Abstract
Results from multiple human studies have continued to spur the development of dendritic cells (DCs) as therapeutic vaccines for the treatment of cancer, chronic viral infections, and autoimmune diseases. The antigen-specific activity of DCs is dependent on the ability of the DCs to take up and process tumor-associated antigens for presentation to the immune system. Although immature DCs have been shown to naturally take up tumor-associated antigens by phagocytosis, approaches that significantly affect antigen delivery need further evaluation, especially if such methodologies can be demonstrated to result in the elicitation of more robust and comprehensive immune responses. We have developed a rapid, robust, scalable, and regulatory-compliant process for loading DCs with whole tumor lysate. The use of whole tumor lysate facilitates the generation of a more robust immune response targeting multiple unique antigenic determinants in patient's tumors and likely reduces the tumor's potential of immune escape. We demonstrate that DCs electroloaded with tumor lysate elicit significantly stronger antitumor responses both in a tumor challenge model and in a therapeutic vaccination model for preexisting metastasic disease. These effects are observed in a processing scheme that requires 20- to 40-fold lower amounts of tumor lysate when compared with the standard coincubation/coculture methods employed in loading DCs.
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References
1. Neumann, E., Schaefer-Ridder, M., Wand, Y., and Hofschneider, P. (1982) Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1, 841–845.
2. Xie, T.D., Sun, L., and Tsong, T.Y. (1990) Study of mechanisms of electric field-induced DNA transfection. I. DNA entry by surface binding and diffusion through membrane pores. Biophys. J. 58, 13–19.
3. Tsong, T.Y. (1991) Electroporation of cell membranes. Biophys. J. 60, 297–306.
4. Salek, A., Schnettler, R. and Zimmermann, U. (1992) Stably inherited killer activity in industrial yeast strains obtained by electrotransformation. FEMS Micro Lett. 75, 103–109.
5. Li, L.H., Shivakumar, R., Feller, S., et al. (2002) Highly efficient, large volume flow electroporation. Tech. Cancer Res. Treat. 1, 341–349.
6. Weiss, J.M., Shivakumar, R., Feller, S., et al. (2004) Rapid, in vivo, evaluation of anti-angiogenic and anti-neoplastic gene products by non-viral transfection of tumor cells. Cancer Gene Ther. 11, 346–353.
7. Li, S. (2004) Electroporation gene therapy: new developments in vivo and in vitro. Curr. Gene Ther. 4, 309–316.
8. Breckpot, K., Heirman, C., Neyns, B., and Thielemans, K. (2004) Exploiting dendritic cells for cancer immunotherapy: genetic modification of dendritic cells. J. Gene Med. 6, 1175–1188.
9. Ribas, A. (2005) Genetically modified dendritic cells for cancer immunotherapy. Curr. Gene Ther. 5, 619–628.
10. Weiss, J.M., Allen, C., Shivakumar, R., Feller, S., Li, L.H., and Liu, L.N. (2005) Efficient responses in a murine renal tumor model by electroloading dendritic cells with whole-tumor lysate. J. Immunother. 28, 542–550.
11. Geiger, J.D., Hutchinson, R.J., Hohenkirk, L.F., et al. (2001) Vaccination of pediatric solid tumor patients with tumor lysate-pulsed dendritic cells can expand specific T cells and mediate tumor regression. Cancer Res. 61, 8513–8519.
12. Thurnher, M., Rieser, C., Holtl, L., Papesh, C., Ramoner, R., and Bartsch, G. (1998) Dendritic cell-based immunotherapy of renal cell carcinoma. Urol. Int. 61, 67–71.
13. Schnurr, M., Galambos, P., Scholz, C., et al. (2001) Tumor cell lysate-pulsed human dendritic cells induce a T-cell response against pancreatic carcinoma cells: an in vitro model for the assessment of tumor vaccines. Cancer Res. 61, 6445–6450.
14. Wen, Y.J., Min, R., Tricot, G., Barlogie, B., and Yi, Q. (2002) Tumor lysate-specific cytotoxic T lymphocytes in multiple myeloma: promising effector cells for immunotherapy. Blood. 99, 3280–3285.
15. Parajuli, P. and Sloan, A.E. (2004) Dendritic cell-based immunotherapy of malignant gliomas. Cancer Invest. 22, 479–480.
16. Berard, F., Blanco, P., Davoust, J., et al. (2000) Cross-priming of naive CD8 T cells against melanoma antigens using dendritic cells loaded with killed allogeneic melanoma cells. J. Exp. Med. 192, 1535–1544.
17. Shibagaki, N. and Udey M.C. (2003) Dendritic cells transduced with TAT protein transduction domain-containing tyrosinase-related protein 2 vaccinate against murine melanoma. Eur. J. Immunol. 33, 850–860.
18. Hadzantonis, M. and O’Neill, H. (1999) Review: dendritic cell immunotherapy for melanoma. Cancer Biother. Radiopharm. 14, 11–22.
19. Fratantoni, J.C., Dzekunov, S., Singh, V., and Liu, L.N. (2003) A non-viral gene delivery system designed for clinical use. Cytotherapy. 5, 208–210.
20. Fratantoni, J.C., Dzekunov, S., Wang, S., and Liu, L.N. (2004) A scalable cell-loading system for non-viral gene delivery and other applications. Bioprocess. J. 3, 49–54.
Acknowledgments
The authors thank Nicholas Chopas for instrumentation assistance.
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© 2008 Humana Press
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Liu, L.N., Shivakumar, R., Allen, C., Fratantoni, J.C. (2008). Delivery of Whole Tumor Lysate into Dendritic Cells for Cancer Vaccination. In: Li, S. (eds) Electroporation Protocols. Methods in Molecular Biology™, vol 423. Humana Press. https://doi.org/10.1007/978-1-59745-194-9_9
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DOI: https://doi.org/10.1007/978-1-59745-194-9_9
Publisher Name: Humana Press
Print ISBN: 978-1-58829-877-5
Online ISBN: 978-1-59745-194-9
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