Identification of novel hepatitis C virus-specific cytotoxic T lymphocyte epiotpe in NS3 region
Identification of novel hepatitis C virus-specific cytotoxic T lymphocyte epiotpe in NS3 region
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DOI：
10.1016/j.hepres.2006.08.011
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发表时间：
2006-12-01
影响因子：
4.2
通讯作者：
Imawari, Michio
中科院分区：
文献类型：
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作者：
Ito, Keiichi;Shiraki, Katsuya;Imawari, Michio
丙型肝炎病毒（HCV）特异性细胞毒性T淋巴细胞（CTL）被认为是有效地限制病毒传播和清除感染期间的病毒。因此，我们试图建立丙型肝炎病毒特异性CTL和确定新的丙型肝炎病毒特异性CTL表位的急性丙型肝炎患者的一种新的筛选方法，使用重组痘苗病毒（rVV）和合成肽。从急性丙型肝炎患者的外周血单个核细胞（PBMC）中分离CD 8（+）CD 45 RA（-）T细胞（记忆T细胞）。以有限稀释度克隆HCV特异性CTL，并使用标准Cr-51释放试验测试HCV特异性CTL活性。用rVV-J表达区和重叠及截短的HCV-J合成肽进行CTL试验。用rVV-HCV通过51 Cr释放试验分离识别NS 3区的CTL。分离的CTL受HLA I类分子B*5603限制。我们证实，分离的CTL识别HCV-J的NS 3区的8-mer氨基酸与重叠和截短的合成肽的51 Cr释放试验。总之，我们分离了HLA-B*5603限制的HCV特异性CTL，并在NS 3区鉴定了新的HCV特异性CTL表位（IPFYGKAI，氨基酸1373 - 1380）。所鉴定的HCV特异性CTL表位为HCV的治疗提供了可能. (c)2006爱思唯尔爱尔兰有限公司保留所有权利。
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 Cr-51 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 51Cr 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 51Cr 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. (c) 2006 Elsevier Ireland Ltd. All rights reserved.