Chronic cigarette smoke exposure and pneumococcal infection induce oropharyngeal microbiota dysbiosis and contribute to long-lasting lung damage in mice Open Access

Abstract

Environmental factors, such as cigarette smoking or lung infections, may influence chronic obstructive pulmonary disease (COPD) progression by modifying the respiratory tract microbiome. However, whether the disease itself induces or maintains dysbiosis remains undefined. In this longitudinal study, we investigated the oropharyngeal microbiota composition and disease progression of mice (in cages of 5–10 mice per cage) before, during and up to 3 months after chronic cigarette smoke exposure or exposure to room air for 6 months. Cigarette smoke exposure induced pulmonary emphysema measurable at the end of exposure for 6 months, as well as 3 months following smoke exposure cessation. Using both classical culture methods and 16S rRNA sequencing, we observed that cigarette smoke exposure altered the relative composition of the oropharyngeal microbiota and reduced its diversity ( <0.001). More than 60 taxa were substantially reduced after 6 months of smoke exposure ( <0.001) However, oropharyngeal microbiota disordering was reversed 3 months after smoke exposure cessation and no significant difference was observed compared to age-matched control mice. The effects of lung infection with on established smoke-induced emphysema and on the oropharyngeal microbiota were also evaluated. Inoculation with induced lung damage and altered the microbiota composition for a longer time compared to control groups infected but not previously exposed to smoke (=0.01). Our data demonstrate effects of cigarette smoke and pneumococcus infection leading to altered microbiota and emphysema development. The reversal of the disordering of the microbiota composition, but not lung damage, following smoke exposure cessation and after clearance of infection suggest that changes in lung structure are not sufficient to sustain a disordered microbiota in mice. Whether changes in the airway microbiota contribute to inducing emphysema requires further investigation.

Keyword(s): COPD , emphysema , microbiota , pneumococcus and smoking
Funding
This study was supported by the:
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (Award 173137)
    • Principle Award Recipient: CharafBenarafa
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (Award 149790)
    • Principle Award Recipient: CharafBenarafa
Loading

Article metrics loading...

/content/journal/mgen/10.1099/mgen.0.000485
2020-12-09
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/mgen/6/12/mgen000485.html?itemId=/content/journal/mgen/10.1099/mgen.0.000485&mimeType=html&fmt=ahah

References

  1. Lokke A, Lange P, Scharling H, Fabricius P, Vestbo J. Developing COPD: a 25 year follow up study of the general population. Thorax 2006; 61:935–939 [View Article][PubMed]
    [Google Scholar]
  2. Fischer F, Kraemer A. Meta-analysis of the association between second-hand smoke exposure and ischaemic heart diseases, COPD and stroke. BMC Public Health 2015; 15:1202 [View Article][PubMed]
    [Google Scholar]
  3. Salvi SS, Barnes PJ. Chronic obstructive pulmonary disease in non-smokers. Lancet 2009; 374:733–743 [View Article][PubMed]
    [Google Scholar]
  4. Shrine N, Guyatt AL, Erzurumluoglu AM, Jackson VE, Hobbs BD et al. New genetic signals for lung function highlight pathways and chronic obstructive pulmonary disease associations across multiple ancestries. Nat Genet 2019; 51:481–493 [View Article][PubMed]
    [Google Scholar]
  5. Sakornsakolpat P, Prokopenko D, Lamontagne M, Reeve NF, Guyatt AL et al. Genetic landscape of chronic obstructive pulmonary disease identifies heterogeneous cell-type and phenotype associations. Nat Genet 2019; 51:494–505 [View Article][PubMed]
    [Google Scholar]
  6. Ragland MF, Benway CJ, Lutz SM, Bowler RP, Hecker J et al. Genetic advances in chronic obstructive pulmonary disease. Insights from COPDGene. Am J Respir Crit Care Med 2019; 200:677–690 [View Article][PubMed]
    [Google Scholar]
  7. Prokopenko D, Sakornsakolpat P, Fier HL, Qiao D, Parker MM et al. Whole-genome sequencing in severe chronic obstructive pulmonary disease. Am J Respir Cell Mol Biol 2018; 59:614–622 [View Article][PubMed]
    [Google Scholar]
  8. Hansel NN, Ruczinski I, Rafaels N, Sin DD, Daley D et al. Genome-wide study identifies two loci associated with lung function decline in mild to moderate COPD. Hum Genet 2013; 132:79–90 [View Article][PubMed]
    [Google Scholar]
  9. Radder JE, Gregory AD, Leme AS, Cho MH, Chu Y et al. Variable susceptibility to cigarette smoke-induced emphysema in 34 inbred strains of mice implicates Abi3bp in emphysema susceptibility. Am J Respir Cell Mol Biol 2017; 57:367–375 [View Article][PubMed]
    [Google Scholar]
  10. Tsuji H, Fujimoto H, Matsuura D, Nishino T, Lee KM et al. Comparison of mouse strains and exposure conditions in acute cigarette smoke inhalation studies. Inhal Toxicol 2011; 23:602–615 [View Article][PubMed]
    [Google Scholar]
  11. Hilty M, Burke C, Pedro H, Cardenas P, Bush A et al. Disordered microbial communities in asthmatic airways. PLoS One 2010; 5:e8578 [View Article][PubMed]
    [Google Scholar]
  12. Mika M, Korten I, Qi W, Regamey N, Frey U et al. The nasal microbiota in infants with cystic fibrosis in the first year of life: a prospective cohort study. Lancet Respir Med 2016; 4:627–635 [View Article][PubMed]
    [Google Scholar]
  13. Erb-Downward JR, Thompson DL, Han MK, Freeman CM, McCloskey L et al. Analysis of the lung microbiome in the "healthy" smoker and in COPD. PLoS One 2011; 6:e16384 [View Article][PubMed]
    [Google Scholar]
  14. Lipinski JH, Moore BB, O'Dwyer DN. The evolving role of the lung microbiome in pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol 2020; 319:L675–L682 [View Article][PubMed]
    [Google Scholar]
  15. Wang Z, Maschera B, Lea S, Kolsum U, Michalovich D et al. Airway host-microbiome interactions in chronic obstructive pulmonary disease. Respir Res 2019; 20:113 [View Article][PubMed]
    [Google Scholar]
  16. Ren L, Zhang R, Rao J, Xiao Y, Zhang Z et al. Transcriptionally active lung microbiome and its association with bacterial biomass and host inflammatory status. mSystems 2018; 3:e00199-18 [View Article][PubMed]
    [Google Scholar]
  17. Droemann D, Goldmann T, Tiedje T, Zabel P, Dalhoff K et al. Toll-like receptor 2 expression is decreased on alveolar macrophages in cigarette smokers and COPD patients. Respir Res 2005; 6:68 [View Article][PubMed]
    [Google Scholar]
  18. Lee J, Taneja V, Vassallo R. Cigarette smoking and inflammation: cellular and molecular mechanisms. J Dent Res 2012; 91:142–149 [View Article][PubMed]
    [Google Scholar]
  19. Minematsu N, Blumental-Perry A, Shapiro SD. Cigarette smoke inhibits engulfment of apoptotic cells by macrophages through inhibition of actin rearrangement. Am J Respir Cell Mol Biol 2011; 44:474–482 [View Article][PubMed]
    [Google Scholar]
  20. Phipps JC, Aronoff DM, Curtis JL, Goel D, O'Brien E et al. Cigarette smoke exposure impairs pulmonary bacterial clearance and alveolar macrophage complement-mediated phagocytosis of Streptococcus pneumoniae . Infect Immun 2010; 78:1214–1220 [View Article][PubMed]
    [Google Scholar]
  21. He Z, Chen Y, Hou C, He W, Chen P. Cigarette smoke extract changes expression of endothelial nitric oxide synthase (eNOS) and p16(INK4a) and is related to endothelial progenitor cell dysfunction. Med Sci Monit 2017; 23:3224–3231 [View Article][PubMed]
    [Google Scholar]
  22. Kulkarni R, Antala S, Wang A, Amaral FE, Rampersaud R et al. Cigarette smoke increases Staphylococcus aureus biofilm formation via oxidative stress. Infect Immun 2012; 80:3804–3811 [View Article][PubMed]
    [Google Scholar]
  23. Mutepe ND, Cockeran R, Steel HC, Theron AJ, Mitchell TJ et al. Effects of cigarette smoke condensate on pneumococcal biofilm formation and pneumolysin. Eur Respir J 2013; 41:392–395 [View Article][PubMed]
    [Google Scholar]
  24. Huang C, Shi G. Smoking and microbiome in oral, airway, gut and some systemic diseases. J Transl Med 2019; 17:225 [View Article][PubMed]
    [Google Scholar]
  25. Morris A, Beck JM, Schloss PD, Campbell TB, Crothers K et al. Comparison of the respiratory microbiome in healthy nonsmokers and smokers. Am J Respir Crit Care Med 2013; 187:1067–1075 [View Article][PubMed]
    [Google Scholar]
  26. Faner R, Sibila O, Agustí A, Bernasconi E, Chalmers JD et al. The microbiome in respiratory medicine: current challenges and future perspectives. Eur Respir J 2017; 49:1602086 [View Article][PubMed]
    [Google Scholar]
  27. Sethi S, Murphy TF. Infection in the pathogenesis and course of chronic obstructive pulmonary disease. N Engl J Med 2008; 359:2355–2365 [View Article][PubMed]
    [Google Scholar]
  28. Mammen MJ, Sethi S. COPD and the microbiome. Respirology 2016; 21:590–599 [View Article][PubMed]
    [Google Scholar]
  29. Polosukhin VV, Richmond BW, Du R-H, Cates JM, Wu P et al. Secretory IgA deficiency in individual small airways is associated with persistent inflammation and remodeling. Am J Respir Crit Care Med 2017; 195:1010–1021 [View Article][PubMed]
    [Google Scholar]
  30. Martinez FJ, Han MK, Allinson JP, Barr RG, Boucher RC et al. At the root: defining and halting progression of early chronic obstructive pulmonary disease. Am J Respir Crit Care Med 2018; 197:1540–1551 [View Article][PubMed]
    [Google Scholar]
  31. Remington LT, Sligl WI. Community-acquired pneumonia. Curr Opin Pulm Med 2014; 20:215–224 [View Article][PubMed]
    [Google Scholar]
  32. Torres A, Blasi F, Dartois N, Akova M. Which individuals are at increased risk of pneumococcal disease and why? Impact of COPD, asthma, smoking, diabetes, and/or chronic heart disease on community-acquired pneumonia and invasive pneumococcal disease. Thorax 2015; 70:984–989 [View Article][PubMed]
    [Google Scholar]
  33. Hathaway LJ, Brugger SD, Morand B, Bangert M, Rotzetter JU et al. Capsule type of Streptococcus pneumoniae determines growth phenotype. PLoS Pathog 2012; 8:e1002574 [View Article][PubMed]
    [Google Scholar]
  34. Troxler LJ, Werren JP, Schaffner TO, Mostacci N, Vermathen P et al. Carbon source regulates polysaccharide capsule biosynthesis in Streptococcus pneumoniae . J Biol Chem 2019; 294:17224–17238 [View Article][PubMed]
    [Google Scholar]
  35. Basilico P, Cremona TP, Oevermann A, Piersigilli A, Benarafa C. Increased myeloid cell production and lung bacterial clearance in mice exposed to cigarette smoke. Am J Respir Cell Mol Biol 2016; 54:424–435 [View Article][PubMed]
    [Google Scholar]
  36. Cremona TP, Tschanz SA, von Garnier C, Benarafa C. SerpinB1 deficiency is not associated with increased susceptibility to pulmonary emphysema in mice. Am J Physiol Lung Cell Mol Physiol 2013; 305:L981–L989 [View Article][PubMed]
    [Google Scholar]
  37. Hsia CC, Hyde DM, Ochs M, Weibel ER. on behalf of the ATS/ERS Joint Task Force on Quantitative Assessment of Lung Structure An official research policy statement of the American Thoracic Society/European Respiratory Society: standards for quantitative assessment of lung structure. Am J Respir Crit Care Med 2010; 181:394–418 [View Article][PubMed]
    [Google Scholar]
  38. Tschanz SA, Burri PH, Weibel ER. A simple tool for stereological assessment of digital images: the STEPanizer. J Microsc 2011; 243:47–59 [View Article][PubMed]
    [Google Scholar]
  39. Kraemer JG, Ramette A, Aebi S, Oppliger A, Hilty M. Influence of pig farming on the human nasal microbiota: key role of airborne microbial communities. Appl Environ Microbiol 2018; 84:e02470-17 [View Article][PubMed]
    [Google Scholar]
  40. Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJ et al. DADA2: high-resolution sample inference from Illumina amplicon data. Nat Methods 2016; 13:581–583 [View Article][PubMed]
    [Google Scholar]
  41. Mika M, Nita I, Morf L, Qi W, Beyeler S et al. Microbial and host immune factors as drivers of COPD. ERJ Open Res 2018; 4:00015-2018 [View Article][PubMed]
    [Google Scholar]
  42. Mayhew D, Devos N, Lambert C, Brown JR, Clarke SC et al. Longitudinal profiling of the lung microbiome in the AERIS study demonstrates repeatability of bacterial and eosinophilic COPD exacerbations. Thorax 2018; 73:422–430 [View Article][PubMed]
    [Google Scholar]
  43. Salter SJ, Cox MJ, Turek EM, Calus ST, Cookson WO et al. Reagent and laboratory contamination can critically impact sequence-based microbiome analyses. BMC Biol 2014; 12:87 [View Article][PubMed]
    [Google Scholar]
  44. Dickson RP, Erb-Downward JR, Falkowski NR, Hunter EM, Ashley SL et al. The lung microbiota of healthy mice are highly variable, cluster by environment, and reflect variation in baseline lung innate immunity. Am J Respir Crit Care Med 2018; 198:497–508 [View Article][PubMed]
    [Google Scholar]
  45. Biesbroek G, Sanders EA, Roeselers G, Wang X, Caspers MP et al. Deep sequencing analyses of low density microbial communities: working at the boundary of accurate microbiota detection. PLoS One 2012; 7:e32942 [View Article][PubMed]
    [Google Scholar]
  46. Charlson ES, Bittinger K, Haas AR, Fitzgerald AS, Frank I et al. Topographical continuity of bacterial populations in the healthy human respiratory tract. Am J Respir Crit Care Med 2011; 184:957–963 [View Article][PubMed]
    [Google Scholar]
  47. Ahmed B, Cox MJ, Cuthbertson L, James PL, Cookson WOC et al. Comparison of the upper and lower airway microbiota in children with chronic lung diseases. PLoS One 2018; 13:e0201156 [View Article][PubMed]
    [Google Scholar]
  48. Sakki T, Knuuttila M. Controlled study of the association of smoking with lactobacilli, mutans streptococci and yeasts in saliva. Eur J Oral Sci 1996; 104:619–622 [View Article][PubMed]
    [Google Scholar]
  49. Parvinen T. Stimulated salivary flow rate, pH and Lactobacillus and yeast concentrations in non-smokers and smokers. Scand J Dent Res 1984; 92:315–318 [View Article][PubMed]
    [Google Scholar]
  50. Einarsson GG, Comer DM, McIlreavey L, Parkhill J, Ennis M et al. Community dynamics and the lower airway microbiota in stable chronic obstructive pulmonary disease, smokers and healthy non-smokers. Thorax 2016; 71:795–803 [View Article][PubMed]
    [Google Scholar]
  51. Shen P, Whelan FJ, Schenck LP, McGrath JJC, Vanderstocken G et al. Streptococcus pneumoniae colonization is required to alter the nasal microbiota in cigarette smoke-exposed mice. Infect Immun 2017; 85:e00434-17 [View Article][PubMed]
    [Google Scholar]
  52. Carney SM, Clemente JC, Cox MJ, Dickson RP, Huang YJ et al. Methods in lung microbiome research. Am J Respir Cell Mol Biol 2020; 62:283–299 [View Article][PubMed]
    [Google Scholar]
  53. Yadava K, Pattaroni C, Sichelstiel AK, Trompette A, Gollwitzer ES et al. Microbiota promotes chronic pulmonary inflammation by enhancing IL-17A and autoantibodies. Am J Respir Crit Care Med 2016; 193:975–987 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/mgen/10.1099/mgen.0.000485
Loading
/content/journal/mgen/10.1099/mgen.0.000485
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed