PrimeFlow™ RNA assay as a method to elucidate the effect of protein transport inhibitors on cytokine gene expression in whole blood (TECH3P.941)

Abstract

Abstract Intracellular antibody staining followed by flow cytometric analysis is commonly used to study cytokine expression in leukocytes. Detection of cytokines typically requires that cells be treated with protein transport inhibitors such as Brefeldin A and Monensin in order to accumulate sufficient levels of intracellular protein. However, it is not known whether the use of these inhibitors affects gene expression. PrimeFlow™ RNA is a new flow cytometric assay that allows the simultaneous interrogation of RNA and protein levels within single cells. We have used this assay to investigate the effect of protein transport inhibitors on the expression of human IL-2, TNF alpha and IFN gamma during the course of a 5 hour stimulation of whole blood with PMA and ionomycin. This represents first time that the effect of protein transport inhibitors on gene expression in whole blood has been examined in single cells.