Adenovirus-mediated gene transfer to the ocular surface epithelium.
Adenovirus-mediated gene transfer to the ocular surface epithelium.
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腺病毒介导的基因转移至眼表上皮。
DOI：
10.1006/exer.1998.0557
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发表时间：
1998
影响因子：
3.4
通讯作者：
I. Saito
中科院分区：
文献类型：
--
作者：
K. Tsubota;H. Inoue;K. Ando;M. Ono;K. Yoshino;I. Saito
眼表上皮的基因转移具有潜在的治疗价值。通过用重组复制缺陷型腺病毒5型处理，确定报告基因是否可以在体外（人细胞系）、离体（人组织）和体内（大鼠）引入眼表面上皮。将人和结膜细胞系与含有报告基因lacZ（1.3-2.0x10（4）PFU ml-1）的腺病毒载体（Ad 5：Adex 1CAlacZ）的各种感染复数（MOI; 3.2x10（-5）-5x10（-1））一起培养。离体研究使用从眼库和手术期间获得的人角膜和结膜组织。检测Ad 5感染的结膜上皮炎性细胞因子的非特异性上调，并检测类固醇对其的抑制。对于体内研究，将Ad 5（5 × 10（5）PFU，5-10微升）施加到8-12周龄棉鼠的眼睛，在24和48小时后将其摘除。在体外，在角膜上皮细胞系中在第7天（1 × 10（-1）MOI）和结膜上皮细胞系中在第2天（4 × 10（-4）MOI）证实了最大lacZ表达。此外，在离体研究中，lacZ也在浅层角膜和结膜上皮中表达。倍他米松（BM）处理可显着抑制Ad 5从结膜上皮表达IL-6、IL-8和ICAM-1。对于体内研究，仅结膜上皮在施用后24和48小时表现出β-Gal活性。这些数据表明腺病毒载体能够直接将基因递送到角膜和结膜上皮，提示了多种可能的基因治疗用途。同时应用类固醇滴眼液可避免炎症。
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; 3.2x10(-5)-5x10(-1)) of adenovirus vector (Ad5:Adex1CAlacZ) containing the reporter gene lacZ (1.3-2.0x10(4) 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 Ad5 was assayed and its suppression by steroids. For the in vivo study, Ad5 (5x10(5) PFU, 5-10 microliter) 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 (1x10(-1) MOI) and conjunctival epithelial cell line at 2 days (4x10(-4) MOI). Furthermore, lacZ was also expressed in the superficial corneal and conjunctival epithelium in the ex vivo study. IL-6, IL-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 beta-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.
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作者：
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DOI：
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发表时间：
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期刊：
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