بيان و خنثي سازي گليكوپروتئين نو تركيب روتا ويروس ميموني (NSP4) با آنتي بادي اختصاصي در نوزاد موش

Expression and Neutralization of Simian Rotavirus Recombinant Glycoprotein (NSP4) with Specific Antibody in Neonatal Mice

Background and Objectives: Rotaviruses are the most important cause of severe diarrhea in infants and young children in both developed and developing countries. Rotaviruses are members of the reoviridae family and contain 11 double-stranded RNA segments. Segment 10 encodes nonstructural glycoprotein NSP4, that can induce diarrhea in newborn mice. It has been suggested that NSP4 may be a key determinant for rotavirus pathogenesis and a target for vaccine development. Hence, the aim of this study was to examine the entire expression of mammalian rotavirus (sa11) NSP4 gene in E.coli and also its biological and immunogenesity properties in animal models. Materials and Methods: In this experimental study, expression of NSP4 was demonstrated by Western blotting. The recombinant Protein from pYS50 was purified by Preparative SDS polyacrylamid gel electrophoresis. Antibody against recombinant NSP4 was raised in rabbit. Intraperitoneal administration of full-length recombinant NSP4 caused diarrhea in 100% of the 4 to 5 days BALB/C neonatal mice. Results: Intraperitoneal and oral inoculation of NSP4 antiserum significantly decreased diarrhea. Interaperitoneal administration of the full-length NSP4 induced diarrhea in 100% (5/5) of 4 to 5 days old BALB/C mice. Diarrhea in mice that received antibody against recombinant NSP4 was significantly decreased (p<0.000). Conclusion: These results indicated successful expression of the biologically active full-length NSP4 in E.coli and confirmed that recombinant NSP4 was able to induced diarrhea in neonatal mice and also had enterotoxigenic activity. The recombinant NSP4 which produced in this study can be applied as antigen for detection of specific antibodies against NSP4 in human sera