Background: Hepatocellular carcinoma (HCC) is an aggressive malignancy with limited response to conventional immunotherapies. Multi-epitope vaccines targeting multiple tumor-associated antigens (TAAs) such as alpha-fetoprotein (AFP), glypican-3 (GPC3), and telomerase reverse transcriptase (TERT) offer a promising strategy to overcome immune evasion. However, peptide-based vaccines suffer from rapid degradation and poor delivery to antigen-presenting cells. This study aimed to develop a nanoliposomal vaccine encapsulating a bioinformatics-designed merged multi-epitope peptide derived from AFP, GPC3, and TERT, and evaluate its ability to modulate inflammatory gene expression in the spleen of immunized mice. Methods: A merged peptide (PGLPDSALDINECLRGKKKDGARGGPPEAFTTSVRKKKKPEGLSPNLNRFLGDR) was designed using IEDB and NetMHCpan servers. Cationic nanoliposomes (DPPC:cholesterol:DSPE-PEG2000, 55:40:5 molar ratio) were prepared by thin-film hydration. Size, zeta potential, and peptide loading were characterized by DLS and BCA assay. Female BALB/c mice (n=24) were divided into four groups: PBS control, empty liposome, free peptide, and NLME vaccine. After three subcutaneous immunizations, splenic expression of IFN-γ, TNF-α, IL-6, IL-1β, and IL-10 was measured by qRT-PCR. Results: Nanoliposomes exhibited mean size of 105±12 nm, zeta potential of +30.4±2.1 mV, and peptide loading efficiency of 89.4±3.2%. NLME vaccination significantly upregulated IFN-γ (5.67-fold, p<0.01), TNF-α (4.23-fold, p<0.01), IL-1β (2.98-fold, p<0.01), and IL-6 (2.12-fold, p<0.05), while downregulating IL-10 (0.42-fold, p<0.01) compared to controls. Conclusion: The NLME vaccine induces a robust Th1-polarized inflammatory response in the spleen with high pro-inflammatory cytokine expression and suppressed IL-10, supporting its potential as an effective immunotherapy for HCC.
Sayyad et al. (Fri,) studied this question.