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Exp Hematol. 2004 Jun;32(6):536-46.

Gene transfer into human T lymphocytes and natural killer cells by Ad5/F35 chimeric adenoviral vectors.

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  • 1Department of Hematology and Oncology, Georg-August-Universitaet Goettingen, Goettingen, Germany. R.Schroers@medizin.uni-goettingen.de



Genetic modification of effector lymphocytes, such as T cells and natural killer (NK) cells, is essential for many approaches to gene-based immunotherapy of cancer. However, transduction of lymphocytes has proven difficult by currently available gene transfer methods. Previous studies have shown that chimeric fiber-modified Ad5/F35 adenoviral vectors are able to efficiently transduce hematopoietic cells including immature progenitors. In this study, we examined the gene transfer into T lymphocytes and NK cells using Ad5/F35 compared with conventional Ad5 adenovectors.


Primary T and NK cells were isolated from healthy donors' peripheral blood leukocytes by immunomagnetic selection. Cell lines and primary lymphocytes were transduced with replication-defective Ad5/F35 and Ad5, both containing a GFP reporter gene under the control of a CMV promoter. Transduction efficiencies were monitored by flow cytometry. The function of transduced lymphocytes was assessed by analysis of proliferative responses to mitogenic agents and in mixed leukocyte reactions.


Transgene expression was detected in up to 45% of primary CD3+ T lymphocytes and in up to 60% of primary NK cells using Ad5/F35. In contrast, conventional Ad5 transduced less than 8% and 5% of primary T cells and NK cells, respectively. Transduction efficiencies were similar in CD4+ and CD8+ T lymphocytes, and transgene expression could be detected for up to seven days. Activation of T cells significantly enhanced the efficiency of Ad5/F35-mediated gene transfer. Adenoviral transduction of lymphocytes did not result in any impairment of proliferative functions.


The results of this study demonstrate that both T lymphocytes and NK cells can be transduced by chimeric Ad5/F35 adenoviral vectors.

[PubMed - indexed for MEDLINE]
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