Prokaryotic Expression of Hepatitis C Virus-NS3 Protein and Preparation of a Monoclonal Antibody

2017 ◽  
Vol 36 (6) ◽  
pp. 251-258 ◽  
Author(s):  
Yun Xi ◽  
Yuming Zhang ◽  
Jianmin Fang ◽  
Kelly Whittaker ◽  
Shuhong Luo ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Prokaryotic Expression ◽  
Ns3 Protein

685 IMMUNIZATION AGAINST HEPATITIS C VIRUS USING A PEPTIDE FROM NS3 PROTEIN LINKED TO MODULINS DERIVED FROM STAPHYLOCOCCUS EPIDERMIDIS

2010 ◽  
Vol 52 ◽  
pp. S267
Author(s):  
M. Durantez ◽  
C. Fayole ◽  
N. Casares ◽  
V. Belsue ◽  
J.I. Riezu-Boj ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Staphylococcus Epidermidis ◽  
Ns3 Protein

scholarly journals The N-terminal helix α0 of hepatitis C virus NS3 protein dictates the subcellular localization and stability of NS3/NS4A complex

Virology ◽  
2012 ◽  
Vol 422 (2) ◽  
pp. 214-223 ◽  
Author(s):  
Ying He ◽  
Leiyun Weng ◽  
Rui Li ◽  
Li Li ◽  
Tetsuya Toyoda ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Subcellular Localization ◽  
Ns3 Protein

scholarly journals Isolation of RNA Aptamers Specific to the NS3 Protein of Hepatitis C Virus from a Pool of Completely Random RNA

Virology ◽  
1997 ◽  
Vol 237 (2) ◽  
pp. 270-282 ◽  
Author(s):  
P.K.R. Kumar ◽  
Keigo Machida ◽  
Petri T. Urvil ◽  
Nobuko Kakiuchi ◽  
Daesety Vishnuvardhan ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Rna Aptamers ◽  
Ns3 Protein

scholarly journals DNA Vaccination Protects Mice against Challenge with Listeria monocytogenes Expressing the Hepatitis C Virus NS3 Protein

2003 ◽  
Vol 71 (11) ◽  
pp. 6372-6380 ◽  
Author(s):  
Benjamin E. Simon ◽  
Kenneth A. Cornell ◽  
Tina R. Clark ◽  
Sunwen Chou ◽  
Hugo R. Rosen ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Listeria Monocytogenes ◽  
Immune Responses ◽  
Bacterial Pathogen ◽  
Cell Responses ◽  
Ns3 Protein ◽  
Plasmid Dna Vaccine ◽  
In Vivo Induction

ABSTRACT The goal of this study was to develop a new surrogate challenge model for use in evaluating protective cell-mediated immune responses against hepatitis C virus (HCV) antigens. The use of recombinant Listeria monocytogenes organisms which express HCV antigens provides novel tools with which to assay such in vivo protection, as expression of immunity against this hepatotropic bacterial pathogen is dependent on antigen-specific CD8+ T lymphocytes. A plasmid DNA vaccine encoding a ubiquitin-NS3 fusion protein was generated, and its efficacy was confirmed by in vivo induction of NS3-specific, gamma interferon-secreting T cells following vaccination of BALB/c mice. These immunized mice also exhibited specific in vivo protection against subsequent challenge with a recombinant L. monocytogenes strain (TC-LNS3) expressing the NS3 protein. Notably, sublethal infection of naive mice with strain TC-LNS3 induced similar NS3-specific T-cell responses. These findings suggest that recombinant strains of L. monocytogenes expressing HCV antigens should prove useful for evaluating, or even inducing, protective immune responses against HCV antigens.

scholarly journals A Point Mutation Leading to Hepatitis C Virus Escape from Neutralization by a Monoclonal Antibody to a Conserved Conformational Epitope

2008 ◽  
Vol 82 (12) ◽  
pp. 6067-6072 ◽  
Author(s):  
Zhen-Yong Keck ◽  
Oakley Olson ◽  
Meital Gal-Tanamy ◽  
Jinming Xia ◽  
Arvind H. Patel ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Vaccine Development ◽  
Human Monoclonal Antibody ◽  
Conformational Epitope ◽  
Single Amino Acid ◽  
Effective Vaccine ◽  
Linear Sequence ◽  
Virus Escape

ABSTRACT A challenge in hepatitis C virus (HCV) vaccine development is defining conserved protective epitopes. A cluster of these epitopes comprises an immunodominant domain on the E2 glycoprotein, designated domain B. CBH-2 is a neutralizing human monoclonal antibody to a domain B epitope that is highly conserved. Alanine scanning demonstrated that the epitope involves residues G523, G530, and D535 that are also contact residues for E2 binding to CD81, a coreceptor required for virus entry into cells. However, another residue, located at position 431 and thus at a considerable distance in the linear sequence of E2, also contributes to the CBH-2 epitope. A single amino acid substitution at this residue results in escape from CBH-2-mediated neutralization in a genotype 1a virus. These results highlight the challenges inherent in developing HCV vaccines and show that an effective vaccine must induce antibodies to both conserved and more invariant epitopes to minimize virus escape.

scholarly journals Monoclonal Antibody AP33 Defines a Broadly Neutralizing Epitope on the Hepatitis C Virus E2 Envelope Glycoprotein

2005 ◽  
Vol 79 (17) ◽  
pp. 11095-11104 ◽  
Author(s):  
Ania Owsianka ◽  
Alexander W. Tarr ◽  
Vicky S. Juttla ◽  
Dimitri Lavillette ◽  
Birke Bartosch ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Envelope Glycoprotein ◽  
Envelope Protein ◽  
Hypervariable Region ◽  
Neutralizing Antibody ◽  
Broadly Neutralizing Antibody ◽  
New Treatments ◽  
Potent Neutralization

ABSTRACT Hepatitis C virus (HCV) remains a significant threat to the general health of the world's population, and there is a pressing need for the development of new treatments and preventative vaccines. Here, we describe the generation of retrovirus-based pseudoparticles (HCVpp) incorporating a panel of full-length E1E2 clones representative of the major genotypes 1 through 6, and their application to assess the reactivity and neutralizing capability of antisera and monoclonal antibodies raised against portions of the HCV E2 envelope protein. Rabbit antisera raised against either the first hypervariable region or ectodomain of E2 showed limited and strain specific neutralization. By contrast, the monoclonal antibody (MAb) AP33 demonstrated potent neutralization of infectivity against HCVpp carrying E1E2 representative of all genotypes tested. The concentration of AP33 required to achieve 50% inhibition of infection by HCVpp of diverse genotypes ranged from 0.6 to 32 μg/ml. The epitope recognized by MAb AP33 is linear and highly conserved across different genotypes of HCV. Thus, identification of a broadly neutralizing antibody that recognizes a linear epitope is likely to be of significant benefit to future vaccine and therapeutic antibody development.

scholarly journals The Arginine-1493 Residue in QRRGRTGR1493G Motif IV of the Hepatitis C Virus NS3 Helicase Domain Is Essential for NS3 Protein Methylation by the Protein Arginine Methyltransferase 1

2001 ◽  
Vol 75 (17) ◽  
pp. 8031-8044 ◽  
Author(s):  
Jaerang Rho ◽  
Seeyoung Choi ◽  
Young Rim Seong ◽  
Joonho Choi ◽  
Dong-Soo Im
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Rna Binding ◽  
Rna Helicase ◽  
Full Length ◽  
Arginine Residue ◽  
Protein Methylation ◽  
Helicase Domain ◽  
Rna Helicases ◽  
Ns3 Protein

ABSTRACT The NS3 protein of hepatitis C virus (HCV) contains protease and RNA helicase activities, both of which are likely to be essential for HCV propagation. An arginine residue present in the arginine-glycine (RG)-rich region of many RNA-binding proteins is posttranslationally methylated by protein arginine methyltransferases (PRMTs). Amino acid sequence analysis revealed that the NS3 protein contains seven RG motifs, including two potential RG motifs in the 1486-QRRGRTGRG-1494 motif IV of the RNA helicase domain, in which arginines are potentially methylated by PRMTs. Indeed, we found that the full-length NS3 protein is arginine methylated in vivo. The full-length NS3 protein and the NS3 RNA helicase domain were methylated by a crude human cell extract. The purified PRMT1 methylated the full-length NS3 and the RNA helicase domain, but not the NS3 protease domain. The NS3 helicase bound specifically and comigrated with PRMT1 in vitro. Mutational analyses indicate that the Arg1493 in the QRR1488GRTGR1493G region of the NS3 RNA helicase is essential for NS3 protein methylation and that Arg1488 is likely methylated. NS3 protein methylation by the PRMT1 was decreased in the presence of homoribopolymers, suggesting that the arginine-rich motif IV is involved in RNA binding. The results suggest that an arginine residue(s) in QRXGRXGR motif IV conserved in the virus-encoded RNA helicases can be posttranslationally methylated by the PRMT1.

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