title: Development of a plant-derived subunit vaccine candidate against hepatitis C virus.
abstract: Hepatitis C virus (HCV) is a major cause of acute and chronic hepatitis with over 180 million cases worldwide. Vaccine development for HCV has been difficult. Presently, the virus cannot be grown in tissue culture and there is no vaccine or effective therapy against this virus. In this research, we describe the development of an experimental plant-derived subunit vaccine against HCV. A tobamoviral vector was engineered to encode a consensus sequence of hypervariable region 1 (HVR1), a potential neutralizing epitope of HCV, genetically fused to the C-terminal of the B subunit of cholera toxin (CTB). This epitope was selected from among the amino acid sequences of HVR1 "mimotopes" previously derived by phage display technology. The nucleotide sequence encoding this epitope was designed utilizing optimal plant codons. This mimotope is capable of inducing cross-neutralizing antibodies against different variants of the virus. Plants infected with recombinant tobacco mosaic virus (TMV) engineered to express the HVR1/CTB chimeric protein, contained intact TMV particles and produced the HVR1 consensus peptide fused to the functionally active, pentameric B subunit of cholera toxin. Plant-derived HVR1/CTB reacted with HVR1-specific monoclonal antibodies and immune sera from individuals infected with virus from four of the major genotypes of HCV. Intranasal immunization of mice with a crude plant extract containing the recombinant HVR1/CTB protein elicited both anti-CTB serum antibody and anti-HVR1 serum antibody which specifically bound to HCV virus-like particles. Using plant-virus transient expression to produce this unique chimeric antigen will facilitate the development and production of an experimental HCV vaccine. A plant-derived recombinant HCV vaccine can potentially reduce expenses normally associated with production and delivery of conventional vaccines.
authors.affiliation: Vaccine and Therapeutic Development Section, Oral Infection and Immunity Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892-4350, USA.
authors.forename: L G
authors.initials: LG
authors.lastname: Nemchinov
authors.forename: T J
authors.initials: TJ
authors.lastname: Liang
authors.forename: M M
authors.initials: MM
authors.lastname: Rifaat
authors.forename: H M
authors.initials: HM
authors.lastname: Mazyad
authors.forename: A
authors.initials: A
authors.lastname: Hadidi
authors.forename: J M
authors.initials: JM
authors.lastname: Keith
mesh_terms: D000281:Administration, Intranasal; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002772:Cholera Toxin / Q000235:genetics / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D005260:Female; D005822:Genetic Vectors; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology; D006526:Hepatitis C / Q000517:prevention & control; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000096:biosynthesis* / Q000276:immunology; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014027:Tobacco Mosaic Virus / Q000235:genetics; D014611:Vaccination; D022223:Vaccines, Subunit / Q000096:biosynthesis / Q000276:immunology; D014761:Viral Hepatitis Vaccines / Q000008:administration & dosage / Q000096:biosynthesis* / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000939:Epitopes; C400562:HVR1 protein, Hepatitis C virus; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D022223:Vaccines, Subunit; D014761:Viral Hepatitis Vaccines; D014764:Viral Proteins; D002772:Cholera Toxin