Sulfated polyanion inhibition of scrapie-associated PrP accumulation in cultured cells

The accumulation of an abnormal, protease-resistant form of the protein PrP (PrP-res) in hosts with scrapie and related transmissible spongiform encephalopathies appears to be important in disease pathogenesis. To gain insight into the mechanism of PrP-res accumulation and the in vivo antiscrapie activity of certain polyanions, we have studied effects of sulfated glycans on PrP metabolism in scrapie-infected neuroblastoma cells. Pentosan polysulfate, like the amyloid-binding dye Congo red, potently inhibited the accumulation of PrP-res in these cells without apparent effects on the metabolism of the normal isoform. The inhibition was due primarily to prevention of new PrP-res accumulation rather than destabilization of preexisting PrP-res. PrP-res accumulation remained depressed in the cultures after removal of the inhibitors. The activities of other sulfated glycans, nonsulfated polyanions, dextran, and DEAE-dextran were compared with those of pentosan polysulfate and Congo red. This comparison provided evidence that the density of sulfation and molecular size are factors influencing anti-PrP-res activity of sulfated glycans. The relative potencies of these compounds corresponded well with their previously determined antiscrapie activities in vivo, suggesting that the prophylactic effects of sulfated polyanions may be due to inhibition of PrP-res accumulation. Since PrP-res amyloid is known to contain sulfated glycosaminoglycans, we reason that these inhibitors may competitively block an interaction between PrP and endogenous glycosaminoglycans that is essential for its accumulation in a protease-resistant, potentially amyloidogenic state.

[1]  K. Takemoto,et al.  Inhibition of Herpes Virus by Natural and Synthetic Acid Polysaccharides , 1964, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[2]  T. Diamantstein,et al.  Stimulation of humoral antibody formation by polyanions. II. The influence of sulfate esters of polymers on the immune response in mice , 1971, European journal of immunology.

[3]  T. Diamantstein,et al.  Stimulation of DNA synthesis in mouse lymphoid cells by polyanions in vitro. II. Relationship between adjuvant activity and stimulation of DNA synthesis by polyanions , 1973, European journal of immunology.

[4]  E. Wecker,et al.  Action of dextran sulfate as a direct and general B cell mitogen , 1974, European journal of immunology.

[5]  P. D. Hart,et al.  Interference with normal phagosome-lysosome fusion in macrophages, using ingested yeast cells and suramin , 1975, Nature.

[6]  G. Glenner.  Amyloid deposits and amyloidosis: the beta-fibrilloses (second of two parts). , 1980, The New England journal of medicine.

[7]  N. Bloksma,et al.  Influence on macrophage functions as a possible basis of immunomodification by polyanions. , 1980, Annales d'immunologie.

[8]  A. Woody,et al.  Spectroscopic studies of Congo Red binding to RNA polymerase. , 1981, Biochimica et biophysica acta.

[9]  S. Prusiner,et al.  Identification of a protein that purifies with the scrapie prion. , 1982, Science.

[10]  S. Prusiner,et al.  Scrapie prions aggregate to form amyloid-like birefringent rods , 1983, Cell.

[11]  H. Gelderblom,et al.  Scrapie infectivity, fibrils and low molecular weight protein , 1983, Nature.

[12]  S. Prusiner,et al.  Antibodies to a scrapie prion protein , 1984, Nature.

[13]  H. Diringer,et al.  The reticuloendothelial system in scrapie pathogenesis. , 1984, The Journal of general virology.

[14]  B. Chesebro,et al.  Identification of scrapie prion protein-specific mRNA in scrapie-infected and uninfected brain , 1985, Nature.

[15]  P. Brown,et al.  Diagnosis of Creutzfeldt-Jakob disease by Western blot identification of marker protein in human brain tissue. , 1986, The New England journal of medicine.

[16]  S. Prusiner,et al.  Linkage of prion protein and scrapie incubation time genes , 1986, Cell.

[17]  R. Kimberlin,et al.  Suppression of scrapie infection in mice by heteropolyanion 23, dextran sulfate, and some other polyanions , 1986, Antimicrobial Agents and Chemotherapy.

[18]  A. Dickinson,et al.  Prolongation of scrapie incubation period by an injection of dextran sulphate 500 within the month before or after infection. , 1986, The Journal of general virology.

[19]  B. Chesebro,et al.  Characterization of scrapie infection in mouse neuroblastoma cells. , 1987, The Journal of general virology.

[20]  S. Prusiner,et al.  Changes in the localization of brain prion proteins during scrapie infection , 1987, Neurology.

[21]  I. Mcconnell,et al.  Linkage of the scrapie-associated fibril protein (PrP) gene and Sinc using congenic mice and restriction fragment length polymorphism analysis. , 1987, The Journal of general virology.

[22]  R. Kisilevsky.  From arthritis to Alzheimer's disease: current concepts on the pathogenesis of amyloidosis. , 1987, Canadian journal of physiology and pharmacology.

[23]  B. Chesebro,et al.  Analyses of frequency of infection, specific infectivity, and prion protein biosynthesis in scrapie-infected neuroblastoma cell clones , 1988, Journal of virology.

[24]  M. Czub,et al.  Replication of the scrapie agent in hamsters infected intracerebrally confirms the pathogenesis of an amyloid-inducing virosis. , 1988, The Journal of general virology.

[25]  D. Henner,et al.  Sequence of the Bacillus thuringiensis phosphatidylinositol specific phospholipase C. , 1988, Nucleic acids research.

[26]  E. Ruoslahti.  Structure and biology of proteoglycans. , 1988, Annual review of cell biology.

[27]  G. Multhaup,et al.  Fibrils from brains of cows with new cattle disease contain scrapie-associated protein , 1988, Nature.

[28]  E. De Clercq,et al.  Pentosan polysulfate, a sulfated oligosaccharide, is a potent and selective anti-HIV agent in vitro. , 1988, Antiviral research.

[29]  R. Flavell,et al.  Cell-specific heterogeneity in sensitivity of phosphatidylinositol-anchored membrane antigens to release by phospholipase C. , 1988, Journal of immunological methods.

[30]  P. McBride,et al.  Precise targeting of the pathology of the sialoglycoprotein, PrP, and vacuolar degeneration in mouse scrapie , 1989, Neuroscience Letters.

[31]  Y. Sakaki,et al.  Pro----leu change at position 102 of prion protein is the most common but not the sole mutation related to Gerstmann-Sträussler syndrome. , 1989, Biochemical and biophysical research communications.

[32]  W. Brown,et al.  Mutations in familial Creutzfeldt-Jakob disease and Gerstmann-Sträussler-Scheinker's syndrome , 1989, Experimental Neurology.

[33]  P. Spear,et al.  Initial interaction of herpes simplex virus with cells is binding to heparan sulfate , 1989, Journal of virology.

[34]  Jurg Ott,et al.  Linkage of a prion protein missense variant to Gerstmann–Sträussler syndrome , 1989, Nature.

[35]  K. Moelling,et al.  Inhibition of human immunodeficiency virus type 1 RNase H by sulfated polyanions , 1989, Journal of virology.

[36]  S. Prusiner,et al.  Spontaneous neurodegeneration in transgenic mice with mutant prion protein , 1990, Science.

[37]  S. Prusiner,et al.  Scrapie and cellular prion proteins differ in their kinetics of synthesis and topology in cultured cells , 1990, The Journal of cell biology.

[38]  B. Chesebro,et al.  Analysis of linkage between scrapie incubation period and the prion protein gene in mice. , 1990, The Journal of general virology.

[39]  J. Aiken,et al.  The search for scrapie agent nucleic acid. , 1990, Microbiological reviews.

[40]  S. Prusiner,et al.  Immunolocalization of heparan sulfate proteoglycans to the prion protein amyloid plaques of Gerstmann-Straussler syndrome, Creutzfeldt-Jakob disease and scrapie. , 1990, Laboratory investigation; a journal of technical methods and pathology.

[41]  K. Beyreuther,et al.  Two alleles of a neural protein gene linked to scrapie in sheep. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[42]  A. Haase,et al.  Scrapie-associated prion protein accumulates in astrocytes during scrapie infection. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[43]  G. J. Raymond,et al.  The scrapie-associated form of PrP is made from a cell surface precursor that is both protease- and phospholipase-sensitive. , 1991, The Journal of biological chemistry.

[44]  S. Prusiner,et al.  Ultrastructural localization of scrapie prion proteins in cytoplasmic vesicles of infected cultured cells. , 1991, Laboratory investigation; a journal of technical methods and pathology.

[45]  B. Ehlers,et al.  Chemoprophylaxis of scrapie in mice. , 1991, The Journal of general virology.

[46]  S. Prusiner,et al.  Molecular biology of prion diseases , 1991, Science.

[47]  R. Rohwer,et al.  The scrapie agent: "a virus by any other name". , 1991, Current topics in microbiology and immunology.

[48]  B. Greenberg,et al.  High affinity interactions between the Alzheimer's beta-amyloid precursor proteins and the basement membrane form of heparan sulfate proteoglycan. , 1991, The Journal of biological chemistry.

[49]  B. Caughey,et al.  Secondary structure analysis of the scrapie-associated protein PrP 27-30 in water by infrared spectroscopy. , 1991, Biochemistry.

[50]  G. J. Raymond,et al.  N-terminal truncation of the scrapie-associated form of PrP by lysosomal protease(s): implications regarding the site of conversion of PrP to the protease-resistant state , 1991, Journal of virology.

[51]  M. Pocchiari,et al.  Amphotericin B treatment dissociates in vivo replication of the scrapie agent from PrP accumulation , 1992, Nature.

[52]  B. Caughey,et al.  Potent Inhibition of Scrapie‐Associated PrP Accumulation by Congo Red , 1992, Journal of neurochemistry.

[53]  M. Salvatore,et al.  Sulphate polyanions prolong the incubation period of scrapie-infected hamsters. , 1992, The Journal of general virology.