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Non-viral gene delivery into primary natural killer lymphocytes

Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich Heine University, Düsseldorf, Germany

¹Correspondence: Institute for Transplantation Diagnostics and Cell Therapeutics, University Clinic of Düsseldorf, Building 14.80, Moorenstrasse 5, Düsseldorf D-40225, Germany. E-mail: uhrberg{at}itz.uni-duesseldorf.de

Goffinet and Keppler reported in The FASEB Journal on the use of nucleofection for gene delivery into primary rodent lymphocytes (1) . Work from us (2) is cited as evidence that "lymphocyte populations other than T cells from humans and rodents have been considered virtually refractory to nonviral gene delivery." This notion is misleading since in this (2) , as well as in several other publications from our group (3 , 4) , it was clearly shown that nucleofection is an effective way to transfect human natural killer cells. The transfection efficiencies observed in Trompeter et al. (2) were more than 50% for the IL-2 dependent NK cell line NK3.3 as measured by surface expression of a H-2K^k reporter construct and about half the efficiency for primary NK cells as measured by luciferase activity. These numbers are actually in the same range as reported by Goffinet and Keppler for rodent NK cells. In addition, it was previously shown by us and others, that nucleofection is not only suitable for transfection of activated lymphocytes, which is recommended by Goffinet and Keppler, but is also efficient in transfection of resting NK and T cells in humans and rodents, respectively (2 , 5) . This observation makes nucleofection an attractive tool for immunotherapeutic use of "untouched" lymphocytes, as it enables non-viral gene delivery into lymphocytes without the need for polyclonal stimulation and thus unspecific activation.

REFERENCES

1. Goffinet, C., Keppler, O. T. (2006) Efficient nonviral gene delivery into primary lymphocytes from rats and mice. FASEB J. 20,500-502[Abstract/Free Full Text]
2. Trompeter, H. I., Weinhold, S., Thiel, C., Wernet, P., Uhrberg, M. (2003) Rapid and highly efficient gene transfer into natural killer cells by nucleofection. J. Immunol. Methods 274,245-256[CrossRef][Medline]
3. Santourlidis, S., Trompeter, H. I., Weinhold, S., Eisermann, B., Meyer, K. L., Wernet, P., Uhrberg, M. (2002) Crucial role of DNA methylation in determination of clonally distributed killer cell Ig-like receptor expression patterns in NK cells. J. Immunol. 169,4253-4261[Abstract/Free Full Text]
4. Trompeter, H. I., Gomez-Lozano, N., Santourlidis, S., Eisermann, B., Wernet, P., Vilches, C., Uhrberg, M. (2005) Three structurally and functionally divergent kinds of promoters regulate expression of clonally distributed killer cell Ig-like receptors (KIR), of KIR2DL4, and of KIR3DL3. J. Immunol. 174,4135-4143[Abstract/Free Full Text]
5. Lai, W., Chang, C. H., Farber, D. L. (2003) Gene transfection and expression in resting and activated murine CD4 T cell subsets. J. Immunol. Methods 282,93-102[CrossRef][Medline]

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