ma J, Kim JH, Sudini K, Consolini N, Cormier SA, Lomnicki S, Hasan F et al (2015) Exposure to electronic cigarettes impairs pulmonary anti-bacterial and anti-viral defenses in a mouse model. PLoS ONE 10(2), e0116861. doi:10.1371/journal.pone.0116861

    Article  PubMed  PubMed Central  Google Scholar 

  46. Wu Q, Jiang D, Minor M, Chu HW (2014) Electronic cigarette liquid increases inflammation and virus infection in primary human airway epithelial cells. PLoS ONE 9(9), e108342. doi:10.1371/journal.pone.0108342

    Article  PubMed  PubMed Central  Google Scholar 

  47. Cheng AG, McAdow M, Kim HK, Bae T, Missiakas DM, Schneewind O (2010) Contribution of coagulases towards Staphylococcus aureus disease and protective immunity. PLoS Pathog 6, e1001036. doi:10.1371/journal.ppat.1001036

    Article  PubMed  PubMed Central  Google Scholar 

  48. Hamilton SM, Bryant AE, Carroll KC, Lockary V, Ma Y, McIndoo E, Miller LG, Perdreau-Remington F, Pullman J, Risi GF et al (2007) In vitro production of panton-valentine leukocidin among strains of methicillin-resistant Staphylococcus aureus causing diverse infections. Clin Infect Dis: Off Publ Infect Dis Soc Am 45:1550–1558

    Article  CAS  Google Scholar 

  49. Montgomery CP, Boyle-Vavra S, Adem PV, Lee JC, Husain AN, Clasen J, Daum RS (2008) Comparison of virulence in community-associated methicillin-resistant Staphylococcus aureus pulsotypes USA300 and USA400 in a rat model of pneumonia. J Infect Dis 198:561–570

    Article  CAS  PubMed  Google Scholar 

  50. Gillet Y, Issartel B, Vanhems P, Fournet JC, Lina G, Bes M, Vandenesch F, Piemont Y, Brousse N, Floret D et al (2002) Association between Staphylococcus aureus strains carrying gene for Panton-Valentine leukocidin and highly lethal necrotising pneumonia in young immunocompetent patients. Lancet 359:753–759

    Article  CAS  PubMed  Google Scholar 

  51. Crotty Alexander LE, Maisey HC, Timmer AM, Rooijakkers SH, Gallo RL, von Kockritz-Blickwede M, Nizet V (2010) M1T1 group A streptococcal pili promote epithelial colonization but diminish systemic virulence through neutrophil extracellular entrapment. J Mol Med 88:371–381

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

This work was supported by the VA Career Development Award (CDA)-2, award no. 1IK2BX001313, PI Crotty Alexander, from the U.S. Department of Veterans Affairs, Biomedical Laboratory Research and Development (BLR&D) Program. Thank you to Drs. Ross Corriden and Simon Döhrmann for their technical support. Thank you to Dr. Atul Malhotra for his guidance and support.

Author contributions

JHH, EM, WMO, SB, SD, and LCA designed the research. JHH, ML, KS, SE, SD, EM, DM, AM, SD, SB, WMO, and LCA performed the research and analyzed the data. JHH, SE, SB, ML, DTP, WMO, and LCA performed statistical analyses and edited the paper. JHH, SD, SB, and LCA wrote the paper.

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Correspondence to Laura E. Crotty Alexander.

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Hwang, J.H., Lyes, M., Sladewski, K. et al. Electronic cigarette inhalation alters innate immunity and airway cytokines while increasing the virulence of colonizing bacteria. J Mol Med 94, 667–679 (2016). https://doi.org/10.1007/s00109-016-1378-3

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  • DOI: https://doi.org/10.1007/s00109-016-1378-3

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