Immuno-monitoring of CD8+ T cells in whole blood versus PBMC samples

Victor Appay*, Severine Reynard, Verena Voelter, Pedro Romero, Daniel E. Speiser, Serge Leyvraz

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

The study of natural T cell responses against pathogens or tumors, as well as the assessment of new immunotherapy strategies aimed at boosting these responses, requires increasingly precise ex vivo analysis of blood samples. For practical reasons, studies are often performed using purified PBMC samples, usually cryopreserved. Here, we report on FACS analyses of peripheral blood T cells, performed by direct antibody staining of non-purified total blood. For comparison, fresh PBMC, purified by Ficoll, were analysed. Our results show that the latter method can induce a bias in subpopulation distribution, in particular of CD8+ T cells, and sometimes lead to inaccurate measurement of antigen specific CD8+ T cell responses. Direct analysis of total blood can be applied to longitudinal immuno-monitoring of T cell-based therapy. While the need to purify and cryopreserve PBMC for subsequent studies is obvious, the use of whole blood has the advantage of providing unbiased results and only small amounts of blood are used.

Original languageEnglish (US)
Pages (from-to)192-199
Number of pages8
JournalJournal of Immunological Methods
Volume309
Issue number1-2
DOIs
StatePublished - Feb 20 2006
Externally publishedYes

Keywords

  • Flow cytometry
  • Lymphocytes
  • Naïve
  • Tetramers

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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