Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon.
White spot syndrome virus (WSSV) is a large dsDNA virus responsible for white spot disease in shrimp and other crustaceans. VP28 is one of the major envelope proteins of WSSV and plays a crucial role in viral infection. In an effort to develop a vaccine against WSSV, we have constructed a recombinant baculovirus with an immediate early promoter 1 which expresses VP28 at an early stage of infection in insect cells. Baculovirus expressed rVP28 was able to maintain its structural and antigenic conformity as indicated by immunofluorescence assay and western blot analysis. Interestingly, our results with confocal microscopy revealed that rVP28 was able to localize on the plasma membrane of insect cells infected with recombinant baculovirus. In addition, we demonstrated with transmission electron microscopy that baculovirus successfully acquired rVP28 from the insect cell membrane via the budding process. Using this baculovirus displaying VP28 as a vaccine against WSSV, we observed a significantly higher survival rate of 86.3% and 73.5% of WSSV-infected shrimp at 3 and 15 days post vaccination respectively. Quantitative real-time PCR also indicated that the WSSV viral load in vaccinated shrimp was significantly reduced at 7 days post challenge. Furthermore, our RT-PCR and immunohistochemistry results demonstrated that the recombinant baculovirus was able to express VP28 in vivo in shrimp tissues. This study will be of considerable significance in elucidating the morphogenesis of WSSV and will pave the way for new generation vaccines against WSSV.
Syed Musthaq
S
S
Animal Health Biotechnology, Temasek Lifesciences Laboratory, 1 Research Link, National University of Singapore, 117604, Singapore.
Madhan
Selvaraj
S
Sahul Hameed
A S
AS
Kwang
Jimmy
J
D000818:Animals; D016367:Baculoviridae / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D005822:Genetic Vectors; D018613:Microscopy, Confocal / Q000379:methods; D016253:Microscopy, Immunoelectron / Q000379:methods; D008954:Models, Biological; D033561:Penaeidae / Q000276:immunology / Q000821:virology*; D018411:Spodoptera; D016019:Survival Analysis; D014759:Viral Envelope Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D019562:Viral Load; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology*; D014777:Virus Diseases / Q000517:prevention & control / Q000662:veterinary*; D046848:White spot syndrome virus 1 / Q000235:genetics / Q000276:immunology*
D014759:Viral Envelope Proteins; D014765:Viral Vaccines