Skip to main content

Identification of Islet Antigen-Specific CD8 T Cells Using MHCI-Peptide Tetramer Reagents in the Non Obese Diabetic (NOD) Mouse Model of Type 1 Diabetes

Part of the Methods in Molecular Biology book series (MIMB,volume 1433)

Abstract

MHCI-peptide tetramer staining is an important technique in order to identify antigen-specific T cells within a heterogeneous cell population. The reagents may be used to isolate antigen-specific T cells and can help identify their role in disease. Here, we describe how to make tetramer from peptide:MHC monomers together with a protocol for staining antigen-specific cell populations with advice on generating a complementary antibody phenotyping panel.

Keywords:

  • MHCI-peptide tetramer staining
  • Antigen-specific
  • T cell receptor
  • Flow cytometry

This is a preview of subscription content, access via your institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/7651_2015_295
  • Chapter length: 7 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
eBook
USD   89.00
Price excludes VAT (USA)
  • ISBN: 978-1-4939-3643-4
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   119.99
Price excludes VAT (USA)
Hardcover Book
USD   169.99
Price excludes VAT (USA)
Fig. 1

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Altman JD, Moss PA, Goulder PJ et al (1996) Phenotypic analysis of antigen-specific T lymphocytes. Science 274(5284):94–96

    CAS  CrossRef  PubMed  Google Scholar 

  2. Wooldridge L, Ekeruche-Makinde J, van den Berg HA et al (2012) A single autoimmune T cell receptor recognizes more than a million different peptides. J Biol Chem 287(2):1168–1177

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  3. Weichsel R, Dix C, Wooldridge L et al (2008) Profound inhibition of antigen-specific T-cell effector functions by dasatinib. Clin Cancer Res 14(8):2484–2491

    CAS  CrossRef  PubMed  Google Scholar 

  4. Wong FS, Karttunen J, Dumont C et al (1999) Identification of an MHC class I-restricted autoantigen in type 1 diabetes by screening an organ-specific cDNA library. Nat Med 5(9):1026–1031

    CAS  CrossRef  PubMed  Google Scholar 

  5. Lieberman SM, Evans AM, Han B et al (2003) Identification of the beta cell antigen targeted by a prevalent population of pathogenic CD8+ T cells in autoimmune diabetes. Proc Natl Acad Sci U S A 100(14):8384–8388

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  6. Wong FS, Siew LK, Scott G et al (2009) Activation of insulin-reactive CD8 T-cells for development of autoimmune diabetes. Diabetes 58(5):1156–1164

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgement

This work was supported by a Medical Research Council (grant number G0901155) to FSW and a Diabetes UK PhD studentship to JAP. We acknowledge the NIH Tetramer Core Facility (contract HHSN272201300006C) for provision of MHC | peptide tetramers.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to F. Susan Wong .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Pearson, J.A., Wong, F.S. (2015). Identification of Islet Antigen-Specific CD8 T Cells Using MHCI-Peptide Tetramer Reagents in the Non Obese Diabetic (NOD) Mouse Model of Type 1 Diabetes. In: Gillespie, K. (eds) Type-1 Diabetes. Methods in Molecular Biology, vol 1433. Humana Press, New York, NY. https://doi.org/10.1007/7651_2015_295

Download citation

  • DOI: https://doi.org/10.1007/7651_2015_295

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3641-0

  • Online ISBN: 978-1-4939-3643-4

  • eBook Packages: Springer Protocols