Immuno-monitoring of CD8+ T cells in whole blood versus PBMC samples.
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.
Appay
Victor
V
Immunology Group, Multidisciplinary Oncology Center, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland. victor.appay@chups.jussieu.fr
Reynard
Severine
S
Voelter
Verena
V
Romero
Pedro
P
Speiser
Daniel E
DE
Leyvraz
Serge
S
D001773:Blood Cells / Q000166:cytology / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D019496:Cancer Vaccines / Q000627:therapeutic use; D002469:Cell Separation; D005434:Flow Cytometry; D006801:Humans; D007113:Immunity, Innate; D007167:Immunotherapy; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D008137:Longitudinal Studies; D008545:Melanoma / Q000276:immunology / Q000628:therapy; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology
D019496:Cancer Vaccines