Advertisement

MAIT Cells pp 179-188 | Cite as

A Flow Chamber Assay for Studying MAIT Cell Trafficking

  • Farhat Parween
  • Hongwei H. Zhang
  • Joshua M. FarberEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2098)

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.

Key words

MAIT cells Transendothelial migration Flow chamber Lymphocytes Chemokines 

Notes

Acknowledgment

This research was supported by the Intramural Research Program of the NIH, National Institute of Allergy and Infectious Diseases.

References

  1. 1.
    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 117:1250–1259CrossRefGoogle Scholar
  2. 2.
    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 11:701–708CrossRefGoogle Scholar
  3. 3.
    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:e32532CrossRefGoogle Scholar
  4. 4.
    Pober JS, Sessa WC (2007) Evolving functions of endothelial cells in inflammation. Nat Rev Immunol 7:803–815CrossRefGoogle Scholar
  5. 5.
    Nourshargh S, Alon R (2014) Leukocyte migration into inflamed tissues. Immunity 41:694–707CrossRefGoogle Scholar
  6. 6.
    Springer TA (1994) Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 76:301–314CrossRefGoogle Scholar
  7. 7.
    Vestweber D (2015) How leukocytes cross the vascular endothelium. Nat Rev Immunol 15:692–704CrossRefGoogle Scholar
  8. 8.
    Cinamon G, Alon R (2003) A real time in vitro assay for studying leukocyte transendothelial migration under physiological flow conditions. J Immunol Methods 273:53–62CrossRefGoogle Scholar
  9. 9.
    Kitayama J, Hidemura A, Saito H, Nagawa H (2000) Shear stress affects migration behavior of polymorphonuclear cells arrested on endothelium. Cell Immunol 203:39–46CrossRefGoogle Scholar
  10. 10.
    Cinamon G, Shinder V, Alon R (2001) Shear forces promote lymphocyte migration across vascular endothelium bearing apical chemokines. Nat Immunol 2:515–522CrossRefGoogle Scholar
  11. 11.
    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 157:326–335PubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Farhat Parween
    • 1
  • Hongwei H. Zhang
    • 1
  • Joshua M. Farber
    • 1
    Email author
  1. 1.Inflammation Biology Section, Laboratory of Molecular ImmunologyNational Institute of Allergy and Infectious Diseases, National Institutes of HealthBethesdaUSA

Personalised recommendations