Cytokine stimulation and the choice of promoter are critical factors for the efficient transduction of mouse T cells with HIV-1 vectors

被引:20
作者
Gilham, David E. [1 ]
Lie-A-Ling, Michael [2 ]
Taylor, Naomi [3 ]
Hawkins, Robert E.
机构
[1] Univ Manchester, Manchester Acad Hlth Sci Ctr, Canc Res UK Dept Med Oncol, Cell Therapy Grp,Paterson Inst Canc Res, Manchester M20 4BX, Lancs, England
[2] Paterson Inst Canc Res, Canc Res UK Gene Therapy Grp, Manchester M20 9BX, Lancs, England
[3] Univ Montpellier, CNRS, Inst Genet Mol Montpellier, IFR,Unite Mixte Rech 5535, Montpellier, France
关键词
cytokines; HIV-1; lentivirus; mouse T cell; promoter; vector; MEDIATED GENE-TRANSFER; RETROVIRAL TRANSDUCTION; LENTIVIRAL VECTORS; IN-VIVO; LYMPHOCYTES; EXPRESSION; REPLICATION; INFECTION; DELIVERY; CD4(+);
D O I
10.1002/jgm.1421
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background HIV-1 fails to successfully infect mouse T cells as a result of several blocks in the viral replication cycle. We investigated whether this also impacted on the use of HIV-1 derived lentiviral vectors for stable gene transfer into mouse T cells. Methods Freshly isolated primary mouse T cells were immediately mixed with lentiviral vectors encoding an enhanced green fluorescent protein marker gene and transduction frequency was determined after 5 days of culture. Results Optimal transduction required both mouse T cell activation and cytokine support. Furthermore, transduction was also dependent upon the promoter chosen, with the rank order of potency being PGK > EF1 > SFFV > CMV. HIV-1 lentiviral vectors also efficiently transduced cytokine-stimulated T cells (in the absence of antibody driven T cell activation), albeit with a lower level of transgene expression compared to fully-activated T cells. Conclusions The present study demonstrates that primary mouse T cells can be efficiently transduced with HIV-1 lentiviral vectors, opening up prospects for their use in mouse models of gene-modified adoptive cellular therapy. Copyright (C) 2009 John Wiley & Sons, Ltd.
引用
收藏
页码:129 / 136
页数:8
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