Main Research Topic:

Immunobiology of Toll-like Receptors (TLRs) recognising nucleic acids

In retrospective, several laboratories have contributed to the discovery of endosomally expressed TLRs that predominantly recognise pathogen-derived nucleic acids. For example, TLR3 senses double stranded (ds) RNA, TLR-7, -8 recognise single stranded (ss) RNA and TLR9 is activated by ss CpG-DNA. By serendipity, Tim Sparwasser observed in 1996/1997 that synthetic Oligonucleotides (ODN) rich in CpG-motifs activated macrophages and dendritic cells. Grayson Lipford noted 1999 that CpG-ODN admixed to exogenous antigen displayed adjuvanticity thus promoting Th-1 responses and induction of cytotoxic T lymphocytes (CTL). Together these studies sparked our interest to define the respective receptors and signalling pathways involved. In tackling this question, Hans Häcker, Martin Vabulas and Parviz Ahmad-Nejad observed that CpG-ODN induced signalling was depending on the cytosolic adaptor protein MyD88, similar to IL-1R signalling. In collaboration with Akira and Hemmi and by genetic complementation studies (Stefan Bauer), this information led 2000-2001 to the discovery that CpG-DNA acts as ligand for TLR9. Subsequently, Parviz Ahmand-Nejad noted 2002 that TLR9 signalling is initiated within endosomes (where TLR-9 is expressed) while LPS initiates TLR4 signalling at the cell membrane. Flavell’s group (USA) had reported that dsRNA is recognized by TLR-3 and Hemmi (Japan) had observed that TLR-7 could be activated by the synthetic compounds imiquimod. Subsequently, Stefan Bauer aimed at defining the natural TLR-7 ligand. In parallel with de Sousa’s group (England) ss U-rich RNA sequences were defined in 2004 as natural ligand for TLR-7.

Antje Heit focussed on the development of vaccination protocols using endosomal TLR ligands as adjuvants such as CpG-DNA. One problem she was facing was to efficiently translocate CpG-DNA (together with exogenous Ag) into endosomes of antigen-presenting cells (APCs). In her experience this occurs best either by chemically linking CpG-DNA to the proteinaceous Ag, or by co-entrapping CpG-DNA and Ag in biodegradable microspheres. Frank Schmitz analysed the molecular basis of how TLR-9 activation leads to ?-interferon production in myeloid dendritic cells as opposed to plasmacytoid cells. Tobias Haas was interested in the question why the immunobiology of natural phospho-di-ester DNA activates TLR-9 rather independently of CpG-motifs while the TLR-9 activating properties of phospho-thioate modified synthetic DNA is CpG-motif dependent. In collaboration with Carsten Kirschning, Stephan Bauer, we recently succeeded to show that the orphan receptor TLR13 (expressed in mice but not in human) senses a ss RNA motif within 23S ribosomal rRNA, provided Adenosin (a.a. 2058) is not modified to confer antibiotic resistance.

Former Scientists of the group:

Parviz Ahmad-Nejad (now Mannheim)

Stefan Bauer (now Marburg)

Tobias Haas (München)

Hans Häcker (now Salt Lake City/USA)

Klaus Heeg (now Heidelberg)

Antje Heit (now Seattle/USA)

Carsten Kirschning (now Essen)

Grayson Lipford (now USA)

Tim Sparwasser (now Hannover)

Frank Schmitz (now Seattle/USA)

Martin Vabulas (now Berlin)

Selected Publications (since 2007)

• Heit, A., Schmitz, F., Haas, T., Busch, D. H., and Wagner, H. (2007) Antigen co-encapsulated with adjuvants efficiently drive protective T cell immunity, Eur J Immunol 37, 2063-2074.
• Kaiser-Schulz, G., Heit, A., Quintanilla-Martinez, L., Hammerschmidt, F., Hess, S., Jennen, L., Rezaei, H., Wagner, H., and Schatzl, H. M. (2007) Polylactide-coglycolide microspheres co-encapsulating recombinant tandem prion protein with CpG-oligonucleotide break self-tolerance to prion protein in wild-type mice and induce CD4 and CD8 T cell responses, J Immunol 179, 2797-2807.
• Lahl, K., Loddenkemper, C., Drouin, C., Freyer, J., Arnason, J., Eberl, G., Hamann, A., Wagner, H., Huehn, J., and Sparwasser, T. (2007) Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease, J Exp Med 204, 57-63.
• Rad, R., Brenner, L., Krug, A., Voland, P., Mages, J., Lang, R., Schwendy, S., Reindl, W., Dossumbekova, A., Ballhorn, W., Wagner, H., Schmid, R. M., Bauer, S., and Prinz, C. (2007) Toll-like receptor-dependent activation of antigen-presenting cells affects adaptive immunity to Helicobacter pylori, Gastroenterology 133, 150-163 e153.
• Schmitz, F., Heit, A., Guggemoos, S., Krug, A., Mages, J., Schiemann, M., Adler, H., Drexler, I., Haas, T., Lang, R., and Wagner, H. (2007) Interferon-regulatory-factor 1 controls Toll-like receptor 9-mediated IFN-beta production in myeloid dendritic cells, Eur J Immunol 37, 315-327.
• Spiller, S., Dreher, S., Meng, G., Grabiec, A., Thomas, W., Hartung, T., Pfeffer, K., Hochrein, H., Brade, H., Bessler, W., Wagner, H., and Kirschning, C. J. (2007) Cellular recognition of trimyristoylated peptide or enterobacterial lipopolysaccharide via both TLR2 and TLR4, J Biol Chem 282, 13190-13198.
• Stemberger, C., Huster, K. M., Koffler, M., Anderl, F., Schiemann, M., Wagner, H., and Busch, D. H. (2007) A single naive CD8+ T cell precursor can develop into diverse effector and memory subsets, Immunity 27, 985-997.
• Weber, A., Paschen, S. A., Heger, K., Wilfling, F., Frankenberg, T., Bauerschmitt, H., Seiffert, B. M., Kirschnek, S., Wagner, H., and Hacker, G. (2007) BimS-induced apoptosis requires mitochondrial localization but not interaction with anti-apoptotic Bcl-2 proteins, J Cell Biol 177, 625-636.
• Cirl, C., Wieser, A., Yadav, M., Duerr, S., Schubert, S., Fischer, H., Stappert, D., Wantia, N., Rodriguez, N., Wagner, H., Svanborg, C., and Miethke, T. (2008) Subversion of Toll-like receptor signaling by a unique family of bacterial Toll/interleukin-1 receptor domain-containing proteins, Nat Med 14, 399-406.
• Gratz, N., Siller, M., Schaljo, B., Pirzada, Z. A., Gattermeier, I., Vojtek, I., Kirschning, C. J., Wagner, H., Akira, S., Charpentier, E., and Kovarik, P. (2008) Group A streptococcus activates type I interferon production and MyD88-dependent signaling without involvement of TLR2, TLR4, and TLR9, J Biol Chem 283, 19879-19887.
• Haas, T., Metzger, J., Schmitz, F., Heit, A., Muller, T., Latz, E., and Wagner, H. (2008) The DNA sugar backbone 2′ deoxyribose determines toll-like receptor 9 activation, Immunity 28, 315-323.
• Heit, A., Gebhardt, F., Lahl, K., Neuenhahn, M., Schmitz, F., Anderl, F., Wagner, H., Sparwasser, T., Busch, D. H., and Kastenmuller, K. (2008) Circumvention of regulatory CD4(+) T cell activity during cross-priming strongly enhances T cell-mediated immunity, Eur J Immunol 38, 1585-1597.
• Kaisho, T., and Wagner, H. (2008) Host-pathogen interactions, Curr Opin Immunol 20, 369-370.
• Rodriguez, N., Lang, R., Wantia, N., Cirl, C., Ertl, T., Durr, S., Wagner, H., and Miethke, T. (2008) Induction of iNOS by Chlamydophila pneumoniae requires MyD88-dependent activation of JNK, J Leukoc Biol 84, 1585-1593.
• Samuelsson, C., Hausmann, J., Lauterbach, H., Schmidt, M., Akira, S., Wagner, H., Chaplin, P., Suter, M., O’Keeffe, M., and Hochrein, H. (2008) Survival of lethal poxvirus infection in mice depends on TLR9, and therapeutic vaccination provides protection, J Clin Invest 118, 1776-1784.
• Schmitz, F., Heit, A., Dreher, S., Eisenacher, K., Mages, J., Haas, T., Krug, A., Janssen, K. P., Kirschning, C. J., and Wagner, H. (2008) Mammalian target of rapamycin (mTOR) orchestrates the defense program of innate immune cells, Eur J Immunol 38, 2981-2992.
• Spiller, S., Elson, G., Ferstl, R., Dreher, S., Mueller, T., Freudenberg, M., Daubeuf, B., Wagner, H., and Kirschning, C. J. (2008) TLR4-induced IFN-gamma production increases TLR2 sensitivity and drives Gram-negative sepsis in mice, J Exp Med 205, 1747-1754.
• Urbonaviciute, V., Furnrohr, B. G., Meister, S., Munoz, L., Heyder, P., De Marchis, F., Bianchi, M. E., Kirschning, C., Wagner, H., Manfredi, A. A., Kalden, J. R., Schett, G., Rovere-Querini, P., Herrmann, M., and Voll, R. E. (2008) Induction of inflammatory and immune responses by HMGB1-nucleosome complexes: implications for the pathogenesis of SLE, J Exp Med 205, 3007-3018.
• Wagner, H. (2008) The sweetness of the DNA backbone drives Toll-like receptor 9, Curr Opin Immunol 20, 396-400.
• Conrad, M. L., Ferstl, R., Teich, R., Brand, S., Blumer, N., Yildirim, A. O., Patrascan, C. C., Hanuszkiewicz, A., Akira, S., Wagner, H., Holst, O., von Mutius, E., Pfefferle, P. I., Kirschning, C. J., Garn, H., and Renz, H. (2009) Maternal TLR signaling is required for prenatal asthma protection by the nonpathogenic microbe Acinetobacter lwoffii F78, J Exp Med 206, 2869-2877.
• Haas, T., Schmitz, F., Heit, A., and Wagner, H. (2009) Sequence independent interferon-alpha induction by multimerized phosphodiester DNA depends on spatial regulation of Toll-like receptor-9 activation in plasmacytoid dendritic cells, Immunology 126, 290-298.
• Rad, R., Ballhorn, W., Voland, P., Eisenacher, K., Mages, J., Rad, L., Ferstl, R., Lang, R., Wagner, H., Schmid, R. M., Bauer, S., Prinz, C., Kirschning, C. J., and Krug, A. (2009) Extracellular and intracellular pattern recognition receptors cooperate in the recognition of Helicobacter pylori, Gastroenterology 136, 2247-2257.
• Wagner, H. (2009) The immunogenicity of CpG-antigen conjugates, Adv Drug Deliv Rev 61, 243-247.
• Werninghaus, K., Babiak, A., Gross, O., Holscher, C., Dietrich, H., Agger, E. M., Mages, J., Mocsai, A., Schoenen, H., Finger, K., Nimmerjahn, F., Brown, G. D., Kirschning, C., Heit, A., Andersen, P., Wagner, H., Ruland, J., and Lang, R. (2009) Adjuvanticity of a synthetic cord factor analogue for subunit Mycobacterium tuberculosis vaccination requires FcRgamma-Syk-Card9-dependent innate immune activation, J Exp Med 206, 89-97.
• Layland, L. E., Mages, J., Loddenkemper, C., Hoerauf, A., Wagner, H., Lang, R., and da Costa, C. U. (2010) Pronounced phenotype in activated regulatory T cells during a chronic helminth infection, J Immunol 184, 713-724.
• Wagner, H. (2010) Hemozoin: malaria’s “built-in” adjuvant and TLR9 agonist, Cell Host Microbe 7, 5-6.
• Wagner, H. (2010) Bone diseases: Interferon regulatory factor-8 suppresses osteoclastogenesis, Nat Rev Rheumatol 6, 73-74.
• Wagner, H. (2011) New vistas on TLR9 activation, Eur J Immunol 41, 2814-2816
• Wantia, N., Rodriguez, N., Cirl, C., Ertl, T., Durr, S., Layland, L. E., Wagner, H., and Miethke, T. (2011) Toll-like receptors 2 and 4 regulate the frequency of IFNgamma-producing CD4+ T-cells during pulmonary infection with Chlamydia pneumoniae, PLoS ONE 6, e26101.
• Wagner, H. (2012) Innate immunity’s path to the Nobel Prize 2011 and beyond, Eur J Immunol 42, 1089-1092.
• Oldenburg, M., Kruger, A., Ferstel, R., Kaufman, A., Sigmund, A., Bathke, B., Lautenbach, H., Suter, M., Dreher, S., Koedel, U., Akira, S., Kawai, T., Buer, J., Wanger, H., Bauer, S., Hochrein, H. and Kirschning, C. (2012) TLR13 recognises bacterias 23S rRNA devoid ot erythromycin resistance-forming modification. Science 337, 1111-1115