title: Development of a Wilms' tumor antigen-specific T-cell receptor for clinical trials: engineered patient's T cells can eliminate autologous leukemia blasts in NOD/SCID mice.
abstract: BACKGROUND
  The Wilms' tumor antigen (WT1) is an attractive target for immunotherapy of leukemia. In the past, we isolated and characterized the specificity and function of a WT1-specific T-cell receptor. The goal of this translational study was to develop a safe and efficient WT1-T-cell receptor retroviral vector for an adoptive immunotherapy trial with engineered T cells.
  
  
  METHODS
  We generated a panel of retroviral constructs containing unmodified or codon-optimized WT1-T-cell receptor alpha and beta genes, linked via internal ribosome entry sites or 2A sequences, with or without an additional inter-chain disulfide bond in the T-cell receptor constant domains. These constructs were functionally analyzed in vitro, and the best one was tested in an autologous primary leukemia model in vivo.
  
  
  RESULTS
  We identified a WT1-T-cell receptor construct that showed optimal tetramer staining, antigen-specific cytokine production and killing activity when introduced into primary human T cells. Fresh CD34(+) cells purified from a patient with leukemia were engrafted into NOD/SCID mice, followed by adoptive immunotherapy with patient's autologous T cells transduced with the WT1-T-cell receptor. This therapeutic treatment evidently decreased leukemia engraftment in mice and resulted in a substantial improvement of leukemia-free survival.
  
  
  CONCLUSIONS
  This is the first report that patient's T cells, engineered to express the WT1-T-cell receptor, can eliminate autologous leukemia progenitor cells in an in vivo model. This study provides a firm basis for the planned WT1-T-cell receptor gene therapy trial in leukemia patients.
authors.affiliation: 1Department of Immunology, Division of Infection & Immunity, University College London, Royal Free Hospital, Rowland Hill Street,London, UK. s.xue@medsch.ucl.ac.uk
authors.forename: Shao-An
authors.initials: SA
authors.lastname: Xue
authors.forename: Liquan
authors.initials: L
authors.lastname: Gao
authors.forename: Sharyn
authors.initials: S
authors.lastname: Thomas
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Hart
authors.forename: John Zhao
authors.initials: JZ
authors.lastname: Xue
authors.forename: Roopinder
authors.initials: R
authors.lastname: Gillmore
authors.forename: Ralf-Holger
authors.initials: RH
authors.lastname: Voss
authors.forename: Emma
authors.initials: E
authors.lastname: Morris
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Stauss
mesh_terms: D000328:Adult; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001752:Blast Crisis / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D005818:Genetic Engineering / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000096:biosynthesis / Q000737:chemistry; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D006801:Humans; D019169:Jurkat Cells; D007938:Leukemia / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000502:physiology / Q000627:therapeutic use*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D014182:Transplantation, Autologous / Q000276:immunology; D009396:Wilms Tumor / Q000276:immunology / Q000473:pathology* / Q000628:therapy
chemical_list: D000951:Antigens, Neoplasm; D011948:Receptors, Antigen, T-Cell
references.citation: Nat Biotechnol. 2004 May;22(5):589-94
references.pmid: 15064769
references.citation: Eur J Haematol. 2009 Jan;82(1):61-8
references.pmid: 18801058
references.citation: Nat Rev Immunol. 2008 Apr;8(4):247-58
references.pmid: 18323851
references.citation: Leukemia. 1995 Jun;9(6):1060-7
references.pmid: 7596170
references.citation: Blood. 1996 Sep 15;88(6):2267-78
references.pmid: 8822948
references.citation: Blood. 2007 Mar 15;109(6):2331-8
references.pmid: 17082316
references.citation: Bone Marrow Transplant. 2004 Oct;34(7):653-4
references.pmid: 15258561
references.citation: Haematologica. 2005 Oct;90(10):1415-21
references.pmid: 16219579
references.citation: Blood. 1997 Feb 15;89(4):1405-12
references.pmid: 9028964
references.citation: Leukemia. 1999 Mar;13(3):393-9
references.pmid: 10086730
references.citation: Blood. 2008 Jan 1;111(1):236-42
references.pmid: 17875804
references.citation: Proc Natl Acad Sci U S A. 2004 Sep 21;101(38):13885-90
references.pmid: 15365188
references.citation: Eur J Haematol. 2008 Mar;80(3):201-7
references.pmid: 18081724
references.citation: Science. 2002 Oct 25;298(5594):850-4
references.pmid: 12242449
references.citation: Blood. 2000 Apr 1;95(7):2198-203
references.pmid: 10733485
references.citation: Blood. 2003 Mar 1;101(5):1698-704
references.pmid: 12406915
references.citation: Blood. 1994 Nov 1;84(9):3071-9
references.pmid: 7949179
references.citation: Blood. 1996 Dec 1;88(11):4396-8
references.pmid: 8943879
references.citation: Blood. 1997 Aug 1;90(3):1217-25
references.pmid: 9242555
references.citation: Blood. 2005 Nov 1;106(9):3062-7
references.pmid: 16020516
references.citation: Leukemia. 2004 Jan;18(1):165-6
references.pmid: 14603333
references.citation: Leukemia. 2008 Sep;22(9):1721-7
references.pmid: 18548092
references.citation: Clin Cancer Res. 2006 Apr 15;12(8):2434-41
references.pmid: 16638849
references.citation: Immunogenetics. 2000 Feb;51(2):99-107
references.pmid: 10663572
references.citation: Br J Haematol. 2004 Jun;125(5):590-600
references.pmid: 15147374
references.citation: Gene Ther. 2005 Jan;12(1):3-4
references.pmid: 15510172
references.citation: Clin Cancer Res. 2005 Dec 15;11(24 Pt 1):8799-807
references.pmid: 16361568
references.citation: Clin Cancer Res. 2006 Jan 1;12(1):34-42
references.pmid: 16397021
references.citation: Int J Cancer. 2006 Sep 15;119(6):1360-7
references.pmid: 16596644
references.citation: Acta Haematol. 2004;112(1-2):79-84
references.pmid: 15179007
references.citation: Clin Immunol. 2006 May;119(2):135-45
references.pmid: 16458072
references.citation: Proc Natl Acad Sci U S A. 2004 Oct 5;101 Suppl 2:14639-45
references.pmid: 15381769
references.citation: J Hepatol. 2001 Apr;34(4):562-9
references.pmid: 11394656
references.citation: ScientificWorldJournal. 2007 May 29;7:649-65
references.pmid: 17619750
references.citation: Science. 2006 Oct 6;314(5796):126-9
references.pmid: 16946036
references.citation: Blood. 2007 Sep 15;110(6):1924-32
references.pmid: 17505014
references.citation: Gene Ther. 2006 Apr;13(7):641-5
references.pmid: 16355114
references.citation: J Immunol. 2007 Nov 1;179(9):5803-10
references.pmid: 17947653
references.citation: Gene Ther. 2008 Apr;15(8):625-31
references.pmid: 18305579