Construction and immunogenicity of recombinant porcine circovirus-like particles displaying somatostatin

摘要

In order to obtain a virus-like particles (VLPs) vaccine both for porcine circovirus type 2 (PCV2) prevention and growth-promotion, somatostatin (SS) gene was fused to the 3'-terminal of ORF2 gene of PCV2 with PCR, and a recombinant baculovirus (rAc-Cap-SS) was constructed. The expression of fusion protein Cap-SS (rCap-SS) with molecular weight of approximately 32 kDa was identified by Western blot and indirect immunofluorescence assay in Sf9 cells. The self-assembled VLPs were observed under electron microscopy, which being morphologically similar to the recombinant Cap protein (rCap) expressed in the same baculovirus expressing system. Ninety four-week-old mice were immunized with the recombinant proteins twice. The results showed that mice immunized with rCap-SS protein developed antibody against Cap, which levels being similar to those immunized with rCap protein. The body weight gain and anti-SS antibody in rCap-SS group was higher than those of rCap and negative control groups during 28 and 42 days post inoculation (dpi). Furthermore, twenty 28-day-old piglets were vaccinated twice subcutaneously with the recombinant proteins. The results indicated that PCV2-specific antibody could be induced after vaccination with rCap-SS or rCap protein. Anti-SS antibody could be induced after rCap-SS vaccination and was higher than other groups at 14 and 28 dpi. The level of somatostatin concentration in the blood of pigs in rCap-SS group was significantly decreased at 14 dpi than other groups (P < 0.05). The relative daily weight gain (RDWG) of pigs in rCap-SS group was obviously higher than that in other groups at 28 dpi. After challenge with PCV2, pigs in the vaccinated groups had no clearly clinical signs, and the RDWG was significantly higher than that in the challenge control group (CC) (P < 0.05). The pathological lesions, viremia and viral load presented in the vaccinated groups were milder than those in challenge control group. It suggested that the recombinant porcine circovirus-like particles displaying somatostatin might be a novel subunit vaccine candidate for preventing PMWS and promoting pig growth