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Journal of Virology, July 2005, p. 8182-8188, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8182-8188.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development of a Novel System To Study Hepatitis Delta Virus Genome Replication

Jinhong Chang, Severin O. Gudima, Chi Tarn, Xingcao Nie, and John M. Taylor*

Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, Pennsylvania 19111-2497

Received 19 November 2004/ Accepted 28 February 2005

Hepatitis delta virus (HDV) genome replication requires the virus-encoded small delta protein ({delta}Ag). During replication, nucleotide sequence changes accumulate on the HDV RNA, leading to the translation of {delta}Ag species that are nonfunctional or even inhibitory. A replication system was devised where all {delta}Ag was conditionally provided from a separate and unchanging source. A line of human embryonic kidney cells was stably transfected with a single copy of cDNA encoding small {delta}Ag, with expression under tetracycline (TET) control. Next, HDV genome replication was initiated in these cells by transfection with a mutated RNA unable to express {delta}Ag. Thus, replication of this RNA was under control of the TET-inducible {delta}Ag. In the absence of TET, there was sufficient {delta}Ag to allow a low level of HDV replication that could be maintained for at least 1 year. When TET was added, both {delta}Ag and genomic RNA increased dramatically within 2 days. With clones of such cells, designated 293-HDV, the burst of HDV RNA replication interfered with cell cycling. Within 2 days, there was a fivefold enhancement of G1/G0 cells relative to both S and G2/M cells, and by 6 days, there was extensive cell detachment and death. These findings and those of other studies that are under way demonstrate the potential applications of this experimental system.

* Corresponding author. Mailing address: Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111-2497. Phone: (215) 728-2436. Fax: (215)728-3105. E-mail: JM_Taylor{at}FCCC.edu.

Journal of Virology, July 2005, p. 8182-8188, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8182-8188.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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