Figure 5.

Vaccinia virus has limited impact on NK cells. (A) NK-CCR5 cells were incubated with OV-ffLuc or OV-ffLuc-CCL5 at MOI=1. Virus proliferation was monitored by the IVIS imaging 24 and 48 hours later. HCT-116 cells infected with OV-ffLuc at MOI=0.1 were used as positive control; PBS-treated NK cells were used as negative control. (B) NK-CCR5 cell viability was determined 48 hours after infection with virus at different MOI values. (C) After incubation with virus at MOI=1 for 48 hours, NK-CCR5 cells were mixed with tumor cells at the indicated E:T ratios for 6 hours and NK cell cytotoxicity determined. (D) The impact of oncolytic vaccinia virus on NK cells was determined; NK cells were pretreated with OV for 48 hours, following which the Transwell assay was performed. After counting the number of migrated cells, the migrating indices were determined relative to NK cells treated with PBS. NK cells were collected from five healthy donors. Data are presented as the means±SD. Statistical analysis was performed with t-tests and one-way ANOVA. ANOVA, analysis of variance; n.d., not detected; E:T, effector to target; IVIS, in vivo imaging system; MOI, multiplicity of infection; NK, natural killer; OV, oncolytic viruse; PBS, phosphate buffered saline.