PMID- 10102690
OWN - NLM
STAT- MEDLINE
DCOM- 19990415
LR  - 20190515
IS  - 0012-1797 (Print)
IS  - 0012-1797 (Linking)
VI  - 48
IP  - 4
DP  - 1999 Apr
TI  - Gene transfer to human pancreatic endocrine cells using viral vectors.
PG  - 745-53
AB  - We have studied the factors that influence the efficiency of infection of human 
      fetal and adult pancreatic endocrine cells with adenovirus, murine retrovirus, 
      and lentivirus vectors all expressing the green fluorescent protein (Ad-GFP, 
      MLV-GFP, and Lenti-GFP, respectively). Adenoviral but not retroviral vectors 
      efficiently infected intact pancreatic islets and fetal islet-like cell clusters 
      (ICCs) in suspension. When islets and ICCs were plated in monolayer culture, 
      infection efficiency with all three viral vectors increased. Ad-GFP infected 
      90-95% of the cells, whereas infection with MLV-GFP and Lenti-GFP increased only 
      slightly. Both exposure to hepatocyte growth factor/scatter factor (HGF/SF) and 
      dispersion of the cells by removal from the culture dish and replating had 
      substantial positive effects on the efficiency of infection with retroviral 
      vectors. Studies of virus entry and cell replication revealed that cell 
      dispersion and stimulation by HGF/SF may be acting through both mechanisms to 
      increase the efficiency of retrovirus-mediated gene transfer. Although HGF/SF and 
      cell dispersion increased the efficiency of infection with MLV-GFP, only rare 
      cells with weak staining for insulin were infected, whereas approximately 25% of 
      beta-cells were infected with Lenti-GFP. We conclude that adenovirus is the most 
      potent vector for ex vivo overexpression of foreign genes in adult endocrine 
      pancreatic cells and is the best vector for applications where high-level but 
      transient expression is desired. Under the optimal conditions of cell dispersion 
      plus HGF/SF, infection with MLV and lentiviral vectors is reasonably efficient 
      and stable, but only lentiviral vectors efficiently infect pancreatic beta-cells.
FAU - Leibowitz, G
AU  - Leibowitz G
AD  - Center for Molecular Genetics, Whittier Institute, UCSD School of Medicine, 
      University of California, San Diego, USA.
FAU - Beattie, G M
AU  - Beattie GM
FAU - Kafri, T
AU  - Kafri T
FAU - Cirulli, V
AU  - Cirulli V
FAU - Lopez, A D
AU  - Lopez AD
FAU - Hayek, A
AU  - Hayek A
FAU - Levine, F
AU  - Levine F
LA  - eng
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  - Diabetes
JT  - Diabetes
JID - 0372763
SB  - IM
MH  - Adenoviridae Infections/pathology
MH  - Cells, Cultured
MH  - Cytological Techniques
MH  - Fetus/physiology
MH  - *Gene Transfer Techniques
MH  - *Genetic Vectors
MH  - Humans
MH  - Islets of Langerhans/embryology/*physiology/virology
MH  - Lentivirus/physiology
MH  - Lentivirus Infections/virology
MH  - Mitosis/physiology
MH  - Moloney murine leukemia virus/physiology
MH  - Retroviridae Infections/pathology/virology
MH  - Rhabdoviridae Infections/virology
MH  - Tumor Virus Infections/virology
MH  - Vesicular stomatitis Indiana virus/classification/physiology
MH  - Viruses/*genetics
EDAT- 1999/04/02 00:00
MHDA- 1999/04/02 00:01
CRDT- 1999/04/02 00:00
PHST- 1999/04/02 00:00 [pubmed]
PHST- 1999/04/02 00:01 [medline]
PHST- 1999/04/02 00:00 [entrez]
AID - 10.2337/diabetes.48.4.745 [doi]
PST - ppublish
SO  - Diabetes. 1999 Apr;48(4):745-53. doi: 10.2337/diabetes.48.4.745.