Development of Mycoplasma pneumoniae biofilms in vitro and the limited role of motility

Feng, Monica; Schaff, Andrew C; Aruguete, Sara A. Cuadra; Riggs, Hailey E.; Distelhorst, Steven L.; Balish, Mitchell F.

doi:10.1016/j.ijmm.2018.01.007

“…First, individual WT M. pneumoniae cells are generated by motility from biofilm towers [29], but the formation of towers of comparable appearance formed by the essentially non-motile, isogenic M. pneumoniae mutant II-3R shows that individual cells are not required for biofilm spread [29].…”

Section: Discussionmentioning

confidence: 99%

“…For biofilm measurements, samples were incubated for 30 min (or in some experiments, 4 h) at 37 °C, after which they were washed with warm PBS to remove non-adherent cells. Crystal violet assays were performed on the remaining biofilms as described previously [29], with absorbance at 600 nm taken to indicate biofilm volume, to determine the effect of each enzyme or chemical on biofilm dispersal. Each condition was performed in triplicate, with three biological replicates.…”

Section: Methodsmentioning

confidence: 99%

Mycoplasma pneumoniae biofilms grown in vitro: traits associated with persistence and cytotoxicity

Feng

¹

,

Schaff

²

,

Balish

³

2020

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The atypical bacterial pathogen Mycoplasma pneumoniae 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 in vitro setting, M. pneumoniae 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 M. pneumoniae biofilms, open questions remain concerning phenotypes associated with persistence, such as what properties might favour the bacteria while minimizing host damage. M. pneumoniae also produces several cytotoxic molecules including community-acquired respiratory distress syndrome (CARDS) toxin, H2S and H2O2, 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 M. pneumoniae 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 H2S and H2O2 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 M. pneumoniae biofilms contribute to both the establishment and propagation of M. pneumoniae infections, and how both biofilm towers and individual cells participate in persistence and chronic disease.

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“…Wild-type M. pneumoniae strain M129 passage B18 was cultured in SP-4 broth as previously described [6]. The human bronchial epithelial cell line BEAS-2B was purchased from the American Type Culture Collection.…”

Section: Methodsmentioning

confidence: 99%

“…To date, the only mechanism potentially associated with M. pneumoniae persistence in hosts that has been described in any detail is biofilm formation [4–7]. In general, biofilms are responsible for up to 80 % of clinical bacterial infections [8], making them an important aspect of human disease.…”

Section: Introductionmentioning

confidence: 99%

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Modelling persistent Mycoplasma pneumoniae biofilm infections in a submerged BEAS-2B bronchial epithelial tissue culture model

Feng

¹

,

Burgess

²

,

Cuellar

³

et al. 2021

Journal of Medical Microbiology

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Introduction. Infections with the respiratory pathogen Mycoplasma pneumoniae are often chronic, recurrent and resistant, persisting after antibiotic treatment. M. pneumoniae grown on glass forms protective biofilms, consistent with a role for biofilms in persistence. These biofilms consist of towers of bacteria interspersed with individual adherent cells. Hypothesis/Gap Statement. A tissue culture model for M. pneumoniae biofilms has not been described or evaluated to address whether growth, development and resistance properties are consistent with persistence in the host. Moreover, it is unclear whether the M. pneumoniae cells in the biofilm towers and individual bacterial cells have distinct roles in disease. Aim. We evaluated the properties of biofilms of M. pneumoniae grown on the immortalized human bronchial epithelial cell line BEAS-2B in relation to persistence in the host. We observed nucleation of biofilm towers and the disposition of individual cells in culture, leading to a model of how tower and individual cells contribute to infection and disease. Methodology. With submerged BEAS-2B cells as a substrate, we evaluated growth and development of M. pneumoniae biofilms using scanning electron microscopy and confocal laser scanning microscopy. We characterized resistance to erythromycin and complement using minimum inhibitory concentration assays and quantification of colony forming units. We monitored biofilm tower formation using time-lapse microscopic analysis of host-cell-free M. pneumoniae cultures. Results. Bacteria grown on host cells underwent similar development to those grown without host cells, including tower formation, rounding and incidence of individual cells outside towers. Erythromycin and complement significantly reduced growth of M. pneumoniae . Towers formed exclusively from pre-existing aggregates of bacteria. We discuss a model of the M. pneumoniae biofilm life cycle in which protective towers derive from pre-existing aggregates, and generate individual cytotoxic cells. Conclusion . M. pneumoniae can form protective biofilms in a tissue culture model, implicating biofilms in chronic infections, with aggregates of M. pneumoniae cells being important for establishing infections.

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Insight into the Pathogenic Mechanism of Mycoplasma pneumoniae

Hu

¹

,

Ye

²

,

Chen

³

et al. 2022

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Mycoplasma pneumoniae, an obligate parasitic pathogen without cell wall, can cause severe upper and lower respiratory tract symptoms. It is the pathogen of human bronchitis and walking pneumonia, and named community-acquired pneumonia. In addition to severe respiratory symptoms, there are clinical extrapulmonary manifestations in the skin, brain, kidney, musculoskeletal, digestive system, and even blood system after M. pneumoniae infection. Hereby, we comprehensively summarized and reviewed the intrapulmonary and extrapulmonary pathogenesis of M. pneumoniae infection. The pathogenesis of related respiratory symptoms caused by M. pneumoniae is mainly adhesion damage, direct damage including nutrient predation, invasion and toxin, cytokine induced inflammation damage and immune evasion effect. The pathogenesis of extrapulmonary manifestations includes direct damage mediated by invasion and inflammatory factors, indirect damage caused by host immune response, and vascular occlusion. The intrapulmonary and extrapulmonary pathogenic mechanisms of M. pneumoniae infection are independent and interrelated, and have certain commonalities. In fact, the pathogenic mechanisms of M. pneumoniae are complicated, and the specific content is still not completely clear, further researches are necessary for determining the detailed pathogenesis of M. pneumoniae. This review can provide certain guidance for the effective prevention and treatment of M. pneumoniae infection. Abbreviations CARDSCommunity-acquired respiratory distress syndrome toxin GlpO L-α-glycerophosphate oxidase KATP channels ATP-sensitive K + channels ROS Reactive oxygen species LAMPS Lipid-associated membrane proteins HDAC 5 Histone deacetylase 5 IbpM Immunoglobulin binding protein of Mycoplasma NET Neutrophil extracellular traps EF-Tu Elongation factor thermo unstable MpEPDs Mycoplasma pneumoniae-related extrapulmonary diseases * Weimin Xie

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Development of Mycoplasma pneumoniae biofilms in vitro and the limited role of motility

Cited by 11 publications

References 54 publications

Mycoplasma pneumoniae biofilms grown in vitro: traits associated with persistence and cytotoxicity

Mycoplasma pneumoniae biofilms grown in vitro: traits associated with persistence and cytotoxicity

Modelling persistent Mycoplasma pneumoniae biofilm infections in a submerged BEAS-2B bronchial epithelial tissue culture model

Insight into the Pathogenic Mechanism of Mycoplasma pneumoniae

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