Skip to main content

The Planar Lipid Bilayer System Serves as a Reductionist Approach for Studying NK Cell Immunological Synapses and Their Functions

  • Protocol
  • First Online:
Natural Killer Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1441))

Abstract

The immunological synapse (IS) is the junction between an immune cell (e.g., a T or NK cell) and another cell (e.g., an antigen-presenting cell (APC), or a tumor cell). The formation of the IS is crucial for cell-mediated immunity, and as such, an understanding of both the composition of the IS and the factors that drive its formation are essential for understanding how and when NK cells eliminate susceptible target cells. The supported lipid bilayer (SLB) system is a highly effective tool for directly studying the IS. SLBs confer three main advantages: (1) they allow for synapse formation on a level horizontal surface, allowing for direct visualization of the IS under high resolution imaging systems, (2) they mimic the surface of a target cell by providing a fluid mosaic into which surface proteins can be embedded while permitting free motion in two dimensions, which is important for studying the dynamics of synapse formation, and (3) they allow investigators to determine the exact composition of the bilayer, thus in turn allowing them to answer very specific questions about the IS. It is our hope that this chapter will furnish readers with an awareness of the applications of the SLB system for studying the IS in NK cells, and also of a basic knowledge of how to use this system for themselves.

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

Access this chapter

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

Institutional subscriptions

References

  1. Grakoui A, Bromley SK, Sumen C et al (1999) The immunological synapse: a molecular machine controlling T cell activation. Science 285(5425):221–227

    Article  CAS  PubMed  Google Scholar 

  2. McConnell HM, Watts TH, Weis RM et al (1986) Supported planar membranes in studies of cell-cell recognition in the immune system. Biochim Biophys Acta 864(1):95–106

    Article  CAS  PubMed  Google Scholar 

  3. Liu D, Bryceson YT, Meckel T et al (2009) Integrin-dependent organization and bidirectional vesicular traffic at cytotoxic immune synapses. Immunity 31(1):99–109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Liu D, Peterson ME, Long EO (2012) The adaptor protein Crk controls activation and inhibition of natural killer cells. Immunity 36(4):600–611

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Abeyweera TP, Merino E, Huse M (2011) Inhibitory signaling blocks activating receptor clustering and induces cytoskeletal retraction in natural killer cells. J Cell Biol 192(4):675–690

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Abeyweera TP, Kaissar M, Huse M (2013) Inhibitory receptor signaling destabilizes immunological synapse formation in primary NK cells. Front Immunol 4:410. doi:10.3389/fimmu.2013.00410

    Article  PubMed  PubMed Central  Google Scholar 

  7. Zheng P, Bertolet G, Chen Y, Huang S, Liu D (2015) Super-resolution imaging of the natural killer cell immunological synapses on a glass-supported planar lipid bilayer. J Vis Exp 96. doi:10.3791/52502. (PMCID: PMC4354632)

  8. Rak GD, Mace EM, Banerjee PP et al (2011) Natural killer cell lytic granule secretion occurs through a pervasive actin network at the immune synapse. PLoS Biol 9(9):e1001151. doi:10.1371/journal.pbio.1001151

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Brown AC, Oddos S, Dobbie IM et al (2011) Remodelling of cortical actin where lytic granules dock at natural killer cell immune synapses revealed by super-resolution microscopy. PLoS Biol 9(9):e1001152. doi:10.1371/journal.pbio.1001152

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Yokosuka T, Takamatsu M, Kobayashi-Imanishi W et al (2012) Programmed cell death 1 forms negative costimulatory microclusters that directly inhibit T cell receptor signaling by recruiting phosphatase SHP2. J Exp Med 209(6):1201–1217

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Dongfang Liu Ph.D. .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Bertolet, G., Liu, D. (2016). The Planar Lipid Bilayer System Serves as a Reductionist Approach for Studying NK Cell Immunological Synapses and Their Functions. In: Somanchi, S. (eds) Natural Killer Cells. Methods in Molecular Biology, vol 1441. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3684-7_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3684-7_13

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3682-3

  • Online ISBN: 978-1-4939-3684-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics