Anti-cancer effect of interleukin-2 fused to flagellin expressed by tumor-targeting Salmonella

Abstract

Interleukin-2 (IL2) treatment has been explored as a potent immunotherapy agent, particularly for cancers, due to its ability to stimulate T cell proliferation and activity. However, significant challenges and limitations are associated with IL2 treatment, including its short half-life, systemic toxicity and side effects, and limited efficacy in solid tumors. In this study, we deployed an attenuated Salmonella Gallinarum (SG), an avian-specific pathogen capable of targeting tumor tissue, to express and secrete the IL2 using a bacterial flagellum type 3 secretion system (T3SS). Since the T3SS is used for the secretion of flagellin monomers (FliC), DNA of the human IL2 gene was fused to the SG fliC gene so that the fusion proteins would be exported together. A superb anti-cancer effect was observed when the SG expressing and secreting the FliC-IL2 fusion protein was injected into a syngeneic tumor mouse model with CT26 colorectal cancer via the tail vein. Within the fusion protein, the FliC moiety led to a selective increase in MHCIIhighCD206- M1-like macrophages, while the IL2 moiety promoted selective expansion of cytotoxic CD8+ T cells and NK cells, without expanding CD4+FoxP3+ regulatory T cells in the tumor microenvironment (TME). It was concluded that the local delivery of IL2 within the TME by cancer-targeting SG could overcome the limitations associated with IL2-based cancer immunotherapy.