TY - JOUR
T1 - Immuno-monitoring of CD8+ T cells in whole blood versus PBMC samples
AU - Appay, Victor
AU - Reynard, Severine
AU - Voelter, Verena
AU - Romero, Pedro
AU - Speiser, Daniel E.
AU - Leyvraz, Serge
N1 - Funding Information:
We thank the donors for study participation and blood donation. This work was sponsored by Fond'action Contre le Cancer, the Ludwig Institute for Cancer Research, the Nelia et Amadeo Barletta Foundation, and by the National Center of Competence in Research (NCCR) Molecular Oncology.
PY - 2006/2/20
Y1 - 2006/2/20
N2 - 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.
AB - 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.
KW - Flow cytometry
KW - Lymphocytes
KW - Naïve
KW - Tetramers
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U2 - 10.1016/j.jim.2005.11.007
DO - 10.1016/j.jim.2005.11.007
M3 - Article
C2 - 16403411
AN - SCOPUS:31944438409
SN - 0022-1759
VL - 309
SP - 192
EP - 199
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -