Purpose: This study aimed to describe a novel cancer vaccine developed using H₂O₂-inactivated Salmonella typhimurium  RE88 [with deletions of AroA (the first enzyme in the aromatic amino acid biosynthesis pathway) and DNA adenine methylase] as the carrier.
Methods: The pVLT33 plasmid was used to engineer an RE88 strain induced to express ovalbumin (OVA) by isopropylthiogalactoside (RE88-pVLT33-OVA). The immune responses and anticancer effects of H₂O₂-inactivated RE88-pVLT33-OVA were compared with those of non-inactivated RE88-pVLT33-OVA and OVA (positive control) in mice carrying OVA-expressing tumors (EG7-OVA) cells.
Results: Anti-ovalbumin IgG (immunoglobulin G) titer following vaccination with H₂O₂-inactivated RE88-pVLT33-OVA was higher for subcutaneous than for intragastric vaccination. When subcutaneous administration was used, H₂O₂-inactivated RE88-pVLT33-OVA (2 × 10⁹ CFU (colony forming units)/mouse) achieved an anti-ovalbumin IgG titer higher than that for the same dose of RE88-pVLT33-OVA and comparable to that for 10 μg ovalbumin (positive control). The binding of mouse serum antibodies to EG7-OVA cells was stronger for H₂O₂-inactivated RE88-pVLT33-OVA (2 × 10⁹ CFU/mouse) than for 10 μg ovalbumin. Furthermore, subcutaneous vaccination with H₂O₂-inactivated RE88-pVLT33-OVA (2 × 10⁹ CFU/mouse) induced greater activation of splenic T cells and more extensive tumor infiltration with CD4⁺/CD8⁺ T cells compared with 10 μg ovalbumin (positive control). The mice vaccinated subcutaneously with H₂O₂-inactivated RE88-pVLT33-OVA at a dose of 2 × 10⁸ or 6 × 10⁸ CFU/mouse had smaller tumors compared with mice in the negative control groups. Tumor weight in mice vaccinated with H₂O₂-inactivated RE88-pVLT33-OVA at a dose of 2 × 10⁹ CFU/mouse was significantly lower than that in both negative control groups (P < 0.05) and decreased with the increasing dose of H₂O₂-inactivated RE88-pVLT33-OVA. H₂O₂-inactivated RE88-pVLT33-OVA was potentially safer than the non-inactivated strain, could carry exogenous antigens, and had specific epitopes that could be exploited as natural adjuvants to facilitate the induction of cellular and humoral immune responses.
Conclusion: It was anticipated that H₂O₂-inactivated RE88-pVLT33-OVA could be used as a novel delivery system for new cancer vaccines.
Keywords: cancer vaccine, H₂O₂ inactivation, RE88-pVLT33-OVA, whole-cell bacterial vaccine