PMID- 15893497
OWN - NLM
STAT- MEDLINE
DCOM- 20060214
LR  - 20201119
IS  - 1286-4579 (Print)
IS  - 1286-4579 (Linking)
VI  - 7
IP  - 9-10
DP  - 2005 Jul
TI  - Vaccinia virus infection and gene transduction in cultured neurons.
PG  - 1087-96
AB  - The study of neurons in culture would benefit from the development of a gene 
      transduction system capable of delivering foreign genes at high efficiency, as 
      transduction of primary neurons with existing systems is inefficient. The 
      efficacy of lytic vaccinia virus (VV) infection of primary retinal cultures and 
      PC12 cells (a model of neuronal differentiation) was examined in order to 
      determine the efficiency of gene transduction using VV in neuronal primary 
      culture. VV was able to infect retinal cells and PC12 cells and express 
      transgenes of Escherichia coli beta-galactosidase (lacZ) and epithelial fatty 
      acid binding protein (E-FABP) in a virus dose-dependent manner. Most (50-100%) of 
      the retinal cells were positive for transgene protein at multiplicities of 
      infection (MOI) between 10 and 100 plaque-forming units (PFU), while over 50% of 
      VV-infected PC12 cells expressed the virus encoded gene at an MOI = 10. The 
      production of foreign mRNA and protein by VV following infection was verified by 
      PCR and Western blot. Because VV is a lytic virus, cytopathic effects were 
      examined. Retinal cultures maintained for 0.5 days in vitro showed greater than 
      90% survival at 24 h post-infection, while 14-day cultures were equally viable 
      for 48 h. Retinal ganglion cells and differentiated PC12 cells appear to be more 
      protected against lytic VV infection than proliferating glial and 
      undifferentiated PC12 cells. These data suggest that VV may be a useful vector 
      for delivering foreign genes to neuronal cells with an efficient transient 
      transgene expression.
FAU - Allen, Gregory A
AU  - Allen GA
AD  - Department of Physiology, School of Medicine, Loma Linda University, Loma Linda, 
      CA, USA.
FAU - Denes, Bela
AU  - Denes B
FAU - Fodor, Istvan
AU  - Fodor I
FAU - De Leon, Marino
AU  - De Leon M
LA  - eng
GR  - R25 GM060507/GM/NIGMS NIH HHS/United States
PT  - Journal Article
DEP - 20050419
PL  - France
TA  - Microbes Infect
JT  - Microbes and infection
JID - 100883508
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (RNA, Bacterial)
RN  - 0 (RNA, Messenger)
RN  - EC 3.2.1.23 (beta-Galactosidase)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Cell Survival
MH  - Cells, Cultured
MH  - Escherichia coli Proteins/analysis/genetics
MH  - Gene Expression
MH  - Genes, Reporter
MH  - Immunohistochemistry
MH  - Microscopy, Confocal
MH  - Neuroglia/chemistry/*virology
MH  - Neurons/*virology
MH  - PC12 Cells
MH  - RNA, Bacterial/analysis
MH  - RNA, Messenger/analysis
MH  - Rats
MH  - Retinal Ganglion Cells/chemistry/*virology
MH  - Transduction, Genetic/*methods
MH  - Vaccinia virus/*genetics/*physiology
MH  - beta-Galactosidase/analysis/genetics
EDAT- 2005/05/17 09:00
MHDA- 2006/02/16 09:00
CRDT- 2005/05/17 09:00
PHST- 2004/06/25 00:00 [received]
PHST- 2004/11/09 00:00 [revised]
PHST- 2005/02/24 00:00 [accepted]
PHST- 2005/05/17 09:00 [pubmed]
PHST- 2006/02/16 09:00 [medline]
PHST- 2005/05/17 09:00 [entrez]
AID - S1286-4579(05)00106-1 [pii]
AID - 10.1016/j.micinf.2005.02.014 [doi]
PST - ppublish
SO  - Microbes Infect. 2005 Jul;7(9-10):1087-96. doi: 10.1016/j.micinf.2005.02.014. 
      Epub 2005 Apr 19.