1887

Abstract

The atypical bacterial pathogen is a leading etiological agent of community-acquired pneumonia in humans; infections are often recalcitrant, recurrent and resistant to antibiotic treatment. These characteristics suggest a mechanism that facilitates long-term colonization in hosts. In an setting, forms biofilms that are unusual in that motility plays no more than a very limited role in their formation and development. Given the unusual nature of biofilms, open questions remain concerning phenotypes associated with persistence, such as what properties might favour the bacteria while minimizing host damage. also produces several cytotoxic molecules including community-acquired respiratory distress syndrome (CARDS) toxin, HS and HO, but how it deploys these agents during growth is unknown. Whereas several biochemical techniques for biofilm disruption were ineffective, sonication was required for disruption of biofilms to generate individual cells for comparative studies, suggesting unusual physical properties likely related to the atypical cell envelope. Nonetheless, like for other bacteria, biofilms were less susceptible to antibiotic inhibition and complement killing than dispersed cells, with resistance increasing as the biofilms matured. CARDS toxin levels and enzymatic activities associated with HS and HO production were highest during early biofilm formation and decreased over time, suggesting attenuation of virulence in connection with chronic infection. Collectively, these findings result in a model of how biofilms contribute to both the establishment and propagation of infections, and how both biofilm towers and individual cells participate in persistence and chronic disease.

Funding
This study was supported by the:
  • Monica Feng , Miami University (US) , (Award DUOS)
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000928
2020-05-18
2020-06-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/10.1099/mic.0.000928/mic000928.html?itemId=/content/journal/micro/10.1099/mic.0.000928&mimeType=html&fmt=ahah

References

  1. Waites KB, Xiao L, Liu Y, Balish MF, Atkinson TP. Mycoplasma pneumoniae from the respiratory tract and beyond. Clin Microbiol Rev 2017; 30:747–809 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  2. Zhao F, Li J, Liu J, Guan X, Gong J et al. Antimicrobial susceptibility and molecular characteristics of Mycoplasma pneumoniae isolates across different regions of China. Antimicrob Resist Infect Control 2019; 8:143 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  3. Balish MF. Mycoplasma pneumoniae, an underutilized model for bacterial cell biology. J Bacteriol 2014; 196:3675–3682 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  4. Großhennig S, Schmidl SR, Schmeisky G, Busse J, Stülke J. Implication of glycerol and phospholipid transporters in Mycoplasma pneumoniae growth and virulence. Infect Immun 2013; 81:896–904 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  5. Baeuerle PA, Henkel T. Function and activation of NF-kappa B in the immune system. Annu Rev Immunol 1994; 12:141–179 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  6. Hames C, Halbedel S, Hoppert M, Frey J, Stülke J. Glycerol metabolism is important for cytotoxicity of Mycoplasma pneumoniae. J Bacteriol 2009; 191:747–753 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  7. Elkhal CK, Kean KM, Parsonage D, Maenpuen S, Chaiyen P et al. Structure and proposed mechanism of L-α-glycerophosphate oxidase from Mycoplasma pneumoniae. Febs J 2015; 282:3030–3042 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  8. Großhennig S, Ischebeck T, Gibhardt J, Busse J, Feussner I et al. Hydrogen sulfide is a novel potential virulence factor of Mycoplasma pneumoniae: characterization of the unusual cysteine desulfurase/desulfhydrase hapE. Mol Microbiol 2016; 100:42–54 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  9. Hutchison CA, Peterson SN, Gill SR, Cline RT, White O et al. Global transposon mutagenesis and a minimal Mycoplasma genome. Science 1999; 286:2165–2169 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  10. Kannan TR, Provenzano D, Wright JR, Baseman JB. Identification and characterization of human surfactant protein A binding protein of Mycoplasma pneumoniae. Infect Immun 2005; 73:2828–2834 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  11. Kannan TR, Baseman JB. Adp-Ribosylating and vacuolating cytotoxin of Mycoplasma pneumoniae represents unique virulence determinant among bacterial pathogens. Proc Natl Acad Sci U S A 2006; 103:6724–6729 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  12. Krishnan M, Kannan TR, Baseman JB. Mycoplasma pneumoniae CARDS toxin is internalized via clathrin-mediated endocytosis. PLoS One 2013; 8:e62706 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  13. Bose S, Segovia JA, Somarajan SR, Chang T-H, Kannan TR et al. ADP-ribosylation of NLRP3 by Mycoplasma pneumoniae CARDS toxin regulates inflammasome activity. mBio 2014; 5:e02186–14 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  14. Segovia JA, Chang T-H, Winter VT, Coalson JJ, Cagle MP et al. NLRP3 is a critical regulator of inflammation and innate immune cell response during Mycoplasma pneumoniae infection. Infect Immun 2018; 86:e00548–17 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  15. Gnarpe J, Lundbäck A, Sundelöf B, Gnarpe H. Prevalence of Mycoplasma pneumoniae in subjectively healthy individuals. Scand J Infect Dis 1992; 24:161–164 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  16. Spuesens EBM, Fraaij PLA, Visser EG, Hoogenboezem T, Hop WCJ et al. Carriage of Mycoplasma pneumoniae in the upper respiratory tract of symptomatic and asymptomatic children: an observational study. PLoS Med 2013; 10:e1001444 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  17. Martin RJ, Kraft M, Chu HW, Berns EA, Cassell GH. A link between chronic asthma and chronic infection. J Allergy Clin Immunol 2001; 107:595–601 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  18. Kraft M, Cassell GH, Pak J, Martin RJ. Mycoplasma pneumoniae and Chlamydia pneumoniae in asthma: effect of clarithromycin. Chest 2002; 121:1782–1788 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  19. Hardy RD, Jafri HS, Olsen K, Hatfield J, Iglehart J et al. Mycoplasma pneumoniae induces chronic respiratory infection, airway hyperreactivity, and pulmonary inflammation: a murine model of infection-associated chronic reactive airway disease. Infect Immun 2002; 70:649–654 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  20. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science 1999; 284:1318–1322 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  21. Lebeaux D, Ghigo J-M, Beloin C. Biofilm-related infections: bridging the gap between clinical management and fundamental aspects of recalcitrance toward antibiotics. Microbiol Mol Biol Rev 2014; 78:510–543 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  22. Whitchurch CB, Tolker-Nielsen T, Ragas PC, Mattick JS. Extracellular DNA required for bacterial biofilm formation. Science 2002; 295:1487 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  23. Steinberger RE, Holden PA. Extracellular DNA in single- and multiple-species unsaturated biofilms. Appl Environ Microbiol 2005; 71:5404–5410 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  24. Kristian SA, Birkenstock TA, Sauder U, Mack D, Götz F et al. Biofilm formation induces C3a release and protects Staphylococcus epidermidis from IgG and complement deposition and from neutrophil-dependent killing. J Infect Dis 2008; 197:1028–1035 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  25. Silva AJ, Benitez JA. Vibrio cholerae biofilms and cholera pathogenesis. PLoS Negl Trop Dis 2016; 10:e0004330 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  26. Sanchez CJ, Kumar N, Lizcano A, Shivshankar P, Dunning Hotopp JC et al. Streptococcus pneumoniae in biofilms are unable to cause invasive disease due to altered virulence determinant production. PLoS One 2011; 6:e28738 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  27. Kornspan JD, Tarshis M, Rottem S. Adhesion and biofilm formation of Mycoplasma pneumoniae on an abiotic surface. Arch Microbiol 2011; 193:833–836 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  28. Simmons WL, Daubenspeck JM, Osborne JD, Balish MF, Waites KB et al. Type 1 and type 2 strains of Mycoplasma pneumoniae form different biofilms. Microbiology 2013; 159:737–747 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  29. Feng M, Schaff AC, Cuadra Aruguete SA, Riggs HE, Distelhorst SL et al. Development of Mycoplasma pneumoniae biofilms in vitro and the limited role of motility. Int J Med Microbiol 2018; 308:324–334 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  30. Hengge R. Principles of c-di-GMP signalling in bacteria. Nat Rev Microbiol 2009; 7:263–273 [CrossRef]
    [Google Scholar]
  31. Totten AH, Xiao L, Crabb DM, Ratliff AE, Dybvig K et al. Shaken or stirred?: Comparison of methods for dispersion of Mycoplasma pneumoniae aggregates for persistence in vivo. J Microbiol Methods 2017; 132:56–62 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  32. Tully JG, Whitcomb RF, Clark HF, Williamson DL. Pathogenic mycoplasmas: cultivation and vertebrate pathogenicity of a new Spiroplasma. Science 1977; 195:892–894 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  33. Hedreyda CT, Lee KK, Krause DC. Transformation of Mycoplasma pneumoniae with Tn4001 by electroporation. Plasmid 1993; 30:170–175 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  34. Simmons WL, Dybvig K. Biofilms protect Mycoplasma pulmonis cells from lytic effects of complement and gramicidin. Infect Immun 2007; 75:3696–3699 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  35. Clinical and Laboratory Standards Institute CLSI Guidelines [Internet]. Wayne, PA: CLSI; 2011 October 27. M43-A: Antimicrobial susceptibility testing of human mycoplasmas: approved guideline. Available from: https://clsi.org/standards/products/microbiology/documents/m43/.
  36. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  37. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 1979; 76:4350–4354 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  38. Kannan TR, Musatovova O, Balasubramanian S, Cagle M, Jordan JL et al. Mycoplasma pneumoniae Community Acquired Respiratory Distress Syndrome toxin expression reveals growth phase and infection-dependent regulation. Mol Microbiol 2010; 76:1127–1141 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  39. Pyrowolakis G, Hofmann D, Herrmann R. The subunit B of the F0F1-type ATPase of the bacterium Mycoplasma pneumoniae is a lipoprotein. J Biol Chem 1998; 273:24792–24796 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  40. Palmgren R, Nielsen PH. Accumulation of DNA in the exopolymeric matrix of activated sludge and bacterial cultures. Water Sci Technol 1996; 34:233–240 [CrossRef]
    [Google Scholar]
  41. Grande R, Di Marcantonio MC, Robuffo I, Pompilio A, Celia C et al. Helicobacter pylori ATCC 43629/NCTC 11639 outer membrane vesicles (OMVs) from biofilm and planktonic phase associated with extracellular DNA (eDNA). Front Microbiol 2015; 6:1369 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  42. Gawande PV, LoVetri K, Yakandawala N, Romeo T, Zhanel GG et al. Antibiofilm activity of sodium bicarbonate, sodium metaperiodate and SDS combination against dental unit waterline-associated bacteria and yeast. J Appl Microbiol 2008; 105:986–992 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  43. Yamamoto T, Kida Y, Sakamoto Y, Kuwano K. Mpn491, a secreted nuclease of Mycoplasma pneumoniae, plays a critical role in evading killing by neutrophil extracellular traps. Cell Microbiol 2017; 19:e12666 [CrossRef]
    [Google Scholar]
  44. Raymond BBA, Jenkins C, Turnbull L, Whitchurch CB, Djordjevic SP. Extracellular DNA release from the genome-reduced pathogen Mycoplasma hyopneumoniae is essential for biofilm formation on abiotic surfaces. Sci Rep 2018; 8:10373 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  45. McAuliffe L, Ayling RD, Ellis RJ, Nicholas RAJ. Biofilm-grown Mycoplasma mycoides subsp. mycoides SC exhibit both phenotypic and genotypic variation compared with planktonic cells. Vet Microbiol 2008; 129:315–324 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  46. Chen S, Hao H, Zhao P, Ji W, Li M et al. Differential immunoreactivity to bovine convalescent serum between Mycoplasma bovis biofilms and planktonic cells revealed by comparative immunoproteomic analysis. Front Microbiol 2018; 9:379 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  47. Lai J-F, Zindl CL, Duffy LB, Atkinson TP, Jung YW et al. Critical role of macrophages and their activation via MyD88-NFκB signaling in lung innate immunity to Mycoplasma pneumoniae. PLoS One 2010; 5:e14417 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  48. Alvarez D, Merino S, Tomás JM, Benedí VJ, Albertí S. Capsular polysaccharide is a major complement resistance factor in lipopolysaccharide O side chain-deficient Klebsiella pneumoniae clinical isolates. Infect Immun 2000; 68:953–955 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  49. Campos MA, Vargas MA, Regueiro V, Llompart CM, Albertí S et al. Capsule polysaccharide mediates bacterial resistance to antimicrobial peptides. Infect Immun 2004; 72:7107–7114 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  50. Fong JCN, Syed KA, Klose KE, Yildiz FH. Role of Vibrio polysaccharide (vps) genes in VPS production, biofilm formation and Vibrio cholerae pathogenesis. Microbiology 2010; 156:2757–2769 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  51. Zhu L, Kreth J. The role of hydrogen peroxide in environmental adaptation of oral microbial communities. Oxid Med Cell Longev 2012; 2012:71784310 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  52. Jordan JL, Chang H-Y, Balish MF, Holt LS, Bose SR et al. Protein P200 is dispensable for Mycoplasma pneumoniae hemadsorption but not gliding motility or colonization of differentiated bronchial epithelium. Infect Immun 2007; 75:518–522 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  53. Güell M, van Noort V, Yus E, Chen W-H, Leigh-Bell J et al. Transcriptome complexity in a genome-reduced bacterium. Science 2009; 326:1268–1271 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  54. Schmidl SR, Otto A, Lluch-Senar M, Piñol J, Busse J et al. A trigger enzyme in Mycoplasma pneumoniae: impact of the glycerophosphodiesterase GlpQ on virulence and gene expression. PLoS Pathog 2011; 7:e1002263 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  55. Maier T, Schmidt A, Güell M, Kühner S, Gavin A-C et al. Quantification of mRNA and protein and integration with protein turnover in a bacterium. Mol Syst Biol 2011; 7:511 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  56. Pritchard RE, Balish MF. Mycoplasma iowae: relationships among oxygen, virulence, and protection from oxidative stress. Vet Res 2015; 46:36 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  57. Muir MT, Cohn SM, Louden C, Kannan TR, Baseman JB. Novel toxin assays implicate Mycoplasma pneumoniae in prolonged ventilator course and hypoxemia. Chest 2011; 139:305–310 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  58. Szczepanek SM, Majumder S, Sheppard ES, Liao X, Rood D et al. Vaccination of BALB/c mice with an avirulent Mycoplasma pneumoniae P30 mutant results in disease exacerbation upon challenge with a virulent strain. Infect Immun 2012; 80:1007–1014 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  59. Prince OA, Krunkosky TM, Krause DC. In vitro spatial and temporal analysis of Mycoplasma pneumoniae colonization of human airway epithelium. Infect Immun 2014; 82:579–586 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  60. Schmidl SR, Gronau K, Hames C, Busse J, Becher D et al. The stability of cytadherence proteins in Mycoplasma pneumoniae requires activity of the protein kinase PrkC. Infect Immun 2010; 78:184–192 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  61. Page CA, Krause DC. Protein kinase/phosphatase function correlates with gliding motility in Mycoplasma pneumoniae. J Bacteriol 2013; 195:1750–1757 [CrossRef][PubMed][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000928
Loading
/content/journal/micro/10.1099/mic.0.000928
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error