NR AXUK
AU Piccardo,P.; Vasilyeva,I.; Yakovleva,O.; Bacik,I.; Cervenak,J.; Maximova,O.; Pomeroy,K.; McKenzie,C.; Akimov,S.; Cervenakova,L.; Asher,D.
TI Potential of Cell Substrates used for Production of Biologics to Propagate Transmissible Spongiform Encephalopathy (TSE) Agents
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.13
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: TSE agents have contaminated a variety of products, including humantissue-
derived therapeutics and animal vaccines. Many biologics are prepared in cell cultures. Although most cell cultures studied have resisted infection with TSE agents, a few have been successfully infected. Susceptibility of cultured cells to infection with TSE agents cannot be predicted from species or tissues of origin or the level of expression of prion protein (PrP).
Aims: We are investigating the susceptibility of several cell lines used or proposed for manufacture of various biologic products to propagate TSE agents.
Methods: We inoculated bacteria-free filtrates of three reference TSE agent inoculabrain suspensions containing the agents of BSE, variant Creutzfeldt-Jakob disease (vCJD) and sporadic CJD into several cell lines important or potentially important in the manufacture of biologic products.
Results: Cell lines studied to date include, Vero (green monkey), CHO (Chinese hamster), MDCK (canine), Rab9 (rabbit), HEK 393 (human), and WI-38 (human diploid). We also studied lines of human-neuroblastoma-derived cells (SH-SY5Y), including lines engineered to overexpress mutations associated with familial TSEs. Cell exposed to TSE agents were serially propagated for 30 passages and selected passages tested for appearance of TSE-associated PrP (PrPTSE) and for persistence of infectivity by intracerebral inoculation into TSE-susceptible transgenic mice and squirrel monkeys (BSE-exposed cells only). Normal cellular PrPc was demonstrated in all cell lines tested (except MDCK). No PrPTSE was found in any exposed cells. Known susceptible cells exposed to a human TSE agent as positive control accumulated detectable PrPTSE. No exposed cell line tested has transmitted TSE to mice or monkeys after more than a year of observation.
Conclusion: We have not yet found evidence that any candidate cell substrate exposed to three TSE agents, including BSE, has sustained or propagated infectivity. Methods with increased sensitivity for detecting PrPTSE in cultures are under development, and additional bioassays in susceptible animals are planned.
AD P. Piccardo, I. Bacik, J. Cervenak, O. Maximova, K. Pomeroy, D. Asher, OBRR, CBER, Food and Drug Administration, Lab Bacterial Parasitic Unconventional Agents, USA; I. Vasilyeva, O. Yakovleva, C. McKenzie, S. Akimov, L. Cervenakova, American Red Cross, J Holland Laboratory, USA
SP englisch
PO Schottland
EA pdf-Datei und Poster (Postertitel: Potential to Propagate TSE Agents by Cell Substrates Used to Produce Biologics)