ADCP was measured as previously described77 (link). Briefly, biotinylated SARS-CoV-2 S stabilized trimer was incubated with red streptavidin-fluorescent beads (Molecular Probes) for 2 h at 37 °C. Ten μl of a 100-fold dilution of beads–protein mixture was incubated for 2 h at 37 °C with 100 μl of monoclonal antibodies diluted at 5 μg ml−1 before addition of THP-1 cells (20,000 cells per well; Millipore). After 19 h incubation at 37 °C, the cells were fixed with 2% formaldehyde solution and fluorescence was evaluated on a LSRII flow cytometer (BD Bioscience). The phagocytic score was calculated by multiplying the percentage of bead-positive cells by the geometric mean fluorescence intensity (MFI) of the bead-positive cells and dividing by 104.
Other organizations :
Walter Reed Army Institute of Research, Henry M. Jackson Foundation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockefeller University, Trudeau Institute, Howard Hughes Medical Institute, Pfizer (United States)
Phagocytic score calculated by multiplying the percentage of bead-positive cells by the geometric mean fluorescence intensity (MFI) of the bead-positive cells and dividing by 10^4
control variables
Incubation time of 2 h at 37 °C for biotinylated SARS-CoV-2 S stabilized trimer with red streptavidin-fluorescent beads
Incubation time of 2 h at 37 °C for bead-protein mixture with monoclonal antibodies
THP-1 cells (20,000 cells per well)
Incubation time of 19 h at 37 °C for bead-protein mixture with THP-1 cells
Fixation of cells with 2% formaldehyde solution
Fluorescence evaluation on a LSRII flow cytometer (BD Bioscience)
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