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Adjuvants are important components of recombinant protein vaccines whichare often poorly immunogenic. For decades, the search for new vaccine adjuvantshas been predominantly empirical. In addition, combinations of more than one adjuvantplus antigen have been systematically studied. Plasmid DNA containing additionaloligodeoxynucleotides with unmethylated CpG motifs (CpG ODN) entrapped in liposomeshas been used as an adjuvant for DNA vaccines and has shown powerful immunostimulatoryfunctions. In our study, the combination of plasmid DNA containing 16 additionalCpG ODNs (pv-16CpG) and aluminum hydroxide (AL) or incomplete Freund's adjuvant(IFA) was used as an adjuvant for a hepatitis B surface antigen (HBsAg) vaccineto immunize C57BL/6J mice. ELISA and ELISPOT assays were used to analyze the immunologicaleffects of the novel vaccine. A significant enhancement of the anti-HBs titerand seroconversion was observed when the CpG plasmid was combined with IFA, butnot with AL. In addition, anti-HBs antibody isotype analysis revealed that thecombination of CpG plasmid and IFA induced a strong HBsAg-specific IgG2a response.Moreover, the ELISPOT assays suggested that pv-16CpG suspended in IFA evoked astrong T helper 1 (Th1) immune response and high IFN-γ production. These resultsdemonstrate that pv-16CpG suspended in IFA is able to induce cellular and humoralimmune responses to HBsAg, and confirm its potential as an adjuvant for use inprotein vaccines.

molecular medicine reports

Plasmid DNA containing multiple CpG motifs triggers a strong immune response to hepatitis B surface antigen when combined with incomplete Freund’s adjuvant but not aluminum hydroxide