LiveCount Assay: concomitant measurement of cytolytic activity and phenotypic characterisation of CD8(+) T-cells by flow cytometry

Journal of Immunological Methods
Estelle DevêvreYolanda D Mahnke

Abstract

Tumour immunologists strive to develop efficient tumour vaccination and adoptive transfer therapies that enlarge the pool of tumour-specific and -reactive effector T-cells in vivo. To assess the efficiency of the various strategies, ex vivo assays are needed for the longitudinal monitoring of the patient's specific immune responses providing both quantitative and qualitative data. In particular, since tumour cell cytolysis is the end goal of tumour immunotherapy, routine immune monitoring protocols need to include a read-out for the cytolytic efficiency of Ag-specific cells. We propose to combine current immune monitoring techniques in a highly sensitive and reproducible multi-parametric flow cytometry based cytotoxicity assay that has been optimised to require low numbers of Ag-specific T-cells. The possibility of re-analysing those T-cells that have undergone lytic activity is illustrated by the concomitant detection of CD107a upregulation on the surface of degranulated T-cells. To date, the LiveCount Assay provides the only possibility of assessing the ex vivo cytolytic activity of low-frequency Ag-specific cytotoxic T-lymphocytes from patient material.

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Citations

Jul 24, 2010·Cancer Immunology, Immunotherapy : CII·Sharmila Pejawar-GaddyIoannis Bossis
May 12, 2011·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Lisa H ButterfieldMary L Disis
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Jun 24, 2011·Cancer Research·Cláudia Trindade NunesStephen Man
Oct 15, 2011·Journal of Immunological Methods·Florentin-Martial Mbitikon-KoboLydie Trautmann

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