Profiling Cytokine Levels Using a Multiplex Human Cytokine ELISA Array (94.26)

Abstract

Abstract The induction of multiple cytokines is a common phenomenon in many immunological responses; therefore, a method to quickly and simultaneously screen in vitro or in vivo samples for multiple cytokine levels is needed. We developed a “multiplex” sandwich ELISA Array on a 96-well plate allowing researchers to screen several human serum, plasma or cell culture supernatant samples for multiple cytokines in one experiment. The ELISA Array is easy to perform and only requires a standard ELISA plate reader. It can be also used in conjunction with corresponding single analyte ELISA kits to perform validation and absolute quantification. As an illustrative example, we monitored the time-dependent (0, 6, 24, and 48 hours) patterns of Th1/Th2 cytokine induction by human peripheral blood mononuclear cells (PBMC) in response to PMA (50 ng/ml) and ionomycin (1 μg/ml). Large amounts of IL-2, IFN-gamma and TNF-alpha are induced after the 6-h stimulation when only more moderate amounts of IL-4, IL-5, IL-10 and IL-13 are secreted. IFN-gamma levels are the highest among the eight cytokines tested after 6 hours, while IL-2 has the highest level after 48 hours. The amount of IL-12 p70 is not detectable even after 48 hours. The ELISA Array thus provides a powerful, easy-to-use solution for profiling the relative levels of several cytokines across multiple experimental conditions at the same time.