1. Miniaturized High-Throughput Multiparameter Flow Cytometry Assays Measuring In Vitro Human Dendritic Cell Maturation and T-Cell Activation in Mixed Lymphocyte Reactions.
- Author
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Fan Y, Naglich JG, Koenitzer JD, Ribeiro H, Lippy J, Blum J, Li X, Milburn C, Barnhart B, Zhang L, and Fereshteh MP
- Subjects
- Cytokines metabolism, Dendritic Cells immunology, Humans, T-Lymphocytes immunology, Dendritic Cells metabolism, Flow Cytometry methods, High-Throughput Screening Assays, Lymphocyte Activation immunology, Lymphocyte Culture Test, Mixed, T-Lymphocytes metabolism
- 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.
- Published
- 2018
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