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Journal of Virology, January 2006, p. 596-604, Vol. 80, No. 2
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.2.596-604.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Inhibition of Protease-Resistant Prion Protein Formation in a Transformed Deer Cell Line Infected with Chronic Wasting Disease{ddagger}

Gregory J. Raymond,1 Emily A. Olsen,1 Kil Sun Lee,1 Lynne D. Raymond,1 P. Kruger Bryant III,2 Gerald S. Baron,1 Winslow S. Caughey,1 David A. Kocisko,1 Linda E. McHolland,2 Cynthia Favara,1 Jan P. M. Langeveld,3 Fred G. van Zijderveld,3 Richard T. Mayer,2 Michael W. Miller,4 Elizabeth S. Williams,5,{dagger} and Byron Caughey1*

Laboratory of Persistent Viral Diseases, NIAID, NIH, Rocky Mountain Laboratories, Hamilton, Montana 59840,1 USDA/ARS/ABADRL, Laramie, Wyoming 82071,2 Department for Bacteriology and TSEs, CIDC-Lelystad, 8203 AA 2004, Lelystad, The Netherlands,3 Colorado Division of Wildlife, Wildlife Research Center, Fort Collins, Colorado 80526-2097,4 Department of Veterinary Sciences, University of Wyoming, Laramie, Wyoming 820705

Received 3 August 2005/ Accepted 17 October 2005

Chronic wasting disease (CWD) is an emerging transmissible spongiform encephalopathy (prion disease) of North American cervids, i.e., mule deer, white-tailed deer, and elk (wapiti). To facilitate in vitro studies of CWD, we have developed a transformed deer cell line that is persistently infected with CWD. Primary cultures derived from uninfected mule deer brain tissue were transformed by transfection with a plasmid containing the simian virus 40 genome. A transformed cell line (MDB) was exposed to microsomes prepared from the brainstem of a CWD-affected mule deer. CWD-associated, protease-resistant prion protein (PrPCWD) was used as an indicator of CWD infection. Although no PrPCWD was detected in any of these cultures after two passes, dilution cloning of cells yielded one PrPCWD-positive clone out of 51. This clone, designated MDBCWD, has maintained stable PrPCWD production through 32 serial passes thus far. A second round of dilution cloning yielded 20 PrPCWD-positive subclones out of 30, one of which was designated MDBCWD2. The MDBCWD2 cell line was positive for fibronectin and negative for microtubule-associated protein 2 (a neuronal marker) and glial fibrillary acidic protein (an activated astrocyte marker), consistent with derivation from brain fibroblasts (e.g., meningeal fibroblasts). Two inhibitors of rodent scrapie protease-resistant PrP accumulation, pentosan polysulfate and a porphyrin compound, indium (III) meso-tetra(4-sulfonatophenyl)porphine chloride, potently blocked PrPCWD accumulation in MDBCWD cells. This demonstrates the utility of these cells in a rapid in vitro screening assay for PrPCWD inhibitors and suggests that these compounds have potential to be active against CWD in vivo.

* Corresponding author. Mailing address: Rocky Mountain Labs, 903 S. 4th St., Hamilton, MT 59840. Phone: (406) 363-9264. Fax: (406) 363-9286. E-mail: bcaughey{at}niaid.nih.gov.

{ddagger} We dedicate this paper to the memory of Elizabeth S. Williams, a pioneer of CWD research.

{dagger} Deceased.

Journal of Virology, January 2006, p. 596-604, Vol. 80, No. 2
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.2.596-604.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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