Mapping of a conformational epitope shared between E1 and E2 on the serum-derived human hepatitis C virus envelope.
Monoclonal antibody D32.10 produced by immunizing mice with a hepatitis C virus (HCV)-enriched pellet obtained from plasmapheresis of a chronically HCV1b-infected patient binds HCV particles derived from serum of different HCV1a- and HCV1b-infected patients. Moreover, this monoclonal has been shown to recognize both HCV envelope proteins E1 and E2. In an attempt to provide novel insight into the membrane topology of HCV envelope glycoproteins E1 and E2, we localized the epitope recognized by D32.10 on the E1 and/or E2 sequence using Ph.D.-12 phage display peptide library technology. Mimotopes selected from the phage display dodecapeptide library by D32.10 shared partial similarities with 297RHWTTQGCNC306 of the HCV E1 glycoprotein and with both 613YRLWHYPCT621 and 480PDQRPYCWHYPPKPC494 of the HCV E2 glycoprotein. Immunoreactivity of D32.10 with overlapping peptides corresponding to these three HCV regions confirmed these localizations and suggested that the three regions identified are likely closely juxtaposed on the surface of serum-derived particles as predicted by the secondary model structure of HCV E2 derived from the tick-borne encephalitis virus E protein. This assertion was supported by the detection of specific antibodies directed against these three E1E2 regions in sera from HCV-infected patients.
Petit
Marie-Anne
MA
INSERM Unité 271, 151 cours Albert Thomas, 69424 Lyon 03, France. petit@lyon.inserm.fr
Jolivet-Reynaud
Colette
C
Peronnet
Estelle
E
Michal
Yvonne
Y
Trépo
Christian
C
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry*; D016174:Hepacivirus / Q000737:chemistry / Q000276:immunology; D006526:Hepatitis C / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000097:blood / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019151:Peptide Library; D011487:Protein Conformation; D014759:Viral Envelope Proteins / Q000276:immunology*; D015678:Viral Structural Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D018937:Hepatitis C Antibodies; D019151:Peptide Library; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; C107842:protein E1, Classical swine fever virus; C091015:glycoprotein E2, Hepatitis C virus