title: Generation of T-cell receptors targeting a genetically stable and immunodominant cytotoxic T-lymphocyte epitope within hepatitis C virus non-structural protein 3.
abstract: Hepatitis C virus (HCV) is a major cause of severe liver disease, and one major contributing factor is thought to involve a dysfunction of virus-specific T-cells. T-cell receptor (TCR) gene therapy with HCV-specific TCRs would increase the number of effector T-cells to promote virus clearance. We therefore took advantage of HLA-A2 transgenic mice to generate multiple TCR candidates against HCV using DNA vaccination followed by generation of stable T-cell-BW (T-BW) tumour hybrid cells. Using this approach, large numbers of non-structural protein 3 (NS3)-specific functional T-BW hybrids can be generated efficiently. These predominantly target the genetically stable HCV genotype 1 NS3(1073-1081) CTL epitope, frequently associated with clearance of HCV in humans. These T-BW hybrid clones recognized the NS3(1073) peptide with a high avidity. The hybridoma effectively recognized virus variants and targeted cells with low HLA-A2 expression, which has not been reported previously. Importantly, high-avidity murine TCRs effectively redirected human non-HCV-specific T-lymphocytes to recognize human hepatoma cells with HCV RNA replication driven by a subgenomic HCV replicon. Taken together, TCR candidates with a range of functional avidities, which can be used to study immune recognition of HCV-positive targets, have been generated. This has implications for TCR-related immunotherapy against HCV.
authors.affiliation: Department of Laboratory Medicine, Stockholm, Sweden.|Department of Dental Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Pasetto
authors.affiliation: Department of Laboratory Medicine, Stockholm, Sweden.
authors.forename: Lars
authors.initials: L
authors.lastname: Frelin
authors.affiliation: Department of Laboratory Medicine, Stockholm, Sweden.
authors.forename: Anette
authors.initials: A
authors.lastname: Brass
authors.affiliation: Department of Dental Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Anila
authors.initials: A
authors.lastname: Yasmeen
authors.affiliation: Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research (A*STAR), Singapore.|Department of Laboratory Medicine, Stockholm, Sweden.
authors.forename: Sarene
authors.initials: S
authors.lastname: Koh
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University of Heidelberg, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Lohmann
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University of Heidelberg, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
authors.affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet and Swedish Institute for Infectious Disease Control, Stockholm, Sweden.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Magalhaes
authors.affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet and Swedish Institute for Infectious Disease Control, Stockholm, Sweden.
authors.forename: Markus
authors.initials: M
authors.lastname: Maeurer
authors.affiliation: Department of Laboratory Medicine, Stockholm, Sweden.
authors.forename: Matti
authors.initials: M
authors.lastname: Sällberg
authors.affiliation: Department of Dental Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Margaret
authors.initials: M
authors.lastname: Chen
mesh_terms: D000818:Animals; D001782:Blood Donors; D002478:Cells, Cultured; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology; D016174:Hepacivirus / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D015789:HLA-A2 Antigen; D016056:Immunodominant Epitopes; D011948:Receptors, Antigen, T-Cell; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
references.citation: Gene Ther. 2003 Apr;10(8):686-99
references.pmid: 12692597
references.citation: Hepatology. 2007 Jun;45(6):1422-32
references.pmid: 17538964
references.citation: J Virol. 2008 Apr;82(7):3466-79
references.pmid: 18216094
references.citation: J Immunol. 2007 Oct 1;179(7):4741-53
references.pmid: 17878373
references.citation: J Exp Med. 2005 Mar 7;201(5):675-80
references.pmid: 15753202
references.citation: Eur J Immunol. 2006 Aug;36(8):2046-60
references.pmid: 16856207
references.citation: J Virol. 2008 Oct;82(20):10017-31
references.pmid: 18667516
references.citation: Nat Rev Immunol. 2006 Dec;6(12):883-94
references.pmid: 17110956
references.citation: Vaccine. 2008 Jul 23;26(31):3818-26
references.pmid: 18582999
references.citation: J Immunol. 2003 Mar 1;170(5):2582-9
references.pmid: 12594285
references.citation: Euro Surveill. 2008 May 22;13(21):
references.pmid: 18761967
references.citation: Hepatology. 2003 Dec;38(6):1437-48
references.pmid: 14647055
references.citation: J Immunol. 2002 Sep 15;169(6):3447-58
references.pmid: 12218168
references.citation: Immunol Lett. 2003 Jan 22;85(2):165-71
references.pmid: 12527224
references.citation: Hepatology. 2001 Dec;34(6):1206-17
references.pmid: 11732011
references.citation: Cell Immunol. 2000 Aug 1;203(2):111-23
references.pmid: 11006009
references.citation: J Transl Med. 2005 Sep 20;3:35
references.pmid: 16174302
references.citation: J Mol Med (Berl). 1996 Oct;74(10):583-8
references.pmid: 8912179
references.citation: Int J Cancer. 1999 Jun 11;81(6):979-84
references.pmid: 10362148
references.citation: J Clin Invest. 2008 Mar;118(3):1143-53
references.pmid: 18246203
references.citation: Eur J Immunol. 2000 Sep;30(9):2479-87
references.pmid: 11009080
references.citation: Expert Opin Biol Ther. 2010 May;10(5):749-62
references.pmid: 20201709
references.citation: J Virol. 2001 Dec;75(23):11392-400
references.pmid: 11689620
references.citation: Hepatology. 2006 Oct;44(4):945-54
references.pmid: 17006905
references.citation: Clin Cancer Res. 2010 Dec 1;16(23):5852-61
references.pmid: 21138872
references.citation: Hepatology. 2006 Feb;43(2 Suppl 1):S230-4
references.pmid: 16447286
references.citation: Immunol Cell Biol. 2011 Mar;89(3):340-5
references.pmid: 21301482
references.citation: Gastroenterology. 2001 Feb;120(2):512-24
references.pmid: 11159892
references.citation: Gene Ther. 2004 Mar;11(6):522-33
references.pmid: 14999224
references.citation: Virus Res. 2007 Aug;127(2):195-207
references.pmid: 17428568
references.citation: Hepatology. 2004 Nov;40(5):1062-71
references.pmid: 15486925
references.citation: J Immunol. 1990 Aug 1;145(3):804-11
references.pmid: 2115542
references.citation: Hepatology. 2005 Jul;42(1):104-12
references.pmid: 15962289
references.citation: Hepatology. 2003 Mar;37(3):577-89
references.pmid: 12601356
references.citation: J Immunol. 2005 Sep 15;175(6):3603-13
references.pmid: 16148104
references.citation: PLoS Pathog. 2010 Jul 29;6(7):e1001018
references.pmid: 20686664
references.citation: J Infect Dis. 2003 Jun 15;187(12):1951-8
references.pmid: 12792872
references.citation: J Viral Hepat. 1999 Jul;6 Suppl 1:36-40
references.pmid: 10760033
references.citation: Eur J Immunol. 2008 Oct;38(10):2665-77
references.pmid: 18958874
references.citation: Hepatology. 2005 Mar;41(3):643-51
references.pmid: 15726647
references.citation: J Exp Med. 2005 Sep 19;202(6):817-28
references.pmid: 16157687
references.citation: J Exp Med. 1997 Jun 16;185(12):2043-51
references.pmid: 9182675
references.citation: Gut. 2006 Feb;55(2):266-74
references.pmid: 16105887
references.citation: J Immunol. 2011 Jan 15;186(2):901-12
references.pmid: 21160049
references.citation: Hepatology. 2008 Sep;48(3):713-22
references.pmid: 18712791
references.citation: J Virol. 2002 Dec;76(24):12735-46
references.pmid: 12438599
references.citation: Blood. 2005 May 1;105(9):3605-14
references.pmid: 15657184
references.citation: J Virol. 2007 Mar;81(6):2545-53
references.pmid: 17182670
references.citation: Aliment Pharmacol Ther. 2009 Apr 1;29(7):689-705
references.pmid: 19183149