title: Identification of novel hepatitis C virus-specific cytotoxic T lymphocyte epiotpe in NS3 region.
abstract: Hepatitis C virus (HCV)-specific cytotoxic T lymphocytes (CTL) are thought to be effective in limiting viral spread and in clearing virus during infection. Therefore, we attempted to establish HCV-specific CTL and identify novel HCV-specific CTL epitopes in a patient with acute hepatitis C by a novel screening method using recombinant vaccinia viruses (rVV) and synthetic peptides. CD8(+)CD45RA(-) T cells (memory T cells) were isolated from peripheral blood mononuclear cells (PBMC) of a patient with acute hepatitis C. HCV-specific CTL were cloned at limited dilutions and tested for HCV-specific CTL activity using a standard (51)Cr release assay. CTL assay was performed using rVV expressing regions of HCV-J, and overlapping and truncated synthetic peptides from HCV-J. CTL recognizing the NS3 region were isolated by (51)Cr release assay with rVV-HCV. Isolated CTL were restricted by HLA class I molecules B(*)5603. We confirmed that isolated CTL recognized 8-mer amino acids in the NS3 region of HCV-J by (51)Cr release assay with overlapping and truncated synthetic peptides. In conclusion, we isolated HCV-specific CTL restricted by HLA-B(*)5603 and identified a novel HCV-specific CTL epitope (IPFYGKAI, amino acids 1373-1380) in the NS3 region. The identified HCV-specific CTL epitope might be useful for HCV therapy.
authors.affiliation: First Department of Internal Medicine, Mie University School of Medicine, 2-174 Edobashi, Tsu, Mie 514-8507, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Ito
authors.forename: Katsuya
authors.initials: K
authors.lastname: Shiraki
authors.forename: Kiyomi
authors.initials: K
authors.lastname: Funatsuki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Ishiko
authors.forename: Kazushi
authors.initials: K
authors.lastname: Sugimoto
authors.forename: Kazumoto
authors.initials: K
authors.lastname: Murata
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nakano
authors.forename: Michio
authors.initials: M
authors.lastname: Imawari