UV-HSV-1 induces metabolic reprogramming of NK cells. (A) PBMCs from 9 healthy donors were exposed to UV-HSV-1 (0.1 pfu/PBMC for 16 hours), and the levels of lactate in the culture supernatant were quantitated as described in “Materials and methods.” Results are expressed as pmol lactate/cell. (B) PBMCs from 4 healthy donors were exposed to UV-HSV-1 (0.1 pfu/PBMC for 16 hours), ±11 mM 2-deoxyglucose and lactate levels determined as above. *P < .01 from control; **P < .01 from UV-HSV-1. (C) NK cells from 2 healthy donors were exposed to UV-HSV-1 (0.1 pfu/PBMC for 16 hours), and the levels of lactate in the culture supernatant were quantitated. *P < .05 from control. (D) Representative flow cytometry plots of oxygen consumption measurements using pimonidazole as an indicator of cellular respiration. (E) PBMCs from 4 healthy donors were exposed to UV-HSV-1 (0.1 pfu/PBMC), ±100 μΜ etomoxir for 4 h in complete medium supplemented with pimonidazole, and the accumulation of pimonidazole adducts was quantitated as described in “Materials and methods.” PBMCs treated with 6 mM sodium cyanide served as a background control, and results are shown as cyanide-corrected values. Each donor was tested in triplicate, and the average value was used for statistical purposes. *P < .05 from control; **P < .05 from UV-HSV-1 alone. (F) EasySep purified CD56⁺ cells were exposed, or not, to 0.1 pfu/NK cell UV-HSV-1 for 4 hours in complete medium supplemented with pimonidazole, and the accumulation of pimonidazole adducts was quantitated as above. *P < .05 from control. (G) PBMCs from 3 healthy donors were exposed to 0.1 pfu/PBMC UV-HSV-1 for 16 hours, and the accumulation of intracellular triglycerides was monitored by flow cytometry as described in “Materials and methods.” Each donor was tested in triplicate, and the average value was used for statistical purposes. *P < .05 from control.