Combining impedence-based viability measurements and flow cytometric analyte quantitation to evaluate effector cell killing of T lymphocytes

Abstract

Abstract Cancer immunotherapy is increasingly being evaluated in the lab, as well as the clinic as an approach for cancer treatment by harnessing and enhancing the immune system to attack cancer cells. Moving this research from the bench to the clinic is critically important, but we need reliable tools to be able to translate the in-vitro protocols and results for use in-vivo. Here, we have used a combination of impedence-based technology and flow cytometry to evaluate both the target and effector cells in an immune-cell based killing assay. The impedence based Real Time Cell Analysis technology, provides us with a continuous readout of target cell viability and flow cytometry allows us to measure responses using specific labels, such as effector cell viability and cytokine production in the supernatant of these assays. Here, we demonstrate cell killing assays measuring the cytotoxic effect of T-lymphocytes on a B-cancer cell line in conjunction with cytokine concentration measurements of IL-2, IL-4, IL-5, IL-10, TNF-a, IFN-g, IL-8, IL-1b, IL-6, IL-10, and IL-12p70 to simultaneously understand the T-lymphocyte response in these assays. Using both of these methods allows us to thoroughly evaluate immune cell killing efficacy and kinetics. The potency assays described in these studies have great potential for use in-vivo models and clinic.