The mechanisms of neuronal degeneration induced by the transformation of normal cellular prion protein (PrP) into disease-associated PrP are not fully understood. Previous reports have demonstrated that cross-linking cellular prion protein by anti-PrP antibodies can promote neuronal apoptosis. In this report, we now show that treatment of neuronal cells with anti-prion antibodies leads to sequestration of free cholesterol in cell membranes, significant overexpression of apolipoprotein E, and to cytoplasmic phospholipase A2 activation as well as to production of prostaglandin. These results confirm the toxic effects and indicate that anti-prion antibody treatment of neurons lead to deleterious effects. Finally, great caution should be exerted when adopting antibody-based therapy for prion diseases.


Article metrics loading...

Loading full text...

Full text loading...



  1. Akiba, S., Yoneda, Y., Ohno, S., Nemoto, M. & Sato, T.(2003). Oxidized LDL activates phospholipase A2 to supply fatty acids required for cholesterol esterification. J Lipid Res 44, 1676–1685.[CrossRef] [Google Scholar]
  2. Allen, J. A., Halverson-Tamboli, R. A. & Rasenick, M. M.(2007). Lipid raft microdomains and neurotransmitter signalling. Nat Rev Neurosci 8, 128–140.[CrossRef] [Google Scholar]
  3. Anstee, D. J., Gardner, B., Spring, F. A., Holmes, C. H., Simpson, K. L., Parsons, S. F., Mallinson, G., Yousaf, S. M. & Judson, P. A.(1991). New monoclonal antibodies in CD44 and CD58: their use to quantify CD44 and CD58 on normal human erythrocytes and to compare the distribution of CD44 and CD58 in human tissues. Immunology 74, 197–205. [Google Scholar]
  4. Bate, C., Marshall, V., Colombo, L., Diomede, L., Salmona, M. & Williams, A.(2008a). Docosahexaenoic and eicosapentaenoic acids increase neuronal death in response to HuPrP82–146 and Aβ1–42. Neuropharmacology 54, 934–943.[CrossRef] [Google Scholar]
  5. Bate, C., Tayebi, M., Diomede, L., Salmona, M. & Williams, A.(2008b). Docosahexaenoic and eicosapentaenoic acids increase prion formation in neuronal cells. BMC Biol 6, 39.[CrossRef] [Google Scholar]
  6. Bate, C., Tayebi, M. & Williams, A.(2008c). Sequestration of free cholesterol in cell membranes by prions correlates with cytoplasmic phospholipase A2 activation. BMC Biol 6, 8.[CrossRef] [Google Scholar]
  7. Baumann, M. H., Kallijarvi, J., Lankinen, H., Soto, C. & Haltia, M.(2000). Apolipoprotein E includes a binding site which is recognized by several amyloidogenic polypeptides. Biochem J 349, 77–84.[CrossRef] [Google Scholar]
  8. Beringue, V., Mallinson, G., Kaisar, M., Tayebi, M., Sattar, Z., Jackson, G., Anstee, D., Collinge, J. & Hawke, S.(2003). Regional heterogeneity of cellular prion protein isoforms in the mouse brain. Brain 126, 2065–2073.[CrossRef] [Google Scholar]
  9. Beringue, V., Vilette, D., Mallinson, G., Archer, F., Kaisar, M., Tayebi, M., Jackson, G. S., Clarke, A. R., Laude, H. & other authors(2004). PrPSc binding antibodies are potent inhibitors of prion replication in cell lines. J Biol Chem 279, 39671–39676.[CrossRef] [Google Scholar]
  10. Brandner, S., Isenmann, S., Raeber, A., Fischer, M., Sailer, A., Kobayashi, Y., Marino, S., Weissmann, C. & Aguzzi, A.(1996). Normal host prion protein necessary for scrapie-induced neurotoxicity. Nature 379, 339–343.[CrossRef] [Google Scholar]
  11. Castaño, E. M. & Frangione, B.(1995). Non-Alzheimer's disease amyloidoses of the nervous system. Curr Opin Neurol 8, 279–285.[CrossRef] [Google Scholar]
  12. Chang, T. Y., Chang, C. C. Y. & Cheng, D.(1997). Acyl-coenzyme A:cholesterol acyltransferase. Annu Rev Biochem 66, 613–638.[CrossRef] [Google Scholar]
  13. Chiesa, R. & Harris, D. A.(2001). Prion diseases: what is the neurotoxic molecule? Neurobiol Dis 8, 743–763.[CrossRef] [Google Scholar]
  14. Cooper, D. M. F. & Crossthwaite, A. J.(2006). Higher-order organization and regulation of adenylyl cyclases. Trends Pharmacol Sci 27, 426–431.[CrossRef] [Google Scholar]
  15. Delton-Vandenbroucke, I., Bouvier, J., Makino, A., Besson, N., Pageaux, J. F., Lagarde, M. & Kobayashi, T.(2007). Anti-bis(monoacylglycero)phosphate antibody accumulates acetylated LDL-derived cholesterol in cultured macrophages. J Lipid Res 48, 543–552. [Google Scholar]
  16. Diedrich, J. F., Minnigan, H., Carp, R. I., Whitaker, J. N., Race, R., Frey, W., II & Haase, A. T.(1991). Neuropathological changes in scrapie and Alzheimer's disease are associated with increased expression of apolipoprotein E and cathepsin D in astrocytes. J Virol 65, 4759–4768. [Google Scholar]
  17. Enari, M., Flechsig, E. & Weissmann, C.(2001). Scrapie prion protein accumulation by scrapie-infected neuroblastoma cells abrogated by exposure to a prion protein antibody. Proc Natl Acad Sci U S A 98, 9295–9299.[CrossRef] [Google Scholar]
  18. Farooqui, A. A. & Horrocks, L. A.(2006). Phospholipase A2-generated lipid mediators in the brain: the good, the bad, and the ugly. Neuroscientist 12, 245–260.[CrossRef] [Google Scholar]
  19. Farooqui, A. A., Ong, W.-Y. & Horrocks, L. A.(2006). Inhibitors of brain phospholipase A2 activity: their neuropharmacological effects and therapeutic importance for the treatment of neurologic disorders. Pharmacol Rev 58, 591–620.[CrossRef] [Google Scholar]
  20. Gao, C., Lei, Y. J., Han, J., Shi, Q., Chen, L., Guo, Y., Gao, Y. J., Chen, J. M., Jiang, H. Y. & other authors(2006). Recombinant neural protein PrP can bind with both recombinant and native apolipoprotein E in vitro. Acta Biochim Biophys Sin (Shanghai) 38, 593–601.[CrossRef] [Google Scholar]
  21. Goritz, C., Mauch, D. H. & Pfrieger, F. W.(2005). Multiple mechanisms mediate cholesterol-induced synaptogenesis in a CNS neuron. Mol Cell Neurosci 29, 190–201.[CrossRef] [Google Scholar]
  22. Graziani, A., Bricko, V., Carmignani, M., Graier, W. F. & Groschner, K.(2004). Cholesterol- and caveolin-rich membrane domains are essential for phospholipase A(2)-dependent EDHF formation. Cardiovasc Res 64, 234–242.[CrossRef] [Google Scholar]
  23. Heeren, J., Grewal, T., Laatsch, A., Rottke, D., Rinninger, F., Enrich, C. & Beisiegel, U.(2003). Recycling of apoprotein E is associated with cholesterol efflux and high density lipoprotein internalization. J Biol Chem 278, 14370–14378.[CrossRef] [Google Scholar]
  24. Heeren, J., Grewal, T., Laatsch, A., Becker, N., Rinninger, F., Rye, K. A. & Beisiegel, U.(2004). Impaired recycling of apolipoprotein E4 is associated with intracellular cholesterol accumulation. J Biol Chem 279, 55483–55492.[CrossRef] [Google Scholar]
  25. Horsburgh, K. & Nicoll, J. A.(1996). Selective alterations in the cellular distribution of apolipoprotein E immunoreactivity following transient cerebral ischaemia in the rat. Neuropathol Appl Neurobiol 22, 342–349.[CrossRef] [Google Scholar]
  26. Horsburgh, K., Fitzpatrick, M., Nilsen, M. & Nicoll, J. A.(1997). Marked alterations in the cellular localisation and levels of apolipoprotein E following acute subdural haematoma in rat. Brain Res 763, 103–110.[CrossRef] [Google Scholar]
  27. Horsburgh, K., Graham, D. I., Stewart, J. & Nicoll, J. A.(1999). Influence of apolipoprotein E genotype on neuronal damage and apoE immunoreactivity in human hippocampus following global ischemia. J Neuropathol Exp Neurol 58, 227–234.[CrossRef] [Google Scholar]
  28. Horsburgh, K., McCulloch, J., Nilsen, M., McCracken, E., Large, C., Roses, A. D. & Nicoll, J. A.(2000). Intraventricular infusion of apolipoprotein E ameliorates acute neuronal damage after global cerebral ischemia in mice. J Cereb Blood Flow Metab 20, 458–462. [Google Scholar]
  29. Jones, D. R., Taylor, W. A., Bate, C., David, M. & Tayebi, M.(2010). A camelid anti-PrP antibody abrogates PrP replication in prion-permissive neuroblastoma cell lines. PLoS ONE 5, e9804.[CrossRef] [Google Scholar]
  30. Kempster, S., Bate, C. & Williams, A.(2007). Simvastatin treatment prolongs the survival of scrapie-infected mice. Neuroreport 18, 479–482.[CrossRef] [Google Scholar]
  31. Kesäniemi, Y. A., Ehnholm, C. & Miettinen, T. A.(1987). Intestinal cholesterol absorption efficiency in man is related to apoprotein E phenotype. J Clin Invest 80, 578–581.[CrossRef] [Google Scholar]
  32. Kim, B. H., Lee, H. G., Choi, J. K., Kim, J. I., Choi, E. K., Carp, R. I. & Kim, Y. S.(2004). The cellular prion protein (PrPC) prevents apoptotic neuronal cell death and mitochondrial dysfunction induced by serum deprivation. Brain Res Mol Brain Res 124, 40–50.[CrossRef] [Google Scholar]
  33. Kitagawa, K., Matsumoto, M., Hori, M. & Yanagihara, T.(2002). Neuroprotective effect of apolipoprotein E against ischemia. Ann N Y Acad Sci 977, 468–475.[CrossRef] [Google Scholar]
  34. Kitatani, K., Nemoto, M., Akiba, S. & Sato, T.(2002). Stimulation by de novo-synthesized ceramide of phospholipase A2-dependent cholesterol esterification promoted by the uptake of oxidized low-density lipoprotein in macrophages. Cell Signal 14, 695–701.[CrossRef] [Google Scholar]
  35. Koh, C. H., Peng, Z. F., Ou, K., Melendez, A., Manikandan, J., Qi, R. Z. & Cheung, N. S.(2007). Neuronal apoptosis mediated by inhibition of intracellular cholesterol transport: microarray and proteomics analyses in cultured murine cortical neurons. J Cell Physiol 211, 63–87.[CrossRef] [Google Scholar]
  36. Korth, C., Streit, P. & Oesch, B.(1999). Monoclonal antibodies specific for the native, disease-associated isoform of the prion protein. Methods Enzymol 309, 106–122. [Google Scholar]
  37. Krugers, H. J., Mulder, M., Korf, J., Havekes, L., de Kloet, E. R. & Joels, M.(1997). Altered synaptic plasticity in hippocampal CA1 area of apolipoprotein E deficient mice. Neuroreport 8, 2505–2510.[CrossRef] [Google Scholar]
  38. Li, S., Couet, J. & Lisanti, M. P.(1996). Src tyrosine kinases, Gα subunits, and H-Ras share a common membrane-anchored scaffolding protein, caveolin. Caveolin binding negatively regulates the auto-activation of Src tyrosine kinases. J Biol Chem 271, 29182–29190.[CrossRef] [Google Scholar]
  39. Lucas, K. K., Svensson, C. I., Hua, X. Y., Yaksh, T. L. & Dennis, E. A.(2005). Spinal phospholipase A2 in inflammatory hyperalgesia: role of group IVA cPLA2. Br J Pharmacol 144, 940–952.[CrossRef] [Google Scholar]
  40. Lung, F. W., Kao, W. T., Shu, B. C., Yen, Y. C. & Tzeng, D. S.(2006). A module map showing interaction between apolipoprotein E and phospholipase A2 polymorphism in lipid profiles. Hum Hered 62, 135–144.[CrossRef] [Google Scholar]
  41. Ma, J., Wollmann, R. & Lindquist, S.(2002). Neurotoxicity and neurodegeneration when PrP accumulates in the cytosol. Science 298, 1781–1785.[CrossRef] [Google Scholar]
  42. Mabile, L., Lefebvre, C., Lavigne, J., Boulet, L., Davignon, J., Lussier-Cacan, S. & Bernier, L.(2003). Secreted apolipoprotein E reduces macrophage-mediated LDL oxidation in an isoform-dependent way. J Cell Biochem 90, 766–776.[CrossRef] [Google Scholar]
  43. Mahley, R. W. & Huang, Y.(1999). Apolipoprotein E: from atherosclerosis to Alzheimer's disease and beyond. Curr Opin Lipidol 10, 207–217.[CrossRef] [Google Scholar]
  44. Mallucci, G., Dickinson, A., Linehan, J., Klohn, P. C., Brandner, S. & Collinge, J.(2003). Depleting neuronal PrP in prion infection prevents disease and reverses spongiosis. Science 302, 871–874.[CrossRef] [Google Scholar]
  45. Maxfield, F. R. & Tabas, I.(2005). Role of cholesterol and lipid organization in disease. Nature 438, 612–621.[CrossRef] [Google Scholar]
  46. Mazzoni, I. E., Ledebur, H. C., Jr, Paramithiotis, E. & Cashman, N.(2005). Lymphoid signal transduction mechanisms linked to cellular prion protein. Biochem Cell Biol 83, 644–653.[CrossRef] [Google Scholar]
  47. Michel, V. & Bakovic, M.(2007). Lipid rafts in health and disease. Biol Cell 99, 129–140.[CrossRef] [Google Scholar]
  48. Mok, S. W. F., Thelen, K. M., Riemer, C., Bamme, T., Gultner, S., Lutjohann, D. & Baier, M.(2006). Simvastatin prolongs survival times in prion infections of the central nervous system. Biochem Biophys Res Commun 348, 697–702.[CrossRef] [Google Scholar]
  49. Mouillet-Richard, S., Ermonval, M., Chebassier, C., Laplanche, J. L., Lehmann, S., Launay, J. M. & Kellermann, O.(2000). Signal transduction through prion protein. Science 289, 1925–1928.[CrossRef] [Google Scholar]
  50. Oh, P. & Schnitzer, J. E.(2001). Segregation of heterotrimeric G proteins in cell surface microdomains. Gq binds caveolin to concentrate in caveolae, whereas Gi and Gs target lipid rafts by default. Mol Biol Cell 12, 685–698.[CrossRef] [Google Scholar]
  51. Pani, A., Norfo, C., Abete, C., Mulas, C., Putzolu, M., Laconi, S., Orru, C. D., Cannas, M. D., Vascellari, S. & other authors(2007). Antiprion activity of cholesterol esterification modulators: a comparative study using ex vivo sheep fibroblasts and lymphocytes and mouse neuroblastoma cell lines. Antimicrob Agents Chemother 51, 4141–4147.[CrossRef] [Google Scholar]
  52. Panini, S. R., Yang, L., Rusinol, A. E., Sinensky, M. S., Bonventre, J. V. & Leslie, C. C.(2001). Arachidonate metabolism and the signaling pathway of induction of apoptosis by oxidized LDL/oxysterol. J Lipid Res 42, 1678–1686. [Google Scholar]
  53. Paramithiotis, E., Pinard, M., Lawton, T., LaBoissiere, S., Leathers, V. L., Zou, W. Q., Estey, L. A., Lamontagne, J., Lehto, M. T. & other authors(2003). A prion protein epitope selective for the pathologically misfolded conformation. Nat Med 9, 893–899.[CrossRef] [Google Scholar]
  54. Peretz, D., Williamson, R. A., Kaneko, K., Vergara, J., Leclerc, E., Schmitt-Ulms, G., Mehlhorn, I. R., Legname, G., Wormald, M. R. & other authors(2001). Antibodies inhibit prion propagation and clear cell cultures of prion infectivity. Nature 412, 739–743.[CrossRef] [Google Scholar]
  55. Pfrieger, F. W.(2003). Role of cholesterol in synapse formation and function. Biochim Biophys Acta 1610, 271–280.[CrossRef] [Google Scholar]
  56. Pike, L. J.(2004). Lipid rafts: heterogeneity on the high seas. Biochem J 378, 281–292.[CrossRef] [Google Scholar]
  57. Pleasure, S. J., Page, C. & Lee, V. M.(1992). Pure, postmitotic, polarized human neurons derived from NTera 2 cells provide a system for expressing exogenous proteins in terminally differentiated neurons. J Neurosci 12, 1802–1815. [Google Scholar]
  58. Prusiner, S. B.(1994). Molecular biology and genetics of prion diseases. Philos Trans R Soc Lond B Biol Sci 343, 447–463.[CrossRef] [Google Scholar]
  59. Prusiner, S. B.(1998). The prion diseases. Brain Pathol 8, 499–513. [Google Scholar]
  60. Puglielli, L., Tanzi, R. E. & Kovacs, D. M.(2003). Alzheimer's disease: the cholesterol connection. Nat Neurosci 6, 345–351.[CrossRef] [Google Scholar]
  61. Riek, R., Hornemann, S., Wider, G., Billeter, M., Glockshuber, R. & Wuthrich, K.(1996). NMR structure of the mouse prion protein domain PrP(121–321). Nature 382, 180–182.[CrossRef] [Google Scholar]
  62. Roediger, B. & Armati, P. J.(2003). Oxidative stress induces axonal beading in cultured human brain tissue. Neurobiol Dis 13, 222–229.[CrossRef] [Google Scholar]
  63. Russelakis-Carneiro, M., Hetz, C., Maundrell, K. & Soto, C.(2004). Prion replication alters the distribution of synaptophysin and caveolin 1 in neuronal lipid rafts. Am J Pathol 165, 1839–1848.[CrossRef] [Google Scholar]
  64. Sathishkumar, K., Murthy, S. N. & Uppu, R. M.(2007). Cytotoxic effects of oxysterols produced during ozonolysis of cholesterol in murine GT1–7 hypothalamic neurons. Free Radic Res 41, 82–88.[CrossRef] [Google Scholar]
  65. Simons, K. & Ikonen, E.(2000). How cells handle cholesterol. Science 290, 1721–1726.[CrossRef] [Google Scholar]
  66. Simons, K. & Toomre, D.(2000). Lipid rafts and signal transduction. Nat Rev Mol Cell Biol 1, 31–39. [Google Scholar]
  67. Solforosi, L., Criado, J. R., McGavern, D. B., Wirz, S., Sanchez-Alavez, M., Sugama, S., DeGiorgio, L. A., Volpe, B. T., Wiseman, E. & other authors(2004). Cross-linking cellular prion protein triggers neuronal apoptosis in vivo. Science 303, 1514–1516.[CrossRef] [Google Scholar]
  68. Sun, G. Y., Xu, J., Jensen, M. D. & Simonyi, A.(2004). Phospholipase A2 in the central nervous system: implications for neurodegenerative diseases. J Lipid Res 45, 205–213. [Google Scholar]
  69. Tabas, I.(2002). Consequences of cellular cholesterol accumulation: basic concepts and physiological implications. J Clin Invest 110, 905–911.[CrossRef] [Google Scholar]
  70. Tayebi, M. & Hawke, S.(2006). Antibody-mediated neuronal apoptosis: therapeutic implications for prion diseases. Immunol Lett 105, 123–126.[CrossRef] [Google Scholar]
  71. Van Everbroeck, B., Croes, E. A., Pals, P., Dermaut, B., Jansen, G., van Duijn, C. M., Cruts, M., Van Broeckhoven, C., Martin, J. J. & other authors(2001). Influence of the prion protein and the apolipoprotein E genotype on the Creutzfeldt–Jakob disease phenotype. Neurosci Lett 313, 69–72.[CrossRef] [Google Scholar]
  72. White, F., Nicoll, J. A. & Horsburgh, K.(2001). Alterations in ApoE and ApoJ in relation to degeneration and regeneration in a mouse model of entorhinal cortex lesion. Exp Neurol 169, 307–318.[CrossRef] [Google Scholar]
  73. White, A. R., Enever, P., Tayebi, M., Mushens, R., Linehan, J., Brandner, S., Anstee, D., Collinge, J. & Hawke, S.(2003). Monoclonal antibodies inhibit prion replication and delay the development of prion disease. Nature 422, 80–83.[CrossRef] [Google Scholar]
  74. Williams, A. E., Van Dam, A. M., Man, A. H. W., Berkenbosch, F., Eikelenboom, P. & Fraser, H.(1994). Cytokines, prostaglandins and lipocortin-1 are present in the brains of scrapie-infected mice. Brain Res 654, 200–206.[CrossRef] [Google Scholar]
  75. Xu, Q., Bernardo, A., Walker, D., Kanegawa, T., Mahley, R. W. & Huang, Y.(2006). Profile and regulation of apolipoprotein E (ApoE) expression in the CNS in mice with targeting of green fluorescent protein gene to the ApoE locus. J Neurosci 26, 4985–4994.[CrossRef] [Google Scholar]
  76. Ye, S. Q., Olson, L. M., Reardon, C. A. & Getz, G. S.(1992). Human plasma lipoproteins regulate apolipoprotein E secretion from a post-Golgi compartment. J Biol Chem 267, 21961–21966. [Google Scholar]
  77. Ye, S. Q., Reardon, C. A. & Getz, G. S.(1993). Inhibition of apolipoprotein E degradation in a post-Golgi compartment by a cysteine protease inhibitor. J Biol Chem 268, 8497–8502. [Google Scholar]
  78. Yokode, M., Kita, T., Kikawa, Y., Ogorochi, T., Narumiya, S. & Kawai, C.(1988). Stimulated arachidonate metabolism during foam cell transformation of mouse peritoneal macrophages with oxidized low density lipoprotein. J Clin Invest 81, 720–729.[CrossRef] [Google Scholar]

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error