display of vp1 on the surface of baculovirus and its immunogenicity against heterologous human enterovirus 71 strains in micevp1杆状病毒表面展示及其免疫原性与异种的人类肠道病毒71株在小鼠身上.pdfVIP

Display of VP1 on the Surface of Baculovirus and Its Immunogenicity against Heterologous Human Enterovirus 71 Strains in Mice 1. 1. 1 2 1,2 Tao Meng , Annasaheb B. Kolpe , Tanja K. Kiener , Vincent T. K. Chow , Jimmy Kwang * 1 Animal Health Biotechnology, Temasek Life Sciences Laboratory, Republic of Singapore, 2 Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Republic of Singapore Abstract Background: Human Enterovirus 71 (EV71) is a common cause of hand, foot and mouth disease (HFMD) in young children. It is often associated with severe neurological diseases and has caused high mortalities in recent outbreaks across the Asia Pacific region. Currently, there is no effective vaccine and antiviral agents available against EV71 infections. VP1 is one of the major immunogenic capsid protein of EV71 and plays a crucial role in viral infection. Antibodies against VP1 are important for virus neutralization. Methodology/Principal Finding: In the present study, infectious EV71 viruses were generated from their synthetic complementary DNA using the human RNA polymerase I reverse genetics system. Secondly, the major immunogenic capsid protein (VP1) of EV71-Fuyang (subgenogroup C4) was displayed on the surface of recombinant baculovirus Bac-Pie1-gp64- VP1 as gp64 fusion protein under a novel White Spot Syndrome Virus (WSSV) immediate early ie1 promoter. Baculovirus expressed VP1 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 VP1 was able to localize on