Skip to main content
SpringerLink
Log in
Book cover

Neutrophil pp 223–233Cite as

Influence of Oxygen on Function and Cholesterol Composition of Murine Bone Marrow-Derived Neutrophils

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2087))

Abstract

During inflammation and infection, invading pathogens as well as infiltrating neutrophils locally consume oxygen and reduce the present oxygen level. Since oxygen is an elementary component of the microenvironment required for cell activity and alters multiple cellular functions, it is important to study neutrophil functionality and phenotype at characteristic pathophysiological oxygen levels that reflect the hypoxic phenotype during infection and inflammation. Here, we describe methods to study murine neutrophils under hypoxic compared to normoxic conditions, including analysis of cholesterol content as a key lipid involved in biological functions.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Carreau A, El Hafny-Rahbi B, Matejuk A et al (2011) Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia. J Cell Mol Med 15:1239–1253

    Article  CAS  Google Scholar 

  2. Lone AG, Atci E, Renslow R et al (2015) Staphylococcus aureus induces hypoxia and cellular damage in porcine dermal explants. Infect Immun 83:2531–2541

    Article  CAS  Google Scholar 

  3. Melican K, Boekel J, Månsson LE et al (2008) Bacterial infection-mediated mucosal signalling induces local renal ischemia as a defense against sepsis. Cell Microbiol 10:1987–1998

    Article  CAS  Google Scholar 

  4. Campbell EL, Bruyninckx WJ, Kelly CJ et al (2014) Transmigrating neutrophils shape the mucosal microenvironment through localized oxygen depletion to influence resolution of inflammation. Immunity 40:66–77

    Article  CAS  Google Scholar 

  5. Schaffer K, Taylor CT (2015) The impact of hypoxia on bacterial infection. FEBS J 282:2260–2266

    Article  CAS  Google Scholar 

  6. Zeitouni NE, Fandrey J, Naim HY et al (2015) Measuring oxygen levels in Caco-2 cultures. Hypoxia 9:53–66

    Google Scholar 

  7. Möllerherm H, Branitzki-Heinemann K, Brogden G et al (2017) Hypoxia modulates the response of mast cells to Staphylococcus aureus infection. Front Immunol 11:541

    Article  Google Scholar 

  8. Branitzki-Heinemann K, Möllerherm H, Völlger L et al (2016) Formation of neutrophil extracellular traps under low oxygen level. Front Immunol 7:518

    Article  Google Scholar 

  9. Brinkmann V, Reichard U, Goosmann C et al (2004) Neutrophil extracellular traps kill bacteria. Science 303:1532–1535

    Article  CAS  Google Scholar 

  10. von Köckritz-Blickwede M, Nizet V (2009) Innate immunity turned inside-out: antimicrobial defense by phagocyte extracellular traps. J Mol Med 87:775–783

    Article  Google Scholar 

  11. Fuchs TA, Abed U, Goosmann C et al (2007) Novel cell death program leads to neutrophil extracellular traps. J Cell Biol 176:231–241

    Article  CAS  Google Scholar 

  12. Stoiber W, Obermayer A, Steinbacher P et al (2015) The role of reactive oxygen species (ROS) in the formation of extracellular traps (ETs) in humans. Biomol Ther 5:702–723

    CAS  Google Scholar 

  13. Neumann A, Brogden G, Jerjomiceva N et al (2014) Lipid alterations in human blood-derived neutrophils lead to formation of neutrophil extracellular traps. Eur J Cell Biol 93:347–354

    Article  CAS  Google Scholar 

  14. Chow OA, von Köckritz-Blickwede M, Bright AT et al (2010) Statins enhance formation of phagocyte extracellular traps. Cell Host Microbe 8:445–454

    Article  CAS  Google Scholar 

  15. Gilk SD, Cockrell DC, Luterbach C et al (2013) Bacterial colonization of host cells in the absence of cholesterol. PLoS Pathog 9:e1003107

    Article  CAS  Google Scholar 

  16. Brogden G, Neumann A, Husein DM et al (2017) Methods to study lipid alterations in neutrophils and the subsequent formation of neutrophil extracellular traps. J Vis Exp (121):54667

    Google Scholar 

  17. Zhao Y, Wu T, Shao S et al (2016) Phenotype, development, and biological function of myeloid-derived suppressor cells. Onco Immunology 5(2)

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by R2N Project funded by the Federal State of Lower Saxony.

Author information

Authors and Affiliations

  1. Department of Physiological Chemistry, University of Veterinary Medicine Hannover, Hannover, Germany

    Katja Branitzki-Heinemann, Graham Brogden & Maren von Köckritz-Blickwede

  2. Research Center for Emerging Infections and Zoonoses (RIZ), University of Veterinary Medicine Hannover, Hannover, Germany

    Katja Branitzki-Heinemann & Maren von Köckritz-Blickwede

Authors
  1. Katja Branitzki-Heinemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Graham Brogden
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maren von Köckritz-Blickwede
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maren von Köckritz-Blickwede .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Immunology, Montana State University, Bozeman, MT, USA

    Mark T. Quinn

  2. Laboratory of Bacteriology, Rocky Mountain Laboratories Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA

    Frank R. DeLeo

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Branitzki-Heinemann, K., Brogden, G., von Köckritz-Blickwede, M. (2020). Influence of Oxygen on Function and Cholesterol Composition of Murine Bone Marrow-Derived Neutrophils. In: Quinn, M., DeLeo, F. (eds) Neutrophil. Methods in Molecular Biology, vol 2087. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0154-9_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0154-9_17

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0153-2

  • Online ISBN: 978-1-0716-0154-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation