Abstract
Human MAIT cells show little expression of the selectin CD62L and the chemokine receptor CCR7, which are important for entering lymph nodes, and high expression of selectin ligands and chemokine receptors that mediate trafficking into inflamed tissue. Extravasation of leukocytes into tissue requires sequential steps including rolling, firm arrest, crawling, and transendothelial migration, and can be modeled using endothelial cell monolayers in flow chambers that approximate the sheer stress found in post-capillary venules. Using MAIT cells purified from elutriated lymphocytes by fluorescence-activated cell sorting, we have used flow chambers to demonstrate roles for individual chemokine receptors in specific steps required for extravasation. These methods provide a general way to study the molecular mechanisms underlying MAIT cell trafficking from blood into tissue.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Dusseaux M, Martin E, Serriari N, Peguillet I, Premel V, Louis D, Milder M, Le Bourhis L, Soudais C, Treiner E, Lantz O (2011) Human MAIT cells are xenobiotic-resistant, tissue-targeted, CD161hi IL-17-secreting T cells. Blood 117:1250–1259
Le Bourhis L, Martin E, Peguillet I, Guihot A, Froux N, Core M, Levy E, Dusseaux M, Meyssonnier V, Premel V, Ngo C, Riteau B, Duban L, Robert D, Huang S, Rottman M, Soudais C, Lantz O (2010) Antimicrobial activity of mucosal-associated invariant T cells. Nat Immunol 11:701–708
Lee CH, Zhang HH, Singh SP, Koo L, Kabat J, Tsang H, Singh TP, Farber JM (2018) C/EBPdelta drives interactions between human MAIT cells and endothelial cells that are important for extravasation. Elife 7:e32532
Pober JS, Sessa WC (2007) Evolving functions of endothelial cells in inflammation. Nat Rev Immunol 7:803–815
Nourshargh S, Alon R (2014) Leukocyte migration into inflamed tissues. Immunity 41:694–707
Springer TA (1994) Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 76:301–314
Vestweber D (2015) How leukocytes cross the vascular endothelium. Nat Rev Immunol 15:692–704
Cinamon G, Alon R (2003) A real time in vitro assay for studying leukocyte transendothelial migration under physiological flow conditions. J Immunol Methods 273:53–62
Kitayama J, Hidemura A, Saito H, Nagawa H (2000) Shear stress affects migration behavior of polymorphonuclear cells arrested on endothelium. Cell Immunol 203:39–46
Cinamon G, Shinder V, Alon R (2001) Shear forces promote lymphocyte migration across vascular endothelium bearing apical chemokines. Nat Immunol 2:515–522
Luscinskas FW, Ding H, Tan P, Cumming D, Tedder TF, Gerritsen ME (1996) L- and P-selectins, but not CD49d (VLA-4) integrins, mediate monocyte initial attachment to TNF-alpha-activated vascular endothelium under flow in vitro. J Immunol 157:326–335
Acknowledgment
This research was supported by the Intramural Research Program of the NIH, National Institute of Allergy and Infectious Diseases.
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
Parween, F., Zhang, H.H., Farber, J.M. (2020). A Flow Chamber Assay for Studying MAIT Cell Trafficking. In: Kaipe, H., Magalhaes, I. (eds) MAIT Cells. Methods in Molecular Biology, vol 2098. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0207-2_12
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0207-2_12
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0206-5
Online ISBN: 978-1-0716-0207-2
eBook Packages: Springer Protocols