T cells/µl were measured using BD Trucount™ Tubes with FITC-conjugated anti-CD3, PE-conjugated anti-CD8, PerCP-conjugated anti-CD45 and APC-conjugated anti-CD4 antibodies . Details of additional antibodies used for analysis of T and B cell subpopulations are provided in. Red blood cells in the whole blood samples were lysed using a commercial BD FACS™ Lysing Solution . The samples were acquired on a FACSCanto II .
After 48 hours of incubation, the plates were washed 4 times with 0.05% Tween 20 in PBS, followed by 2 hours of incubation at 37°C with biotinylated anti-human IFN-γ monoclonal antibodies diluted at 1μg/ml in PBS. Subsequently, the plates were washed 4 times with 0.05% Tween 20 in PBS, and then were incubated with 50μl/well Avidin-Peroxidase-Complex at 37°C for an hour. The Avidin-Peroxidase Complex was prepared and left to stand for 30 min at room temperature before use.
Our hospitalized COVID-19 cohort represents the infection in wider society in terms of biological sex distribution, encompassing two thirds of males and one third of females (
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