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Flt3-L gene therapy enhances immunocytokine-mediated antitumor effects and induces long-term memory

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Abstract

Therapeutic treatment with hu14.18-IL-2 immunocytokine (IC) or Flt3-L (FL) protein is initially effective at resolving established intradermal NXS2 neuroblastoma tumors in mice. However, many treated animals develop recurrent disease. We previously found that tumors recurring following natural killer (NK) mediated IC treatment show augmented MHC class I expression, while the tumors that recurred following T cell dependent Flt3-L treatment exhibited decreased MHC class I expression. We hypothesized that this divergent MHC modulation on recurrent tumors was due to therapy-specific immunoediting. We further postulated that combining IC and Flt3-L treatments might decrease the likelihood of recurrent disease by preventing MHC modulation as a mechanism for immune escape. We now report that combinatorial treatment of FL plus hu14.18-IL-2 IC provides greater antitumor benefit than treatment with either alone, suppressing development of recurrent disease. We administered FL by gene therapy using a clinically relevant approach: hydrodynamic limb vein (HLV) delivery of DNA for transgene expression by myofibers. Delivery of FL DNA by HLV injection in mice resulted in systemic expression of >10 ng/ml of FL in blood at day 3, and promoted up to a fourfold and tenfold increase in splenic NK and dendritic cells (DCs), respectively. Furthermore, the combination of FL gene therapy plus suboptimal IC treatment induced a greater expansion in the absolute number of splenic NK and DCs than achieved by individual component treatments. Mice that received combined FL gene therapy plus IC exhibited complete and durable resolution of established NXS2 tumors, and demonstrated protection from subsequent rechallenge with NXS2 tumor.

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Abbreviations

CTL:

cytolytic T-lymphocyte

DC:

dendritic cell

Flt3-L:

fms-like tyrosine kinase 3 ligand

IC:

immunocytokine

NB:

neuroblastoma

NK:

natural killer

TAA:

tumor associated antigen

TEV:

tumor escape variants

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Acknowledgments

The authors thank Kathy Schell (Department of Human Oncology, University of Wisconsin-Madison, WI) for assistance with the flow cytometric analyses of excised tumors, and Drs. Jackie Hank, Ilia Buhtoiarov, Alexander Rakhmilevich, Ralph Reisfeld and Jon Wolff for helpful discussions. This work was supported in part by grants R01-063285 and R01-087025 from the National Institutes of Health and research support from The Midwest Athletes Against Childhood Cancer (MACC) Fund (PI, P.M. Sondel).

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Authors and Affiliations

  1. Mirus Bio Corporation, 505 S. Rosa Road, Madison, WI, 53719, USA

    Zane C. Neal, Mary Kay Bates & Hans Herweijer

  2. Paul P. Carbone Comprehensive Cancer Center, University of Wisconsin-Madison, Madison, WI, USA

    Zane C. Neal & Paul M. Sondel

  3. Departments of Pediatrics and Human Oncology, University of Wisconsin-Madison, Madison, WI, USA

    Paul M. Sondel

  4. EMD Lexigen Research Center, Billerica, MA, USA

    Stephen D. Gillies

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  1. Zane C. Neal
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  2. Paul M. Sondel
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  3. Mary Kay Bates
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  4. Stephen D. Gillies
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  5. Hans Herweijer
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Correspondence to Hans Herweijer.

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Neal, Z.C., Sondel, P.M., Bates, M.K. et al. Flt3-L gene therapy enhances immunocytokine-mediated antitumor effects and induces long-term memory. Cancer Immunol Immunother 56, 1765–1774 (2007). https://doi.org/10.1007/s00262-007-0320-5

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  • DOI: https://doi.org/10.1007/s00262-007-0320-5

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