@Article{ecn.2015.0369,
AUTHOR = {Julie G. Burel, Simon H. Apte, Denise L. Doolan},
TITLE = {Development of a cytokine-secreting-based assay for the identification, sorting and transcriptomic analysis of polyfunctional human T cells},
JOURNAL = {European Cytokine Network},
VOLUME = {26},
YEAR = {2015},
NUMBER = {4},
PAGES = {67--72},
URL = {http://www.techscience.com/ECN/v26n4/65544},
ISSN = {1952-4005},
ABSTRACT = {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.},
DOI = {10.1684/ecn.2015.0369}
}