Advertisement

Cellular and Molecular Neurobiology

, Volume 27, Issue 3, pp 271–284 | Cite as

Antibodies to a Nonconjugated Prion Protein Peptide 95-123 Interfere with PrP Sc Propagation in Prion-Infected Cells

  • Maria B. Oboznaya
  • Sabine Gilch
  • Maia A. Titova
  • Dmitry O. Koroev
  • Tatyana D. Volkova
  • Olga M. Volpina
  • Hermann M. Schätzl
Article

Summary

1. Vaccination-induced anti-prion protein antibodies are presently regarded as a promising approach toward treatment of prion diseases. Here, we investigated the ability of five peptides corresponding to three different regions of the bovine prion protein (PrP) to elicit antibodies interfering with PrPSc propagation in prion-infected cells.

2. Rabbits were immunized with free nonconjugated peptides. Obtained immune sera were tested in enzyme-linked immunosorbent assay (ELISA) and immunoblot for their binding to recombinant PrP and cell-derived pathogenic isoform (PrPSc) and normal prion protein (PrPc), respectively. Sera positive in all tests were chosen for PrPSc inhibition studies in cell culture.

3. All peptides induced anti-peptide antibodies, most of them reacting with recombinant PrP. Moreover, addition of the serum specific to peptide 95–123 led to a transient reduction of PrPSc levels in persistently prion-infected cells.

4. Thus, anti-PrP antibodies interfering with PrPSc propagation were induced with a prion protein peptide nonconjugated to a protein carrier. These results point to the potential application of the nonconjugated peptide 95–123 for the treatment of prion diseases.

KEY WORDS

prion protein prion peptide nonconjugated peptide anti-peptide antibodies 

Notes

ACKNOWLEDGMENTS

We thank Elke Maas and Dr. Alexa Ertmer for their help and advice, Dr. Max Nunziante, Gunnar Schulz and Dr. Ina Vorberg for helpful discussions and ideas. Maria Oboznaya was an exchange scientist sponsored by the DAAD. This work was supported in part by the European Union (NoE NeuroPrion), the Bavarian Government (LMU5*), and by the BMBF (01KO0108).

REFERENCES

  1. Aguzzi, A., and Polymenidou, M. (2004). Mammalian prion biology: One century of evolving concepts. Cell 116:1–15.CrossRefGoogle Scholar
  2. Brennan, F. R., and Dougan, G. (2005). Non-clinical safety evaluation of novel vaccines and adjuvants: new products, new strategies. Vaccine 23:3210–3222.PubMedCrossRefGoogle Scholar
  3. Enari, M., Flechsig, E., and Weissmann, C. (2001). Scrapie prion protein accumulation by scrapie infected neuroblastoma cells abrogated by exposure to a prion protein antibody. Proc. Natl. Acad. Sci. USA 98:9295–9299.PubMedCrossRefGoogle Scholar
  4. Ertmer, A., Gilch, S., Yun, S. W., Flechsig, E., Klebl, B., Stein-Gerlach, M., Klein, M. A., and Schatzl, H. M. (2004). The tyrosine kinase inhibitor STI571 induces cellular clearance of PrPSc in prion-infected cells. J. Biol. Chem. 279:41918–41927.PubMedCrossRefGoogle Scholar
  5. Feraudet, C., Morel, N., Simon, S., Volland, H., Frobert, Y., Creminon, C., Vilette, D., Lehmann, S., and Grassi, J. (2005). Screening of 145 anti-PrP monoclonal antibodies for their capacity to inhibit PrPSc replication in infected cells. J. Biol. Chem. 280:11247–11258.PubMedCrossRefGoogle Scholar
  6. Fernandez-Escamilla, A.-M., Rousseau, F., Schymkowitz, J., and Serrano, L. (2004). Prediction of sequence-dependent and mutational effects on the aggregation of peptides and proteins. Nat. Biotech. 22:1302–1306.CrossRefGoogle Scholar
  7. Forloni, G., Angeretti, N., Chiesa, R., Monzani, E., Salmona, M., Bugiani, O., and Tagliavini, F. (1993). Neurotoxicity of a prion protein fragment. Nature 362(6420):543–546.PubMedCrossRefGoogle Scholar
  8. Gilch, S., Winklhofer, K. F., Groschup, M. H., Nunziante, M., Lucassen, R., Spielhaupter, C., Muranyi, W., Riesner, D., Tatzelt, J., and Schatzl, H. M. (2001). Intracellular re-routing of prion protein prevents propagation of PrPSc and delays onset of prion disease. EMBO J. 20:3957–3966.PubMedCrossRefGoogle Scholar
  9. Gilch, S., Wopfner, F., Renner-Muller, I., Kremmer, E., Bauer, C., Wolf, E., Brem, G., Groschup, M. H., and Schatzl, H. M. (2003). Polyclonal anti-PrP auto-antibodies induced with dimeric PrP interfere efficiently with PrPSc propagation in prion infected cells. J. Biol. Chem. 278:18524–18531.PubMedCrossRefGoogle Scholar
  10. Kretzschmar, H. A., Prusiner, S. B., Stowring, L. E., and DeArmond, S. J. (1986). Scrapie prion proteins are synthesized in neurons. Am. J. Pathol. 122:1–5.PubMedGoogle Scholar
  11. Long, J. R., Tang, L. M., Qiu, X. P., Zeng, F. T., Xie, X. H., and Zhang, Y. P. (2004). Sequence variation in the rabbit major histocompatibility complex DQA gene. Proceedings of the 8th World Rabbit Congress, Puebla, Mexico: 96–101.Google Scholar
  12. Magri, G., Clerici, M., Dall’Ara, P., Biasin, M., Caramelli, M., Casalone, C., Giannino, M. L., Longhi, R., Piacentini, L., Della Bella, S., Gazzuola, P., Martino, P. A., Della Bella, S., Pollera, C., Puricelli, M., Servida, F., Crescio, I., Boasso, A., Ponti, W., and Poli, G. (2005). Decrease in pathology and progression of scrapie after immunization with synthetic prion protein peptides in hamsters. Vaccine 23:2862–2868.PubMedCrossRefGoogle Scholar
  13. Nunziante, M., Gilch, S., and Schatzl, H. M. (2003). Prion diseases: From molecular biology to intervention strategies. Chem. Bio. Chem. 4:1268–1284.PubMedGoogle Scholar
  14. Peretz, D., Williamson, R. A., Kaneko, K., Vergara, J., Leclerc, E., Schmitt-Ulms, G., Mehlhorn, I. R., Legname, G., Wormald, M. R., Rudd, P. M., Dwek, R. A., Burton, D. R., and Prusiner, S. B. (2001). Antibodies inhibit prion propagation and clear cell cultures of prion infectivity. Nature 412:739–743.PubMedCrossRefGoogle Scholar
  15. Perrier, V., Solassol, J., Crozet, C., Frobert, Y., Mourton-Gilles, C., Grassi, J., and Lehmann, S. (2004). Anti-PrP antibodies block PrPSc replication in prion-infected cell cultures by accelerating PrPc degradation. J. Neurochem. 89:454–463.PubMedCrossRefGoogle Scholar
  16. Polymenidou, M., Heppner, F. L., Pellicioli, E. C., Urich, E., Miele, G., Braun, N., Wopfner, F., Schatzl, H. M., Becher, B., and Aguzzi, A. (2004). Humoral immune response to native eukaryotic prion protein correlates with anti-prion protection. Proc. Natl. Acad. Sci. USA 101:14670–14676.PubMedCrossRefGoogle Scholar
  17. Proske, D., Gilch, S., Wopfner, F., Schatzl, H. M., Winnacker, E. L., and Famulok, M. (2002). Prion-Protein-Specific Aptamer Reduces PrPSc Formation. Chem. Bio. Chem. 3:717–725.PubMedGoogle Scholar
  18. Prusiner, S. B. (1997). Prion diseases and the BSE crisis. Science 278:245–251.PubMedCrossRefGoogle Scholar
  19. Rymer, D. L., Good, T. A. (2000). The role of prion peptide structure and aggregation in toxicity and membrane binding. J. Neurochem. 75(6):2536–2545.PubMedCrossRefGoogle Scholar
  20. Schwarz, A., Kratke, O., Burwinkel, M., Riemer, C., Schultz, J., Henkleinb, P., Bammea, T., and Baier, M. (2003). Immunization with a synthetic prion protein-derived peptide prolongs survival times of mice orally exposed to the scrapie agent. Neurosci. Lett. 350:187–189.PubMedCrossRefGoogle Scholar
  21. Sette, A., and Fikes, J. (2003). Epitope-based vaccines: an update on epitope identification, vaccine design and delivery. Curr. Opin. Immunol. 15:461–470.PubMedCrossRefGoogle Scholar
  22. Sigurdsson, E. M., Brown, D. R., Daniels, M., Kascsak, R. J., Kascsak, R., Carp, R., Meeker, H. C., Frangione, B., and Wisniewski, T. (2002). Immunization delays the onset of prion disease in mice. Am. J. Pathol. 161:13–17.PubMedGoogle Scholar
  23. Souan, L., Tal, Y., Felling, Y., Cohen, I. R., Taraboulos, A., and Mor, F. (2001). Modulation of proteinase-K resistant prion protein by prion peptide immunization. Eur. J. Immunol. 31:2338–2346.PubMedCrossRefGoogle Scholar
  24. Tagliavini, F., Prelli, F., Verga, L., Giaccone, G., Sarma, R., Gorevic, P., Ghetti, B., Passerini, F., Ghibaudi, E., and Forloni, G. (1993). Synthetic peptides homologous to prion protein residues 106-147 form amyloid-like fibrils in vitro. Proc. Natl. Acad. Sci. 90(20):9678–9682.PubMedCrossRefGoogle Scholar
  25. Tal, Y., Souan, L., Cohen, I. R., Meiner, Z., Taraboulos, A., and Mor, F. (2003). Complete Freund's adjuvant immunization prolongs survival in experimental prion disease in mice. J. Neurosci. Res. 71:286–290.PubMedCrossRefGoogle Scholar
  26. Udenfriend, S., and Meienhoferm, J. (1987). The peptides: Analysis, synthesis and biology, vol. 9, Academic Press, London.Google Scholar
  27. White, A. R., Enever, P., Tayebi, M., Mushens, R., Linehan, J., Brandner, S., Anstee, D., Collinge, J., and Hawke, S. (2003). Monoclonal antibodies inhibit prion replication and delay the development of prion disease. Nature 422:80–83.PubMedCrossRefGoogle Scholar
  28. Ziegler, J., Viehrig, C., Geimer, S., Rosch, P., and Schwarzinger, S. (2006). Putative aggregation initiation sites in prion protein. FEBS Lett. 580(8):2033–2040.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Maria B. Oboznaya
    • 1
  • Sabine Gilch
    • 2
  • Maia A. Titova
    • 1
  • Dmitry O. Koroev
    • 1
  • Tatyana D. Volkova
    • 1
  • Olga M. Volpina
    • 1
  • Hermann M. Schätzl
    • 2
  1. 1.Shemyakin-Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesGSP MoscowRussia
  2. 2.Technical University of MunichInstitute of Virology/Prion Research GroupMunichGermany

Personalised recommendations