Abstract
Gene transfer to the ocular surface epithelium is of potential therapeutic value. It was determined whether a reporter gene can be introduced into the ocular surface epithelium in vitro (human cell lines), ex vivo (human tissues), and in vivo (rats) by treating with a recombinant, replication-deficient, adenovirus type 5. Human and conjunctival cell lines were cultured with various multiplicities of infection (MOI; 3.2 x 10-5-5 x 10-1) of adenovirus vector (Ad5:Adex1CAlacZ) containing the reporter gene lacZ (1-3-2.0 x 104 PFU ml-1). The ex vivo study used human corneal and conjunctival tissues obtained from an eye bank and during surgery. Non- specific upregulation of inflammatory cytokines of conjunctival epithelium infected by AdS was assayed and its suppression by steroids. For the in vivo study, Ad5 (5 x 105 PFU, 5-10 μl) was applied to the eyes of 8-12-week-old cotton rats, which were enucleated 24 and 48 hr later. The maximum lacZ expression in vitro was demonstrated in the corneal epithelial cell line at 7 days (1 x 10- MOI) and conjunctival epithelial cell line at 2 days (4 x 10- 4 MOI). Furthermore, lacZ was also expressed in the superficial corneal and conjunctival epithelium in the ex vivo study. IL-6, 11-8, and ICAM-1 expression from conjunctival epithelium by Ad5 was significantly inhibited by treatment with betamethasone (BM). For the in vivo study, only the conjunctival epithelium demonstrated β-Gal activity at 24 and 48 hr after application. These data indicate that adenovirus vector is capable of directly delivering gene to the corneal and conjunctival epithelium, suggesting a variety of possible gene therapy uses. The concomitant application of steroid eye drops may avoid inflammation.
| Original language | English |
|---|---|
| Pages (from-to) | 531-538 |
| Number of pages | 8 |
| Journal | Experimental Eye Research |
| Volume | 67 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 1998 Nov |
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Keywords
- β-galactosidase
- Conjunctiva
- Dry eye
- Epithelium
- Ocular surface
ASJC Scopus subject areas
- Ophthalmology
- Sensory Systems
Cite this
Adenovirus-mediated gene transfer to the ocular surface epithelium. / Tsubota, Kazuo; Inoue, Hiroko; Ando, Kiyoshi; Ono, Masafumi; Yoshino, Kenichi; Saito, Ichiro.
In: Experimental Eye Research, Vol. 67, No. 5, 11.1998, p. 531-538.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Adenovirus-mediated gene transfer to the ocular surface epithelium
AU - Tsubota, Kazuo
AU - Inoue, Hiroko
AU - Ando, Kiyoshi
AU - Ono, Masafumi
AU - Yoshino, Kenichi
AU - Saito, Ichiro
PY - 1998/11
Y1 - 1998/11
N2 - Gene transfer to the ocular surface epithelium is of potential therapeutic value. It was determined whether a reporter gene can be introduced into the ocular surface epithelium in vitro (human cell lines), ex vivo (human tissues), and in vivo (rats) by treating with a recombinant, replication-deficient, adenovirus type 5. Human and conjunctival cell lines were cultured with various multiplicities of infection (MOI; 3.2 x 10-5-5 x 10-1) of adenovirus vector (Ad5:Adex1CAlacZ) containing the reporter gene lacZ (1-3-2.0 x 104 PFU ml-1). The ex vivo study used human corneal and conjunctival tissues obtained from an eye bank and during surgery. Non- specific upregulation of inflammatory cytokines of conjunctival epithelium infected by AdS was assayed and its suppression by steroids. For the in vivo study, Ad5 (5 x 105 PFU, 5-10 μl) was applied to the eyes of 8-12-week-old cotton rats, which were enucleated 24 and 48 hr later. The maximum lacZ expression in vitro was demonstrated in the corneal epithelial cell line at 7 days (1 x 10- MOI) and conjunctival epithelial cell line at 2 days (4 x 10- 4 MOI). Furthermore, lacZ was also expressed in the superficial corneal and conjunctival epithelium in the ex vivo study. IL-6, 11-8, and ICAM-1 expression from conjunctival epithelium by Ad5 was significantly inhibited by treatment with betamethasone (BM). For the in vivo study, only the conjunctival epithelium demonstrated β-Gal activity at 24 and 48 hr after application. These data indicate that adenovirus vector is capable of directly delivering gene to the corneal and conjunctival epithelium, suggesting a variety of possible gene therapy uses. The concomitant application of steroid eye drops may avoid inflammation.
AB - Gene transfer to the ocular surface epithelium is of potential therapeutic value. It was determined whether a reporter gene can be introduced into the ocular surface epithelium in vitro (human cell lines), ex vivo (human tissues), and in vivo (rats) by treating with a recombinant, replication-deficient, adenovirus type 5. Human and conjunctival cell lines were cultured with various multiplicities of infection (MOI; 3.2 x 10-5-5 x 10-1) of adenovirus vector (Ad5:Adex1CAlacZ) containing the reporter gene lacZ (1-3-2.0 x 104 PFU ml-1). The ex vivo study used human corneal and conjunctival tissues obtained from an eye bank and during surgery. Non- specific upregulation of inflammatory cytokines of conjunctival epithelium infected by AdS was assayed and its suppression by steroids. For the in vivo study, Ad5 (5 x 105 PFU, 5-10 μl) was applied to the eyes of 8-12-week-old cotton rats, which were enucleated 24 and 48 hr later. The maximum lacZ expression in vitro was demonstrated in the corneal epithelial cell line at 7 days (1 x 10- MOI) and conjunctival epithelial cell line at 2 days (4 x 10- 4 MOI). Furthermore, lacZ was also expressed in the superficial corneal and conjunctival epithelium in the ex vivo study. IL-6, 11-8, and ICAM-1 expression from conjunctival epithelium by Ad5 was significantly inhibited by treatment with betamethasone (BM). For the in vivo study, only the conjunctival epithelium demonstrated β-Gal activity at 24 and 48 hr after application. These data indicate that adenovirus vector is capable of directly delivering gene to the corneal and conjunctival epithelium, suggesting a variety of possible gene therapy uses. The concomitant application of steroid eye drops may avoid inflammation.
KW - β-galactosidase
KW - Conjunctiva
KW - Dry eye
KW - Epithelium
KW - Ocular surface
UR - http://www.scopus.com/inward/record.url?scp=0032413204&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032413204&partnerID=8YFLogxK
U2 - 10.1006/exer.1998.0557
DO - 10.1006/exer.1998.0557
M3 - Article
C2 - 9878215
AN - SCOPUS:0032413204
VL - 67
SP - 531
EP - 538
JO - Experimental Eye Research
JF - Experimental Eye Research
SN - 0014-4835
IS - 5
ER -