PMID- 24453365
OWN - NLM
STAT- MEDLINE
DCOM- 20140508
LR  - 20211021
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 88
IP  - 7
DP  - 2014 Apr
TI  - Maintenance of large numbers of virus genomes in human cytomegalovirus-infected 
      T98G glioblastoma cells.
PG  - 3861-73
LID - 10.1128/JVI.01166-13 [doi]
AB  - After infection, human cytomegalovirus (HCMV) persists for life. Primary 
      infections and reactivation of latent virus can both result in congenital 
      infection, a leading cause of central nervous system birth defects. We previously 
      reported long-term HCMV infection in the T98G glioblastoma cell line (1). HCMV 
      infection has been further characterized in T98Gs, emphasizing the presence of 
      HCMV DNA over an extended time frame. T98Gs were infected with either HCMV Towne 
      or AD169-IE2-enhanced green fluorescent protein (eGFP) strains. Towne infections 
      yielded mixed IE1 antigen-positive and -negative (Ag(+)/Ag(-)) populations. 
      AD169-IE2-eGFP infections also yielded mixed populations, which were sorted to 
      obtain an IE2(-) (Ag(-)) population. Viral gene expression over the course of 
      infection was determined by immunofluorescent analysis (IFA) and reverse 
      transcription-PCR (RT-PCR). The presence of HCMV genomes was determined by PCR, 
      nested PCR (n-PCR), and fluorescence in situ hybridization (FISH). Compared to 
      the HCMV latency model, THP-1, Towne-infected T98Gs expressed IE1 and 
      latency-associated transcripts for longer periods, contained many more HCMV 
      genomes during early passages, and carried genomes for a greatly extended period 
      of passaging. Large numbers of HCMV genomes were also found in purified Ag(-) 
      AD169-infected cells for the first several passages. Interestingly, latency 
      transcripts were observed from very early times in the Towne-infected cells, even 
      when IE1 was expressed at low levels. Although AD169-infected Ag(-) cells 
      expressed no detectable levels of either IE1 or latency transcripts, they also 
      maintained large numbers of genomes within the cell nuclei for several passages. 
      These results identify HCMV-infected T98Gs as an attractive new model in the 
      study of the long-term maintenance of virus genomes in the context of neural cell 
      types. IMPORTANCE: Our previous work showed that T98G glioblastoma cells were 
      semipermissive to HCMV infection; virus trafficked to the nucleus, and yet only a 
      proportion of cells stained positive for viral antigens, thus allowing continual 
      subculturing and passaging. The cells eventually transitioned to a state where 
      viral genomes were maintained without viral antigen expression or virion 
      production. Here we report that during long-term T98G infection, large numbers of 
      genomes were maintained within all of the cells' nuclei for the first several 
      passages (through passage 4 [P4]), even in the presence of continual cellular 
      division. Surprisingly, genomes were maintained, albeit at a lower level, through 
      day 41. This is decidedly longer than in any other latency model system that has 
      been described to date. We believe that this system offers a useful model to aid 
      in unraveling the cellular components involved in viral genome maintenance (and 
      presumably replication) in cells carrying long-term latent genomes in a neural 
      context.
FAU - Duan, Ying-Liang
AU  - Duan YL
AD  - State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of 
      Sciences, Wuhan, China.
FAU - Ye, Han-Qing
AU  - Ye HQ
FAU - Zavala, Anamaria G
AU  - Zavala AG
FAU - Yang, Cui-Qing
AU  - Yang CQ
FAU - Miao, Ling-Feng
AU  - Miao LF
FAU - Fu, Bi-Shi
AU  - Fu BS
FAU - Seo, Keun Seok
AU  - Seo KS
FAU - Davrinche, Christian
AU  - Davrinche C
FAU - Luo, Min-Hua
AU  - Luo MH
FAU - Fortunato, Elizabeth A
AU  - Fortunato EA
LA  - eng
GR  - R01 AI051463/AI/NIAID NIH HHS/United States
GR  - R01-AI51463/AI/NIAID NIH HHS/United States
GR  - P20 GM103408/GM/NIGMS NIH HHS/United States
GR  - P20 RR016454/RR/NCRR NIH HHS/United States
GR  - P20 RR015587/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20140122
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
SB  - IM
MH  - Adenoviridae/physiology
MH  - Cell Line, Tumor
MH  - Cytomegalovirus/*physiology
MH  - Gene Expression Profiling
MH  - Gene Expression Regulation, Viral
MH  - Humans
MH  - Neuroglia/*virology
MH  - *Virus Latency
PMC - PMC3993548
EDAT- 2014/01/24 06:00
MHDA- 2014/05/09 06:00
PMCR- 2014/10/01
CRDT- 2014/01/24 06:00
PHST- 2014/01/24 06:00 [entrez]
PHST- 2014/01/24 06:00 [pubmed]
PHST- 2014/05/09 06:00 [medline]
PHST- 2014/10/01 00:00 [pmc-release]
AID - JVI.01166-13 [pii]
AID - 01166-13 [pii]
AID - 10.1128/JVI.01166-13 [doi]
PST - ppublish
SO  - J Virol. 2014 Apr;88(7):3861-73. doi: 10.1128/JVI.01166-13. Epub 2014 Jan 22.