Abstract
It is unknown if surface bound toll-like-receptor (TLR) agonists activate cells via density or total molecular number. To answer this question, we developed a TLR agonist surface conjugated polystyrene microparticle (MP) system. Using a library of MPs with varying TLR agonist density and number, we simultaneously observed innate immune cell MP uptake and TNFα expression using ImageStream flow cytometry on a cell by cell basis. The data shows that total TLR number and not density drives cellular activation with a threshold of approximately 105-106 TLR agonists. We believe that this information will be crucial for the design of particulate vaccine formulations.
Keywords:
In vitro quantification; activation threshold; innate immunity; microparticle; toll-like-receptor.
Copyright © 2020 Deak, Kimani, Cassaidy and Esser-Kahn.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Animals
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Cell Adhesion
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Cells, Cultured
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Humans
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Immunity, Innate
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Immunoassay / methods*
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Lipid A / analogs & derivatives
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Lipid A / chemistry
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Lipid A / metabolism
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Lymphocytes / metabolism*
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Microplastics / chemistry
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Oligopeptides / chemistry
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Oligopeptides / metabolism
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Polystyrenes / chemistry
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Toll-Like Receptor 2 / agonists*
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Toll-Like Receptor 2 / chemistry
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Toll-Like Receptor 2 / metabolism
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Toll-Like Receptor 4 / agonists*
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Toll-Like Receptor 9 / agonists
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Toll-Like Receptor 9 / chemistry
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Toll-Like Receptor 9 / metabolism
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Toll-Like Receptors / agonists*
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Tumor Necrosis Factor-alpha / metabolism
Substances
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Lipid A
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Microplastics
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Oligopeptides
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Polystyrenes
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Toll-Like Receptor 2
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Toll-Like Receptor 4
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Toll-Like Receptor 9
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Toll-Like Receptors
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Tumor Necrosis Factor-alpha
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PAM2-CSK4
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monophosphoryl lipid A