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- W3004914911 abstract "Phagocytosis, the process by which immune cells engulf and neutralize foreign particles, is essential to human health. However, the mechanistic relationship between chemical cues, mechanics, and intracellular signaling during phagocytosis remains poorly understood. Here, we vary the density of immobilized IgG antibody on model targets for phagocytosis and examine the effects on human neutrophil spreading dynamics and intracellular calcium signaling. We first validate our ability to tune the IgG density on targets by quantifying the final surface concentration via flow cytometry or confocal microscopy. To test for phagocytic efficiency in bulk, we incubate cells and IgG-coated fluorescent microspheres for one hour and then quantify the amount of phagocytosis via flow cytometry. Alternatively, to measure spreading dynamics of single cells, we image frustrated phagocytic spreading of neutrophils on an IgG-coated glass coverslip. In this experiment, we can also monitor calcium signaling by pre-loading the cells with a calcium-sensitive fluorophore. We find that the phagocytic activity in our bulk assays generally increases with IgG surface concentration. In the cell spreading experiments, higher IgG densities correlate with larger maximum contact areas, more uniform, concentric spreading, and earlier induction of a global calcium burst. In contrast, the speed of spreading remains constant on different densities of IgG, suggesting that active protrusion rather than passive adhesion primarily drives frustrated phagocytic spreading. We further test this interpretation using a computational model of cell spreading, in which we treat the neutrophil as an axisymmetric, highly viscous fluid body surrounded by an elastic cortex and experiencing adhesive and/or protrusive forces. Our model confirms that adhesion-driven spreading is not consistent with the conservation of spreading speeds across vastly different ligand densities." @default.
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- W3004914911 date "2020-02-01" @default.
- W3004914911 modified "2023-09-26" @default.
- W3004914911 title "Density of Immobilized Antibodies Modulates Neutrophil Biophysical Behavior and Calcium Dynamics During Phagocytic Spreading" @default.
- W3004914911 doi "https://doi.org/10.1016/j.bpj.2019.11.3269" @default.
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