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Infection of Human Neutrophils to Study Virulence Properties of Mycobacterium tuberculosis

  • Tobias Dallenga
  • Björn Corleis
  • Ulrich E. SchaibleEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1285)

Abstract

Polymorphonuclear neutrophils (PMN) are professional phagocytes and the first line of defense against invading microbes. Upon infection with Mycobacterium tuberculosis, PMN are attracted to the site of infection along an interleukin 8 gradient. In patients with active tuberculosis, PMN comprise the predominant population in the lung and carry the main mycobacterial load suggesting a minor role for PMN in protective host defense against M. tuberculosis but rather in pathology. Therefore, better understanding of PMN biology in tuberculosis is of pivotal importance to develop novel immune modulating measures and host directed therapies. Virulent M. tuberculosis escape the otherwise microbicidal armamentarium of PMNs by inducing necrotic cell death through the PMN’s own reactive oxygen species. Studying the interactions between PMN and different M. tuberculosis strains, and virulence factors thereof, is vital to comprehend tuberculosis pathogenesis. Working with PMN is challenging as these cells are non-adherent, motile and—with a half-life of 6–12 h in vitro—rather short-lived. Here, we provide an isolation and infection protocol that is tailored to study mycobacterial infection in human PMN regarding the intracellular fate of mycobacteria and host cell responses, such as cell death and release of microbicidal effectors.

Key words

Polymorphonuclear Neutrophil Phagocytosis Cell death Mycobacterium tuberculosis 

Notes

Acknowledgements

This work was supported by DFG Priority Programme 1580 “Intracellular Compartments as Places of Pathogen-Host-Interaction” and the Research Center Borstel.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Tobias Dallenga
    • 1
  • Björn Corleis
    • 2
    • 3
  • Ulrich E. Schaible
    • 1
    • 3
    • 4
    Email author
  1. 1.Cellular Microbiology, Priority Area InfectionsResearch Center BorstelBorstelGermany
  2. 2.The Ragon Institute of MGH, MIT and HarvardCambridgeUSA
  3. 3.Department of ImmunologyLondon School of Hygiene and Tropical MedicineLondonUK
  4. 4.German Centre of Infection Research, TTU-TBBorstelGermany

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