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Neutrophil pp 79-91 | Cite as

Analysis of Neutrophil Transmigration Through Epithelial Cell Monolayers

  • Liliya N. Kirpotina
  • Douglas J. Kominsky
  • Mark T. Quinn
  • Steve D. SwainEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2087)

Abstract

Transmigration of neutrophils through an epithelial layer, such as in the intestine or lung, is a necessary response to a perceived attack at the mucosal surface of that tissue. This process is dynamically regulated by a number of interactive events between the neutrophil and other cell types and allows for an effective and localized neutrophil response. However, in certain inflammatory diseases, including inflammatory bowel disease and chronic obstructive pulmonary disease (COPD), persistent neutrophil accumulation can contribute to disease pathology. Elucidating the mechanisms of this aberrant neutrophil accumulation is crucial for understanding and ameliorating these disease processes. The method we describe here is a controlled model system that allows for the investigation of the interactive signals involved in neutrophil transmigration through epithelial barriers, and possible mechanisms of deregulation of this process.

Key words

Neutrophil Epithelium Transmigration Inflammation 

References

  1. 1.
    Bekkering A, Torensma R (2013) Another look at the life of a neutrophil. World J Hematol 2:44–58CrossRefGoogle Scholar
  2. 2.
    Wéra O, Lancellotti P, Oury C (2016) The dual role of neutrophils in inflammatory bowel diseases. J Clin Med 5:118CrossRefGoogle Scholar
  3. 3.
    Linden A, Laan M, Anderson GP (2005) Neutrophils, interleukin-17A and lung disease. Eur Respir J 25:159–172CrossRefGoogle Scholar
  4. 4.
    Bekkering A, Torensma R (2013) Another look at the life of a neutrophil. World J Hematol 2:44-58.CrossRefGoogle Scholar
  5. 5.
    Liu Y, Shaw SK, Ma S et al (2004) Regulation of leukocyte transmigration: cell surface interactions and signaling events. J Immunol 172:7–13CrossRefGoogle Scholar
  6. 6.
    Szabady RL, McCormick BA (2013) Control of neutrophil inflammation at mucosal surfaces by secreted epithelial products. Front Immunol 4:220CrossRefGoogle Scholar
  7. 7.
    Vogt KL, Summers C, Chilvers ER et al (2018) Priming and de-priming of neutrophil responses in vitro and in vivo. Eur J Clin Investig 48(Suppl 2):e12967CrossRefGoogle Scholar
  8. 8.
    Parkos CA (2016) Neutrophil-epithelial interactions: a double-edged sword. Am J Pathol 186:1404–1416CrossRefGoogle Scholar
  9. 9.
    Kusek ME, Pazos MA, Pirzai W et al (2014) In vitro coculture assay to assess pathogen induced neutrophil trans-epithelial migration. J Vis Exp:e50823Google Scholar
  10. 10.
    Colgan SP, Parkos CA, Delp C et al (1993) Neutrophil migration across cultured intestinal epithelial monolayers is modulated by epithelial exposure to IFN-gamma in a highly polarized fashion. J Cell Biol 120:785–798CrossRefGoogle Scholar
  11. 11.
    Devriese S, Van den Bossche L, Van Welden S et al (2017) T84 monolayers are superior to Caco-2 as a model system of colonocytes. Histochem Cell Biol 148:85–93CrossRefGoogle Scholar
  12. 12.
    Parkos CA, Delp C, Arnaout MA et al (1991) Neutrophil migration across a cultured intestinal epithelium. Dependence on a CD11b/CD18-mediated event and enhanced efficiency in physiological direction. J Clin Invest 88:1605–1612CrossRefGoogle Scholar
  13. 13.
    Ren H, Birch NP, Suresh V (2016) An optimised human cell culture model for alveolar epithelial transport. PLoS One 11:e0165225CrossRefGoogle Scholar
  14. 14.
    Yamaura Y, Chapron BD, Wang Z et al (2016) Functional comparison of human colonic carcinoma cell lines and primary small intestinal epithelial cells for investigations of intestinal drug permeability and first-pass metabolism. Drug Metab Dispos 44:329–335CrossRefGoogle Scholar
  15. 15.
    Rajan N, Habermehl J, Coté M-F et al (2006) Preparation of ready-to-use, storable and reconstituted type I collagen from rat tail tendon for tissue engineering applications. Nat Protoc 1:2753–2758CrossRefGoogle Scholar
  16. 16.
    Millius A, Weiner OD (2009) Chemotaxis in neutrophil-like HL-60 cells. Methods Mol Biol 571:167–177CrossRefGoogle Scholar
  17. 17.
    Carrigan SO, Weppler AL, Issekutz AC et al (2005) Neutrophil differentiated HL-60 cells model Mac-1 (CD11b/CD18)-independent neutrophil transepithelial migration. Immunology 115:108–117CrossRefGoogle Scholar
  18. 18.
    Srinivasan B, Kolli AR, Esch MB et al (2015) TEER measurement techniques for in vitro barrier model systems. J Lab Autom 20:107–126CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Liliya N. Kirpotina
    • 1
  • Douglas J. Kominsky
    • 1
  • Mark T. Quinn
    • 1
  • Steve D. Swain
    • 1
    Email author
  1. 1.Department of Microbiology and ImmunologyMontana State UniversityBozemanUSA

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