Abstract
Porcine rotavirus is a major cause of acute viral gastroenteritis in suckling piglets, and vaccination is considered to be an effective measure to control these infections. The development of a live mucosal vaccine using Bacillus subtilis spores as an antigen delivery vehicle is a convenient and attractive vaccination strategy against porcine rotavirus. In this study, a shuttle vector was constructed for the spore surface display of the spike protein VP8* from porcine rotavirus (the genotype was G5P[7]). A successful display of the CotB-VP8* fusion protein on the spore surface was confirmed by Western blot and immunofluorescence microscopy analysis. The capacity for immune response generated after immunization with the recombinant strain was evaluated in a mouse model. The intestinal fecal IgA and serum IgG were detected by enzyme-linked-immunosorbent serologic assay (ELISA). Importantly, recombinant strain spores could elicit strong specific mucosal and humoral immune responses. These encouraging results suggest that recombinant B. subtilis BV could provide a strategy for a potential novel application approach to the development of a new and safe mucosal subunit vaccine against porcine rotavirus.
Highlights
Rotaviruses are members of the Reoviridae family of double-stranded RNA viruses
Electrophoresis and sequencing results showed that the VP8* gene of porcine rotavirus was successfully inserted into the prokaryotic expression vector pET-32a, and the prokaryotic expression plasmid pET-32a-VP8* was successfully constructed (Figure 1)
The serum of the mice was assayed by enzyme-linked-immunosorbent serologic assay (ELISA) and the antibody titer was 1:12800
Summary
The virus genome is composed of 11 segments encoding six structural viral proteins and six nonstructural proteins [1,2,3]. Rotavirus is classified into multiple groups by the inner capsid protein (VP6) and the outer capsid proteins (VP4 and VP7), which form the base of the G and p dual typing system [4,5]. The main symptom of porcine rotavirus is severe diarrhea, which results in huge economic losses in the pig industry worldwide [6]. Pigs of all ages can be infected with rotavirus, and nursing piglets have more severe symptoms. Infection of weaned piglets is characterized by mild to moderate or no clinical manifestations, but they can continue to be exposed to infectious viruses in the environment [7,8,9]
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