Presentation of the hydrophilic domains of hepatitis C viral E2 envelope glycoprotein on hepatitis B surface antigen particles.
Subviral particles of hepatitis B virus have been used to present foreign epitopes. We attempted to present the hydrophilic domains of E2 envelope protein of hepatitis C virus (HCV) as a fusion protein with hepatitis B virus surface antigen (HBsAg). The five hydrophilic domains of HCV E2 antigen were inserted into HBsAg such that the inserted hydrophilic domains were presented on the outer surface of HBV subviral particles. In addition, a fusion encoding the hypervariable region (HVR) of E2 antigen was also made. Cell lysate and culture medium were analyzed for the synthesis and secretion of the fusion proteins by immunoprecipitation with polyclonal anti-HBsAg antibody using recombinant vaccinia virus system. The results showed that the fusion proteins containing these six E2 domains were made in the cell, but only two out of six fusion proteins were secreted into culture medium. Further, cesium chloride density gradient analysis and electron microscopy revealed that these fusions were secreted into culture media as particles. It will be of interest to test immunogenicity of the HBsAg fusion particles containing the HCV E2 domains in animal model.
Lee
I H
IH
Biotech Research Institute, LG Chemical Ltd/Research Park, Taejon, Korea.
Kim
C H
CH
Ryu
W S
WS
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D006224:Cricetinae; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D016174:Hepacivirus / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism*; D018936:Hepatitis C Antigens / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion
D006023:Glycoproteins; D006514:Hepatitis B Surface Antigens; D018936:Hepatitis C Antigens; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus