Kliniken & Institute … Institute Zentrum für… Medizinische… Forschung Research

CpG-DNA and Toll-like Receptors

We have studied Toll-like receptor 9 activation through synthetic CpG-Oligonucleotides (CpG-ODN). This substance class has been proven to be a powerful immunostimulatory agent. CpG-ODNs are now under investigation for treatment of Th2 mediated diseases like allergy and asthma, for therapeutical usage in cancer models and most important as adjuvants mediating cellular immunity in vaccination trials.

We have investigated the structure-activity relationships as well as general immunopharmacology of CpG-ODNs. In this respect we have defined the importance of poly-guanosine motifs for the cellular uptake of unmodified phosphodiester CpG-ODN. Furthermore we were able to show that CpG-DNA needs to have a free 5' end in order to being recognized and this is probably mediated in the double-stranded but not the single-stranded form as formerly suggested. This was a project in close collaboration with the group of Prof. C. Richert, University Stuttgart.

We also found that the stimulatory potential of bacterial DNA correlates with its CG-content. Moreover we defined a new class of oligonucleotides which are able to act as inhibitor of CpG/TLR9 signaling.

The research in CpG-DNA has given us insights in recognition, signal transduction and effector cascades induced by TLR-ligands. The results are fundamental for the studies we now perform in the biological consequences of TLR-induced signals (see: Differentiation of dendritic cells and B cells).

Selected publications

  1. Hildebrand, D., A. Sähr, S. J. Wölfle, K. Heeg, and K. F. Kubatzky. 2012. Regulation of Toll-like receptor 4-mediated immune responses through Pasteurella multocida toxin-induced G protein signalling. Cell Communication and Signaling 10: 22.
  2. Bekeredjian-Ding, I., S. Foermer, C. J. Kirschning, M. Parcina, and K. Heeg. 2012. Poke Weed Mitogen Requires Toll-Like Receptor Ligands for Proliferative Activity in Human and Murine B Lymphocytes. PLoS ONE 7: e29806.
  3. Ulrich, K., S. Wölfle, A. Mayer, K. Heeg, T. Braunbeck, L. Erdinger, and H. Bartz. 2010. Extractable organic matter of standard reference material 1649a influences immunological response induced by pathogen-associated molecular patterns. Environ Sci Pollut Res Int 17: 1257–1267.
  4. Bode, K. A., F. Schmitz, L. Vargas, K. Heeg, and A. H. Dalpke. 2009. Kinetic of RelA Activation Controls Magnitude of TLR-Mediated IL-12p40 Induction. J.Immunol. 182: 2176–2184.
  5. Zimmermann, S., A. Dalpke, and K. Heeg. 2008. CpG oligonucleotides as adjuvant in therapeutic vaccines against parasitic infections. Int. J. Med. Microbiol. 298: 39–44.
  6. Peter, M., K. Bode, G. B. Lipford, F. Eberle, K. Heeg, and A. H. Dalpke. 2008. Characterization of suppressive oligodeoxynucleotides that inhibit Toll-like receptor-9-mediated activation of innate immunity. Immunology 123: 118–128.
  7. Mrabet-Dahbi, S., A. H. Dalpke, M. Niebuhr, M. Frey, C. Draing, S. Brand, K. Heeg, T. Werfel, and H. Renz. 2008. The Toll-like receptor 2 R753Q mutation modifies cytokine production and Toll-like receptor expression in atopic dermatitis. J. Allergy Clin. Immunol. 121: 1013–1019.
  8. Heeg, K., A. Dalpke, M. Peter, and S. Zimmermann. 2008. Structural requirements for uptake and recognition of CpG oligonucleotides. Int. J. Med. Microbiol. 298: 33–38.
  9. Dalpke, A., K. Heeg, H. Bartz, and A. Baetz. 2008. Regulation of innate immunity by suppressor of cytokine signaling (SOCS) proteins. Immunobiology 213: 225–235.
  10. Bekeredjian-Ding, I., A. Doster, M. Schiller, P. Heyder, H. M. Lorenz, B. Schraven, U. Bommhardt, and K. Heeg. 2008. TLR9-activating DNA up-regulates ZAP70 via sustained PKB induction in IgM+ B cells. J.Immunol. 181: 8267.
  11. Mayer, A. K., M. Muehmer, J. Mages, K. Gueinzius, C. Hess, K. Heeg, R. Bals, R. Lang, and A. H. Dalpke. 2007. Differential recognition of TLR-dependent microbial ligands in human bronchial epithelial cells. J. Immunol. 178: 3134–3142.
  12. Heyder, P., I. Bekeredjian-Ding, M. Parcina, N. Blank, A. D. Ho, M. Herrmann, H.-M. Lorenz, K. Heeg, and M. Schiller. 2007. Purified apoptotic bodies stimulate plasmacytoid dendritic cells to produce IFN-alpha. Autoimmunity 40: 331–332.
  13. He, G., A. Patra, K. Siegmund, M. Peter, K. Heeg, A. Dalpke, and C. Richert. 2007. Immunostimulatory CpG oligonucleotides form defined three-dimensional structures: results from an NMR study. ChemMedChem 2: 549–560.
  14. Bode, K. A., K. Schroder, D. A. Hume, T. Ravasi, K. Heeg, M. J. Sweet, and A. H. Dalpke. 2007. Histone deacetylase1 inhibitors decrease Toll-like receptor-mediated activation of proinflammatory gene expression by impairing transcription factor recruitment. Immunology 122: 596–606.
  15. Tietze, K., A. Dalpke, S. Morath, R. Mutters, K. Heeg, and C. Nonnenmacher. 2006. Differences in innate immune responses upon stimulation with gram-positive and gram-negative bacteria. J. Periodont. Res. 41: 447–454.
  16. Dalpke, A., J. Frank, M. Peter, and K. Heeg. 2006. Activation of toll-like receptor 9 by DNA from different bacterial species. Infect. Immun. 74: 940–946.
  17. Hegele, A., A. Dalpke, K. Heeg, P. Barth, Z. Varga, R. Hofmann, and P. Olbert. 2005. Immunostimulatory CpG oligonucleotides reduce tumor burden after intravesical administration in an orthotopic murine bladder cancer model. Tumour Biol. 26: 274–280.
  18. 1Dalpke, A. H., M. D. Lehner, T. Hartung, and K. Heeg. 2005. Differential effects of CpG-DNA in Toll-like receptor-2/-4/-9 tolerance and cross-tolerance. Immunology 116: 203–212.
  19. Dalpke, A., and K. Heeg. 2004. CpG DNA as immune response modifier. Int.J.Med.Microbiol. 294: 345–354.
  20. Bartz, H., Y. Mendoza, M. Gebker, T. Fischborn, K. Heeg, and A. Dalpke. 2004. Poly-guanosine strings improve cellular uptake and stimulatory activity of phosphodiester CpG oligonucleotides in human leukocytes. Vaccine 23: 148–155.
  21. Albrecht, I., T. Tapmeier, S. Zimmermann, M. Frey, K. Heeg, and A. Dalpke. 2004. Toll-like receptor (TLR) selective nucleosomal remodeling at the IL-12p40 promoter. EMBO Rep. 5: 172–177.
  22. Zimmermann, S., A. Dalpke, and K. Heeg. 2003. Immunostimulatory DNA as adjuvants: Efficacy of phosphodiester CpG oligonucleotides is enhanced by 5’ sequence modifications. Vaccine 21: 990–995.
  23. Narayanan, S., A. H. Dalpke, K. Siegmund, K. Heeg, and C. Richert. 2003. CpG oligonucleotides with modified termini and nicked dumbbell structure show enhanced immunostimulaty activity. J.Med.Chem. 46: 5031–5044.
  24. Dalpke, A., and K. Heeg. 2003. Synergistic and antagonistic interactions between LPS and superantigens. J.Endotox.Res. 9: 51–54.
  25. Dalpke, A. H., M. Frey, S. Morath, T. Hartung, and K. Heeg. 2002. Interaction of lipoteichoic acid and CpG-DNA during activation of innate immune cells. Immunobiology 206: 392–407.
  26. Dalpke, A. H., K. H. Sch�fer, M. Frey, S. Zimmermann, J. Tebbe, E. Weihe, and K. Heeg. 2002. Immunostimulatory CpG-DNA activates murine microglia. J.Immunol. 168# #: 4854–4863.
  27. Dalpke, A., S. Zimmermann, and K. Heeg. 2002. Immunopharmacology of CpG DNA. Biol.Chem. 383: 1491–1500.
  28. Dalpke, A., S. Zimmermann, and K. Heeg. 2002. CpG-DNA in the prevention and treatment of infections. BioDrugs 16: 419–431.
  29. Dalpke, A., S. Zimmermann, T. Hartung, and K. Heeg. 2002. Collaboration of lipoteichoic acid and CpG-DNA during activation of innate immune cells. Immunobiology 206: 392–407.
  30. Dalpke, A., S. Zimmermann, I. Albrecht, and K. Heeg. 2002. Phosphodiester CpG oligonucleotides as adjuvants: Poly-guanosine runs enhance cellular uptake and improve immunostimulative activity of phosphodiester CpG# #oligonucleotides in vitro and in vivo# #. Immunology 106: 102–112.
  31. Dalpke, A., S. Zimmermann, and K. Heeg. 2001. CpG oligonucleotides in vaccination: Signaling and mechanisms of action. Immunobiology 204: 667–676.
  32. Lipford, G. B., T. Sparwasser, S. Zimmermann, K. Heeg, and H. Wagner. 2000. CpG-DNA mediated transient lymphoadenopathy is associated with a state of Th1 predisposition to antige driven responses. J.Immunol. 165: 1228–1235.
  33. Lipford, G. B., S. Bendigs, K. Heeg, and H. Wagner. 2000. Poly-guanosine motifs costimulate antigen reactive CD8 T cells while bacterial CpG-DNA affect T-cell activation via antigen-presenting cell-derived cytokines. Immunology 101: 46–52.
  34. Kranzer, K., M. Bauer, G. B. Lipford, K. Heeg, H. Wagner, and R. Lang. 2000. CpG oligodeoxynucleotides enhance T cell receptor-triggered IFN-gamma production and up-regulation of CD69 via induction of antigen-presenting cell-derived interferon type I and IL-12. Immunology 99: 170–178.
  35. Heeg, K., and S. Zimmermann. 2000. CpG DNA as a Th1 trigger. Int.Arch.Allergy Immunol. 121: 87–97.
  36. Heeg, K. 2000. CpG-DNA costimulates antigen reactive T cells. Curr.Top.Microbiol.Immunol. 247: 93–105.
  37. Bendigs, S., U. Salzer, G. B. Lipford, H. Wagner, and K. Heeg. 1999. CpG-oligodeoxynucleotides costimulate primary T cells in the absence of APC. Eur.J.Immunol. 29: 1209–1218.
  38. Bauer, M., K. Heeg, H. Wagner, and G. B. Lipford. 1999. DNA activates human immune cells through a CpG sequence dependent manner. Immunology 97: 699–705.
  39. Zimmermann, S., O. Egeter, S. Hausmann, G. B. Lipford, M. Röcken, H. Wagner, and K. Heeg. 1998. CpG oligodeoxynucleotides trigger protective and curative Th1 responses in lethal murine leishmaniasis. J.Immunol. 160: 3627–3630.
  40. Lipford, G. B., K. Heeg, and H. Wagner. 1998. Bacterial DNA as immune cell activator. Trends.Microbiol. 6: 496–500.
  41. Heeg, K., T. Sparwasser, G. B. Lipford, H. Häcker, S. Zimmermann, and H. Wagner. 1998. Bacterial DNA as an evolutinary conserved ligand signalling danger of infection to immune cells. Eur.J.Clin.Microbiol.Infect.Dis. 17: 464–469.
  42. Sparwasser, T., T. Miethke, G. B. Lipford, K. Borschert, H. Häcker, K. Heeg, and H. Wagner. 1997. Bacterial DNA cause septic shock. Nature. 386: 336–337.
  43. Lipford, G. B., T. Sparwasser, M. Bauer, S. Zimmermann, E. S. Koch, K. Heeg, and H. Wagner. 1997. Immunostimulatory DNA: sequence dependent production of potentially harmful or useful cytokines. Eur.J.Immunol. 27: 3420–3426.
  44. Lipford, G. B., M. Bauer, C. Blank, R. Reiter, H. Wagner, and K. Heeg. 1997. CpG-containing synthetic oligonucleotides promote B and cytotoxic T cell responses to protein antigen: a new class of vaccine adjuvants. Eur.J.Immunol. 27: 2340–250344.