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Ming-Ling Tsai, Show-Li Chen, Ping-I Chou, Liang-Yen Wen, Ray Jui-Fang Tsai, Yeou-Ping Tsao; Inducible Adeno-Associated Virus Vector–Delivered Transgene Expression in Corneal Endothelium. Invest. Ophthalmol. Vis. Sci. 2002;43(3):751-757.
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purpose. To investigate whether recombinant adeno-associated virus (rAAV)
vector–mediated transgene expression is induced by inflammation in
corneal endothelial cells in vivo.
methods. The ocular anterior chamber of New Zealand White rabbits was injected
with rAAV-LacZ (107 units of infection).
Transient ocular anterior segment inflammation was induced by an
intravitreal injection of lipopolysaccharide (LPS). The effect of
inflammation on LacZ gene expression in corneal
endothelial cells was evaluated by histochemical staining and reverse
transcription–polymerase chain reaction (RT-PCR). The influence of
rAAV on endothelial cell function was monitored by measuring corneal
results. Inflammatory reaction peaked at 1 day after LPS treatment and, at the
same time, most of the endothelial cells (91.3% ± 7.2%) showed
prominent LacZ gene expression. The transgene expression
gradually diminished to basal level (3.4% ± 2.1%) when the
inflammation subsided at 15 days after LPS treatment. The diminished
transgene expression was efficiently reactivated to a high level
(86.1% ± 8.7%) by a second LPS injection 60 days later. Moreover,
the transgene expression remained low for a long period (60 days) in
the absence of LPS treatment, but was increased to high levels (87.3%±
8.1%) 1 day after LPS treatment. Throughout the observation period,
endothelial cell function remained intact.
conclusions. The rAAV vector can deliver genes into endothelial cells, and transgene
expression is dramatically induced by inflammation. The rAAV-delivered
transgene is stable and does not compromise endothelial cell function.
Inducible rAAV-mediated transgene expression in corneal endothelial
cells is a potential strategy in the treatment and prevention of ocular
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