PMID- 9765435
OWN - NLM
STAT- MEDLINE
DCOM- 19981105
LR  - 20240214
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 72
IP  - 11
DP  - 1998 Nov
TI  - Factors influencing adeno-associated virus-mediated gene transfer to human cystic 
      fibrosis airway epithelial cells: comparison with adenovirus vectors.
PG  - 8904-12
AB  - Adeno-associated virus (AAV) vectors appear promising for use in gene therapy in 
      cystic fibrosis (CF) patients, yet many features of AAV-mediated gene transfer to 
      airway epithelial cells are not well understood. We compared the transduction 
      efficiencies of AAV vectors and adenovirus (Ad) vectors in immortalized cell 
      lines from CF patients and in nasal epithelial primary cultures from normal 
      humans and CF patients. Similar dose-dependent relationships between the vector 
      multiplicities of infection and the efficiencies of lacZ gene transfer were 
      observed. However, levels of transduction for both Ad and recombinant AAV (rAAV) 
      were significantly lower in the airway epithelial cell than in the control cell 
      lines HeLa and HEK 293. Transduction efficiencies differed among cultured 
      epithelial cell types, with poorly differentiated cells transducing more 
      efficiently than well-differentiated cells. A time-dependent increase in gene 
      expression was observed after infection for both vectors. For Ad, but not for 
      AAV, this increase was dependent on prolonged incubation of cells with the 
      vector. Furthermore, for rAAV (but not for rAd), the delay in maximal 
      transduction could be abrogated by wild-type Ad helper infection. Thus, although 
      helper virus is not required for maximal transduction, it increases the kinetics 
      by which this is achieved. Expression of Ad E4 open reading frame 6 or addition 
      of either hydroxyurea or camptothecin resulted in increased AAV transduction, as 
      previously demonstrated for nonairway cells (albeit to lower final levels), 
      suggesting that second-strand synthesis may not be the sole cause of inefficient 
      transduction. Finally, the efficiency of AAV-mediated ex vivo gene transfer to 
      lung cells was similar to that previously described for Ad vectors in that 
      transduction was limited to regions of epithelial injury and preferentially 
      targeted basal-like cells. These studies address the primary factors influencing 
      rAAV infection of human airway cells and should impact successful gene delivery 
      in CF patients.
FAU - Teramoto, S
AU  - Teramoto S
AD  - CF/Pulmonary Research and Treatment Center, University of North Carolina at 
      Chapel Hill, Chapel Hill, North Carolina, USA.
FAU - Bartlett, J S
AU  - Bartlett JS
FAU - McCarty, D
AU  - McCarty D
FAU - Xiao, X
AU  - Xiao X
FAU - Samulski, R J
AU  - Samulski RJ
FAU - Boucher, R C
AU  - Boucher RC
LA  - eng
GR  - P01 HL051818/HL/NHLBI NIH HHS/United States
GR  - HL 42384/HL/NHLBI NIH HHS/United States
GR  - HL 51818/HL/NHLBI NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (CFTR protein, human)
RN  - 126880-72-6 (Cystic Fibrosis Transmembrane Conductance Regulator)
SB  - IM
MH  - Adenoviruses, Human/*genetics
MH  - Cell Line
MH  - Cells, Cultured
MH  - Cystic Fibrosis/*genetics/*therapy
MH  - Cystic Fibrosis Transmembrane Conductance Regulator/genetics
MH  - Dependovirus/*genetics
MH  - Epithelial Cells/metabolism/virology
MH  - *Gene Transfer Techniques
MH  - *Genetic Vectors
MH  - Humans
MH  - Lac Operon
MH  - Nasal Mucosa/metabolism/virology
MH  - Trachea/metabolism/virology
MH  - Transduction, Genetic
PMC - PMC110307
EDAT- 1998/10/10 00:00
MHDA- 1998/10/10 00:01
PMCR- 1998/11/01
CRDT- 1998/10/10 00:00
PHST- 1998/10/10 00:00 [pubmed]
PHST- 1998/10/10 00:01 [medline]
PHST- 1998/10/10 00:00 [entrez]
PHST- 1998/11/01 00:00 [pmc-release]
AID - 0784 [pii]
AID - 10.1128/JVI.72.11.8904-8912.1998 [doi]
PST - ppublish
SO  - J Virol. 1998 Nov;72(11):8904-12. doi: 10.1128/JVI.72.11.8904-8912.1998.