Genetische Impfstoffe - eine Revolution in der Vakzinforschung

DNA oder RNA Vakzine, die einfachste Form der genetischen Impfstoffe, ermöglichen völlig neue Wege in der Bekämpfung von unterschiedlichen Krankheiten. Auf Grund ihrer einzigartigen immunologischen Mechanismen können sie nicht nur als Schutzimpfung, sondern auch als therapeutische Impfung, z.B. gegen Tumor oder Allergie, eingesetzt werden. Zahlreiche experimentelle Modelle und erste klinische Studien beweisen die grundsätzliche Wirksamkeit dieses neuen Impfstofftyps.

Unsere Gruppe gehört zu den Europäischen Pionieren auf dem Gebiet der DNA und RNA Vakzinforschung. Gemeinsam mit nationalen und internationalen Partnern aus wissenschaftlichen Instituten und industriellen Forschungslabors haben wir genetische Impfstoffe gegen Malaria, Borreliose, Tumorantigene und Allergene entwickelt. Letztere wurden zu unserem Lieblingsgebiet und zur Zeit liegt unser Forschungsschwerpunkt auf der Entwicklung von neuen Impfstrategien mit niedrig dosierten aber hoch effizienten genetischen Vakzinen für protektive und therapeutische Anwendungen gegen allergische Erkrankungen.

Publikationen

2010 Thalhamer, T., Dobias, H., Stepanoska, T., Pröll, M., Stutz, H., Dissertori, O., Lackner, P., Ferreira, F., Wallner, M., Thalhamer, J., Hartl, A. (2010) Designing Hypoallergenic Derivatives for Allergy Treatment by In Silico Mutation and Screening. J. Allergy Clin. Immunol. (in press) 2009 Dullnig, V., Weiss, R., Amon, S., Rizzi. A., Stutz, H., (2009). Confirmation of immuno-reactivity of the recombinant major birch pollen allergen Bet v 1a by affinity-CIEF. Electrophoresis 30(13): 2337-46. Danzer, M., Polin, H., Proll, J., Haunschmid, R., Hofer, K., Stabentheiner, S., Hackl, C., Kasparu, H., Konig, J., Hauser, H., Binder, M., Weiss, R., Gabriel C., Krieger, O., (2009). Clinical significance of HLA-E*0103 homozygosity on survival after allogeneic hematopoietic stem-cell transplantation. Transplantation 88(4): 528-32. Roesler, E., Weiss, R., Weinberger, E.E., Fruehwirth, A., Stoecklinger, A., Mostböck, S., Ferreira, F., Thalhamer, J., Scheiblhofer, S., (2009) "Immunize and Disappear” – Safety Optimized mRNA Vaccination with a Panel of 29 Allergens. J.Allergy Clin. Immunol. 124(5): 1070-7. Leb, V.M., Jahn-Schmid, B., Kueng, H.J., Schmetterer, K.G., Haiderer, D., Neunkirchner, A., Fischer, G.F., Hartl, A., Thalhamer, J., Steinberger, P., Bohle, B., Seed, B., Pickl, W.F. (2009) Modulation of Allergen-Specific T-Lymphocate Function by Virus-Like Particles Decorated with HLA Class II Molecules. J.Allergy Clin.Immunol. 124(1): 121-8. Gruber, P., Gadermaier, G., Bauer, R., Weiss, R., Wagner, S., Leonard, R., Breiteneder, H., Ebner, C., Ferreira, F., Egger, M. (2009) Role of the polypeptide backbone and post-translational modifications in cross-reactivity of Art v 1, the major mugwort pollen allergen. Biol Chem. 390 (5-6): 445-51. Schulmeister, U., Hochwallner, H., Swoboda, I., Focke-Tejkl, M., Geller, B., Nystrand, M., Harlin, A., Thalhamer, J., Scheiblhofer, S., Keller, W., Niggemann, B., Quirce, S., Ebner, C., Mari, A., Pauli, G., Herz, U., Valenta, R., Spitzauer, S. (2009) Cloning, Expression, and Mapping of Allergenic Determinants of _S1-Casein, a Major Cow’s Milk Allergen. J.Immunol. 182(11): 7019-29. Edlmayer, J., Niespodziana, K., Linhart, B., Focke-Tejkl, M., Westritschnig, K., Scheiblhofer, S., Stoecklinger, A., Kneidinger, M., Valent, P., Campana, R., Thalhamer, J., Popow-Kraupp, T., Valenta, R. (2009) A Combination Vaccine for Allergy and Rhinovirus Infections Based on Rhinovirus-Derived Surface Protein VP1 and a Nonallergenic Peptide of the Major Timothy Grass Pollen Allergen Phl p 11. J.Immunol. 182(10): 6298-306. Wallmann, J., Proell, M., Stepanoska, T., Hantusch, B., Pali-Schöll, I., Thalhamer, T., Thalhamer, J., Jensen-Jarolim, E., Hartl, A. (2009) A mimotope gene vaccine encoding the major IgE epitope of grass pollen allergen Phl p 5 for epitope-specific treatment of mice. Immunol Lett. 122(1):68-75. Ball, T., Linhart, B., Sonneck, K., Blatt, K., Herrmann, H., Valent, P., Stoecklinger, A., Lupinek, C., Thalhamer, J., Fedorov, A.A., Almo, S.C., Valenta, R. (2009) Reducing allergenicity by altering allergen fold: A mosaic protein of Phl p 1 for allergy vaccination. Allergy. 64:569-80. 2008 Jensen-Jarolim E, Wallmann J, Szalai K, Epstein M, Singh P, Hartl A, Thalhamer J, Pali-Scholl I. (2008) Update on vaccines for specific immunotherapy. Drugs Today (Barc). Suppl B:97-8. Constantin, C., Quirce, S., Grote, M., Touraev, A., Swoboda, I., Stoecklinger, A., Mari, A., Thalhamer, J., Heberle-Bors, E., Valenta, R. (2008) Molecular and Immunological Characterization of a Wheat Serine Proteinase Inhibitor as a Novel Allergen in Baker's Asthma. J. Immunol. 180:7451-7460. Weghofer, M., Dall'Antonia, Y., Grote, M., Stoecklinger, A., Kneidinger, M., Balic, N., Krauth, M.T., Fernández-Caldas, E., Thomas, W.R., van Hage, M., Vieths, S., Spitzauer, S., Horak, F., Svergun, D.I., Konarev, P.V., Valent, P., Thalhamer, J., Keller, W., Valenta, R., Vrtala, S. (2008) Characterization of Der p 21, a new important allergen derived from the gut of house dust mites. Allergy 63:758-767. Leb, V.M., Jahn-Schmid, B., Schmetterer, K.G., Kueng, H.J., Haiderer, d., Neunkirchner, A., Fischer, G.F., Nissler, K., Hartl, A., Thalhamer, J., Bohle, B., Seed, B., Pickl, W.F. (2008) Molecular and functional analysis of the antigen receptor of Art v 1-specific helper T lymphocytes. J. Allergy Clin. Immunol. 121:64-71. Wopfner, N., Bauer, R., Thalhamer, J., Ferreira, F., Chapman, M. (2008) Immunologic Analysis of Monoclonal and IgE Antibody Epitopes on Natural and Recombinant Amb a 1. Clin. Exp. Allergy 38:219-226. 2007 Westritschnig, K., Linhart, B., Focke-Tejkl, M., Pavkov, T., Keller, W., Ball, T., Mari, A., A., Hartl, A., Stöcklinger, A., Scheiblhofer, S., Thalhamer, J., Ferreira, F., Vieths, S., Vogel, L., Böhm, A., Valent, P., Valenta, R. (2007) A hypoallergenic vaccine obtained by tail-to-head restructuring of timothy grass pollen profilin, Phl p 12, for the treatment of pollen-plant food cross-sensitization. J. Immunol. 179:7624-7634. Vrtala, S., Focke, M., Kopec, J., Verdino, P., Hartl, A., Sperr, W.R., Fedorov, A.A., Ball, T., Almo, S., Valent, P., Thalhamer, J., Keller, W., Valenta, R. (2007) Genetic Engineering of the Major Timothy Grass Pollen Allergen, Phl p 6, to Reduce Allergenic Activity and Preserve Immunogenicity. J Immunol. 179:1730-1739. Stoecklinger, A., Grieshuber, I., Scheiblhofer, S., Weiss, R., Ritter, U., Kissenpfennig, A., Malissen, B., Koch, F., Romani, N., Ferreira, F., Thalhamer, J., Hammerl, P. (2007) Epidermal Langerhans Cells are Dispensable for Humoral and Cell-mediated Immunity elicited by Gene-Gun Immunization. J Immunol. 179:886-893. Wallner, M., Stoecklinger, A., Thalhamer, T., Bohle, B., Vogel. L., Briza, P., Breiteneder, H., Vieths, S., Hartl, A., Mari, A., Ebner, C., Lackner, P., Hammerl, P., Thalhamer, J., Ferreira, F. (2007) Allergy multi-vaccines created by DNA shuffling of tree pollen allergens. J. Allergy Clin. Immunol. Jul 9 (Epub ahead of print). Wallner, M., Briza, P., Thalhamer, J., Ferreira, F. (2007) Specific immunotherapy in pollen allergy. Curr. Opin. Mol. Ther. 9:160-167. Scheiblhofer, S., Weiss, R., Thalhamer, J. (2007) Genetic immunization: New ways for protective and therapeutic vaccines against allergic diseases. Wien. Med. Wochenschr. 157:111-115. Linhart, B., Bigenzahn, S., Hartl, A., Lupinek, C., Thalhamer, J., Valenta, R., Wekerle, T. (2007) Co-stimulation blockade inhibits allergic sensitization but does not affect established allergy in a murine model of grass pollen allergy. J. Immunol. 178:3924-3931. Gieras, A., Focke-Tejkl, M., Ball, T., Verdino, P., Hartl, A., Thalhamer, J., Valenta, R. (2007) Molecular determinants of allergen-induced effector cell degranulation. J. Allergy Clin. Immunol. 119:384-390. Gehwolf, R., Weiss, R., Gabler, M., Hurst, A.C., Bertl, A., Thalhamer, J., Obermeyer, G. (2007) From sequence to antibody: genetic immunisation is suitable to generate antibodies against a rare plant membrane protein, the KAT 1 channel. FEBS Lett. 16 [Epub ahead of print]. Weiss, R., Scheiblhofer, S., Thalhamer, J., Bickert, T., Richardt, U., Fleischer, B., Ritter, U. (2007) Epidermal inoculation of Leishmania antigen by gold bombardment results in a chronic form of leishmaniasis. Vaccine 25:25-33. Scheiblhofer, S., Stoecklinger, A., Gruber, C., Hauser-Kronberger, C., Alinger, B., Hammerl, P., Thalhamer, J., Weiss, R. (2007) Gene gun immunization with clinically relevant allergens aggravates allergen induced pathology and is contraindicated for allergen immunotherapy. Mol. Immunol. 44:1889-1897. 2006 Scheiblhofer, S., Weiss, R., Thalhamer, J. (2006) Genetic vaccination approaches against malaria based on the circumsporozoite protein. Wien. Klin. Wochenschr. 118:9-17. Gabler, M., Scheiblhofer, S., Kern, K., Leitner, W.W., Hauser-Kronberger, C, Alinger, Beate., Valenta, R., Thalhamer, J., Weiss, R. (2006). Immunization with nanogram doses of a replicon DNA vaccine encoding Phl p 5 effectively prevents from allergic reactions. J. Allergy Clin. Immunol. 118:734-741. Radauer, C., Willerroider, M., Fuchs, F., Hoffmann-Sommergruber, K., Thalhamer, J., Ferreira, F., Scheiner, O., Breiteneder, H. (2006) Cross-reactive and species-specific immunoglobulin E epitopes of plant profilins: an experimental and structure-based analysis. Clin. Exp. Allergy 36:920-929. Bauer, R., Scheiblhofer, S., Kern, K., Gruber, C., Stepanoska, T., Thalhamer, T., Hauser-Kronberger, C., Alinger, B., Zögg, T., Gabler, M., Ferreira, F., Hartl, A., Thalhamer, J., Weiss, R. (2006) Generation of hypoallergenic DNA vaccines by forced ubiquitination: Preventive and therapeutic effects in a mouse model of allergy. J. Allergy Clin. Immunol. 118:269-76. Scheiblhofer, S., Gabler, M., Leitner, W.W., Bauer, R., Zögg, T., Ferreira, F., Thalhamer, J., Weiss, R. (2006) Effective inhibition of type I allergic responses with nanogram doses of replicon-based DNA vaccines. Allergy 61:828-35. Weiss, R., Scheiblhofer, S., Gabler, M., Ferreira, F., Leitner, W.W., Thalhamer, J. (2006) Is genetic vaccination against allergy possible? Int. Arch. Allergy Immunol. 139:332-345. Balenga, N.A., Zahedifard, F., Weiss, R., Sarbolouki, M.N., Thalhamer, J., Rafati, S. (2006) Protective efficiency of dendrosomes as novel nano-sized adjuvants for DNA vaccination against birch pollen allergy. J Biotechnol. 124:602-614. Balenga, N.A., Thalhamer, J., Weiss, R. (2006) Bicistronic expression plasmid encoding allergen and anti-IgE single chain variable fragment antibody as a novel DNA vaccine for allergy therapy and prevention. Med Hypotheses. 67:71-74. Ferreira. F., Briza, P., Inführ; D., Schmidt, G., Wallner, W., Wopfner, N., Thalhamer, J., Achatz, G. (2006) Modified Recombinant Allergens for Safer Immunotherapy. Inflamm. Allergy - Drug Targets 5:5-14. Scheiblhofer, S., Weiss, R., Leitner , W.W., Thalhamer, J. (2006). Replicase-based DNA vaccines for allergy treatment. Methods Mol. Med., 127:221-36. Weiss, R., Scheiblhofer, S., Thalhamer, J. (2006). DNA vaccines for allergy treatment. Methods Mol. Med., 127:253-268. 2005 Linhart, B., Hartl, A., Jahn-Schmid, B., Verdino, P., Keller, W., Krauth, MT., Valent, P., Horak, F., Wiedermann, U., Thalhamer, J., Ebner, C., Kraft, D., Valenta, R. (2005) A hybrid molecule resembling the epitope spectrum of grass pollen for allergy vaccination. J. Allergy Clin. Immunol. 115:1010-1016. Bergmann-Leitner, E.S., Scheiblhofer, S., Weiss, R., Duncan, E.H., Leitner, W.W., Chen, D., Angov. E., Khan, F., Williams, J.L., Winter, D.B., Thalhamer, J., Lyon, J.A., Tsokos, G.C. (2005) C3d Binding to the Circumsporozoite Protein Carboxy-Terminus Deviates Immunity Against Malaria. Int. Immunol. 17:245-255. Weiss, R., Hammerl, P., Hartl, A., Hochreiter R., Leitner, W.W., Scheiblhofer, S. and Thalhamer, J. (2005) Design of protective and therapeutic DNA vaccines for the treatment of allergic diseases. Curr. Drug Targets Inflamm. Allergy 4:585-597. 2004 Focke, M., Linhart, B., Hartl, A., Wiedermann, U., Sperr, W.R., Valent P., Thalhamer, J., Kraft, D., Valenta, R. (2004) Non-anaphylactic surface-exposed peptides of the major birch pollen allergen, Bet v 1, for preventive vaccination. Clin. Exp. Allergy 34:1525-33. Scheiblhofer, S., Weiss, R., Hammerl, P., Hartl, A., Ferreira, F., Leitner, W.W., Thalhamer, J. (2004) DNA Vaccines and Allergy: New approaches against a pandemic civilization disease. In: Recent Research Developments in Allergy & Clinical Immunology 5:27, ed. S.G.Pandalai. Research Signpost, Kerala, India. Ferreira, F., Wallner, M., Thalhamer, J. (2004) Customized antigens and immunomodulations for desensitizing allergic patients. Adv. Immunol. 84:79-129. Westritschnig, K., Focke, M., Verdino, P., Goessler, W., Keller, W., Twardosz, A., Mari, A., Horak, F., Wiedermann, U., Hartl, A., Thalhamer, J., Sperr, W.R., Valent, P., Valenta, R. (2004) Generation of an Allergy Vaccine by Disruption of the Three-Dimensional Structure of the Cross-Reactive Calcium-Binding Allergen, Phl p 7. J. Immunol. 172:5684-5692. Dedic, A., Bauer, R., Hartl, A., Van Ree, R., Thalhamer, J., Ebner, C., Ferreira, F. (2004) Art v 1 isoforms: Cloning, E.coli expression and immunological analysis. J. Allergy Clin. Immunol. 113: 299S. Hartl, A., Weiss, R., Hochreiter, R., Scheiblhofer, S., Thalhamer, J. (2004) DNA vaccines for allergy treatment. Methods. 3:328-339. Mahler, V., Vrtala, S., Kuss, O., Diepgen, T., Suck, R., Cromwell, O., Fiebig, H., Hartl, A., Thalhamer, J., Schuler, G., Kraft, D., Valenta, R. (2004) Vaccines for birch pollen allergy based on genetically engineered hypoallergenic derivatives of the major birch pollen allergen, Bet v 1. Clin Exp Allergy. 34: 115-122. Holler, C., Simon-Nobbe, B., Wally, V., Crameri, R., Richter, K., Hartl, A., Thalhamer, J., Ebner C. (2004) Histamine releasing factor (TCTP) is an auto-reactive IgE-binding antigen in patients allergic to Cladosporium herbarum. J. Allergy Clin. Immunol. 113: 299S. Egger, M., Wopfner, N., Himly, M., Dedic, A., Bauer, R., Mari, A.,Van Ree, r., Hartl, A., Thalhamer, J., Ferreira, F. (2004) Art v 1, the major mugwort allergen, cross-reacts with proteins originating from compositae and grass pollen. J. Allergy Clin. Immunol. 113, 300S. Hartl, A., Hochreiter, R., Stepanoska, T., Ferreira, F. and Thalhamer J. (2004) Characterization of the protective and therapeutic efficiency of a DNA vaccine encoding the major birch pollen allergen Bet v 1a. Allergy 59:65-73. Oertel, A., Aichinger, N., Hochreiter, R., Thalhamer, J., Lütz-Meindl, U. (2004) Analysis of mucilage secretion and excretion in Micrasterias (Chlorophyta) by means of immunoelectron microscopy and digital time lapse video microscopy. J. Phycol. 40: 711-720. 2003 Leitner, W., Thalhamer, J. (2003) DNA vaccines for non-infectious disease. Expert Opin. Biol. Ther. 3:627-638. Hochreiter,R., Stepanoska, T., Ferreira, F., Valenta, R., Vrtala, S., Thalhamer, J. and Hartl, A. (2003) Prevention of Allergen-specific IgE production and suppression of an established Th2-type response by immunization with DNA encoding hypoallergenic allergen derivatives of Bet v 1, the major birch pollen allergen. Eur. J. Immunol. 33:1667-1676. Hartl, A., Weiss, R., Hochreiter, R., Scheiblhofer, S., Bauer, R., Valenta, R., Ferreira, F., Leitner, W. and Thalhamer, J. (2003) Strategies for the development of safe and effective DNA vaccines for allergy treatment. Arb. Paul-Ehrlich Inst. Bundesamt Sera Impfstoffe Frankf A M. 94: 279-98. Mostböck, S., Macejova, D., Baranova, M., Weiss, R., Scheiblhofer, S., Thalhamer, J., Brtko, J. (2003) Reduction of MNU-induced tumor burden with DNA vaccines encoding mutated ras epitopes and the costimulatory molecule B7.1. Gen. Physiol. Biophys. 22:561-565. Repa, A., Grangette, C., Daniel, C., Hochreiter, R., Hoffmann-Sommergruber, K., Thalhamer, J., Kraft, D., Breiteneder, H., Mercenier, A., Wiedermann, U. (2003) Mucosal co-application of lactic bacteria and allergens induces counter-regulatory immune responses in a murine model of birch pollen allergy. Vaccine 22:87-95. Brtko, J. and Thalhamer, J. (2003) Renaissance of the biologically active vitamin A derivatives: Established and novel directed therapies for cancer and chemoprevention. Curr. Pharm. Des. 9:2067-2077. Liska, J., Macejova, D., Galbavy, S., Baranova, M., Zlatos, J., Stvrtina S., Mostbock, S., Weiss, R., Scheiblhofer, S., Thalhamer, J. and Brtko, J.(2003) Treatment of 1-methyl-1-nitrosourea-induced mammary gland tumours with immunostimulatory CpG motifs and 13-cis retinoic acid in female rats:Histopathological study. Exp. Toxicol. Pathol. 55: 173-180. Bauer, R., Himly, M., Dedic, A., Ferreira, F., Thalhamer, J. and Hartl, A. (2003) Optimization of Codon Usage is required for effective genetic immunization against Art v 1, the major allergen of mugwort pollen. Allergy 58:1003-1010. Hochreiter, R., Ferreira, F., Thalhamer, J. and Hammerl, P. (2003) Th1-promoting DNA immunization against allergens modulates the ratio of IgG1/IgG2a but does not affect the anaphylactic potential of IgG1 antibodies: no evidence for the synthesis of non-anaphylactic IgG1. J. Allergy Clin. Immunol. 112:579-584. Scheiblhofer, S., Weiss, R., Dürnberger, H., Mostböck, S., Breitenbach, M., Livey, I., Thalhamer, J. (2003) A DNA vaccine encoding the outer surface protein C from Borrelia burgdorferi is able to induce protective immune responses. Microbes Infect. 5:939-946. Lindinger, P., Mostböck, S., Hammerl, P., Hartl, A., Thalhamer, J., Abrams, S.I. (2003) Induction of Murine ras Oncogene Peptide-Specific T Cell Responses by Immunization with Plasmid DNA-Based Minigene Vectors. Vaccine 21:4285-4296. Willerroider, M., Fuchs, H., Ballmer-Weber, H., Focke, M, Susani, M., Thalhamer, J., Ferreira, F., Wüthrich, B., Scheiner, O., Breiteneder, H. and Hoffmann-Sommer-gruber, K. (2003) Cloning and Molecular and Immunological Characterization of Two New Food Allergens, Lyc e 1 and Cap a 2, Profilins From Bell Pepper and Tomato. Int. Arch. Allergy Immunol. 131:245-255. Hufnagl, K., Wagner, B., Winkler, B., Baier, W., Hochreiter, R., Thalhamer, J., Kraft, D., Scheiner, O., Breiteneder, H., Wiedermann, U. (2003) Induction of mucosal tolerance with recombinant Hev b 1 and recombinant Hev b 3 for treatment of latex allergy in BALB/c mice. Clin.Exp.Immunol. 133:170-176. Mostböck, S.; Macejova, D.; Baranova, M.; Weiss, R.; Scheiblhofer, S.; Verwanger, T.; Krammer, B.; Thalhamer, J. and Brtko, J. (2003) Analysis of altered gene expression profiles in retinoic acid or CpG treated Sprague-Dawley rats with MNU-induced mammary adenocarcinoma by cDNA macro array. Gen. Physiol. Biophys. 22:129-133 Dedic, A., Bauer, R., Hartl., A., van Ree, R., Thalhamer, J., Ebner, C., Ferreira, F. (2003). Cloning, expression and immunological analysis of isoforms of Art v 1, the major allergen of mugwort pollen. Allergy 58 (Suppl 74): 89, 2003 Egger, M., Himly, M., Dedic, A., Bauer, R., Mari, A., van Ree, R., Hartl, A., Thalhamer, J., Wopfner, N., Ferreira, F. (2003). Is Art v 1, the major allergen of mugwort pollen, a cross-reactive allergen? Allergy 58 (Suppl 74): 89, 2003 2002 Thalhamer, J. (2002) DNA vaccination in allergy. Immunology 107: 67, Suppl.1. Nelde, A., Hartl A., Thalhamer, J., Duschl, A. (2002). IL-4 receptor inhibition and specific immunotherapy in a mouse model for immediate type allergies. Immunobiol. 206:214-215 Mostböck, S., Ficova, M., Macejova, D., Baranova, M., Kotyzova, D., Mickova, V., Eybl, V., Thalhamer, J., Brtko, J. (2002) Effect of Cadmium and Mercury on the nuclear retinoic acid receptors. Gen. Physiol. Biophys. 21:443-456. Pittertschatscher, K., Hochreiter, R., Thalhamer, J., Hammerl, P.(2002) Quantification of histamine in blood plasma and cell culture supernatants: A validated one-step GC-MS method. Anal. Biochem. 15:300-306. Wopfner, N., Willeroider, M., Hebenstreit, D., van Ree, R., Aalbers, M., Briza, P., Thalhamer, J., Ebner, C., Richter, K., Ferreira, F. (2002). Molecular and immunological characterization of profilin from mugwort pollen. Biol. Chem. 383:1779-89. Hammerl P., Hartl A., Freund J., Thalhamer J. (2002) Detection of serological crossreactions by Western Cross Blotting. In: The Protein Protocols Handbook 2nd edition, editor John M. Walker, Humana Press Inc.USA. Ferreira F., Wallner M., Breiteneder H., Hartl A., Thalhamer J., Ebner C. (2002) Genetic Engineering of Allergens: Future Therapeutic Products. Int. Arch. Allergy Immunol. 128:171-178. Weiss, R., Scheiblhofer, S., Freund, J., Ferreira F., Livey, I., Thalhamer, J. (2002) Gene gun bombardment with gold particles displays a particular Th2-promoting signal that overrules the Th1-inducing effect of immunostimulatory CpG motifs in DNA vaccines. Vaccine, 20:3148. Wallner, M.F., Nestelbacher, R., Breiteneder, H., Hoffmann-Sommergruber, K., Thalhamer, J., Ferreira, F. (2002) In vitro evolution of the Bet v 1 family by gene shuffling. J. Allergy Clin. Immunology, 109(1), S164. Macejova, D., Dvorcakova, R., Weiss, R., Scheiblhofer S., Mostböck S., Liska, J., Zorad, J., Thalhamer, J., Brtko, J. (2002) Reduction of 1-methyl-1-nitrosourea-induced mammary gland carcinoma by in vivo application of immunostimulatory CpG motifs in Sprague-Dawley rats. Gen. Physiol. Biophys. 20:445-448. 2001 Leitner W. W., Hammerl P., Thalhamer J. (2001) Nucleic acid for the treatment of cancer: Genetic vaccines and DNA adjuvants. Curr. Pharm. Des. 7:1639-1665. Thalhamer, J., Leitner, W.W., Hammerl, P., Brtko. J. (2001) Designing immune responses with genetic immunization and immunostimulatory DNA sequences. Endocr. Regul. 35:143-166. Hochreiter, R., Hartl A., Freund J., Valenta R., Ferreira F., Thalhamer J. (2001) The influence of CpG motifs on a protein or DNA-based Th2 type immune response against major pollen allergens Bet v 1a, Phl p 2 and E-coli derived ß-galactosidase. Int. Arch. Allergy Immunol. 124: 406-10. Scheiblhofer, S., Chen D., Weiss R., Khan F., Mostboeck S., Fegeding K., Leitner W., Thalhamer J., Lyon J.A. (2001) Removal of the circumsporozoite protein glycophosphatidylinositol signal sequence from a CSP DNA vaccine enhances induction of CSP-specific Th2 type immune responses and improves protection against malaria infection. Eur. J. Immunol. 31: 692-698. 2000 Bristol J.A., Orsini C., Lindinger P., Thalhamer J., Abrams S.I. (2000) Identification of a ras oncogene peptide that contains both CD4+ and CD8+ T cell epitopes in a nested configuration and elicits both T cell subset responses by peptide or DNA immunization. Cell. Immunol. 205: 73-83. Webersinke G., Bauer H.C., Danniner C., Krizbai I.A., Schittny J.C., Thalhamer J., Bauer H. (2000) The use of genetically modified glial cells overexpressing laminin ?1-chain peptides in neurite outgrowth studies. Cell Mol Neurobiol. 20: 605-621. Thalhamer J., Hammerl P., Hartl A. (2000) Nitrocellulose-based Immunoaffinity Chromatography. In Methods in Molecular Biology: Affinity Chromatography, Methods and Protocols. Eds P.Bailon, W.Berthold and WJ Fung, Humana Press Inc., Vol. 147, pg. 89-95. Hammerl P., Hartl A., Freund J., Thalhamer J. (2000) Western Cross Blot. In Methods in Molecular Biology: Affinity Chromatography, Methods and Protocols. Eds P.Bailon, W.Berthold and WJ Fung, Humana Press Inc., Vol. 147, pg. 163-175. Thalhamer J., Hammerl P., Hartl A. (2000) Nitrocellulose-based Immunoaffinity Chromatography. In Methods in Molecular Biology: Affinity Chromatography, Methods and Protocols. Eds P.Bailon, W.Berthold and WJ Fung, Humana Press Inc., Vol. 147, pg. 89-95. Hammerl P., Hartl A., Freund J., Thalhamer J. (2000) Western Cross Blot. In Methods in Molecular Biology: Affinity Chromatography, Methods and Protocols. Eds P.Bailon, W.Berthold and WJ Fung, Humana Press Inc., Vol. 147, pg. 163-175. Brtko J., Hartl A., Weiss R., Scheiblhofer S., Mostböck S., Thalhamer J. (2000) Retinoic acid receptor status in mouse spleen during a primary immune response against ß-galactosidase. Endocr. Regul. 34: 113-118. Weiss R., Leitner W.W., Scheiblhofer S., Chen D., Bernhaupt A., Mostböck S., Thalhamer J., Lyon J.A. (2000) Genetic vaccination against malaria infection by intradermal and epidermal injection of a plasmid containing the gene encoding the Plasmodium berghei circumsporozoite protein. Infect. Immun. 68: 5914-5919. Brtko J., Hartl A., Weiss R., Bernhaupt A., Scheiblhofer S., Mostböck S., Thalhamer J. (2000) DNA immunization in vivo down-regulates nuclear all-trans retinoic acid receptors in mouse spleen cells. Mol. Cell. Endocrinol. 165:107-113 Thalhamer J., Hartl A (2000). DNA Immunisierung: Eine neue Form der Immuntherapie zur Allergiebehandlung. Allergologie 23: 589.

The Gene Vaccine Revolution

Genetic vaccination is destined to change the way in which we prevent and treat a variety of human diseases. Numerous studies have demonstrated that this novel technology can have significant advantages over traditional methods of vaccination.

Our group belongs to the European pioneers of DNA and RNA vaccine research. Together with national and international collaboration partners from scientific institutes and companies, we have developed vaccines against malaria, borreliosis, tumor antigens and type I allergy. During the past six years, the latter became our pet subject. At present we focus on genetic vaccination strategies for highly effective and safety-optimized vaccines for protective and therapeutic approaches against allergic diseases.

Ongoing Projects

Title International PhD Program "Immunity in Cancer and Allergy" funded by the Austrian Science Fund (FWF project number W1213) Description of the Project The mammalian immune system acts as a complex surveillance system ensuring to distinguish between self and non-self, and between harmless and dangerous events approaching from outside or inside of the organism. Moreover, it interacts with growth, differentiation and death of cells and tissues, and thus maintains the homeostasis and the integrity of our body. The doctoral college is focused on two pathologies of the immune system, i.e. the overwhelming allergic immune response and the inefficient immune response against certain tumors. Both diseases are a growing concern and there is an urgent medical need to elucidate the underlying mechanisms for the development of new therapies. Unraveling the cellular and molecular immunological mechanisms and pathways enables to develop rational and molecule-based strategies for the treatment of these diseases. The aim of the doctoral college is to attract and select excellent graduate students from all over the world, to provide an intellectually stimulating environment, an excellent instrumental and methodological infrastructure and ambitious scientific projects, and to prepare for a successful career in basic as well as translational and applied science. The college comprises ten research groups. Their track records, experience with national and international programs, and their excellent infrastructure guarantees high quality research and training. Furthermore, the college structure ensures that students benefit from the collective experience of the researchers. Period October 2008 - Title Christian Doppler Laboratory for Allergy Diagnosis and Therapy Module 5 - Development of gene vaccines for allergy treatment (funded by CDG and Biomay) Description of the Project At present, the major restriction of genetic intradermal or intramuscular needle immunization concerning clinical application is the requirement for large doses of DNA and the relatively weak immunogenicity. Alternative, low-dose and immunogenic injection methods such as the gene gun or powderject™ are not suitable for the treatment of allergy because of inducing a Th2-biased type of immune response. The first approach of the project will be to overcome these hurdles with replicase-based gene vaccines. Immunization with these constructs enables to trigger strong humoral and cellular Th1-biased immune responses with nanogram quantities of needleinjected plasmid DNA. The second aspect will cover the development of strategies for minimizing the risk of anaphylactic side effects resulting from the potential expression of biologically active allergens following genetic vaccination. We will develop gene vaccines encoding allergen variants or derivatives, which still retain the induction of T-cell responsiveness but display no allergenic reactivity upon sensitization with the wildtype allergen. For this purpose, forced ubiquitination of DNA vaccines will serve to develop a routine approach for destroying IgE-binding epitopes on allergens in order to avoid recognition by pre-existing IgE antibodies. Simultaneously, T cell epitopes of the allergen will be preserved. Period 2006-2012 Title Mapping of helper T-cell epitopes of the grass pollen allergen Phl p 5 (industrial project, funded by Biomay) Description of the project Allergen gene vaccines as well as allergen protein vaccines (e.g. as used in specific immunotherapy-SIT) display their anti-allergic activity via specific induction of T-cells. In the case of gene vaccines, both, the prophylactic and therapeutic efficacy is based on CD+ T-cells of the Th1 type. On the other hand, successful immunotherapy with protein vaccines is assumed to be based on a combination of regulatory T-cells (inducing a tolerant state of the immune system), IFN-gamma-producing T-cells (deviating the Th2 type immune response) and IgG antibodies binding to the allergen and thus blocking the crosslinking of IgE on mast cells and basophils. A necessary prerequisite for an anti-allergic vaccine would be that all three mentioned mechanisms work in an allergen-specific way, which requires the induction of allergen-specific T-cells. Therefore, the knowledge of T-cell epitopes is crucial for the development and testing of gene and protein vaccines for the treatment of type I allergy. The project will strive for the characterization of mouse and human helper T-cell epitopes of the grass pollen allergen Phl p 5. Period November 2007 – December 2009 Title Systematic evaluation of up-to-date genetic vaccination approaches for protective and therapeutic treatment of type I allergy: Preparation of optimized vaccines for phase I clinical trials (FWF project number L181 B13) Description of the Project The prevalence of allergic diseases has increased substantially over the past few decades in the industrialized world despite the introduction of new and effective drugs for their treatment. At present, allergic diseases affect more than one quarter of the population. Allergy can be defined as a disorder of the immune system, leading to inappropriate responses against commonly encountered substances that are otherwise harmless. Clinically, the most common allergic diseases are asthma, rhinitis/conjunctivitis (hayfever) and eczema. Allergic immune responses are characterized by the synthesis of allergen-specific IgE antibodies and the production of immunomodulatory molecules such as interleukin (IL)-4, IL-5 and IL-13. Today, most anti-allergic treatments only control the symptoms of allergy via immunosuppressive and anti-inflammatory agents such as antihistamines, corticosteroids and beta-agonists. The only curative approach targeting the disease is allergen-specific immunotherapy (SIT). Although SIT has already been introduced by Noon and Freeman in 1911, the immunological mechanisms underlying this treatment are still unclear. Although SIT has proven its suitability and clinical efficacy especially for mono-or oligosensitized patients and for treatment of hypersensitivities against stinging insects, only 30-50% of allergic rhinitis patients respond and SIT is even less effective for asthmatics. Another major disadvantage of classical SIT is the risk of anaphylactic side effects caused by systemic application of large amounts of allergen via subcutaneous injections. In summary, there is an urgent need for improvement and/or alternatives to conventional SIT. Over the last decade, the genetic vaccine revolution has provided researchers with exciting possibilities to design new advanced vaccines. Genetic vaccination employs the allergen in its purest form - the genetic information. Following transfection with the genetic material, cells of the vaccinated individual will translate the information into the respective allergen. This type of vaccine specifically induces a type of immune response dominated by the immunomodulator interferon gamma, which suppresses IL-4/IL-5 dependent allergic immune responses. Meanwhile, a panel of genetic vaccines has been successfully administered in animal models, including plasmid DNA, purified RNA, and "self-replicating" DNA/RNA. Also several different delivery methods such as intramuscular or intradermal injections, application via "gene gun", in vivo electroporation, or intranodal and mucosal routes have been investigated in animal models. However, results from early clinical studies (for infectious diseases and cancer) have often been disappointing due to suboptimal utilization of the latest feasibilities of modern genetic vaccines. Although animal studies with anti-allergic genetic vaccines have been promising, a systematic evaluation of different vaccines, doses, immunization methods and schedules will be of crucial importance for the success of future clinial trials. For the present study, we will establish novel anti-allergic RNA vaccines and bacterial ghost-delivered genetic and recombinant vaccines and evaluate them together with the present state-of-the-art genetic vaccines, application methods, adjuvants and immunization protocols as a basis for preparing clinical application. Simultaneously with providing the urgently necessary comparative information about different vaccination approaches, an objective evaluation will also exert gentle pressure on planning of phase I trials, thus helping to speed up the process of developing rational anti-allergic genetic vaccines for human use. Period 1.1.2006 – 31.12.2008 Title Immune responses to antigen application on bare skin (In collaboration with the Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany) Description of the Project The skin represents an important target for gene gun vaccination strategies. However the exact mechanisms that underline the induction of an adaptive immune response induced by that technique are poorly understood. In the present project we analyze different cutaneous dendritic cell subsets accounting for efficient vaccination. This basic research will help us to improve current vaccination strategies. Furthermore, novel concepts are tested whether epicutaneous incorporation of different antigens is able to modulate ongoing immune responses. Period Since 1.1.2005 Title Bacterial ghosts as delivery system for genetic vaccines (In collaboration with the Institute for Microbiology and Genetics of the University of Vienna) Bacterial ghosts are empty gram negative bacterial cellular envelopes with retained morphological, structural and antigenic features of the cell wall. Bacterial ghosts can be used as vaccine candidate per se. Alternatively, they can be loaded with DNA vaccines and recombinant protein vaccines and have been demonstrated to effectively induce cellular and humoral responses, thus introducing a promising technology for the development of novel vaccine types. The mode of application of DNA vaccines is known to be decisive for the immunogenicity of the construct and the induced type of immune response and bacterial danger signals transmitted via pattern recognition receptors (e.g., TLRs) represent strong triggers of immune reactions. In vitro studies with Mannheimia haemolytica ghosts carrying plasmid DNA revealed efficient uptake by APCs, leading to transfection rates up to 60 %. In addition to targeting the DNA vaccine construct to APCs, bacterial ghosts promote maturation and activation of dendritic cells as the encoded antigen is delivered in the context of an adequate danger signal. However, the endotoxic effects of free LPS are not observed, because LPS is associated with the ghost envelopes. In the present project the application of DNA vaccines with, or recombinant antigens fused to bacterial ghosts is evaluated in comparison with conventional protein immunization procedures and the state-of-the-art gene vaccination methods. Period Since 1.1.2005