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Tracking Antigen-Experienced Effector T Cells In Vitro and In Vivo

  • Claire L. Gorman
  • Claudia Monaco
  • Enrico Ammiratti
  • Anna-Chiara Vermi
  • Federica M. Marelli-Berg
  • Andrew P. Cope
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 616)

Abstract

The TCR complex is a multisubunit complex, comprising at least eight transmembrane units. The clonotypic TCR α and β chains are responsible for antigen recognition, whilst the invariant chains of the CD3 complex (δ, ɛ and γ) and two zeta (ζ) polypeptides couple antigen recognition to downstream signal transduction pathways. TCRζ (CD247) functions as an amplification module in the TCR signalling cascade and is also essential for the assembly and surface expression of the TCR/CD3 complex. Loss of TCRζ expression is common in chronic infectious and inflammatory diseases, as well as in cancer. Previous work has indicated that TCRζlow-expressing cells phenotypically resemble antigen-experienced effector T cells. Here, we describe the derivation of a flow cytometry-based TCRζ expression index for the purpose of more precisely defining TCRζ expression, in addition to utilising a simple transmigration assay in the demonstration that TCRζdim T cells have intrinsic migratory properties that may explain their accumulation at sites of inflammation.

Key words

T cells T-cell receptor zeta flow cytometry cell migration 

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Claire L. Gorman
    • 1
  • Claudia Monaco
    • 1
  • Enrico Ammiratti
    • 2
  • Anna-Chiara Vermi
    • 2
  • Federica M. Marelli-Berg
    • 3
  • Andrew P. Cope
    • 4
  1. 1.Faculty of MedicineImperial College London, The Kennedy Institute of RheumatologyLondonUK
  2. 2.Clinical Cardiovascular Biology Research Centre, Vita-Salute San Raffaele University and San Raffaele Scientific InstituteMilanItaly
  3. 3.Division of Medicine, Department of Immunology, Faculty of MedicineImperial College London, Hammersmith Hospital CampusLondonUK
  4. 4.The Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College LondonLondonUK

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