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Stable human lymphoblastoid cell lines constitutively expressing hepatitis C virus proteins

Benno Wölk^1,,

Christel Gremion^2,

Natalia Ivashkina^1,

¹ Department of Medicine II, University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany
² Clinic for Rheumatology and Clinical Immunology/Allergology, Inselspital, University of Bern, CH-3010 Bern, Switzerland
³ Department of Medicine, Ospedale Regionale di Lugano, Via Tesserete 46, CH-6903 Lugano, Switzerland

Correspondence
Darius Moradpour
Darius.Moradpour{at}hospvd.ch

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-lymphoblastoidxT 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.

These authors contributed equally to this work.

Present address: Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10021, USA.

Present address: Department of Immunology, University of Göttingen, D-37075 Göttingen, Germany.

||Present address: Division of Gastroenterology and Hepatology, Centre Hospitalier Universitaire Vaudois, Rue du Bugnon 44, CH-1011 Lausanne, Switzerland.

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