TY  - EJOU
AU  - Burel, Julie G. 
AU  - Apte, Simon H. 
AU  - Doolan, Denise L. 

TI  - Development of a cytokine-secreting-based assay for the identification, sorting and transcriptomic analysis of polyfunctional human T cells
T2  - European Cytokine Network

PY  - 2015
VL  - 26
IS  - 4
SN  - 1952-4005

AB  - Polyfunctional T cells that simultaneously produce the cytokines IFN-γ, IL-2 and TNF have been
correlated with better clinical outcomes in various diseases. To date, cytokine polyfunctionality within T cells has
been exclusively studied by intracellular cytokine staining coupled with flow cytometric analysis. Thus, further
downstream interrogation of polyfunctional T cell characteristics such as transcriptomic analysis has not been
possible. Here, we report the use of a flow cytometric method based on cytokine secretion assay technology to detect
and isolate, for the first time, viable human polyfunctional T cells directly from in vitro stimulated whole blood
samples. We demonstrate the successful application of this method to sort polyfunctional T cells obtained from
human volunteers, which can be then used for downstream applications such as transcriptomic analysis using RT-qPCR.
This assay will facilitate in-depth investigations of T cells with distinct cytokine polyfunctionality, including
defining their molecular profile and understanding the mechanisms regulating their generation and function.
KW  - polyfunctional T cells
KW  -  cytokine secretion
KW  -  novel assay
KW  -  cell sorting
KW  -  transcriptomic analysis

DO  - 10.1684/ecn.2015.0369