Kupffer cells and not liver sinusoidal endothelial cells prevent lentiviral transduction of hepatocytes.
Lentiviral vectors can stably transduce dividing and nondividing cells in vivo and are best suited to long-term correction of inherited liver diseases. Intraportal administration of lentiviral vectors expressing green fluorescent protein (Lenti-GFP) in mice resulted in a higher transduction of nonparenchymal cells than hepatocytes (7.32 +/- 3.66% vs 0.22 +/- 0.08%, respectively). Therefore, various treatments were explored to increase lentiviral transduction of hepatocytes. Lenti-GFP was injected into the common bile duct, which led to transduction of biliary epithelium and hepatocytes at low efficiency. Transient removal of the sinusoidal endothelial cell layer by cyclophosphamide to increase accessibility to hepatocytes did not improve hepatocyte transduction (0.42 +/- 0.36%). Inhibition of Kupffer cell function by gadolinium chloride led to a significant decrease in GFP-positive nonparenchymal cells (2.15 +/- 3.14%) and a sevenfold increase in GFP-positive hepatocytes compared to nonpretreated mice (1.48 +/- 2.01%). These findings suggest that sinusoidal endothelial cells do not significantly limit lentiviral transduction of hepatocytes, while Kupffer cells sequester lentiviral particles thereby preventing hepatocyte transduction. Therefore, the use of agents that inhibit Kupffer cell function may be important for lentiviral vector treatment of liver disease.
van Til
Niek P
NP
AMC Liver Center, S1-172, Meibergdreef 69, 1105 BK Amsterdam, The Netherlands. n.p.vantil@amc.uva.nl
Markusic
David M
DM
van der Rijt
Roos
R
Kunne
Cindy
C
Hiralall
Johan K
JK
Vreeling
Heleen
H
Frederiks
Wilma M
WM
Oude-Elferink
Ronald P J
RP
Seppen
Jurgen
J
D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D001652:Bile Ducts / Q000821:virology; D002460:Cell Line; D003520:Cyclophosphamide / Q000494:pharmacology; D042783:Endothelial Cells / Q000473:pathology / Q000821:virology; D005682:Gadolinium / Q000494:pharmacology; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism / Q000821:virology*; D007728:Kupffer Cells / Q000473:pathology / Q000502:physiology*; D016086:Lentivirus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D008099:Liver / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D010587:Phagocytosis; D016133:Polymerase Chain Reaction; D011169:Portal Vein / Q000821:virology; D014161:Transduction, Genetic / Q000379:methods*
D003520:Cyclophosphamide; D005682:Gadolinium; D000410:Alanine Transaminase; C038958:gadolinium chloride