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Pancreatic ductal adenocarcinoma is poorly immunogenic, and immune suppression prevents T cell activation. We developed a microbial-based immunotherapeutic concept for selective delivery of immunogenic tetanus toxoid (TT856-1313) antigen into tumor cells by attenuated Listeria monocytogenes, and reactivation of TT-specific memory T cells to kill infected tumor cells. Treatment of KPC mice with Listeria-TT resulted in TT accumulation inside tumor cells, and attraction of TT-specific CD4 and CD8 T cells. Lymph node-like structures were frequently observed in contact with pancreatic tumors of treated mice, and exhibited CD4 and CD8 T cells producing IFNgamma. Gemcitabine combined with Listeria-TT significantly improved migration of CD4 T cells into tumors, and enhanced perforin and granzyme B production. The combination treatment also significantly reduced pancreatic tumors and metastases in Panc-02 and KPC mouse models, with minimal side effects, turning cold tumors into hot tumors. This study provides insight into mechanisms for improving immunotherapy for pancreatic cancer.

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