Stable human lymphoblastoid cell lines constitutively expressing hepatitis C virus proteins.
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.
Wölk
Benno
B
Department of Medicine II, University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany.
Gremion
Christel
C
Clinic for Rheumatology and Clinical Immunology/Allergology, Inselspital, University of Bern, CH-3010 Bern, Switzerland.
Ivashkina
Natalia
N
Department of Medicine II, University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany.
Engler
Olivier B
OB
Clinic for Rheumatology and Clinical Immunology/Allergology, Inselspital, University of Bern, CH-3010 Bern, Switzerland.
Grabscheid
Benno
B
Clinic for Rheumatology and Clinical Immunology/Allergology, Inselspital, University of Bern, CH-3010 Bern, Switzerland.
Bieck
Elke
E
Department of Medicine II, University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany.
Blum
Hubert E
HE
Department of Medicine II, University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany.
Cerny
Andreas
A
Department of Medicine, Ospedale Regionale di Lugano, Via Tesserete 46, CH-6903 Lugano, Switzerland.
Moradpour
Darius
D
Department of Medicine II, University of Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany.
D000818:Animals; D045744:Cell Line, Tumor / Q000378:metabolism; D002999:Clone Cells / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D015789:HLA-A2 Antigen / Q000235:genetics; D016174:Hepacivirus / Q000378:metabolism*; D006526:Hepatitis C / Q000276:immunology; D051379:Mice; D008822:Mice, Transgenic; D020815:Polyproteins / Q000378:metabolism; D014761:Viral Hepatitis Vaccines / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000378:metabolism*
D015789:HLA-A2 Antigen; D020815:Polyproteins; D014761:Viral Hepatitis Vaccines; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins