title: Transgene expression levels determine the immunogenicity of transduced hematopoietic grafts in partially myeloablated mice.
abstract: We investigated whether transgene expression levels influence the immunogenicity of transduced hematopoietic grafts upon transplantation into partially myeloablated mice. To this aim, bone marrow cells (BMCs) transduced with retroviral vectors driving green fluorescent protein (GFP) expression either at high (high-EGFP) or low levels (low-EGFP) were transplanted into congenic recipients conditioned with sublethal doses of total body irradiation (TBI) or busulfan. Virtually all recipients showed evidence of donor engraftment 4 weeks after transplantation. However, as opposed to recipients receiving low-EGFP transduced grafts, the risk of rejecting the EGFP(+) cells by 30 days after transplantation was significantly higher in mice conditioned with busulfan and receiving high-EGFP transduced grafts. Anti-EGFP cellular immune responses were demonstrated in high-EGFP-treated mice conditioned with busulfan by interferon-gamma (IFN-gamma), enzyme-linked immunospot assay (ELISPOT), and cytotoxic T lymphocyte (CTL) assays, in contrast to that observed in mice transplanted with low-EGFP BMC. These results show for the first time that transgene expression levels can be critical for the immunogenicity of gene-modified hematopoietic grafts, especially in immunocompetent or in partially immunosuppressed recipients. These results have profound implications in vector choice and in the design of gene therapy (GT) protocols.
authors.affiliation: Banc de Sang i Teixits, Institut de Recerca Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: Herena
authors.initials: H
authors.lastname: Eixarch
authors.forename: Alba
authors.initials: A
authors.lastname: Gómez
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kádár
authors.forename: Mónica
authors.initials: M
authors.lastname: George
authors.forename: Nuria
authors.initials: N
authors.lastname: Martínez
authors.forename: Carmen
authors.initials: C
authors.lastname: Espejo
authors.forename: Jordi
authors.initials: J
authors.lastname: Pétriz
authors.forename: Ramon
authors.initials: R
authors.lastname: Gimeno
authors.forename: Jordi
authors.initials: J
authors.lastname: Barquinero
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation / Q000009:adverse effects / Q000379:methods; D002066:Busulfan / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000276:immunology*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects* / Q000379:methods*; D007111:Immunity, Cellular / Q000187:drug effects / Q000276:immunology*; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007371:Interferon-gamma / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D019172:Transplantation Conditioning; D014916:Whole-Body Irradiation
chemical_list: D007166:Immunosuppressive Agents; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma; D002066:Busulfan
references.citation: Blood. 2001 Apr 1;97(7):1951-9
references.pmid: 11264158
references.citation: Blood. 2002 Apr 15;99(8):2670-6
references.pmid: 11929752
references.citation: Gene Ther. 2008 Feb;15(4):239-46
references.pmid: 18046427
references.citation: Hum Gene Ther. 2002 Sep 1;13(13):1611-20
references.pmid: 12228016
references.citation: J Immunol Methods. 2005 May;300(1-2):100-14
references.pmid: 15899496
references.citation: Curr Gene Ther. 2007 Dec;7(6):411-20
references.pmid: 18045101
references.citation: Gene Ther. 2002 Nov;9(21):1472-9
references.pmid: 12378410
references.citation: Blood. 2004 May 1;103(9):3300-2
references.pmid: 14695227
references.citation: Gene Ther. 2002 Dec;9(23):1576-88
references.pmid: 12424610
references.citation: Mol Ther. 2009 May;17(5):897-905
references.pmid: 19277013
references.citation: J Virol. 1999 May;73(5):4083-9
references.pmid: 10196304
references.citation: Blood. 1997 Nov 1;90(9):3316-21
references.pmid: 9345013
references.citation: Blood. 2004 Jan 15;103(2):492-9
references.pmid: 14512305
references.citation: Mol Ther. 2000 Jun;1(6):533-44
references.pmid: 10933978
references.citation: Nat Med. 2006 Apr;12(4):401-9
references.pmid: 16582916
references.citation: Hematology. 2005 Aug;10(4):289-95
references.pmid: 16085541
references.citation: Blood. 2007 Jun 1;109(11):4708-15
references.pmid: 17327417
references.citation: Blood. 1991 Dec 15;78(12):3312-6
references.pmid: 1683798
references.citation: Gene Ther. 2000 Dec;7(23):2036-40
references.pmid: 11175316
references.citation: Semin Thromb Hemost. 2004 Apr;30(2):215-26
references.pmid: 15118933
references.citation: Science. 2002 Jun 28;296(5577):2410-3
references.pmid: 12089448
references.citation: Front Biosci. 2002 May 01;7:d1331-7
references.pmid: 11991829
references.citation: Blood. 2003 Jun 1;101(11):4305-12
references.pmid: 12576326
references.citation: Blood. 2002 Jun 15;99(12):4394-9
references.pmid: 12036867
references.citation: Blood. 2001 Jun 15;97(12):3733-7
references.pmid: 11389010
references.citation: Gene Ther. 2008 May;15(9):700-1
references.pmid: 18288211
references.citation: Mol Ther. 2005 Dec;12(6):1034-42
references.pmid: 16226058
references.citation: J Virol. 2005 Jul;79(14):8828-34
references.pmid: 15994776
references.citation: Exp Hematol. 2006 Feb;34(2):132-9
references.pmid: 16459181
references.citation: J Cancer Res Clin Oncol. 2000 Jul;126(7):391-9
references.pmid: 10929761
references.citation: Hum Gene Ther. 2001 Sep 1;12(13):1663-72
references.pmid: 11535169
references.citation: Blood. 2007 Sep 15;110(6):1788-96
references.pmid: 17495135
references.citation: Blood. 1999 Mar 15;93(6):1895-905
references.pmid: 10068662