Wölk Benno, Gremion Christel, Ivashkina Natalia, Engler Olivier B, Grabscheid Benno, Bieck Elke, Blum Hubert E, Cerny Andreas, Moradpour Darius
Department of Medicine II, University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany.
Clinic for Rheumatology and Clinical Immunology/Allergology, Inselspital, University of Bern, CH-3010 Bern, Switzerland.
J Gen Virol. 2005 Jun;86(Pt 6):1737-1746. doi: 10.1099/vir.0.80853-0.
The cellular immune response plays a central role in virus clearance and pathogenesis of liver disease in hepatitis C. The study of hepatitis C virus (HCV)-specific immune responses is limited by currently available cell-culture systems. Here, the establishment and characterization of stable human HLA-A2-positive B-lymphoblastoid x T hybrid cell lines constitutively expressing either the NS3-4A complex or the entire HCV polyprotein are reported. These cell lines, termed T1/NS3-4A and T1/HCVcon, respectively, were maintained in continuous culture for more than 1 year with stable characteristics. HCV structural and non-structural proteins were processed accurately, indicating that the cellular and viral proteolytic machineries are functional in these cell lines. Viral proteins were found in the cytoplasm in dot-like structures when expressed in the context of the HCV polyprotein or in a perinuclear fringe when the NS3-4A complex was expressed alone. T1/NS3-4A and T1/HCVcon cells were lysed efficiently by HCV-specific cytotoxic T lymphocytes from patients with hepatitis C and from human HLA-A2.1 transgenic mice immunized with a liposomal HCV vaccine, indicating that viral proteins are processed endogenously and presented efficiently via the major histocompatibility complex class I pathway. In conclusion, these cell lines represent a unique tool to study the cellular immune response, as well as to evaluate novel vaccine and immunotherapeutic strategies against HCV.
细胞免疫反应在丙型肝炎病毒清除及肝脏疾病发病机制中起核心作用。丙型肝炎病毒（HCV）特异性免疫反应的研究受到现有细胞培养系统的限制。本文报道了稳定的人HLA - A2阳性B淋巴母细胞x T杂交细胞系的建立及特性，这些细胞系组成性表达NS3 - 4A复合物或完整的HCV多聚蛋白。这些细胞系分别称为T1/NS3 - 4A和T1/HCVcon，连续培养超过1年，特性稳定。HCV结构蛋白和非结构蛋白被准确加工，表明细胞和病毒蛋白水解机制在这些细胞系中发挥功能。当在HCV多聚蛋白背景下表达时，病毒蛋白在细胞质中呈点状结构，而单独表达NS3 - 4A复合物时，病毒蛋白呈核周边缘分布。来自丙型肝炎患者和用脂质体HCV疫苗免疫的人HLA - A2.1转基因小鼠的HCV特异性细胞毒性T淋巴细胞可有效裂解T1/NS3 - 4A和T1/HCVcon细胞，这表明病毒蛋白通过内源性加工并通过主要组织相容性复合体I类途径有效呈递。总之，这些细胞系是研究细胞免疫反应以及评估针对HCV的新型疫苗和免疫治疗策略的独特工具。
