Recombinant invasive expressing fibronectin binding protein A induce specific humoral immune response by stimulating differentiation of dendritic cells.

Recombinant lactic acid bacteria (LAB), especially , have been genetically engineered to express heterogeneous invasion proteins, such as the fibronectin binding protein A (FnBPA) from , to increase the invasion ability of the host strains, indicating a promising approach for DNA vaccine delivery. The presence of FnBPA has been also shown to be an adjuvant for co-delivered antigens, however, the underlying mechanisms are still not clear. To explore the above underlying mechanisms, in this study, we constructed a novel strain with surface displayed FnBPA, which could significantly improve the adhesion and invasion ratios of strain on a porcine intestinal epithelial cell line (IPEC-J2) about two-fold compared with the empty vector. At the same time, the presence of FnBPA significantly stimulated the differentiation of bone marrow-derived dendritic cells (DCs) and increased the secretion of interleukin (IL)-6 and mRNA level of IL-6 gene, which were proved by flow cytometry, enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription PCR (qRT-PCR). With regard to study, the presence of FnBPA significantly stimulated the differentiation of DCs in the Peyer's patch (PP) and the percentages of IL-4 and IL-17A T helper (Th) cells of splenocytes in flow cytometry assay. In consistent with these results, the levels of IL-4 and IL-17A in serum as measured via ELISA also increased in mice treated with FnBPA . Finally, the FnBPA strain increased the production of B220 B cells in mesenteric lymph node (MLN) and PP and the levels of FnBPA-specific IgG and sIgA antibodies, indicating the its possible application in vaccine field. This study demonstrated that the invasive with surface displayed FnBPA could modulate host immune response by stimulating the differentiation of DCs and Th cells which could possibly be responsive for the adjuvant effects of FnBPA.