HaCat cells were infected with HSV-2(333)ZAG (0.1 PFU/cell) in 6-well cell culture plates (Costar-Corning, Kennebunk, ME, USA) for 1 h and then overlaid with media for 9 h. Cells were washed with PBS, and then heat-inactivated immune sera (1:50 dilution in incomplete DMEM) was added to cells. Plates were kept on ice for 30 min before adding 20% v/v rabbit complement. After a 4 h incubation at 37 °C in 5% CO2, cells were detached with 500 µL/well Accutase (Thermo Fisher Scientific, Waltham, MA, USA), washed twice with PBS and stained with Zombie-NIR (BioLegend, San Diego, CA, USA) for 20 min at RT. Stained cells were then fixed with 2% paraformaldehyde and read with a Cytek Aurora 5 Laser System Flow cytometer (Cytek Bioscience, Freemont, CA, USA). Data were analyzed with FlowJo Software, version 10 (FlowJo-BD, Franklin Lakes, NJ, USA).

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