In line with that, microscopic photographs at 48 and 72 h post-infection (magnification 10×), show changes in the morphology of cells, confirming the anticancer and cytopathic effect (CPE) on cells treated with the different groups (infected with AdV-D24 or AdV-D24-ICOSL-CD40L only or with pembrolizumab or only treated with pembrolizumab) (Figure 3B,C), respectively), thus suggesting that AdV5/3-D24-ICOSL-CD40L alone and or in combination with pembrolizumab could exert an antitumor effect against melanoma. This evidence concerns the gene CD40LG and melanoma.