Profiling T cell activation using single-molecule fluorescence in situ hybridization and flow cytometry.

ABSTRACT: Flow cytometric characterization of Ag-specific T cells typically relies on detection of protein analytes. Shifting the analysis to detection of RNA would provide several significant advantages, which we illustrate by developing a new host immunity-based platform for detection of infections. Cytokine mRNAs synthesized in response to ex vivo stimulation with pathogen-specific Ags are detected in T cells with single-molecule fluorescence in situ hybridization followed by flow cytometry. Background from pre-existing in vivo analytes is lower for RNAs than for proteins, allowing greater sensitivity for detection of low-frequency cells. Moreover, mRNA analysis reveals kinetic differences in cytokine expression that are not apparent at the protein level but provide novel insights into gene expre

SUBMITTER: Bushkin Y 

PROVIDER: S-EPMC4350577 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature