Flow-Cytometry Analysis
Multiparameter Flow Cytometry analysis was performed on semen samples from COVID-19 patients using a BD FACSymphony A5 instrument (BD Biosciences) and analyzed with FlowJo (FlowJo LLC, Ashland, OR). Gates were done on live cells and the following primary antibodies were used for immune cell characterization: CD163 (BD Horizon, Franklin Lakes, NJ, clone MAC2-158), CD45 (Biolegend, San Diego, CA, clone HI30), CD56 (BD Biosciences, clone MY31), CD19 (Biolegend, clone HIB19), CD16 (Biolegend, clone 3G8), CD25 (BD Biosciences, clone 2A3), CD123 (BD Biosciences, clone 7G3), HLA-DR (BD Biosciences, clone G46-6), PD-1 (Biolegend clone EH12.2H7), CD8 (BD Biosciences, clone SK1), CD95 (BD Biosciences, clone DX2), CD4 (BD Biosciences, clone M-T477), CD11c (Biolegend, clone Bu15), CD127 (eBioscience, clone eBioRDR5), CD45RO(BD Biosciences, clone UCHL1), CD3 (BD Biosciences, clone UCHT1), CD14 (Biolegend, clone M5E2), CD68 (BD Biosciences, clone Y1/82A), CD27 (BD Biosciences, clone M-T271), CCR7 (Bd Biosciences, clone 3D12).