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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

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, and Byron Caughey^1*

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 (PrP^CWD) was used as an indicator of CWD infection. Although no PrP^CWD was detected in any of these cultures after two passes, dilution cloning of cells yielded one PrP^CWD-positive clone out of 51. This clone, designated MDB^CWD, has maintained stable PrP^CWD production through 32 serial passes thus far. A second round of dilution cloning yielded 20 PrP^CWD-positive subclones out of 30, one of which was designated MDB^CWD2. The MDB^CWD2 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 PrP^CWD accumulation in MDB^CWD cells. This demonstrates the utility of these cells in a rapid in vitro screening assay for PrP^CWD inhibitors and suggests that these compounds have potential to be active against CWD in vivo.

 

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* 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@niaid.nih.gov .

 

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

Deceased.

http://jvi.asm.org/cgi/content/abstract/80/2/596