Since the treatment with oncolytic viruses is a promising approach for targeting the immune response against the tumor by combining the cytolytic activity together with the ability of the viruses to activate the immune system [41,42], in the current study, we aimed to design and engineer a novel chimeric oncolytic adenovirus vector AdV-D24-ICOSL-CD40L armed with two potent co-stimulatory molecules, ICOSL and CD40L, selectively expressing in cancer cells such as melanoma. This evidence concerns the gene CD40LG and cancer.