Enhancement of mucosal immune response against the M2eHBc+antigen in mice with the fusion expression products of LTB and M2eHBc+through mucosal immunization route

M2e is the external domain of M2 protein, a conservative transmembrane protein of the avian influenza A virus. Previous research had shown that the vaccine of the formation particle of M2e and hepatitis B virus core antigen (HBcAg) can fully protect mice against a lethal H5N1 subtype avian influenza virus (AIV) infection. As an effective approach against mucosal tissue infectious agent, mucosal vaccination requires effective and safe adjuvants. Here we have first fused two M2e peptide to the N terminal and the major immunodominant region (MIR) of the HBcAg protein simultaneously to create a fusion gene, named as M2eHBc+, and then inserted B subunit of Escherichia coli heat labile enterotoxin (LTB) into the N terminal of M2eHBc+ to construct the second fusion gene, named as LBM2eHBc+. These two fusion genes can be efficiently expressed in Escherichia coli cell and the yield peptide can self-assemble into virus-like particles (VLP). The mice immunization with two types of the purified particles by intranasal dropping and oral routes revealed that LTB can significantly enhance the mucosal immune responses of mice to co-expression M2eHBc+ particle form antigen.