Abstract
Dendritic cells (DC) are the most powerful antigen-presenting cells that induce and maintain primary immune responses in vitro and in vivo. The development of protocols for the ex vivo generation of DC provided a rationale to design and develop DC-based vaccination studies for the treatment of infectious and malignant diseases. The efficacy of antigen loading and delivery into DC is pivotal for the optimal induction of T-cell-mediated immune responses. Recently it was shown that DC transfected with RNA coding for a tumor-associated antigen (TAA) or whole-tumor RNA are able to induce potent antigen- and tumor-specific T-cell responses directed against multiple epitopes. The latter technique does not require the definition of the TAA or HLA haplotype of the patients and has the potential of broad clinical application. Such a polyvalent vaccine might be able to reduce the probability of clonal tumor escape and to elicit CTL responses directed against naturally processed and presented immuno-dominant tumor antigens. Additional targeting of HLA class II restricted epitopes may further amplify and prolong the induced T-cell responses.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Banchereau, J. and Steinman, R. M. (1998) Dendritic cells and the control of immunity. Nature 392, 245–252.
Cella, M., Sallusto, F., and Lanzavecchia, A. (1997) Origin, maturation and antigen presenting function of dendritic cells. Curr. Opin. Immunol. 9, 10–16.
Sallusto, F. and Lanzavecchia, A. (1994) Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony stimulating factor plus interleukin 4 and down regulated by tumour necrosis factor. J. itExp. Med. 179, 1109–1118.
Zhou, L. and Tedder, T. F. (1996) CD14 blood monocytes can differentiate into functionally mature CD83+dendritic cells. Proc. Natl. Acad. Sci. USA 93, 2588–2592.
Brossart, P., Grünebach, F., Stuhler, G., et al. (1998) Generation of functional human dendritic cells from adherent peripheral blood monocytes by CD40 ligation in the absence of granulocyte-macrophage colony-stimulating factor. Blood 92, 4238–4247.
Brossart, P., Wirths, S., Brugger, W., and Kanz, L. (2001) Dendritic cells in cancer vaccines. Exp. Hematol. 29, 1247–1255.
Brossart P., Goldrath, A. W., Butz, E. A., Martin, S., and Bevan, M. J. (1997) Virus-mediated delivery of antigenic epitopes into dendritic cells as a means to induce CTL. J. Immunol. 158, 3270–3276.
Reeves, M. E., Royal, R. E., Lam, J. S., Rosenberg, S. A., and Hwu, P. (1996) Retroviral transduction of human dendritic cells with a tumor-associated antigen gene. Cancer Res. 56, 5672–5677.
Van Tendeloo, V. F., Snoeck, H. W., Lardon, F., et al. (1998) Nonviral transfection of distinct types of human dendritic cells: high-efficiency gene transfer by electroporation into hematopoietic progenitor-but not monocyte-derived dendritic cells. Gene Ther. 5, 700–707.
Boczkowski, D., Nair, S. K., Snyder, D., and Gilboa, E. (1996) Dendritic cells pulsed with RNA are potent antigen-presenting cells in vitro and in vivo. J. Exp. Med. 184, 465–472.
Nair, S. K., Boczkowski, D., Morse, M., Cumming, R. I., Lyerly, H. K., and Gilboa, E. (1998) Induction of primary carcinoembryonic antigen (CEA)-specific cytotoxic T lymphocytes in vitro using human dendritic cells transfected with RNA. Nat. Biotechnol. 16, 364–369.
Müller, M. R., Grünebach, F., Nencioni, A., and Brossart, P. (2003) Transfection of dendritic cells with RNA induces CD4-and CD8-mediated T cell immunity against breast carcinomas and reveals the immunodominance of presented T cell epitopes. J. Immunol.. 5892–5896.
Grünebach, F., Müller, M. R., Nencioni, A., and Brossart, P. (2003) Delivery of tumor-derived RNA for the induction of cytotoxic T-lymphocytes. Gene Ther. 10, 367–374.
Andreason, G. L. and Evans, G. A. (1989) Optimization of electroporation for transfection of mammalian cell lines. Anal. Biochem. 180, 269–275.
Zhang, G., Gurtu, V., and Kain, S. R. (1996) An enhanced green fluorescent protein allows sensitive detection of gene transfer in mammalian cells. Biochem. Biophys. Res. Commun. 227, 707–711.
Van Tendeloo, V. F., Ponsaerts, P., Lardon, F., et al. (2001) Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells. Blood 98, 49–56.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Grünebach, F., Müller, M.R., Brossart, P. (2005). RNA Transfection of Dendritic Cells. In: Ludewig, B., Hoffmann, M.W. (eds) Adoptive Immunotherapy: Methods and Protocols. Methods in Molecular Medicine™, vol 109. Humana Press. https://doi.org/10.1385/1-59259-862-5:047
Download citation
DOI: https://doi.org/10.1385/1-59259-862-5:047
Publisher Name: Humana Press
Print ISBN: 978-1-58829-406-7
Online ISBN: 978-1-59259-862-5
eBook Packages: Springer Protocols