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Various cancer vaccines have been developed to generate and amplify antigen‐specific T cell responses against malignancy. Among them, in situ vaccination is one of the most practical types as it can trigger immune responses without previous antigen identification. Here we reported a novel in situ vaccine by intratumoral injection of imiquimod and OX40 agonist. In mice bearing hepatic carcinoma, both the injected tumor and the noninjected tumor in the distant lesion of the same mice were suppressed after vaccination. Further studies found that this in situ vaccine triggered systemic tumor‐specific responses, with one‐fold increase of effector memory T cells properties and stronger toxicity of lymphocytes in spleen. Besides, we found that imiquimod upregulated the expression of OX40 on CD4 +  T cells and thus enhanced the effectiveness of OX40 agonist. Five immune‐positive‐related pathways were activated after vaccination. This in situ vaccine caused little harm to normal organs and provided long‐term protection against the same syngeneic tumor rechallenge. Due to its effectiveness, feasibility and safety, this strategy could potentially be applied to various types of late‐stage solid tumors and worthy of further clinical research.

Tumor eradicated by combination of imiquimod and OX40 agonist for in situ vaccination