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Priv.-Doz. Dr. rer. nat. Niels Schaft
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Major research topics:
The RNA-group-members are: Dr. J. Dörrie, Dr. N. Schaft, Technician I. Müller, Technician V. Beck.
The main focus of the RNA-group is RNA-transfection of dendritic cells (DC), for the purpose of HLA class I presentation of the transfected antigens (Ag). The cytoplasmic expression within the DC allows the Ag to be processed like endogenous proteins, leading to classical HLA class I presentation. The final goal is on the one hand a functional therapeutic vaccine against melanoma and other types of cancer, and on the other hand, to better understand the processes involved in presentation of cellular Ags and the conditions of efficient T-cell priming. But also the manipulation of DC-function, for example by expression of surface receptors, is a strategy embarked by us.
Current research projects:
Comparison of different transfection strategies with respect to stimulatory capacity
Different means of transfection lead to very different Ag expression profiles. While electroporation results in >90% of the cells expressing intermediate levels of Ag, lipofection reaches only a small portion of the DCs (5-20%) but results in high expression levels per cell. We examine how these two parameters – the number of Ag-presenting DCs and the Ag-quantity per cell – contribute to the DCs capability of T-cell stimulation.
T-cell stimulation and migration: Two actions exerted by the same DC?
DC which are used for anti-tumour vaccination need to migrate from the site of injection to the secondary lymphoid tissue. Therefore, we examine the ability of differently generated DC to migrate towards chemokines and the capability of the migrated cells to stimulate specific T-cells.
Functional manipulation of DC by electroporation with RNA coding for surface proteins
By expression of functional proteins we want to equip the DCs with beneficial features, for example empowering them to transmigrate from the bloodstream into the peripheral lymph-nodes.
Selected References:
Original articles:
Hofmann Christian; Hoefflin Sandra; Hueckelhoven Angela; et al. Human T cells expressing two additional receptors (TETARs) specific for HIV-1 recognize both epitopes. BLOOD 2011,118,19,5174-5177
Knippertz I, Stein MF, Dörrie J, Schaft N, Müller I, Deinzer A, Steinkasserer A, Nettelbeck DM. Mild hyperthermia enhances human monocyte-derived dendritic cell functions and offers potential for applications in vaccination strategies. Int J Hyperthermia. 2011;27(6):591-603.
Birkholz K, Schwenkert M, Kellner C, Gross S, Fey G, Schuler-Thurner B, Schuler G, Schaft N, Dörrie J. Targeting of DEC-205 on human dendritic cells results in efficient MHC class II-restricted antigen presentation. Blood. 2010;116(13): 2277-85
Schreibelt G, Benitez-Ribas D, Schuurhuis D, Lambeck AJ, van Hout-Kuijer M, Schaft N, Punt CJ, Figdor CG, Adema GJ, de Vries IJ. Commonly used prophylactic vaccines as an alternative for synthetically produced TLR ligands to mature monocyte-derived dendritic cells.
Blood. 2010;116(4): 564-74
Krug, C ; Birkholz, K ; Hombach, A ; Reuter, S ; Kershaw, M ; Kampgen, E ; Schwenkert, M ; Fey, G ; Schuler, G ; Abken, H ; Dorrie, J ; Schaft, N. Reprogramming T cells with cancer specificity by non-viral transfer of mRNA encoding recombinant immunoreceptors. HUMAN GENE THERAPY, 20 (11): 1531-1531 NOV 2009
Birkholz, K, Hombach, A, Krug, C, Reuter, S, Kershaw, M, Kampgen, E, Schuler, G, Abken, H, Dorrie, J, Schaft, N. Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4+and CD8+T cells for the use in adoptive immunotherapy of cancer. EXPERIMENTAL DERMATOLOGY, 18 (3): 316-316 MAR 2009
Dorrie, J, Schaft, N, Wohn, CT, Wellner, V, Schunder, T, Muller, I, Kaempgen, E, Schuler, G. Lipofection or electroporation of RNA - consequences of different antigen-distribution on immunogenicity. EXPERIMENTAL DERMATOLOGY, 18 (3): 298-298 MAR 2009
Birkholz, K, Hombach, A, Krug, C, Reuter, S, Kershaw, M, Kampgen, E, Schuler, G, Abken, H, Schaft, N, Dorrie, J. Transfer of mRNA encoding recombinant immunoreceptors reprograms CD4(+) and CD8(+) T cells for use in the adoptive immunotherapy of cancer. GENE THERAPY, 16 (5): 596-604 MAY 2009
Birkholz, K, Hofmann, C, Hoyer, S, Schulz, B, Harrer, T, Kampgen, E, Schuler, G, Dorrie, J, Schaft, N. A fast and robust method to clone and functionally validate T-cell receptors. JOURNAL OF IMMUNOLOGICAL METHODS, 346 (1-2): 45-54 JUL 31 2009
Birkholz, K., Dorrie, J., Schaft, N., Schwenkert, M., Kellner, C., Fey, G. H., and Schuler, G. (2008). DC cancer vaccines: development of new strategies for antigen loading. Experimental Dermatology 17, 259.
Birkholz, K., Hoyer, S., Hofmann, C., Schulz, B., Kaempgen, E., Schuler, G., Schaft, N., and Dorrie, J. (2008). A fast and robust method to clone and test the function of T cell receptors. Experimental Dermatology 17, 258.
Dorrie, J., Schaft, N., Muller, I., Wellner, V., Schunder, T., Hanig, J., Oostingh, G. J., Schon, M. P., Robert, C., Kampgen, E., and Schuler, G. (2008). Introduction of functional chimeric E/L-selectin by RNA electroporation to target dendritic cells from blood to lymph nodes. Cancer Immunology Immunotherapy 57, 467-477.
Dorrie, J., Schaft, N., Wellner, V., Wohn, C., Schunder, T., Muller, I., Kampgen, E., and Schuler, G. (2008). Priming and boosting of melanA-specific T cells - in vitro studies on the importance of antigen-presentation density. Wiener Klinische Wochenschrift 120, 19.
Haendle, I., Schmitt-Haendle, M., Voland, S., Erdmann, M., Schliep, S., Schaft, N., Doerrie, J., Kaempgen, E., Schuler-Thurner, B., and Schuler, G. (2008). T-cell responses in stage IV melanoma patients vaccinated with dendritic cells electroporated with defined RNA. Experimental Dermatology 17, 263.
Heise, D., Schmid, M., Schaft, N., Doerrie, J., Schuler, G., and Kaempgen, E. (2008). Are TLR agonists better maturation stimuli for clinically applicable dendritic cell vaccines ? Experimental Dermatology 17, 264.
Hofmann, C., Harrer, T., Eismann, K., Bergmann, S., Schmitt-Haendle, M., Schuler, G., Dorrie, J., and Schaft, N. (2008). Reprogramming CD8+T cells with a HIV-1-specificity by electroporation of TCR-encoding RNA. Experimental Dermatology 17, 259.
Hofmann, C., Harrer, T., Kubesch, V., Maurer, K., Metzner, K. J., Eismann, K., Bergmann, S., Schmitt-Haendle, M., Schuler, G., Dorrie, J., and Schaft, N. (2008). Generation of HIV-1-specific T cells by electroporation of T-cell receptor RNA. Aids 22, 1577-1582.
Hoyer, S., Birkholz, K., Schuler, G., Dorrie, J., and Schaft, N. (2008). Providing specific T-cell help by electroporation of TCR-encoding RNA intoCD4+T cells. Experimental Dermatology 17, 258.
Mueller, I., Schaft, N., Doerrie, J., Grelle, I., Reiners, C., Becker, J. C., Schuler, G., and Kaempgen, E. (2008). Antigen specific abrogation of immunity using Indium111 labelled dendritic cells. Experimental Dermatology 17, 264.
Schaft, N., Schunder, T., Krug, C., Reuter, S., Kosmides, D., Abken, H., Kershaw, M., Kampgen, E., Schuler, G., and Dorriee, J. (2008). Reprogramming T cells to become MHC independent - electroporation of RNA encoding chimeric antigen receptors. Wiener Klinische Wochenschrift 120, 8.
Schaft, N., Krug, C., Reuter, S., Kosmides, D., Abken, H., Kershaw, M. H., Kaempgen, E., Schuler, G., and Dorrie, I. (2008). Making T cell independent of MHC - electroporation of RNA encoding chimeric T cell receptors. Experimental Dermatology 17, 258.
Schwenkert, M., Birkholz, K., Schwemmlein, M., Kellner, C., Kuegler, M., Peipp, M., Nettelbeck, D. M., Schuler-Thurner, B., Schaft, N., Dorrie, J., Ferrone, S., Kampgen, E., and Fey, G. H. (2008). A single chain immunotoxin, targeting the melanoma-associated chondroitin sulfate proteoglycan, is a potent inducer of apoptosis in cultured human melanoma cells. Melanoma Research 18, 73-84.
Dorrie, J., Birkholz, K., and Schaft, N. (2007). Strategies for immune intervention; from bench to bedside. Cancer Immunology Immunotherapy 56, 1677-1685.
Erdmann, M., Dorrie, J., Schaft, N., Strasser, E., Hendelmeier, M., Kampgen, E., Schuler, G., and Schuler-Thurner, B. (2007). Effective clinical-scale production of dendritic cell vaccines by monocyte elutriation directly in medium, subsequent culture in bags and final antigen loading using peptides or RNA transfection. Journal of Immunotherapy 30, 663-674.
Hendelmeier, M., Oehring, A., Erdmann, M., Doerrie, J., Schaft, N., Kohler, E., Zimmermann, R., Strasser, E., and Eckstein, R. (2007). Vaccination with dendritic cells: Cellular immune modulation by leukocytapheresis in patients with metastatic malignant melanoma. Haematologica-the Hematology Journal 92, 67.
Schaft, N., Doerrie, J., Birkholz, K., Hoyer, S., Wellner, V., Mueller, I., Kaempgen, E., and Schuler, G. (2007). Electroporation of TCR encoding RNA into T cells as a new approach for anti-tumor immunotherapy. Experimental Dermatology 16, 220.
Dorrie, J., Erdmann, M., Schaft, N., Muller, I., Stoica, D., Hirsch, U., Kampgen, E., Schuler-Thurner, B., and Schuler, G. (2007). Electroporation of ELUTRA generated dendritic cells with RNA - Towards a closed system. Experimental Dermatology 16, 230-231.
Schaft, N., Dorrie, J., and Schuler, G. (2007). Endowing T lymphocytes with a new specificity by electroporation of RNA coding for a T cell receptor. Cancer Immunology Immunotherapy 56, 408-409.
Dorrie, J., Schaft, N., Hanig, J., Oostingh, G. J., Schon, M. P., Robert, C., Kampgen, E., and Schuler, G. (2007). Enhanced immunogenicity of DC targeted from blood to lymph nodes by introduction of a chimeric E/L-selectin molecule. Cancer Immunology Immunotherapy 56, 409.
Dorrie J, Wellner V, Kampgen E, Schuler G, Schaft N.
An improved method for RNA isolation and removal of melanin contamination from melanoma tissue: Implications for tumor antigen detection and amplification.
J Immunol Methods. 2006 Jun 30;313(1-2):119-28. Epub 2006 May 16.
Schaft N, Dorrie J.
Strategies for immune intervention.
Cancer Immunol Immunother. 2005 Dec 16;:1-11 [Epub ahead of print] No abstract available.
Schaft N, Dorrie J, Muller I, Beck V, Baumann S, Schunder T, Kampgen E,
Schuler G.
A new way to generate cytolytic tumor-specific T cells: electroporation of RNA coding for a T cell receptor into T lymphocytes.
Cancer Immunol Immunother. 2005 Dec 13;:1-10 [Epub ahead of print]
Niels Schaft, Jan Dörrie, Peter Thumann, Verena E. Beck, Ina Müller, Erwin S. Schultz, Eckhart Kämpgen, Detlef Dieckmann, and Gerold Schuler
Generation of an Optimized Polyvalent Monocyte-Derived Dendritic Cell Vaccine by Transfecting Defined RNAs after Rather Than before Maturation
J Immunol 2005 174: 3087-3097.
N.S., J.D., and P.T. contributed equally to this study
Schaft N, Lankiewicz B, Gratama JW, Bolhuis RL, Debets R.
Flexible and sensitive method to functionally validate tumor-specific receptors via activation of NFAT.
J Immunol Methods. 2003 Sep;280(1-2):13-24.
Schaft N, Willemsen RA, de Vries J, Lankiewicz B, Essers BW, Gratama JW, Figdor CG, Bolhuis RL, Debets R, Adema GJ.
Peptide fine specificity of anti-glycoprotein 100 CTL is preserved following transfer of engineered TCR alpha beta genes into primary human T lymphocytes.
J Immunol. 2003 Feb 15;170(4):2186-94.
Sebo P, Moukrim Z, Kalhous M, Schaft N, Dadaglio G, Sheshko V, Fayolle C, Leclerc C.
In vivo induction of CTL responses by recombinant adenylate cyclase of Bordetella pertussis carrying multiple copies of a viral CD8(+) T-cell epitope.
FEMS Immunol Med Microbiol. 1999 Nov;26(2):167-73.
Wolthers KC, Bea G, Wisman A, Otto SA, de Roda Husman AM, Schaft N, de Wolf F, Goudsmit J, Coutinho RA, van der Zee AG, Meyaard L, Miedema F.
T cell telomere length in HIV-1 infection: no evidence for increased CD4+ T cell turnover.
Science. 1996 Nov 29;274(5292):1543-7.
