recombinant outer capsid glycoprotein (vp7) of rotavirus expressed in insect cells induces neutralizing antibodies in rabbits

Authors

m khodabandehloo

m shamsi shahrabadi

h keyvani

b bambai

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. conclusion: recombinant outer capsid glycoprotein (vp7) of rotavirus expressed in insect cells induces neutralizing antibodies in rabbits and may be a candidate of rotavirus vaccine.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

Recombinant Outer Capsid Glycoprotein (VP7) of Rotavirus Expressed in Insect Cells Induces Neutralizing Antibodies in Rabbits

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 st...

full text

Cloning and Expression of Simian Rotavirus Spike Protein (VP4) in Insect Cells by Baculovirus Expression System

Background: VP4 protein is as spikes on rotavirus outer capsid shell which is responsible for virus attachment to the host. VP4 induces production of neutralizing antibodies which could be used for serotyping of different isolates. Methods: Simian rotavirus SA11 gene 4 cDNA was cloned into a cloning plasmid pDONRTM by recombination reaction using clonase II enzyme mix. The resulting clone was c...

full text

Purified recombinant rotavirus VP7 forms soluble, calcium-dependent trimers.

Rotavirus is a major cause of severe, dehydrating childhood diarrhea. VP7, the rotavirus outer capsid glycoprotein, is a target of protective antibodies and is responsible for the calcium-dependent uncoating of the virus during cell entry. We have purified, characterized, and crystallized recombinant rhesus rotavirus VP7, expressed in insect cells. A critical aspect of the purification is the e...

full text

Cross-linking of rotavirus outer capsid protein VP7 by antibodies or disulfides inhibits viral entry.

Antibodies that neutralize rotavirus infection target outer coat proteins VP4 and VP7 and inhibit viral entry. The structure of a VP7-Fab complex (S. T. Aoki, et al., Science 324:1444-1447, 2009) led us to reclassify epitopes into two binding regions at inter- and intrasubunit boundaries of the calcium-dependent trimer. It further led us to show that antibodies binding at the intersubunit bound...

full text

Two proline residues are essential in the calcium-binding activity of rotavirus VP7 outer capsid protein.

Rotavirus maturation and stability of the outer capsid are calcium-dependent processes. It has been shown previously that the concentration of Ca2+-solubilizing outer capsid proteins from rotavirus particles is dependent on the virus strain. This property of viral particles has been associated with the gene coding for VP7 (gene 9). In this study the correlation between VP7 and resistance to low...

full text

My Resources

Save resource for easier access later


Journal title:
iranian journal of public health

جلد ۴۱، شماره ۵، صفحات ۷۳-۸۴

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023