title: HLA engineering of human pluripotent stem cells.
abstract: The clinical use of human pluripotent stem cells and their derivatives is limited by the rejection of transplanted cells due to differences in their human leukocyte antigen (HLA) genes. This has led to the proposed use of histocompatible, patient-specific stem cells; however, the preparation of many different stem cell lines for clinical use is a daunting task. Here, we develop two distinct genetic engineering approaches that address this problem. First, we use a combination of gene targeting and mitotic recombination to derive HLA-homozygous embryonic stem cell (ESC) subclones from an HLA-heterozygous parental line. A small bank of HLA-homozygous stem cells with common haplotypes would match a significant proportion of the population. Second, we derive HLA class I-negative cells by targeted disruption of both alleles of the Beta-2 Microglobulin (B2M) gene in ESCs. Mixed leukocyte reactions and peptide-specific HLA-restricted CD8(+) T cell responses were reduced in class I-negative cells that had undergone differentiation in embryoid bodies. These B2M(-/-) ESCs could act as universal donor cells in applications where the transplanted cells do not express HLA class II genes. Both approaches used adeno-associated virus (AAV) vectors for efficient gene targeting in the absence of potentially genotoxic nucleases, and produced pluripotent, transgene-free cell lines.
authors: Department of Medicine, University of Washington, Seattle, Washington 98195, USA.
authors: Laura
authors: L
authors: Riolobos
authors: Roli K
authors: RK
authors: Hirata
authors: Cameron J
authors: CJ
authors: Turtle
authors: Pei-Rong
authors: PR
authors: Wang
authors: German G
authors: GG
authors: Gornalusse
authors: Maja
authors: M
authors: Zavajlevski
authors: Stanley R
authors: SR
authors: Riddell
authors: David W
authors: DW
authors: Russell
mesh_terms: D000483:Alleles; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D018390:Gene Targeting; D005818:Genetic Engineering; D005822:Genetic Vectors; D006680:HLA Antigens / Q000235:genetics* / Q000378:metabolism; D006239:Haplotypes; D006648:Histocompatibility / Q000235:genetics; D006720:Homozygote; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011995:Recombination, Genetic; D001613:beta 2-Microglobulin / Q000235:genetics / Q000378:metabolism
chemical_list: D006680:HLA Antigens; D001613:beta 2-Microglobulin
references: Stem Cells. 2004;22(4):448-56
references: 15277692
references: Nat Genet. 1996 Aug;13(4):399-408
references: 8696333
references: Cell Stem Cell. 2010 Oct 8;7(4):521-31
references: 20887957
references: J Immunol. 2008 Nov 1;181(9):6635-43
references: 18941254
references: Blood. 2004 Aug 15;104(4):1075-82
references: 15039282
references: Nat Genet. 2004 Jul;36(7):767-73
references: 15208627
references: N Engl J Med. 2011 Nov 3;365(18):1673-83
references: 22047558
references: Nat Methods. 2011 May;8(5):409-12
references: 21460823
references: Stem Cells. 2006 Dec;24(12):2677-84
references: 16916927
references: Cell Stem Cell. 2012 Nov 2;11(5):615-9
references: 23084023
references: Transplantation. 2001 Apr 15;71(7):982-5
references: 11349735
references: Tissue Antigens. 1978 Feb;11(2):96-112
references: 77067
references: Nat Genet. 2001 Mar;27(3):257-8
references: 11242104
references: Mol Ther. 2010 Jun;18(6):1192-9
references: 20407427
references: J Immunol. 1992 Aug 15;149(4):1319-25
references: 1354233
references: Hastings Cent Rep. 2003 Nov-Dec;33(6):13-27
references: 14983554
references: Immunology. 1996 May;88(1):124-9
references: 8707337
references: Immunol Today. 1990 Jul;11(7):237-44
references: 2201309
references: Dev Dyn. 2004 Feb;229(2):259-74
references: 14745951
references: J Clin Invest. 1998 Jun 15;101(12):2720-9
references: 9637706
references: Science. 1998 Nov 6;282(5391):1145-7
references: 9804556
references: Transplantation. 1990 May;49(5):991-5
references: 2186527
references: Blood. 2012 Jan 5;119(1):72-82
references: 22031866
references: N Engl J Med. 2010 Oct 7;363(15):1429-37
references: 20925545
references: Transplantation. 1993 Apr;55(4):940-6
references: 8475570
references: Nature. 1992 Jul 2;358(6381):66-70
references: 1614533
references: Nat Biotechnol. 2010 Apr;28(4):371-7
references: 20351689
references: Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):5199-204
references: 9560253
references: Clin Exp Immunol. 2007 Apr;148(1):1-16
references: 17349009
references: Methods Mol Biol. 2011;767:449-61
references: 21822895
references: Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9864-9
references: 12114532
references: Gene. 2000 Feb 22;244(1-2):47-54
references: 10689186
references: Lancet. 2005 Dec 10;366(9502):2019-25
references: 16338451
references: Mol Ther. 2012 Jan;20(1):204-13
references: 22031238
references: Nature. 1991 Jan 24;349(6307):329-31
references: 1987491
references: J Immunol. 1993 Jul 1;151(1):425-35
references: 8326135
references: Nat Protoc. 2011 Apr;6(4):482-501
references: 21455185
references: Exp Cell Res. 2008 Oct 1;314(16):2930-40
references: 18692044
references: QJM. 2005 Oct;98(10):719-27
references: 16087697
references: J Immunol. 2010 May 1;184(9):4587-91
references: 20410495
references: J Virol. 1996 Nov;70(11):7569-79
references: 8892876
references: Ann Med. 2009;41(5):360-70
references: 19165643
references: Lancet. 2004 Jul 10-16;364(9429):163-71
references: 15246729
references: Cell. 2007 Nov 30;131(5):861-72
references: 18035408
references: Science. 1996 Nov 1;274(5288):792-5
references: 8864122
references: Stem Cells. 2003;21(5):521-6
references: 12968106
references: Mol Ther. 2007 Oct;15(10):1827-33
references: 17622243
references: Nature. 2010 Sep 16;467(7313):285-90
references: 20644535
references: Science. 2007 Dec 21;318(5858):1917-20
references: 18029452
references: J Exp Med. 1991 Oct 1;174(4):875-80
references: 1919440