Citations : Multi-parametric Human Cell-Based Inflammation Assay for Cytokines and Cell Surface Antigens

Dated: May 01, 2018

Publication Name: The Journal of Immunology

We present results from a multiparametric cell-based assay that uses immunoassays for secreted cytokines and fluorescence read-outs of cell surface markers to evaluate the effect of different mediators on inflammatory response. Primary human umbilical vein endothelial cells (HUVEC) were stimulated with inflammation cytokines (TNF-α, IFN-γ, and IL-1β) for 20 hours. Then, a newly developed microfluidic-based assay system, PuMA, was used to quantify amount of MCP-1, IL-8, and IL-6 in cell supernatants. Expression of adhesion molecules VCAM and HLADR was quantified by measurement of total fluorescence intensity after staining cells with directly conjugated antibodies using an ImageXpress Nano Automated Imaging System. Concentration dependent inhibition of the inflammation responses by the compounds AG126 and MG132 were measured. Clear differences in cytokine expression were seen between the compounds consistent with their reported mechanisms of action. The combination of imaging and microfluidic-based assays provides an efficient multiparametric assay system that can be used to test the efficacy of anti-inflammatory compounds versus toxicity and also provide significant insight into the mechanism of action by selective inhibition of markers triggered by different signaling pathways.

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Contributors: Evan Cromwell, Ori Hoxha, Matthew Hammer and Oksana Sirenko