Fig 1: Therapeutic effects of mono virus and 5voHSV2 treatment in a CT26 Model. Balb/c mice were subcutaneously implanted with CT26 tumor cells and intratumorally inoculated with OVs on days 0, 3, 6 (n = 8–10 per group). A Schematic depiction of the experiment. B The tumor volume growth curves of the OVs and mock-treated groups, the data are shown as mean + SD. ***P < 0.001, ****P < 0.0001 (vs 5voHSV2 group) by repeated-measures 2-way ANOVA with Bonferroni’s correction. C Mouse PBMC killing of CT26-GFP tumor cells. CT26-GFP cells were seeded in the 96-well plate overnight. Then infected with blank control, oHSV2-IL12, -PD1v, -IL15, -IL7 × CCL19, GM-CSF, and 5voHSV2 (MOI = 0.1). PBMC were added with effector cells:target cells ratio being 2:1 for another 60 h. The killing activity was recorded with the PerkinElmer Operetta imaging system
Fig 2: Schematic of the construction and characterization of oHSV2-IL12, -PD1v, -IL15, -IL7 × CCL19, GM-CSF. A The oHSV2 vectors were constructed based on the HG52 strain. Modifications include ICP47 and ICP34.5 genes deletion and the insertion of the related transgenes. IL7 × CCL19 means that IL7 and CCL19 were linked through the 2A peptide. B PCR identification of the inserted sequences. Lane M: marker, lane 1: negative control, lane 2–6 are oHSV2-IL7 × CCL19, GM-CSF, IL12, PD1v and IL15, respectively. C Western blot identification of PD1v (approximately 33 kDa) expression in oHSV2-PD1v infected Vero cells. Lane 1, mock-treated control, lane 2, oHSV2-GFP treated control, lane 3, oHSV2-PD1v infected Vero cells
Fig 3: Infectious and oncolytic activities of oHSV2-IL12, -PD1v, -IL15, -IL7 × CCL19, GM-CSF in vitro. A the recombinant viruses were used to infect tumor cell lines, containing B16R, A549, and CT26 at MOI = 1 for 48 h. Photographs were collected at 24 h and 48 h post-infection. B real-time xCELLigence cell analyzer was used to monitor the kinetics of tumor cell lysis induced by the recombinant viruses. Means of cell index from duplicate wells are presented. Data are representative of three experiments
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