Miniaturized High-Throughput Multiparameter Flow Cytometry Assays Measuring In Vitro Human Dendritic Cell Maturation and T-Cell Activation in Mixed Lymphocyte Reactions. (August 2018)

• Record Type: Journal Article
• Authors: Fan, Yi; Naglich, Joseph G.; Koenitzer, Jennifer D.; Ribeiro, Humberto; Lippy, Jonathan; Blum, Jordan; Li, Xin; Milburn, Christina; Barnhart, Bryan; Zhang, Litao; Fereshteh, Mark P.
• Other Names: Ding Mei guest-editor.; Edwards Bruce S. guest-editor.
• Is Part Of: SLAS discovery. Volume 23:Number 7(2018)
• Page Start: 742
• Page End: 750
• Publication Date: 2018-08
• Subjects: MLR -- mixed lymphocyte reaction -- flow cytometry -- dendritic cell -- T cell -- IntelliCyt; Drugs -- Analysis -- Periodicals; Drugs -- Testing -- Periodicals; Biomolecules -- Analysis -- Periodicals; Biomolecules -- Analysis; Drugs -- Analysis; Drugs -- Testing; Drug Evaluation, Preclinical; Molecular Biology -- methods; Periodicals; Periodicals; 615.1
• DOI: 10.1177/2472555218775409
• Languages: English
• ISSNs: 2472-5552

Abstract

Enhancing antitumor activities of the human immune system is a clinically proven approach with the advent of monoclonal antibodies recognizing programmed cell death protein-1 (PD1) receptors on immune cell surfaces. Historically, using flow cytometry as a means to assess next-generation agent activities was underused, largely due to limits on cell number and assay sensitivity. Here, we leveraged an IntelliCyt high-throughput flow cytometry platform to monitor human dendritic cell maturation and lymphocyte proliferation in mixed lymphocyte reactions. Specifically, we established flow cytometry–based immunophenotyping and screening methodologies capable of measuring T-cell activation as a result of cell-associated antigens presented on dendritic cell surfaces, as indicated by cell proliferation, cytokine secretion, and surface marker expression. Together, the overall novelty of this 384-well platform is its capability to measure multiple functional readouts in one well and consistently evaluate large numbers of compounds in a single study, as well as its ability to show increased assay sensitivity requiring considerably fewer primary cells and less reagents compared to more traditional 96-well flow cytometry methods.