转导(生物物理学)
生物
病毒载体
遗传增强
分子生物学
转基因
细胞因子
细胞生物学
T细胞
病毒学
细胞毒性T细胞
基因
免疫学
免疫系统
体外
遗传学
重组DNA
生物化学
作者
David E. Gilham,Michael Lie‐A‐Ling,Naomi Taylor,Robert E. Hawkins
摘要
Abstract 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 © 2009 John Wiley & Sons, Ltd.
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