Porcine circovirus type 2(PCV2)is the main causative agent of porcine circovirus-associated diseases,and it causes substantial economic losses in the swine industry each year.It is crucial to develop an efective vacci...Porcine circovirus type 2(PCV2)is the main causative agent of porcine circovirus-associated diseases,and it causes substantial economic losses in the swine industry each year.It is crucial to develop an efective vaccine against the circulating strain PCV2d,which is prone to substantial degrees of mutation.In this study,a truncated form of fagellin(tFlic:85-111 aa)was inserted into the C-terminal sequence of 2dCap,and Western blotting results showed that recombinant Cap-tFlic VLPs were successfully expressed.Transmission electron microscopy(TEM)and dynamic light scattering(DLS)data indicated that purifed recombinant Cap-tFlic fusion proteins existed in the form of polymers and that tFlic could not afect the formation and internalization of VLPs.Integrated Cap-tFlic VLPs induced the expression of antigen presentation-related factors(MHC-II and CD86)by bone marrow-derived dendritic cells(BM-DCs),and the expression of TLR5-related factors(TNF-α)was dramatically elevated.Mice intramuscularly immunized with Cap-tFlic VLPs exhibited signifcantly higher levels of Cap-specifc antibodies and neutralizing antibodies than mice immunized with wild-type Cap VLPs.The data obtained in the current study indicate that Cap-tFlic may be a candidate for a subunit vaccine against PCV2 in the future.展开更多
基金This work was supported by the Key Laboratory of Veterinary Biological Products and Chemicals,Ministry of Agriculture,China Animal Husbandry Industry Co.,Ltd.Project name:Novel Adjuvanted Chimeric Porcine Circovirus Type 2 CAP Virus-Like Particle Vaccine and project number:20201612(2020-2024).
文摘Porcine circovirus type 2(PCV2)is the main causative agent of porcine circovirus-associated diseases,and it causes substantial economic losses in the swine industry each year.It is crucial to develop an efective vaccine against the circulating strain PCV2d,which is prone to substantial degrees of mutation.In this study,a truncated form of fagellin(tFlic:85-111 aa)was inserted into the C-terminal sequence of 2dCap,and Western blotting results showed that recombinant Cap-tFlic VLPs were successfully expressed.Transmission electron microscopy(TEM)and dynamic light scattering(DLS)data indicated that purifed recombinant Cap-tFlic fusion proteins existed in the form of polymers and that tFlic could not afect the formation and internalization of VLPs.Integrated Cap-tFlic VLPs induced the expression of antigen presentation-related factors(MHC-II and CD86)by bone marrow-derived dendritic cells(BM-DCs),and the expression of TLR5-related factors(TNF-α)was dramatically elevated.Mice intramuscularly immunized with Cap-tFlic VLPs exhibited signifcantly higher levels of Cap-specifc antibodies and neutralizing antibodies than mice immunized with wild-type Cap VLPs.The data obtained in the current study indicate that Cap-tFlic may be a candidate for a subunit vaccine against PCV2 in the future.