Context Cancer immunotherapy in combination with nanomedicine is of high interest to achieve a systemic immune response. Additional physical methods can bring the specificity related to immunogenic cell death. In the literature, interleukin-12 (IL-12) encoding plasmid associated with sonoporation has been shown to enhance antitumoral activity. Moreover, non-viral carriers and high-frequency ultrasound have both been shown to promote immune response activation. Finally, in situ activation of dendritic cells post-radiofrequency was shown to enhance the immune response. Materials and Methods Cytokines or IL-12 encoding pFAR4-plasmid have been used in these studies. Local hydrogel or non-viral carrier stimulating the immune response were locally delivered. Thermal ablation by radiofrequency or focused ultrasound were applied to release tumor antigens. The concept was evaluated on cloned colorectal mice tumor, which tumor growth was followed by bioluminescence imaging. Results Conditions of thermal deposition using HIFU were optimized. Lipid-mediated cell transfection and intratumoral injection into CT26 tumor mice using pFAR4-IL-12 led to the secretion of the IL-12 cytokine into cell supernatant and mice sera, respectively. The plasmid encoding pFAR4-IL-12 or TLR2 agonist alone had no impact on tumor growth compared with control mice, whereas the complete treatment consisting of pFAR4-IL-12, TLR2 lipid agonist, and HIFU limited tumor growth. Conclusion HIFU conditions can be modulated to stop tumor growth. The combined therapy was the most efficient in terms of IL-12 and IFN-γ production and mice survival. The study showed the interest of the combination of physical methods and nanocarriers to induce a potent immune response.