title: Display of VP1 on the surface of baculovirus and its immunogenicity against heterologous human enterovirus 71 strains in mice.
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.
  
  
  RESULTS
  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 the plasma membrane of insect cells infected with recombinant baculovirus. In addition, we demonstrated with transmission electron microscopy that baculovirus successfully acquired VP1 from the insect cell membrane via the budding process. After two immunizations in mice, Bac-Pie1-gp64-VP1 elicited neutralization antibody titer of 1∶64 against EV71 (subgenogroup C4) in an in vitro neutralization assay. Furthermore, the antisera showed high cross-neutralization activities against all 11 subgenogroup EV71 strains.
  
  
  CONCLUSIONS
  Our results illustrated that Bac-Pie1-gp64-VP1 retained native epitopes of VP1 and acted as an effective EV71 vaccine candidate which would enable rapid production without any biosafety concerns.
authors: Animal Health Biotechnology, Temasek Life Sciences Laboratory, Republic of Singapore.
authors: Tao
authors: T
authors: Meng
authors: Annasaheb B
authors: AB
authors: Kolpe
authors: Tanja K
authors: TK
authors: Kiener
authors: Vincent T K
authors: VT
authors: Chow
authors: Jimmy
authors: J
authors: Kwang
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D029821:Enterovirus A, Human / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity; D005260:Female; D005818:Genetic Engineering; D006801:Humans; D056724:Immunity, Humoral / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D012318:RNA Polymerase I / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology / Q000235:genetics; D014612:Vaccines / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D018076:DNA, Complementary; D004274:DNA, Recombinant; D014612:Vaccines; D012318:RNA Polymerase I