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Visualizing Endogenous Effector T Cell Egress from the Lymph Nodes

Part of the Methods in Molecular Biology book series (MIMB,volume 1591)

Abstract

Local anatomy of lymphoid tissues during infection has emerged as a critical regulator of immunity; thus, studying the cellular choreography in the context of an intact tissue environment in situ is crucial. Following an infection, the local pathogen-specific T cell migration and the subsequent egress of effector T cells from the draining lymph nodes are important and complex biological processes. The mechanisms that regulate this complex process can now be investigated by directly visualizing T cell dynamics in vivo using intravital two-photon (2P) microscopy. In addition, static whole-mount imaging technique can provide us with a comprehensive assessment of global changes in the distribution of cellular populations within an intact tissue. Thus, in this chapter, we detail methods to visualize the migration and egress of endogenous antigen-specific CD8 T cells following viral infection using two methods—intravital 2P microscopy and multicolor whole-mount in situ tetramer staining.

Keywords

  • T cell egress
  • Sphingosine-1-phosphate receptor
  • Intravital microscopy
  • In situ tetramer staining

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  • DOI: 10.1007/978-1-4939-6931-9_5
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References

  1. Cyster JG, Schwab SR (2012) Sphingosine-1-phosphate and lymphocyte egress from lymphoid organs. Annu Rev Immunol 30:69–94

    CAS  CrossRef  PubMed  Google Scholar 

  2. Schulz O, Hammerschmidt SI, Moschovakis GL, Forster R (2016) Chemokines and chemokine receptors in lymphoid tissue dynamics. Annu Rev Immunol 34:203–242

    CrossRef  PubMed  Google Scholar 

  3. Grigorova IL, Schwab SR, Phan TG, Pham TH, Okada T, Cyster JG (2009) Cortical sinus probing, S1P1-dependent entry and flow-based capture of egressing T cells. Nat Immunol 10:58–65

    CAS  CrossRef  PubMed  Google Scholar 

  4. Pham TH, Okada T, Matloubian M, Lo CG, Cyster JG (2008) S1P1 receptor signaling overrides retention mediated by G alpha i-coupled receptors to promote T cell egress. Immunity 28:122–133

    CAS  CrossRef  PubMed  Google Scholar 

  5. Benechet AP, Menon M, Xu D, Samji T, Maher L, Murooka TT, Mempel TR, Sheridan BS, Lemoine FM, Khanna KM (2016) T cell-intrinsic S1PR1 regulates endogenous effector T-cell egress dynamics from lymph nodes during infection. Proc Natl Acad Sci U S A 113:2182–2187

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  6. Mionnet C, Sanos SL, Mondor I, Jorquera A, Laugier JP, Germain RN, Bajenoff M (2011) High endothelial venules as traffic control points maintaining lymphocyte population homeostasis in lymph nodes. Blood 118:6115–6122

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  7. Schulz O, Ugur M, Friedrichsen M, Radulovic K, Niess JH, Jalkanen S, Krueger A, Pabst O (2014) Hypertrophy of infected Peyer's patches arises from global, interferon-receptor, and CD69-independent shutdown of lymphocyte egress. Mucosal Immunol 7:892–904

    CAS  PubMed  Google Scholar 

  8. Marzo AL, Klonowski KD, Le Bon A, Borrow P, Tough DF, Lefrancois L (2005) Initial T cell frequency dictates memory CD8+ T cell lineage commitment. Nat Immunol 6:793–799

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  9. Brinkmann V, Cyster JG, Hla T (2004) FTY720: sphingosine 1-phosphate receptor-1 in the control of lymphocyte egress and endothelial barrier function. Am J Transplant 4:1019–1025

    CAS  CrossRef  PubMed  Google Scholar 

  10. Bannard O, Kraman M, Fearon DT (2009) Secondary replicative function of CD8+ T cells that had developed an effector phenotype. Science 323:505–509

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  11. Mempel TR (2010) Single-cell analysis of cytotoxic T cell function by intravital multiphoton microscopy. Methods Mol Biol 616:181–192

    CAS  CrossRef  PubMed  Google Scholar 

  12. Murooka TT, Mempel TR (2012) Multiphoton intravital microscopy to study lymphocyte motility in lymph nodes. Methods Mol Biol 757:247–257

    CrossRef  PubMed  Google Scholar 

  13. Nolz JC, Starbeck-Miller GR, Harty JT (2011) Naive, effector and memory CD8 T-cell trafficking: parallels and distinctions. Immunotherapy 3:1223–1233

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  14. Sarkisyan G, Cahalan SM, Gonzalez-Cabrera PJ, Leaf NB, Rosen H (2012) Real-time differential labeling of blood, interstitium, and lymphatic and single-field analysis of vasculature dynamics in vivo. Am J Physiol Cell Physiol 302:C1460–C1468

    CAS  CrossRef  PubMed  Google Scholar 

  15. Benechet AP, Menon M, Khanna KM (2014) Visualizing T cell migration in-situ. Front Immunol 5:363

    CrossRef  PubMed  PubMed Central  Google Scholar 

  16. Khanna KM, Bonneau RH, Kinchington PR, Hendricks RL (2003) Herpes simplex virus-specific memory CD8+ T cells are selectively activated and retained in latently infected sensory ganglia. Immunity 18:593–603

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  17. Khanna KM, McNamara JT, Lefrancois L (2007) In situ imaging of the endogenous CD8 T cell response to infection. Science 318:116–120

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  18. Chiba K, Adachi K (2012) Discovery of fingolimod, the sphingosine 1-phosphate receptor modulator and its application for the therapy of multiple sclerosis. Future Med Chem 4:771–781

    CAS  CrossRef  PubMed  Google Scholar 

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Correspondence to Kamal M. Khanna .

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Menon, M., Benechet, A.P., Khanna, K.M. (2017). Visualizing Endogenous Effector T Cell Egress from the Lymph Nodes. In: Rainger, G., Mcgettrick, H. (eds) T-Cell Trafficking. Methods in Molecular Biology, vol 1591. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6931-9_5

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  • DOI: https://doi.org/10.1007/978-1-4939-6931-9_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6929-6

  • Online ISBN: 978-1-4939-6931-9

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