Immunotherapy has become an essential component in cancer treatment. However, the majority of solid metastatic cancers, such as pheochromocytoma, are resistant to this approach. Therefore, understanding immune cell composition in primary and distant metastatic tumors is important for therapeutic intervention and diagnostics. Combined mannan-BAM, TLR ligand, and anti-CD40 antibody-based intratumoral immunotherapy (MBTA therapy) previously resulted in the complete eradication of murine subcutaneous pheochromocytoma and demonstrated a systemic antitumor immune response in a metastatic model. Here, we further evaluated this systemic effect using a bilateral pheochromocytoma model, performing MBTA therapy through injection into the primary tumor and using distant (non-injected) tumors to monitor size changes and detailed immune cell infiltration. MBTA therapy suppressed the growth of not only injected but also distal tumors and prolonged MBTA-treated mice survival. Our flow cytometry analysis showed that MBTA therapy led to increased recruitment of innate and adaptive immune cells in both tumors and the spleen. Moreover, adoptive CD4+ T cell transfer from successfully MBTA-treated mice (i.e., subcutaneous pheochromocytoma) demonstrates the importance of these cells in long-term immunological memory. In summary, this study unravels further details on the systemic effect of MBTA therapy and its use for tumor and metastasis reduction or even elimination.

Department of Medical Biology Faculty of Science University of South Bohemia 37005 Ceske Budejovice Czech Republic

Endoscopy Center and Endoscopy Research Institute Zhongshan Hospital Fudan University Shanghai 200032 China

Laboratory of Tumor Immunology and Biology Center for Cancer Research National Cancer Institute NIH Bethesda MD 20892 USA

Neuro Oncology Branch National Cancer Institute NIH Bethesda MD 20892 USA

Section on Medical Neuroendocrinology Eunice Kennedy Shriver National Institute of Child Health and Human Development NIH Bethesda MD 20892 USA

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Role of B cells in intratumoral MBTA immunotherapy of murine pheochromocytoma model

Intratumoral immunotherapy of murine pheochromocytoma shows no age-dependent differences in its efficacy

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