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Sequential mutations associated with adaptation of human cytomegalovirus to growth in cell culture

Dargan, D. J., Douglas, E., Cunningham, C., Jamieson, F., Stanton, Richard James ORCID:, Baluchova, K., McSharry, Brian, Tomasec, Peter, Emery, V. C., Percivalle, E., Sarasini, A., Gerna, G., Wilkinson, Gavin William Grahame ORCID: and Davison, A. J. 2010. Sequential mutations associated with adaptation of human cytomegalovirus to growth in cell culture. Journal of General Virology 91 (6) , pp. 1535-1546. 10.1099/vir.0.018994-0

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Mutations that occurred during adaptation of human cytomegalovirus to cell culture were monitored by isolating four strains from clinical samples, passaging them in various cell types and sequencing ten complete virus genomes from the final passages. Mutational dynamics were assessed by targeted sequencing of intermediate passages and the original clinical samples. Gene RL13 and the UL128 locus (UL128L, consisting of genes UL128, UL130 and UL131A) mutated in all strains. Mutations in RL13 occurred in fibroblast, epithelial and endothelial cells, whereas those in UL128L were limited to fibroblasts and detected later than those in RL13. In addition, a region containing genes UL145, UL144, UL142, UL141 and UL140 mutated in three strains. All strains exhibited numerous mutations in other regions of the genome, with a preponderance in parts of the inverted repeats. An investigation was carried out on the kinetic growth yields of viruses derived from selected passages that were predominantly non-mutated in RL13 and UL128L (RL13+UL128L+), or that were largely mutated in RL13 (RL13−UL128L+) or both RL13 and UL128L (RL13−UL128L−). RL13−UL128L− viruses produced greater yields of infectious progeny than RL13−UL128L+ viruses, and RL13−UL128L+ viruses produced greater yields than RL13+UL128L+ viruses. These results suggest strongly that RL13 and UL128L exert at least partially independent suppressive effects on growth in fibroblasts. As all isolates proved genetically unstable in all cell types tested, caution is advised in choosing and monitoring strains for experimental studies of vulnerable functions, particularly those involved in cell tropism, immune evasion or growth temperance.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Systems Immunity Research Institute (SIURI)
Subjects: Q Science > QR Microbiology
R Medicine > R Medicine (General)
Publisher: Society for General Microbiology
ISSN: 0022-1317
Last Modified: 20 Oct 2022 08:40

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