Analysis of PBMC Activation Using Parallel Bead and Cellular Assays

Abstract

Abstract Activation of human peripheral blood mononuclear cells (PBMCs) leads to expression of cytokine receptors, cytokine secretion and the expression of specific cell surface molecules resulting in divergent immune responses. The correlation of cell surface activation and cellular health markers from the lymphocyte subpopulations with cytokine levels can vary across donors and can provide valuable information to evaluate secretion levels. Bead-based multiplexed assays using Luminex xMAP® platforms provide a powerful tool for quantifying and studying several secreted cytokine levels in a single assay. Performing cellular assays performed in with flow cytometry provide information in immune cell population percentages, counts, and overall health. In this study, we present the combined cellular and secreted cytokine data on human PBMCs stimulated by the lectin phytohaemagglutinin (PHA) or the endotoxin lipopolysaccharides (LPS). Time course data from multiple donors will be presented. Combining data from these two approaches gives a comprehensive picture of the impact of stimulation on the various PBMC populations and the correlation of the secreted cytokine levels to the number of cells.