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Lentiviral Gene Transduction in Human and Mouse NK Cell Lines

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Book cover Natural Killer Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 612))

Abstract

Natural killer (NK) cells play a vital role in the control of cancer and microbial infections. A major hinderance in studying NK cells is the resistance of these cells to gene transfer. Considering over-expression and gene knockdown studies are crucial tools to study the biology of cells, technologies suitable for transfering genes into NK cells are invaluable. Among various technologies available for gene transfer, lentiviral-mediated transduction has been successful in introducing genes into NK cells. We have standardized methods of lentiviral infection in human and mouse NK cell lines. We obtain transduction efficiencies of 15% in the NK-92 cell line and 30–40% in LNK, YT, and DERL7 cell lines. This method allows efficient and stable introduction of genes and shRNAs into NK cell lines.

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References

  1. Trinchieri, G. (1989) Biology of natural killer cells. Adv Immunol 47, 187–376.

    Article  CAS  PubMed  Google Scholar 

  2. Grund, E. M. and Muise-Helmericks, R. C. (2005) Cost efficient and effective gene transfer into the human natural killer cell line, NK92. J Immunol Methods 296, 31–36.

    Article  CAS  PubMed  Google Scholar 

  3. Maasho, K., Marusina, A., Reynolds, N. M., Coligan, J. E., and Borrego, F. (2004) Efficient gene transfer into the human natural killer cell line, NKL, using the Amaxa nucleofection system. J Immunol Methods 284, 133–140.

    Article  CAS  PubMed  Google Scholar 

  4. Schoenberg, K., Trompeter, H. I., and Uhrberg, M. (2008) Delivery of DNA into natural killer cells for immunotherapy. Methods Mol Biol 423, 165–172.

    Article  CAS  PubMed  Google Scholar 

  5. Jiang, W., Zhang, J., and Tian, Z. (2008) Functional characterization of interleukin-15 gene transduction into the human natural killer cell line NKL. Cytotherapy 10, 265–274.

    Article  CAS  PubMed  Google Scholar 

  6. Kirwan, S., Merriam, D., Barsby, N., McKinnon, A., and Burshtyn, D. N. (2006) Vaccinia virus modulation of natural killer cell function by direct infection. Virology 347, 75–87.

    Article  CAS  PubMed  Google Scholar 

  7. Tran, J., and Kung, S. K. (2007) Lentiviral vectors mediate stable and efficient gene delivery into primary murine natural killer cells. Mol Ther 15, 1331–1339.

    Article  CAS  PubMed  Google Scholar 

  8. Schroers, R., Hildebrandt, Y., Hasenkamp, J., Glass, B., Lieber, A., Wulf, G., and Piesche, M. (2004) Gene transfer into human T lymphocytes and natural killer cells by Ad5/F35 chimeric adenoviral vectors. Exp Hematol 32, 536–546.

    Article  CAS  PubMed  Google Scholar 

  9. Goding, S., Yang, Q., Mi, Z., Robbins, P. D., and Basse, P. H. (2007) Targeting of products of genes to tumor sites using adoptively transferred A-NK and T-LAK cells. Cancer Gene Ther 14, 441–450.

    Article  CAS  PubMed  Google Scholar 

  10. Goding, S. R., Yang, Q., Knudsen, K. B., Potter, D. M., and Basse, P. H. (2007) Cytokine gene therapy using adenovirally transduced, tumor-seeking activated natural killer cells. Hum Gene Ther 18, 701–711.

    Article  CAS  PubMed  Google Scholar 

  11. Guven, H., Konstantinidis, K. V., Alici, E., Aints, A., Abedi-Valugerdi, M., Christensson, B., Ljunggren, H. G., and Dilber, M. S. (2005) Efficient gene transfer into primary human natural killer cells by retroviral transduction. Exp Hematol 33, 1320–1328.

    Article  CAS  PubMed  Google Scholar 

  12. Gong, J. H., Maki, G., and Klingemann, H. G. (1994) Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia 8, 652–658.

    CAS  PubMed  Google Scholar 

  13. Yodoi, J., Teshigawara, K., Nikaido, T., Fukui, K., Noma, T., Honjo, T., Takigawa, M., Sasaki, M., Minato, N., Tsudo, M., et al. (1985) TCGF (IL 2)-receptor inducing factor(s). I. Regulation of IL 2 receptor on a natural killer-like cell line (YT cells). J Immunol 134, 1623–1630.

    CAS  PubMed  Google Scholar 

  14. Robertson, M. J., Cochran, K. J., Cameron, C., Le, J. M., Tantravahi, R., and Ritz, J. (1996) Characterization of a cell line, NKL, derived from an aggressive human natural killer cell leukemia. Exp Hematol 24, 406–415.

    CAS  PubMed  Google Scholar 

  15. Di Noto, R., Pane, F., Camera, A., Luciano, L., Barone, M., Lo Pardo, C., Boccuni, P., Intrieri, M., Izzo, B., Villa, M. R., Macri, M., Rotoli, B., Sacchetti, L., Salvatore, F., and Del Vecchio, L. (2001) Characterization of two novel cell lines, DERL-2 (CD56+/CD3+/Tcry5+) and DERL-7 (CD56+/CD3-/TCRgammadelta-), derived from a single patient with CD56+ non-Hodgkin's lymphoma. Leukemia 15, 1641–1649.

    Article  PubMed  Google Scholar 

  16. Hodge, D. L., Schill, W. B., Wang, J. M., Blanca, I., Reynolds, D. A., Ortaldo, J. R., and Young, H. A. (2002) IL-2 and IL-12 alter NK cell responsiveness to IFN-gamma-inducible protein 10 by down-regulating CXCR3 expression. J Immunol 168, 6090–6098.

    CAS  PubMed  Google Scholar 

  17. Tsutsui, H., Nakanishi, K., Matsui, K., Higashino, K., Okamura, H., Miyazawa, Y., and Kaneda, K. (1996) IFN-gamma-inducing factor up-regulates Fas ligand-mediated cytotoxic activity of murine natural killer cell clones. J Immunol 157, 3967–3973.

    CAS  PubMed  Google Scholar 

  18. Salmon, P., and Trono, D. (2006) Production and titration of lentiviral vectors. Curr Prot Neurosci Chapter 4, Unit 4 21.

    Google Scholar 

  19. Sastry, L., Johnson, T., Hobson, M. J., Smucker, B., and Cornetta, K. (2002) Titering lentiviral vectors: comparison of DNA, RNA and marker expression methods. Gene Ther 9, 1155–1162.

    Article  CAS  PubMed  Google Scholar 

  20. Gismondi, A., Morrone, S., Humphries, M. J., Piccoli, M., Frati, L., and Santoni, A. (1991) Human natural killer cells express VLA-4 and VLA-5, which mediate their adhesion to fibronectin. J Immunol 146, 384–392.

    CAS  PubMed  Google Scholar 

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Savan, R., Chan, T., Young, H.A. (2010). Lentiviral Gene Transduction in Human and Mouse NK Cell Lines. In: Campbell, K. (eds) Natural Killer Cell Protocols. Methods in Molecular Biology, vol 612. Humana Press. https://doi.org/10.1007/978-1-60761-362-6_14

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  • DOI: https://doi.org/10.1007/978-1-60761-362-6_14

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-361-9

  • Online ISBN: 978-1-60761-362-6

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