Expression of HCV envelope proteins and the serological utility of the anti-E2 immune response.
The 5' end of the hepatitis C virus (HCV) genome encodes structural proteins of the virion. The first gene encodes a highly basic core protein. Immediately downstream of the core gene are regions which encode the envelope proteins (E1 and E2) of the virus. Artificial expression and secretion of immunologically active envelope proteins have proven to be a substantial challenge due to the high degree of glycosylation and the existence of certain hydrophobic domains contained within these sequences. Bacterial cell expression of recombinant HCV envelope proteins results in products that are not glycosylated and are poorly immunogenic. Emphasis has shifted to the use of mammalian cell lines (human embryonic kidney [HEK] and Chinese hamster ovary [CHO] cells) for the expression of glycosylated, immunologically active envelope proteins. Using HEK cells, E1 is expressed intracellularly but is not secreted from the cells. When E1 is cloned in fusion with a C-terminal truncated E2 protein, both proteins are detected intracellularly; however, only E2 is secreted. When the E1/E2 processing site is interrupted by constructing deletion mutants, the unprocessed E1/E2 fusion protein can be secreted from the cells. Quantifiable expression and secretion of a truncated E2 protein is now possible using CHO cells and SV40-based vectors. The HCV E2 glycoprotein expressed from CHO cells is highly antigenic; a strong humoral response to this antigen develops in persons infected with HCV. Antibodies to E2 are found in 95% of patients with detectable HCV RNA in their sera. The presence of antibodies to E2 is not indicative of viral clearance and therefore the role these antibodies play in protective immunity, if any, is unclear.
Lesniewski
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Hepatitis Research and Development, Abbott Laboratories, Abbott Park, Illinous 60064, USA.
Watanabe
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Devare
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D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D016174:Hepacivirus / Q000276:immunology / Q000378:metabolism*; D018937:Hepatitis C Antibodies / Q000097:blood* / Q000276:immunology; D006801:Humans; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000276:immunology
C087035:E1 protein, Hepatitis C virus; D018937:Hepatitis C Antibodies; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus