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.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Bekkering A, Torensma R (2013) Another look at the life of a neutrophil. World J Hematol 2:44–58
Wéra O, Lancellotti P, Oury C (2016) The dual role of neutrophils in inflammatory bowel diseases. J Clin Med 5:118
Linden A, Laan M, Anderson GP (2005) Neutrophils, interleukin-17A and lung disease. Eur Respir J 25:159–172
Bekkering A, Torensma R (2013) Another look at the life of a neutrophil. World J Hematol 2:44-58.
Liu Y, Shaw SK, Ma S et al (2004) Regulation of leukocyte transmigration: cell surface interactions and signaling events. J Immunol 172:7–13
Szabady RL, McCormick BA (2013) Control of neutrophil inflammation at mucosal surfaces by secreted epithelial products. Front Immunol 4:220
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):e12967
Parkos CA (2016) Neutrophil-epithelial interactions: a double-edged sword. Am J Pathol 186:1404–1416
Kusek ME, Pazos MA, Pirzai W et al (2014) In vitro coculture assay to assess pathogen induced neutrophil trans-epithelial migration. J Vis Exp:e50823
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 120:785–798
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 148:85–93
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 88:1605–1612
Ren H, Birch NP, Suresh V (2016) An optimised human cell culture model for alveolar epithelial transport. PLoS One 11:e0165225
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 44:329–335
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 1:2753–2758
Millius A, Weiner OD (2009) Chemotaxis in neutrophil-like HL-60 cells. Methods Mol Biol 571:167–177
Carrigan SO, Weppler AL, Issekutz AC et al (2005) Neutrophil differentiated HL-60 cells model Mac-1 (CD11b/CD18)-independent neutrophil transepithelial migration. Immunology 115:108–117
Srinivasan B, Kolli AR, Esch MB et al (2015) TEER measurement techniques for in vitro barrier model systems. J Lab Autom 20:107–126
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kirpotina, L.N., Kominsky, D.J., Quinn, M.T., Swain, S.D. (2020). Analysis of Neutrophil Transmigration Through Epithelial Cell Monolayers. 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_6
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0154-9_6
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