Abstract The immune response raised by oncolytic Vaccinia Virus after the infection and destruction of cancer cells is believed to be a key factor in the promising recent clinical results seen with these viruses. However, Vaccinia Virus primordially activates TLR2, which is associated with Th2 conversion and strong humoral responses. For cancer therapy, elicitation of cellular responses is thought to be more effective for the direct destruction of tumor cells. In this work, we switched the TLR response that a modified oncolytic Vaccinia Virus elicits. Firstly, we looked to activate the TLR3 pathway after infection with Vaccinia Virus, which is more associated with cellular responses. For this purpose, we constructed an oncolytic Vaccinia Virus that codifies for the TRIF protein, a key adaptor of the TLR3 pathway. The resulting virus increased the level of cytokines, chemokines and DAMPs released after infection, resulting in improved cytotoxicity in cancer cells and significantly enhanced therapeutic effects in mouse tumor models. Concurrently, we demonstrated that enzymatic deglycosylation of the surface of the virus is able to ablate the activation of cell surface TLRs, increasing the level of delivery and early viral gene expression in tumors in vivo. Finally, combination of both strategies demonstrated increased amounts of CTLs targeting the tumor, reduced levels of anti-viral neutralizing antibody and a further increase in antitumor activity in orthotopic mouse models.