Development and characterisation of recombinant hepatitis delta virus-like particles

Ward, Scott Matthew, Macnaughton, Thomas Bernard and Gowans, Eric James (2001) Development and characterisation of recombinant hepatitis delta virus-like particles. Virus Genes, 23 1: 97-104. doi:10.1023/A:1011195715747


Author Ward, Scott Matthew
Macnaughton, Thomas Bernard
Gowans, Eric James
Title Development and characterisation of recombinant hepatitis delta virus-like particles
Journal name Virus Genes   Check publisher's open access policy
ISSN 0920-8569
1572-994X
Publication date 2001-08
Sub-type Article (original research)
DOI 10.1023/A:1011195715747
Volume 23
Issue 1
Start page 97
End page 104
Total pages 8
Editor Yechiel Becker (Editor in Chief)
Place of publication Dordrecht, The Netherlands
Publisher Kluwer Academic Publishers
Collection year 2001
Language eng
Subject C1
270303 Virology
730101 Infectious diseases
Abstract Injection of particulate hepatitis B virus surface antigen (HBsAg) in mice leads to the induction of a HBsAg-specific class-I-restricted cytotoxic T lymphocyte (CTL) response. It is proposed that any protein internal to HBsAg will also be able to elicit a specific CTL response. In this study, several carboxy-terminal truncations of hepatitis C virus (HCV) core protein were fused to varying lengths of amino-terminal truncated large hepatitis delta antigen (L-HDAg). These constructs were analysed for their ability to be expressed and the particles secreted in the presence of HBsAg after transfection into HuH-7 cells. The secretion efficiency of the various HCV core-HDAg chimeric proteins was generally poor. Constructs containing full length HDAg appeared to be more stable than truncated versions and the length of the inserted protein was restricted to around 40 amino acids. Thus, the use of L-HDAg as a chimera to package foreign proteins is limited. Consequently, a polyepitope (polytope) containing a B-cell epitope from human papillomavirus (HPV 16) and multiple T-cell epitopes from the HCV polyprotein was used to create the construct, L-HDAg-polyB. This chimeric protein was shown to be reliant on the co-expression of HBsAg for secretion into the cell culture fluid and was secreted more efficiently than the previous HCV core-HDAg constructs. These L-HDAg-polyB virus-like particles (VLPs) had a buoyant density of similar to 1.2 g/cm(3) in caesium chloride and similar to 1.15 g/cm(3) in sucrose. The VLPs were also immunoprecipitated using an anti-HBs but not an anti-HD antibody. Thus, these recombinant VLPs have similar biophysical properties to L-HDAg VLPs.
Keyword Genetics & Heredity
Virology
Hepatitis B Surface Antigen
Hepatitis Delta Antigen
Virus-like Particles
Cytotoxic T-cells
B Surface-antigen
Mhc Class-i
C Virus
Protein
Replication
Peptides
Epitopes
Identification
Molecules
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collection: Clinical Medical Virology Centre Publications
 
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Created: Tue, 14 Aug 2007, 16:07:52 EST