Stability of the prion protein-encoding (PRNP) gene in HeLa cells.
To assess the risk of the de novo emergence of the agent of transmissible spongiform encephalopathies in cultured cells, we examined the stability of the prion protein-encoding (PRNP) gene in HeLa cells and in cultures contaminated with HeLa cells that have been passaged extensively for over 50 years. Various sub-lineages of HeLa cells showed that some contained a mixture of a truncated PRNP gene (R3-R4 deletion) and a full-length PRNP gene, while others were homozygous for the R3-R4 deletion. That finding suggests that the progenitor of several popular sub-lineages of HeLa must have lost part or all of chromosome 20 early in the history of HeLa cells. No mutations were found in the PRNP genes. We conclude that the spontaneous appearance of mutations leading to expression of abnormal prion proteins in continuously passaged heteroploid cell lines is unlikely to pose a substantial risk for the safe production of biologicals in such cells.
Amexis
Georgios
G
Center for Biologics Evaluation and Research, United States Food and Drug Administration, 1401 Rockville Pike, HFM-313, Rockville, MD 20852-1448, USA.
Ridge
Jeanette
J
Cervenakova
Larisa
L
Enterline
Joan C
JC
Chumakov
Konstantin M
KM
Asher
David M
DM
D001483:Base Sequence; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D016133:Polymerase Chain Reaction; D011328:Prions / Q000235:genetics*
D017931:DNA Primers; D011328:Prions