Advertisement

The Detroit 562 Pharyngeal Immortalized Cell Line Model for the Assessment of Infectivity of Pathogenic Neisseria sp.

  • Emily A. Kibble
  • Mitali Sarkar-Tyson
  • Geoffrey W. Coombs
  • Charlene M. KahlerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1969)

Abstract

Neisseria meningitidis and Neisseria gonorrhoeae are obligate pathogens of the human host. Due to their adaptation to the human host, many factors required for infection are specialized for the human host to the point that natural infection processes are difficult to replicate in animal models. Immortalized human cell lines have been used to identify the host factors necessary for successful colonization of human mucosal surfaces. One such model is the Detroit 562 pharyngeal immortalized cell monolayer model which is used to measure the rate of attachment to and invasion of N. meningitidis and N. gonorrhoeae into epithelial cells. The methodology of this assay, as well as the maintenance of Detroit 562 cells necessary for the experiment, will be described.

Key words

Detroit 562 Pharyngeal Meningococcal Gonococcal Host receptors 

Notes

Acknowledgments

This work is supported by the Australian Defence Material Transfer Center (Project Agreement 10.44) and North Atlantic Treaty Organization (SfP-984235) awarded to MST. EK is supported by a Research Training Program (RTP) PhD scholarship from Murdoch University. CK is supported by the Amanda Young Foundation, a not-for-profit organisation, for improved community awareness of meningococcal disease.

References

  1. 1.
    Islam EA, Gray-Owen SD (2014) Modelling infection by the pathogenic Neisseria. In: Davies JK, Kahler CM (eds) Pathogenic Neisseria: Genomics, Molecular Biology and Disease Prevention. Caister Academic Press, NorfolkGoogle Scholar
  2. 2.
    Edwards JL, Jennings MP, Apicella MA, Seib KL (2016) Is gonococcal disease preventable? The importance of understanding immunity and pathogenesis in vaccine development. Crit Rev Microbiol 42:928–941CrossRefGoogle Scholar
  3. 3.
    Soriani M (2017) Unraveling Neisseria meningitidis pathogenesis: from functional genomics to experimental models. F1000Res 6:1228CrossRefGoogle Scholar
  4. 4.
    Simonis A, Schubert-Unkmeir A (2016) Interactions of meningococcal virulence factors with endothelial cells at the human blood-cerebrospinal fluid barrier and their role in pathogenicity. FEBS Lett 590:3854–3867CrossRefGoogle Scholar
  5. 5.
    Quillin SJ, Seifert HS (2018) Neisseria gonorrhoeae host adaptation and pathogenesis. Nat Rev Microbiol 16:226–240CrossRefGoogle Scholar
  6. 6.
    Mubaiwa TD, Semchenko EA, Hartley-Tassell LE, Day CJ, Jennings MP, Seib KL (2017) The sweet side of the pathogenic Neisseria: the role of glycan interactions in colonisation and disease. Pathog Dis 75.  https://doi.org/10.1093/femspd/ftx063
  7. 7.
    Weyand NJ (2017) Neisseria models of infection and persistence in the upper respiratory tract. Pathog Dis 75.  https://doi.org/10.1093/femspd/ftx031
  8. 8.
    Plant L, Sundqvist J, Zughaier S, Lovkvist L, Stephens DS, Jonsson AB (2006) Lipooligosaccharide structure contributes to multiple steps in the virulence of Neisseria meningitidis. Infect Immun 74:1360–1367CrossRefGoogle Scholar
  9. 9.
    Bartley SN, Tzeng YL, Heel K, Lee CW, Mowlaboccus S, Seemann T, Lu W, Lin YH, Ryan CS, Peacock C, Stephens DS, Davies JK, Kahler CM (2013) Attachment and invasion of Neisseria meningitidis to host cells is related to surface hydrophobicity, bacterial cell size and capsule. PLoS One 8:e55798CrossRefGoogle Scholar
  10. 10.
    Reimer A, Seufert F, Weiwad M, Ebert J, Bzdyl NM, Kahler CM, Sarkar-Tyson M, Holzgrabe U, Rudel T, Kozjak-Pavlovic V (2016) Inhibitors of macrophage infectivity potentiator-like PPIases affect neisserial and chlamydial pathogenicity. Int J Antimicrob Agents 48:401–408CrossRefGoogle Scholar
  11. 11.
    Peterson WD Jr, Stulberg CS, Simpson WF (1971) A permanent heteroploid human cell line with type B glucose-6-phosphate dehydrogenase. Proc Soc Exp Biol Med 136:1187–1191CrossRefGoogle Scholar
  12. 12.
    Stephens DS, McGee ZA, Melly MA, Hoffman LH, Gregg CR (1982) Attachment of pathogenic Neisseria to human mucosal surfaces: role in pathogenesis. Infection 10:192–195CrossRefGoogle Scholar
  13. 13.
    Matijevic T, Marjanovic M, Pavelic J (2009) Functionally active toll-like receptor 3 on human primary and metastatic cancer cells. Scand J Immunol 70:18–24CrossRefGoogle Scholar
  14. 14.
    Rydberg C, Mansson A, Uddman R, Riesbeck K, Cardell LO (2009) Toll-like receptor agonists induce inflammation and cell death in a model of head and neck squamous cell carcinomas. Immunology 128:e600–e611CrossRefGoogle Scholar
  15. 15.
    Gould JM, Weiser JN (2001) Expression of C-reactive protein in the human respiratory tract. Infect Immun 69:1747–1754CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Emily A. Kibble
    • 1
    • 2
  • Mitali Sarkar-Tyson
    • 1
  • Geoffrey W. Coombs
    • 2
  • Charlene M. Kahler
    • 1
    Email author
  1. 1.Marshall Centre for Infectious Diseases Research and Training, School of Biomedical SciencesUniversity of Western AustraliaPerthAustralia
  2. 2.School of Veterinary and Life SciencesMurdoch UniversityPerthAustralia

Personalised recommendations