Fig. 5: ORFV recombinants trigger, replicate, and cause pyroptosis in tumor cells.
From: Oncolytic Parapoxvirus induces Gasdermin E-mediated pyroptosis and activates antitumor immunity
a Images acquired with a fluorescence microscope display the replication of ORFV-∆120-EGFP (MOI = 1) and ORFV-∆120-121-EGFP (MOI = 1) in B16 cells for 12 h through the EGFP signal. Pyroptotic cell death were displayed through PI uptake signal for 26 h (scale bar: 20 μm). b Six weeks old male C57BL/6 mice were s.c. engrafted with B16 cells and i.t. treated with vehicle, ORFV (1 × 105 TCID50/mouse), ORFV-∆120-EGFP (1 × 105 TCID50/mouse) and ORFV-∆120-121-EGFP (1 × 105 TCID50/mouse) (n = 7–8). Tumor volume were measured as indicated time points (P < 0.0001, P < 0.0001, P < 0.0001, P = 0.4428, P = 0.6302, and P = 0.2260). c Images acquired with a fluorescence microscope display ORFV-∆120-EGFP replication and PI uptake in in situ B16 tumor tissues (scale bar: 50 μm). d Images acquired with a fluorescence microscope display the replication of ORFV recombinants (ORFV-∆120-EGFP and ORFV-∆120-121-EGFP) and PI uptake in metastatic tumor cells in the lungs (scale bar: 50 μm). e Schematic illustration of experimental design. Tumor-bearing mice were tail intravenous injection (i.v.) treated with ORFV-∆120-EGFP. Lungs were collected for the image. f Six weeks old male C57BL/6 mice were i.v. engrafted with B16-F10 cells (1 × 105 cells/mouse) and i.v. treated with ORFV-∆120-EGFP (1 × 104 TCID50) (n = 3–5). Photographs of lungs and metastatic lesions were counted (P = 0.018). The above experiments were successfully repeated two to three times. ns not significant; ****P < 0.0001. One-way ANOVA with Tukey’s test was performed in (b). Two-tailed unpaired Student’s t-tests were performed for the statistical analyses in (f), and the results are presented as the mean ± SD.
