Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Resistance to leukocytes ties benefits of quorum sensing dysfunctionality to biofilm infection


Social interactions play an increasingly recognized key role in bacterial physiology1. One of the best studied is quorum sensing (QS), a mechanism by which bacteria sense and respond to the status of cell density2. While QS is generally deemed crucial for bacterial survival, QS-dysfunctional mutants frequently arise in in vitro culture. This has been explained by the fitness cost an individual mutant, a ‘quorum cheater’, saves at the expense of the community3. QS mutants are also often isolated from biofilm-associated infections, including cystic fibrosis lung infection4, as well as medical device infection and associated bacteraemia5,6,7. However, despite the frequently proposed use of QS blockers to control virulence8, the mechanisms underlying QS dysfunctionality during infection have remained poorly understood. Here, we show that in the major human pathogen Staphylococcus aureus, quorum cheaters arise exclusively in biofilm infection, while in non-biofilm-associated infection there is a high selective pressure to maintain QS control. We demonstrate that this infection-type dependence is due to QS-dysfunctional bacteria having a significant survival advantage in biofilm infection because they form dense and enlarged biofilms that provide resistance to phagocyte attacks. Our results link the benefit of QS-dysfunctional mutants in vivo to biofilm-mediated immune evasion, thus to mechanisms that are specific to the in vivo setting. Our findings explain why QS mutants are frequently isolated from biofilm-associated infections and provide guidance for the therapeutic application of QS blockers.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: QS in biofilm- and non-biofilm-associated infection.
Fig. 2: QS-dysfunctional S. aureus forms compact biofilms that prevent neutrophil penetration.
Fig. 3: QS-dysfunctional biofilms prevent killing by leukocytes.
Fig. 4: Benefits of QS mutants in biofilm-associated infection are due to interaction with leukocytes.

Data availability

All data generated or analysed during this study are included in this published article (and its supplementary information files) or available from the corresponding author upon request.


  1. Asfahl, K. L. & Schuster, M. Social interactions in bacterial cell–cell signaling. FEMS Microbiol. Rev. 41, 92–107 (2017).

    Article  CAS  Google Scholar 

  2. Miller, M. B. & Bassler, B. L. Quorum sensing in bacteria. Annu. Rev. Microbiol. 55, 165–199 (2001).

    Article  CAS  Google Scholar 

  3. Dandekar, A. A., Chugani, S. & Greenberg, E. P. Bacterial quorum sensing and metabolic incentives to cooperate. Science 338, 264–266 (2012).

    Article  CAS  Google Scholar 

  4. Bjarnsholt, T. et al. Quorum sensing and virulence of Pseudomonas aeruginosa during lung infection of cystic fibrosis patients. PLoS ONE 5, e10115 (2010).

    Article  Google Scholar 

  5. Vuong, C., Kocianova, S., Yao, Y., Carmody, A. B. & Otto, M. Increased colonization of indwelling medical devices by quorum-sensing mutants of Staphylococcus epidermidis in vivo. J. Infect. Dis. 190, 1498–1505 (2004).

    Article  Google Scholar 

  6. Traber, K. E. et al. agr function in clinical Staphylococcus aureus isolates. Microbiology 154, 2265–2274 (2008).

    Article  CAS  Google Scholar 

  7. Fowler, V. G. et al. Persistent bacteremia due to methicillin-resistant Staphylococcus aureus infection is associated with agr dysfunction and low-level in vitro resistance to thrombin-induced platelet microbicidal protein. J. Infect. Dis. 190, 1140–1149 (2004).

    Article  CAS  Google Scholar 

  8. Dickey, S. W., Cheung, G. Y. C. & Otto, M. Different drugs for bad bugs: antivirulence strategies in the age of antibiotic resistance. Nat. Rev. Drug Discov. 16, 457–471 (2017).

    Article  CAS  Google Scholar 

  9. Rumbaugh, K. P. et al. Quorum sensing and the social evolution of bacterial virulence. Curr. Biol. 19, 341–345 (2009).

    Article  CAS  Google Scholar 

  10. Pollitt, E. J., West, S. A., Crusz, S. A., Burton-Chellew, M. N. & Diggle, S. P. Cooperation, quorum sensing, and evolution of virulence in Staphylococcus aureus. Infect. Immun. 82, 1045–1051 (2014).

    Article  Google Scholar 

  11. Gordon, R. J. & Lowy, F. D. Pathogenesis of methicillin-resistant Staphylococcus aureus infection. Clin. Infect. Dis. 46, S350–S359 (2008).

    Article  CAS  Google Scholar 

  12. Le, K. Y. & Otto, M. Quorum-sensing regulation in staphylococci—an overview. Front. Microbiol. 6, 1174 (2015).

    Article  Google Scholar 

  13. Otto, M. Staphylococcal infections: mechanisms of biofilm maturation and detachment as critical determinants of pathogenicity. Annu. Rev. Med. 64, 175–188 (2013).

    Article  CAS  Google Scholar 

  14. Periasamy, S. et al. How Staphylococcus aureus biofilms develop their characteristic structure. Proc. Natl Acad. Sci. USA 109, 1281–1286 (2012).

    Article  CAS  Google Scholar 

  15. Peschel, A. & Otto, M. Phenol-soluble modulins and staphylococcal infection. Nat. Rev. Microbiol. 11, 667–673 (2013).

    Article  CAS  Google Scholar 

  16. Wang, R. et al. Identification of novel cytolytic peptides as key virulence determinants for community-associated MRSA. Nat. Med. 13, 1510–1514 (2007).

    Article  CAS  Google Scholar 

  17. Painter, K. L., Krishna, A., Wigneshweraraj, S. & Edwards, A. M. What role does the quorum-sensing accessory gene regulator system play during Staphylococcus aureus bacteremia?. Trends Microbiol. 22, 676–685 (2014).

    Article  CAS  Google Scholar 

  18. Collins, J. et al. Offsetting virulence and antibiotic resistance costs by MRSA. ISME J. 4, 577–584 (2010).

    Article  Google Scholar 

  19. Paulander, W. et al. Antibiotic-mediated selection of quorum-sensing-negative Staphylococcus aureus. mBio 3, e00459–00412 (2013).

  20. Vuong, C., Saenz, H. L., Gotz, F. & Otto, M. Impact of the agr quorum-sensing system on adherence to polystyrene in Staphylococcus aureus. J. Infect. Dis. 182, 1688–1693 (2000).

    Article  CAS  Google Scholar 

  21. Shopsin, B. et al. Mutations in agr do not persist in natural populations of methicillin-resistant Staphylococcus aureus. J. Infect. Dis. 202, 1593–1599 (2010).

    Article  CAS  Google Scholar 

  22. Vuong, C., Kocianova, S., Yu, J., Kadurugamuwa, J. L. & Otto, M. Development of real-time in vivo imaging of device-related Staphylococcus epidermidis infection in mice and influence of animal immune status on susceptibility to infection. J. Infect. Dis. 198, 258–261 (2008).

    Article  Google Scholar 

  23. Li, M. et al. Evolution of virulence in epidemic community-associated methicillin-resistant Staphylococcus aureus. Proc. Natl Acad. Sci. USA 106, 5883–5888 (2009).

    Article  CAS  Google Scholar 

  24. Carrel, M., Perencevich, E. N. & David, M. Z. USA300 methicillin-resistant Staphylococcus aureus, United States, 2000–2013. Emerg. Infect. Dis. 21, 1973–1980 (2015).

    Article  Google Scholar 

  25. Kourbatova, E. V. et al. Emergence of community-associated methicillin-resistant Staphylococcus aureus USA 300 clone as a cause of health care-associated infections among patients with prosthetic joint infections. Am. J. Infect. Control 33, 385–391 (2005).

    Article  Google Scholar 

  26. Foster, T. J. Immune evasion by staphylococci. Nat. Rev. Microbiol. 3, 948–958 (2005).

    Article  CAS  Google Scholar 

  27. Cheung, G. Y., Wang, R., Khan, B. A., Sturdevant, D. E. & Otto, M. Role of the accessory gene regulator agr in community-associated methicillin-resistant Staphylococcus aureus pathogenesis. Infect. Immun. 79, 1927–1935 (2011).

    Article  CAS  Google Scholar 

  28. Butterfield, J. M. et al. Predictors of agr dysfunction in methicillin-resistant Staphylococcus aureus (MRSA) isolates among patients with MRSA bloodstream infections. Antimicrob. Agents Chemother. 55, 5433–5437 (2011).

    Article  CAS  Google Scholar 

  29. Nowakowska, J., Landmann, R. & Khanna, N. Foreign body infection models to study host–pathogen response and antimicrobial tolerance of bacterial biofilm. Antibiotics (Basel) 3, 378–397 (2014).

    Article  CAS  Google Scholar 

  30. Flannagan, R. S., Heit, B. & Heinrichs, D. E. Antimicrobial mechanisms of macrophages and the immune evasion strategies of staphylococcus aureus. Pathogens 4, 826–868 (2015).

    Article  CAS  Google Scholar 

  31. Joo, H. S., Cheung, G. Y. & Otto, M. Antimicrobial activity of community-associated methicillin-resistant Staphylococcus aureus is caused by phenol-soluble modulin derivatives. J. Biol. Chem. 286, 8933–8940 (2011).

    Article  CAS  Google Scholar 

  32. Diep, B. A., Carleton, H. A., Chang, R. F., Sensabaugh, G. F. & Perdreau-Remington, F. Roles of 34 virulence genes in the evolution of hospital- and community-associated strains of methicillin-resistant Staphylococcus aureus. J. Infect. Dis. 193, 1495–1503 (2006).

    Article  CAS  Google Scholar 

  33. Somerville, G. A. et al. In vitro serial passage of Staphylococcus aureus: changes in physiology, virulence factor production, and agr nucleotide sequence. J. Bacteriol. 184, 1430–1437 (2002).

    Article  CAS  Google Scholar 

  34. Voyich, J. M. et al. Is Panton–Valentine leukocidin the major virulence determinant in community-associated methicillin-resistant Staphylococcus aureus disease? J. Infect. Dis. 194, 1761–1770 (2006).

  35. Wang, R. et al. Staphylococcus epidermidis surfactant peptides promote biofilm maturation and dissemination of biofilm-associated infection in mice. J. Clin. Invest. 121, 238–248 (2011).

    Article  CAS  Google Scholar 

  36. Cox, P. J., Phillips, B. J. & Thomas, P. The enzymatic basis of the selective action of cyclophosphamide. Cancer Res. 35, 3755–3761 (1975).

    CAS  PubMed  Google Scholar 

Download references


This study was supported by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases (NIAID), U.S. National Institutes of Health (NIH) (project no. 1 ZIA AI000904) to M.O., The National Natural Science Foundation of China (grant no. 81501804) to L.H. and the General Program of Shanghai Municipal Commission of Health and Family Planning (grant no. 20154Y0014) to L.H.

Author information

Authors and Affiliations



L.H., K.Y.L. and R.L.H. performed confocal microscopy. B.A.K. performed in vitro quorum cheating assays. L.H., T.H.N., G.Y.C.C., J.S.B., R.L.H. and Y.Z. performed animal experiments. L.H., T.H.N. and R.L.H. performed leukocyte killing assays. L.H. performed all other experiments. L.H., K.Y.L., G.Y.C.C., J.K., J.S.B. and M.O. analysed the data. M.L and M.O. designed and supervised experiments. M.O. wrote the paper.

Corresponding author

Correspondence to Michael Otto.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary Tables 1 and 2, and Supplementary Figures 1–4.

Reporting Summary

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

He, L., Le, K.Y., Khan, B.A. et al. Resistance to leukocytes ties benefits of quorum sensing dysfunctionality to biofilm infection. Nat Microbiol 4, 1114–1119 (2019).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing