title: Recombinant outer capsid glycoprotein (VP7) of rotavirus expressed in insect cells induces neutralizing antibodies in rabbits.
abstract: BACKGROUND
  Rotaviruses cause diarrhea in infants and young children worldwide. Rotavirus outer capsid protein, VP7 is major neutralizing antigen that is important component of subunit vaccine to prevent rotavirus infection. Many efforts have been done to produce recombinant VP7 that maintain native characteristics. We used baculovirus expression system to produce rotavirus VP7 protein and to study its immunogenicity.
  
  
  METHODS
  Simian rotavirus SA11 full-length VP7 ORF was cloned into a cloning plasmid and then the cloned gene was inserted into the linear DNA of baculovirus Autographa californica Nuclear Polyhedrosis Virus (AcNPV) downstream of the polyhedrin promoter by in vitro recombination reactions. The expressed VP7 in the insect cells was recognized by rabbit hyperimmune serum raised against SA11 rotavirus by Immunofluorescence and western blotting assays. Rabbits were immunized subcutaneously by cell extracts expressing VP7 protein.
  
  
  RESULTS
  Reactivity with anti-rotavirus antibody suggested that expressed VP7 protein had native antigenic determinants. Injection of recombinant VP7 in rabbits elicited the production of serum antibodies, which were able to recognize VP7 protein from SA11 rotavirus by Western blotting test and neutralized SA11 rotavirus in cell culture.
  
  
  CONCLUSIONS
  Recombinant outer capsid glycoprotein (VP7) of rotavirus expressed in insect cells induces neutralizing antibodies in rabbits and may be a candidate of rotavirus vaccine.
authors.affiliation: Dept. of Microbiology, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran ; Cellular and Molecular Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran.
authors.forename: M
authors.initials: M
authors.lastname: Khodabandehloo
authors.forename: M Shamsi
authors.initials: MS
authors.lastname: Shahrabadi
authors.forename: H
authors.initials: H
authors.lastname: Keyvani
authors.forename: B
authors.initials: B
authors.lastname: Bambai
authors.forename: Za
authors.initials: Z
authors.lastname: Sadigh
keywords: Baculovirus; Expression; Insect cell; Rotavirus; VP7