Abstract
INTRODUCTION: Hepatocellular carcinoma (HCC) represents a significant health challenge, with immunotherapy serving as a crucial component of its complex treatment regimen. This study investigates the use of TP53Y220C as a preferred antigen to induce cytotoxic T lymphocytes (CTLs) for cytotoxic effects against HCC. METHODS: The TP53(Y220C) mRNA (mTP53(Y220C)) was synthesized through an in vitro transcription method and subsequently introduced into dendritic cells (DCs) using bacterial outer membrane vesicles expressing L7Ae and Listeria monocytogenes lysin O (OMV-LL), electroporation, and lipid nanoparticles, respectively. Co-culture of differently treated DCs with initial T cells induces CTLs. The cytotoxic effects of CTLs on hepatocellular carcinoma were evaluated through experiments such as flow cytometry and mouse tumour models. RESULTS: We assessed the therapeutic efficacy of CTLs, activated by mTP53(Y220C)-loaded DCs, in a murine model of HCC. Results demonstrate that CTLs, activated by DCs loaded with mTP53(Y220C) via OMV-LL or electroporation, effectively initiated immune responses against HCC. While OMV-LL were less efficient than electroporation in mRNA delivery, they induced a significant pro-inflammatory response and activated the innate immune system. CONCLUSION: This study highlights OMV-LL as an innovative mRNA delivery approach to DCs for CTLs activation and demonstrates their potential in CTLs-based therapy for HCC.