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Tracking Antigen-Specific CD8+ T Cells Using MHC Class I Multimers

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Antigen Processing

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 960))

Abstract

The tracking of epitope-specific T cells is a useful approach for the study of adaptive immune responses. This protocol describes how Major Histocompatibility Complex Class I (MHC-I) multimers can be used to stain, enrich, and enumerate (rare) populations of CD8+ T cells specific for a given antigen. It provides the detailed steps for multimer labeling, magnetic enrichment, and cytometric analysis. Additionally, it provides informations for multiplexing experiments in order to achieve simultaneous detection of multiple antigenic specificities, and strategies for coupling the protocol with functional assays (e.g., intracellular cytokine staining). Future developments in cytometric systems (e.g., mass spectroscopy-based cytometry) and gene expression studies (e.g., single cell PCR) will extend these approaches and provide an unprecedented assessment of the immune repertoire.

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References

  1. Arens R, Schoenberger SP (2010) Plasticity in programming of effector and memory CD8 T-cell formation. Immunol Rev 235(1):190–205. doi:10.1111/j.0105-2896.2010.00899.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Schumacher TNM, Gerlach C, Heijst JW (2010) Mapping the life histories of T cells. Nat Pub Group 10(9):621–631. doi:10.1038/nri2822

    CAS  Google Scholar 

  3. Coulie PG, Karanikas V, Lurquin C, Colau D, Connerotte T, Hanagiri T, Van Pel A, Lucas S, Godelaine D, Lonchay C, Marchand M, Van Baren N, Boon T (2002) Cytolytic T-cell responses of cancer patients vaccinated with a MAGE antigen. Immunol Rev 188:33–42

    Article  CAS  PubMed  Google Scholar 

  4. Lamoreaux L, Roederer M, Koup R (2006) Intracellular cytokine optimization and standard operating procedure. Nat Protoc 1(3):1507–1516. doi:10.1038/nprot.2006.268

    Article  CAS  PubMed  Google Scholar 

  5. Sun Y, Iglesias E, Samri A, Kamkamidze G, Decoville T, Carcelain G, Autran B (2003) A systematic comparison of methods to measure HIV-1 specific CD8 T cells. J Immunol Methods 272(1–2):23–34

    Article  CAS  PubMed  Google Scholar 

  6. Altman JD, Moss PA, Goulder PJ, Barouch DH, McHeyzer-Williams MG, Bell JI, McMichael AJ, Davis MM (1996) Phenotypic analysis of antigen-specific T lymphocytes. Science 274(5284):94–96

    Article  CAS  PubMed  Google Scholar 

  7. Altman JD, Davis MM (2003) MHC-peptide tetramers to visualize antigen-specific T cells. Curr Protoc Immunol Chapter 17:Unit 17.13. 10.1002/0471142735.im1703s53

  8. Davis MM, Altman JD, Newell EW (2011) Interrogating the repertoire: broadening the scope of peptide–MHC multimer analysis. Nat Rev Immunol 11(8):551–558. doi:10.1038/nri3020

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Klenerman P, Cerundolo V, Dunbar PR (2002) Tracking T cells with tetramers: new tales from new tools. Nat Rev Immunol 2(4):263–272. doi:10.1038/nri777

    Article  CAS  PubMed  Google Scholar 

  10. Romero P, Valmori D, Pittet MJ, Zippelius A, Rimoldi D, Lévy F, Dutoit V, Ayyoub M, Rubio-Godoy V, Michielin O, Guillaume P, Batard P, Luescher IF, Lejeune F, Liénard D, Rufer N, Dietrich P-Y, Speiser DE, Cerottini J-C (2002) Antigenicity and immunogenicity of Melan-A/MART-1 derived peptides as targets for tumor reactive CTL in human melanoma. Immunol Rev 188:81–96

    Article  CAS  PubMed  Google Scholar 

  11. Baitsch L, Baumgaertner P, Devevre E, Raghav SK, Legat A, Barba L, Wieckowski S, Bouzourene H, Deplancke B, Romero P, Rufer N, Speiser DE (2011) Exhaustion of tumor-specific CD8+ T cells in metastases from melanoma patients. J Clin Invest 121(6):2350–2360. doi:10.1172/JCI46102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Kanodia S, Wieder E, Lu S, Talpaz M, Alatrash G, Clise-Dwyer K, Molldrem JJ (2010) PR1-specific T cells are associated with unmaintained cytogenetic remission of chronic myelogenous leukemia after interferon withdrawal. PLoS One 5(7):e11770. doi:10.1371/journal.pone.0011770

    Article  PubMed  PubMed Central  Google Scholar 

  13. Rosenberg SA, Dudley ME, Restifo NP (2008) Cancer immunotherapy. N Engl J Med 359(10):1072. doi:10.1056/NEJMc081511

    Article  CAS  PubMed  Google Scholar 

  14. Appay V, Douek DC, Price DA (2008) CD8+ T cell efficacy in vaccination and disease. Nat Med 14(6):623–628. doi:10.1038/nm.f.1774

    Article  CAS  PubMed  Google Scholar 

  15. Ma Q, Wang C, Jones D, Quintanilla KE, Li D, Wang Y, Wieder ED, Clise-Dwyer K, Alatrash G, Mj Y, Munsell MF, Lu S, Qazilbash MH, Molldrem JJ (2010) Adoptive transfer of PR1 cytotoxic T lymphocytes associated with reduced leukemia burden in a mouse acute myeloid leukemia xenograft model. Cytotherapy 12(8):1056–1062. doi:10.3109/14653249.2010.506506

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Wooldridge L, Lissina A, Cole DK, van den Berg HA, Price DA, Sewell AK (2009) Tricks with tetramers: how to get the most from multimeric peptide-MHC. Immunology 126(2):147–164. doi:10.1111/j.1365-2567.2008.02848.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Bakker AH, Schumacher TNM (2005) MHC multimer technology: current status and future prospects. Curr Opin Immunol 17(4):428–433. doi:10.1016/j.coi.2005.06.008

    Article  CAS  PubMed  Google Scholar 

  18. Toebes M, Coccoris M, Bins A, Rodenko B, Gomez R, Nieuwkoop NJ, Van De Kasteele W, Rimmelzwaan GF, Haanen JBAG, Ovaa H, Schumacher TNM (2006) Design and use of conditional MHC class I ligands. Nat Med 12(2):246–251. doi:10.1038/nm1360

    Article  CAS  PubMed  Google Scholar 

  19. Hadrup SR, Toebes M, Rodenko B, Bakker AH, Egan DA, Ovaa H, Schumacher TNM (2009) High-throughput T-cell epitope discovery through MHC peptide exchange. Methods Mol Biol 524:383–405. doi:10.1007/978-1-59745-450-6_28

    Article  CAS  PubMed  Google Scholar 

  20. Newell EW, Klein LO, Yu W, Davis MM (2009) Simultaneous detection of many T-cell specificities using combinatorial tetramer staining. Nat Pub Group 6(7):497–499. doi:10.1038/nmeth.1344

    CAS  Google Scholar 

  21. Hadrup SR, Bakker AH, Shu CJ, Andersen RS, van Veluw J, Hombrink P, Castermans E, Thor Straten P, Blank C, Haanen JB, Heemskerk MH, Schumacher TN (2009) Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers. Nat Methods 6(7):520–526. doi:10.1038/nmeth.1345

    Article  CAS  PubMed  Google Scholar 

  22. Chattopadhyay PK, Melenhorst JJ, Ladell K, Gostick E, Scheinberg P, Barrett AJ, Wooldridge L, Roederer M, Sewell AK, Price DA (2008) Techniques to improve the direct ex vivo detection of low frequency antigen-specific CD8+ T cells with peptide-major histocompatibility complex class I tetramers. Cytometry A 73(11):1001–1009. doi:10.1002/cyto.a.20642

    Article  PubMed  PubMed Central  Google Scholar 

  23. Alanio C, Lemaitre F, Law HKW, Hasan M, Albert ML (2010) Enumeration of human antigen-specific naive CD8+ T cells reveals conserved precursor frequencies. Blood 115(18):3718–3725. doi:10.1182/blood-2009-10-251124

    Article  CAS  PubMed  Google Scholar 

  24. Day CL (2003) Ex vivo analysis of human memory CD4 T cells specific for hepatitis C virus using MHC class II tetramers. J Clin Invest 112(6):831–842. doi:10.1172/JCI200318509

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Moon J, Chu H, Pepper M, Mcsorley S, Jameson S, Kedl R, Jenkins M (2007) Naive CD4+ T cell frequency varies for different epitopes and predicts repertoire diversity and response magnitude. Immunity 27(2):203–213. doi:10.1016/j.immuni.2007.07.007

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Moon JJ, Chu HH, Hataye J, Pagán AJ, Pepper M, Mclachlan JB, Zell T, Jenkins MK (2009) Tracking epitope-specific T cells. Nat Protoc 4(4):565–581. doi:10.1038/nprot.2009.9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Obar J, Khanna K, Lefrancois L (2008) Endogenous naive CD8+ T cell precursor frequency regulates primary and memory responses to infection. Immunity 28(6):859–869. doi:10.1016/j.immuni.2008.04.010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Legoux F, Debeaupuis E, Echasserieau K, De La Salle H, Saulquin X, Bonneville M (2010) Impact of TCR reactivity and HLA phenotype on naive CD8 T cell frequency in humans. J Immunol 184(12):6731–6738. doi:10.4049/jimmunol.1000295

    Article  CAS  PubMed  Google Scholar 

  29. Jenkins MK, Chu HH, Mclachlan JB, Moon JJ (2010) On the composition of the preimmune repertoire of T cells specific for peptide–major histocompatibility complex ligands. Annu Rev Immunol 28(1):275–294. doi:10.1146/annurev-immunol-030409-101253

    Article  CAS  PubMed  Google Scholar 

  30. Pittet MJ, Valmori D, Dunbar PR, Speiser DE, Liénard D, Lejeune F, Fleischhauer K, Cerundolo V, Cerottini JC, Romero P (1999) High frequencies of naive Melan-A/MART-1-specific CD8(+) T cells in a large proportion of human histocompatibility leukocyte antigen (HLA)-A2 individuals. J Exp Med 190(5):705–715

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Roederer M (2008) How many events is enough? Are you positive? Cytometry 73A(5):384–385. doi:10.1002/cyto.a.20549

    Article  Google Scholar 

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Authors and Affiliations

  1. Immunobiologie des cellules dendritiques, Unité Mixte Pasteur/Inserm U818, Institut Pasteur and Centre d’Immunologie Humaine, Institut Pasteur, Paris, France

    Cécile Alanio, Isabelle Bouvier, Hélène Jusforgues-Saklani & Matthew L. Albert

Authors
  1. Cécile Alanio
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  2. Isabelle Bouvier
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  3. Hélène Jusforgues-Saklani
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  4. Matthew L. Albert
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Corresponding author

Correspondence to Matthew L. Albert .

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Editors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale, Unité 1013, Université Paris Descartes, Faculté de médecine, Paris, France

    Peter van Endert

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Alanio, C., Bouvier, I., Jusforgues-Saklani, H., Albert, M.L. (2013). Tracking Antigen-Specific CD8+ T Cells Using MHC Class I Multimers. In: van Endert, P. (eds) Antigen Processing. Methods in Molecular Biology™, vol 960. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-218-6_23

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  • DOI: https://doi.org/10.1007/978-1-62703-218-6_23

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-217-9

  • Online ISBN: 978-1-62703-218-6

  • eBook Packages: Springer Protocols

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