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Immunogenetics pp 339-349 | Cite as

Detection of Allo-HLA Cross-Reactivity by Virus-specific Memory T-Cell Clones Using Single HLA-Transfected K562 Cells

  • Lloyd J. D’Orsogna
  • Ellen M. W. van der Meer-Prins
  • Yvonne M. Zoet
  • Dave L. Roelen
  • Ilias I. N. Doxiadis
  • Frans H. J. Claas
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 882)

Abstract

The ability to directly measure virus-specific lymphocytes using fluorochrome-labeled tetrameric complexes has proven a great advancement for the transplantation field. Viral peptide/HLA tetrameric complexes allow the rapid generation of virus-specific clones using single cell sorting apparatus, permitting the determination of alloreactivity from a single TCR with known specificity. When combined with new target “detector” cells called single HLA antigen-transfected K562 cells (SALs), the human alloresponse can for the first time be examined specifically and reliably. Here we describe a method for detection of “heterologous immunity” from virus-specific memory T-cells using single HLA expressing cell lines as allogeneic targets.

Key words

Heterologous immunity Memory T-cells Viral immunity Alloreactivity SALs 

References

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Lloyd J. D’Orsogna
    • 1
    • 2
  • Ellen M. W. van der Meer-Prins
    • 1
  • Yvonne M. Zoet
    • 1
  • Dave L. Roelen
    • 1
  • Ilias I. N. Doxiadis
    • 1
  • Frans H. J. Claas
    • 1
  1. 1.Department of Immunohematology and Blood TransfusionLeiden University Medical CentreLeidenThe Netherlands
  2. 2.PathWest Laboratory MedicinePerthAustralia

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