Transfer of scrapie prion infectivity by cell contact in culture

被引:108
作者
Kanu, N
Imokawa, Y
Drechsel, DN
Williamson, RA
Birkett, CR
Bostock, CJ
Brockes, JP
机构
[1] UCL, Dept Biochem & Mol Biol, London WC1E 6BT, England
[2] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany
[3] Scripps Res Inst, Dept Immunol, La Jolla, CA 92037 USA
[4] Inst Anim Hlth, Compton Lab, Newbury RG20 7NN, Berks, England
基金
英国医学研究理事会; 英国生物技术与生命科学研究理事会;
关键词
D O I
10.1016/S0960-9822(02)00722-4
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Background: When a cell is infected with scrapie prions, newly synthesized molecules of the prion protein PrPC are expressed at the cell surface and may subsequently be converted to the abnormal form PrPSc. In an experimental scrapie infection of an animal, the initial innoculum of PrPsc is cleared relatively rapidly, and the subsequent propagation of the infection depends on the ability of infected cells to convert uninfected target cells to stable production of PrPSc. The mechanism of such cell-based infection is not understood. Results: We have established a system in dissociated cell culture in which scrapie-infected mouse SMB cells are able to stably convert genetically marked target cells by coculture. After coculture and rigorous removal of SMB cells, the target cells express PrPSc and also incorporate [S-35]methionine into PrPSc. The extent of conversion was sensitive to the ratio of the two cell types, and conversion by live SMB required 2500-fold less PrPSc than conversion by a cell-free prion preparation. The conversion activity of SMB cells is not detectable in conditioned medium and apparently depends on close proximity or contact, as evidenced by culturing the SMB and target cells on neighboring but separate surfaces. SMB cells were killed by fixation in aldehydes, followed by washing, and were found to retain significant activity at conversion of target cells. Conclusions: Cell-mediated infection of target cells in this culture system is effective and requires significantly less PrPSc than infection by a prion preparation. Several lines of evidence indicate that it depends on cell contact, in particular, the activity of aldehyde-fixed infected cells.
引用
收藏
页码:523 / 530
页数:8
相关论文
共 31 条
[21]   Successful transmission of three mouse-adapted scrapie strains to murine neuroblastoma cell lines overexpressing wild-type mouse prion protein [J].
Nishida, N ;
Harris, DA ;
Vilette, D ;
Laude, H ;
Frobert, Y ;
Grassi, J ;
Casanova, D ;
Milhavet, O ;
Lehmann, S .
JOURNAL OF VIROLOGY, 2000, 74 (01) :320-325
[22]   RESISTANCE OF SCRAPIE AGENT TO FORMALIN [J].
PATTISON, IH .
JOURNAL OF COMPARATIVE PATHOLOGY AND THERAPEUTICS, 1965, 75 (02) :159-&
[23]   A SINGLE HAMSTER PRP AMINO-ACID BLOCKS CONVERSION TO PROTEASE-RESISTANT PRP IN SCRAPIE-INFECTED MOUSE NEUROBLASTOMA-CELLS [J].
PRIOLA, SA ;
CHESEBRO, B .
JOURNAL OF VIROLOGY, 1995, 69 (12) :7754-7758
[24]  
Prusiner SB, 1999, COLD SPRING HARBOR M, V38, P349
[25]   Entry versus blockade of brain infection following oral or intraperitoneal scrapie administration: Role of prion protein expression in peripheral nerves and spleen [J].
Race, R ;
Oldstone, M ;
Chesebro, B .
JOURNAL OF VIROLOGY, 2000, 74 (02) :828-833
[26]   ANALYSES OF FREQUENCY OF INFECTION, SPECIFIC INFECTIVITY, AND PRION PROTEIN-BIOSYNTHESIS IN SCRAPIE-INFECTED NEURO-BLASTOMA CELL CLONES [J].
RACE, RE ;
CAUGHEY, B ;
GRAHAM, K ;
ERNST, D ;
CHESEBRO, B .
JOURNAL OF VIROLOGY, 1988, 62 (08) :2845-2849
[27]  
Schatzl HM, 1997, J VIROL, V71, P8821
[28]   THE CONSTRUCTION OF A HIGHLY EFFICIENT AND VERSATILE SET OF MAMMALIAN EXPRESSION VECTORS [J].
STEPHENS, PE ;
COCKETT, MI .
NUCLEIC ACIDS RESEARCH, 1989, 17 (17) :7110-7110
[29]   Subcellular colocalization of the cellular and scrapie prion proteins in caveolae-like membranous domains [J].
Vey, M ;
Pilkuhn, S ;
Wille, H ;
Nixon, R ;
Dearmond, SJ ;
Smart, EJ ;
Anderson, RGW ;
Taraboulos, A ;
Prusiner, SB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (25) :14945-14949
[30]   Mapping the prion protein using recombinant antibodies [J].
Williamson, RA ;
Peretz, D ;
Pinilla, C ;
Ball, H ;
Bastidas, RB ;
Rozenshteyn, R ;
Houghten, RA ;
Prusiner, SB ;
Burton, DR .
JOURNAL OF VIROLOGY, 1998, 72 (11) :9413-9418