The Adjuvant Effect of CpG Oligodeoxynucleotide Linked to the Non‐Toxic B Subunit of Cholera Toxin for Induction of Immunity Against H. pylori in Mice

Abstract

The present study was carried out to test the immunostimulatory and adjuvant effects of the non-toxic B subunit of cholera toxin (CTB), CpG oligodeoxynucleotide (ODN) and CpG ODN linked to CTB (CTB-CpG) for generation of immunity against H. pylori in mice. Herein, we showed that CTB-CpG induces more potent proinflammatory cytokine and chemokine responses in the cervical and the mesenteric lymph nodes (CLN and MLN, respectively) cells in vitro compared with those of CTB and CpG ODN. The adjuvant effects of these agents were examined following intranasal immunization of C57Bl/6 mice with H. pylori lysate in combination with CpG ODN, CTB or CTB-CpG. All three immunization regimes resulted in high H. pylori-specific IgG antibody responses; however, only the CTB-CpG and, to some extent, the CpG ODN immunized mice mounted a sustainable IgG2c antibody response. Importantly, mice immunized with H. pylori antigen and CTB-CpG or CpG ODN, but not CTB, developed strong H. pylori-specific proliferative and IFN-gamma responses in their MLN CD4+ T cells upon recall antigen stimulation in vitro. These mice also had significantly lower bacterial load compared with the control-infected mice. Furthermore, the CTB-CpG and the CpG ODN immunized mice developed increased specific IgA antibody responses in their gastrointestinal tracts following H. pylori challenge. These results imply that CTB-CpG and CpG ODN, but not CTB, could serve as nasal adjuvants for induction of a H. pylori-specific Th1 type immunity in MLN and also a specific mucosal IgA antibody response in the gastrointestinal tract upon H. pylori challenge.