In:
Journal of Virology, American Society for Microbiology, Vol. 80, No. 2 ( 2006-01-15), p. 596-604
Abstract:
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.
Type of Medium:
Online Resource
ISSN:
0022-538X
,
1098-5514
DOI:
10.1128/JVI.80.2.596-604.2006
Language:
English
Publisher:
American Society for Microbiology
Publication Date:
2006
detail.hit.zdb_id:
1495529-5
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