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. 1996 Dec 1;320(Pt 2):563–570. doi: 10.1042/bj3200563

Activation effects of a prion protein fragment [PrP-(106-126)] on human leucocytes.

L Diomede 1, S Sozzani 1, W Luini 1, M Algeri 1, L De Gioia 1, R Chiesa 1, P M Lievens 1, O Bugiani 1, G Forloni 1, F Tagliavini 1, M Salmona 1
PMCID: PMC1217966  PMID: 8973567

Abstract

Prion-related encephalopathies are characterized by the intracerebral accumulation of an abnormal isoform of the cellular prion protein (PrPC) named scrapie prion protein (PrPSc). The pathological forms of this protein and its cellular precursor are not only expressed in the brain but also, at lower concentrations, in peripheral tissues. We recently showed that a synthetic peptide corresponding to residues 106-126 [PrP-(106-126)] of the human PrP is toxic to neurons and trophic to astrocytes in vitro. Our experiments were aimed at verifying whether PrP-(106-126) and other peptides corresponding to fragments of the amyloid protein purified from brains of patients with Gerstmann-Sträussler-Scheinker disease-namely PrP-(89-106), PrP-(106-114), PrP-(127-147)-were capable of stimulating circulating leucocytes. Native PrP expression in human lymphocytes, monocytes and neutrophils was first confirmed using PCR amplification of total RNA, after reverse transcription, and immunoblot analysis of cell extracts with anti-PrP antibodies. PrP-(106-126), but not the other peptides, increased membrane microviscosity, intracellular Ca2+ concentration and cell migration in circulating leucocytes, and O2-. production in monocytes and neutrophils. Membrane microviscosity was determined by the fluorescence polarization technique, using diphenylhexatriene as a probe, 300 s after the addition of PrP-(106-126) to the cell suspension in the concentration range 5-50 microM. The increase in intracellular Ca2+ elicited by PrP-(106-126) was dose-dependent in the range 5-500 microM. PrP-(106-126) stimulated O2-. production in monocytes and neutrophils in a dose- (10-300 microM) and time-(5-30 min) dependent manner in the presence of 10 microM dihydrocytochalasin B. Both the increase in Ca2+ concentration and the O2-. production were partially sensitive to pertussis toxin. PrP-(106-126) stimulated leucocyte migration in a dose-dependent (30-300 microM) manner and, at the highest concentration used, this migration was comparable with that elicited by 2.5 nM interleukin 8 or 10 nM fMet-Leu-Phe peptide.

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Selected References

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  1. Allavena P., Paganin C., Zhou D., Bianchi G., Sozzani S., Mantovani A. Interleukin-12 is chemotactic for natural killer cells and stimulates their interaction with vascular endothelium. Blood. 1994 Oct 1;84(7):2261–2268. [PubMed] [Google Scholar]
  2. Baggiolini M., Dewald B., Moser B. Interleukin-8 and related chemotactic cytokines--CXC and CC chemokines. Adv Immunol. 1994;55:97–179. [PubMed] [Google Scholar]
  3. Berg L. J. Insights into the role of the immune system in prion diseases. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):429–432. doi: 10.1073/pnas.91.2.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bolton D. C., McKinley M. P., Prusiner S. B. Identification of a protein that purifies with the scrapie prion. Science. 1982 Dec 24;218(4579):1309–1311. doi: 10.1126/science.6815801. [DOI] [PubMed] [Google Scholar]
  5. Brown D. R., Herms J., Kretzschmar H. A. Mouse cortical cells lacking cellular PrP survive in culture with a neurotoxic PrP fragment. Neuroreport. 1994 Oct 27;5(16):2057–2060. doi: 10.1097/00001756-199410270-00017. [DOI] [PubMed] [Google Scholar]
  6. Cashman N. R., Loertscher R., Nalbantoglu J., Shaw I., Kascsak R. J., Bolton D. C., Bendheim P. E. Cellular isoform of the scrapie agent protein participates in lymphocyte activation. Cell. 1990 Apr 6;61(1):185–192. doi: 10.1016/0092-8674(90)90225-4. [DOI] [PubMed] [Google Scholar]
  7. Chou S. M., Payne W. N., Gibbs C. J., Jr, Gajdusek D. C. Transmission and scanning electron microscopy of spongiform change in Creutzfeldt-Jakob disease. Brain. 1980 Dec;103(4):885–904. doi: 10.1093/brain/103.4.885. [DOI] [PubMed] [Google Scholar]
  8. DeArmond S. J., Mobley W. C., DeMott D. L., Barry R. A., Beckstead J. H., Prusiner S. B. Changes in the localization of brain prion proteins during scrapie infection. Neurology. 1987 Aug;37(8):1271–1280. doi: 10.1212/wnl.37.8.1271. [DOI] [PubMed] [Google Scholar]
  9. DeArmond S. J., Mobley W. C., DeMott D. L., Barry R. A., Beckstead J. H., Prusiner S. B. Changes in the localization of brain prion proteins during scrapie infection. Neurology. 1987 Aug;37(8):1271–1280. doi: 10.1212/wnl.37.8.1271. [DOI] [PubMed] [Google Scholar]
  10. DeArmond S. J., Prusiner S. B. The neurochemistry of prion diseases. J Neurochem. 1993 Nov;61(5):1589–1601. doi: 10.1111/j.1471-4159.1993.tb09792.x. [DOI] [PubMed] [Google Scholar]
  11. Falk W., Goodwin R. H., Jr, Leonard E. J. A 48-well micro chemotaxis assembly for rapid and accurate measurement of leukocyte migration. J Immunol Methods. 1980;33(3):239–247. doi: 10.1016/0022-1759(80)90211-2. [DOI] [PubMed] [Google Scholar]
  12. Forloni G., Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O., Tagliavini F. Neurotoxicity of a prion protein fragment. Nature. 1993 Apr 8;362(6420):543–546. doi: 10.1038/362543a0. [DOI] [PubMed] [Google Scholar]
  13. Forloni G., Del Bo R., Angeretti N., Chiesa R., Smiroldo S., Doni R., Ghibaudi E., Salmona M., Porro M., Verga L. A neurotoxic prion protein fragment induces rat astroglial proliferation and hypertrophy. Eur J Neurosci. 1994 Sep 1;6(9):1415–1422. doi: 10.1111/j.1460-9568.1994.tb01003.x. [DOI] [PubMed] [Google Scholar]
  14. Gerard C., McPhail L. C., Marfat A., Stimler-Gerard N. P., Bass D. A., McCall C. E. Role of protein kinases in stimulation of human polymorphonuclear leukocyte oxidative metabolism by various agonists. Differential effects of a novel protein kinase inhibitor. J Clin Invest. 1986 Jan;77(1):61–65. doi: 10.1172/JCI112302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ghetti B., Piccardo P., Spillantini M. G., Ichimiya Y., Porro M., Perini F., Kitamoto T., Tateishi J., Seiler C., Frangione B. Vascular variant of prion protein cerebral amyloidosis with tau-positive neurofibrillary tangles: the phenotype of the stop codon 145 mutation in PRNP. Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):744–748. doi: 10.1073/pnas.93.2.744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gibbons R. A., Hunter G. D. Nature of the scrapie agent. Nature. 1967 Sep 2;215(5105):1041–1043. doi: 10.1038/2151041a0. [DOI] [PubMed] [Google Scholar]
  17. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  18. Hanukoglu I., Tanese N., Fuchs E. Complementary DNA sequence of a human cytoplasmic actin. Interspecies divergence of 3' non-coding regions. J Mol Biol. 1983 Feb 5;163(4):673–678. doi: 10.1016/0022-2836(83)90117-1. [DOI] [PubMed] [Google Scholar]
  19. Hope J., Morton L. J., Farquhar C. F., Multhaup G., Beyreuther K., Kimberlin R. H. The major polypeptide of scrapie-associated fibrils (SAF) has the same size, charge distribution and N-terminal protein sequence as predicted for the normal brain protein (PrP). EMBO J. 1986 Oct;5(10):2591–2597. doi: 10.1002/j.1460-2075.1986.tb04539.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hunter G. D. The size and intracellular location of the scrapie agent. Biochem J. 1969 Sep;114(2):22P–23P. doi: 10.1042/bj1140022p. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kretzschmar H. A., Prusiner S. B., Stowring L. E., DeArmond S. J. Scrapie prion proteins are synthesized in neurons. Am J Pathol. 1986 Jan;122(1):1–5. [PMC free article] [PubMed] [Google Scholar]
  22. Kretzschmar H. A., Stowring L. E., Westaway D., Stubblebine W. H., Prusiner S. B., Dearmond S. J. Molecular cloning of a human prion protein cDNA. DNA. 1986 Aug;5(4):315–324. doi: 10.1089/dna.1986.5.315. [DOI] [PubMed] [Google Scholar]
  23. Kristensson K., Feuerstein B., Taraboulos A., Hyun W. C., Prusiner S. B., DeArmond S. J. Scrapie prions alter receptor-mediated calcium responses in cultured cells. Neurology. 1993 Nov;43(11):2335–2341. doi: 10.1212/wnl.43.11.2335. [DOI] [PubMed] [Google Scholar]
  24. Lehmann S., Harris D. A. A mutant prion protein displays an aberrant membrane association when expressed in cultured cells. J Biol Chem. 1995 Oct 13;270(41):24589–24597. doi: 10.1074/jbc.270.41.24589. [DOI] [PubMed] [Google Scholar]
  25. Meda L., Cassatella M. A., Szendrei G. I., Otvos L., Jr, Baron P., Villalba M., Ferrari D., Rossi F. Activation of microglial cells by beta-amyloid protein and interferon-gamma. Nature. 1995 Apr 13;374(6523):647–650. doi: 10.1038/374647a0. [DOI] [PubMed] [Google Scholar]
  26. Meiner Z., Halimi M., Polakiewicz R. D., Prusiner S. B., Gabizon R. Presence of prion protein in peripheral tissues of Libyan Jews with Creutzfeldt-Jakob disease. Neurology. 1992 Jul;42(7):1355–1360. doi: 10.1212/wnl.42.7.1355. [DOI] [PubMed] [Google Scholar]
  27. Müller W. E., Ushijima H., Schröder H. C., Forrest J. M., Schatton W. F., Rytik P. G., Heffner-Lauc M. Cytoprotective effect of NMDA receptor antagonists on prion protein (PrionSc)-induced toxicity in rat cortical cell cultures. Eur J Pharmacol. 1993 Aug 15;246(3):261–267. doi: 10.1016/0922-4106(93)90040-g. [DOI] [PubMed] [Google Scholar]
  28. Perini F., Frangione B., Prelli F. Prion protein released by platelets. Lancet. 1996 Jun 8;347(9015):1635–1636. doi: 10.1016/s0140-6736(96)91128-9. [DOI] [PubMed] [Google Scholar]
  29. Prusiner S. B. Molecular biology of prion diseases. Science. 1991 Jun 14;252(5012):1515–1522. doi: 10.1126/science.1675487. [DOI] [PubMed] [Google Scholar]
  30. Selvaggini C., De Gioia L., Cantù L., Ghibaudi E., Diomede L., Passerini F., Forloni G., Bugiani O., Tagliavini F., Salmona M. Molecular characteristics of a protease-resistant, amyloidogenic and neurotoxic peptide homologous to residues 106-126 of the prion protein. Biochem Biophys Res Commun. 1993 Aug 16;194(3):1380–1386. doi: 10.1006/bbrc.1993.1977. [DOI] [PubMed] [Google Scholar]
  31. Shinitzky M., Barenholz Y. Fluidity parameters of lipid regions determined by fluorescence polarization. Biochim Biophys Acta. 1978 Dec 15;515(4):367–394. doi: 10.1016/0304-4157(78)90010-2. [DOI] [PubMed] [Google Scholar]
  32. Sipe J. D. Amyloidosis. Crit Rev Clin Lab Sci. 1994;31(4):325–354. doi: 10.3109/10408369409084679. [DOI] [PubMed] [Google Scholar]
  33. Sozzani S., Agwu D. E., Ellenburg M. D., Locati M., Rieppi M., Rojas A., Mantovani A., McPhail L. C. Activation of phospholipase D by interleukin-8 in human neutrophils. Blood. 1994 Dec 1;84(11):3895–3901. [PubMed] [Google Scholar]
  34. Sozzani S., Luini W., Molino M., Jílek P., Bottazzi B., Cerletti C., Matsushima K., Mantovani A. The signal transduction pathway involved in the migration induced by a monocyte chemotactic cytokine. J Immunol. 1991 Oct 1;147(7):2215–2221. [PubMed] [Google Scholar]
  35. Tagliavini F., Prelli F., Verga L., Giaccone G., Sarma R., Gorevic P., Ghetti B., Passerini F., Ghibaudi E., Forloni G. Synthetic peptides homologous to prion protein residues 106-147 form amyloid-like fibrils in vitro. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9678–9682. doi: 10.1073/pnas.90.20.9678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. White M. F., Kahn C. R. The insulin signaling system. J Biol Chem. 1994 Jan 7;269(1):1–4. [PubMed] [Google Scholar]

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