Suppr超能文献

阳离子脂质体介导的基因在体外和体内向大鼠唾液腺上皮细胞的转移。

Cationic liposome-mediated gene transfer to rat salivary epithelial cells in vitro and in vivo.

作者信息

Baccaglini L, Shamsul Hoque A T, Wellner R B, Goldsmith C M, Redman R S, Sankar V, Kingman A, Barnhart K M, Wheeler C J, Baum B J

机构信息

Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892-1190, USA.

出版信息

J Gene Med. 2001 Jan-Feb;3(1):82-90. doi: 10.1002/1521-2254(2000)9999:9999<::AID-JGM151>3.0.CO;2-X.

Abstract

BACKGROUND

Previously we have shown that gene transfer to salivary gland epithelial cells readily occurs via recombinant adenoviruses, although the response is short-lived and results in a potent host immune response. The aim of the present study was to assess the feasibility of using cationic liposomes to mediate gene transfer to rat salivary cells in vitro and in vivo.

METHODS

Initially, for transfection in vitro, we used two cationic liposome formulations (GAP-DLRIE/DOPE and DOSPA/DOPE) complexed with plasmid encoding human growth hormone (hGH) as a reporter gene. Thereafter, using GAP-DLRIE/DOPE, plasmids were transferred to rat salivary glands in vivo, and hGH levels measured in saliva, serum and gland extracts.

RESULTS

Under optimal conditions, transfection of rat submandibular glands (SMGs) was consistently observed. Approximately 95% of the cells transfected with a plasmid encoding beta-galactosidase were acinar cells. Maximal hGH expression was obtained during the first 48 h post-transfection using a plasmid encoding the hGH cDNA and complexed with GAP-DLRIE/DOPE. hGH was detected in gland extracts and saliva, and occasionally in serum. No systemic or local gland pathology was consistently or significantly observed.

CONCLUSIONS

The levels of the reporter gene product, hGH, obtained after GAP-DLRIE/DOPE-mediated gene transfer are considerably lower (<0.5%) than those achieved with adenoviral vectors (10(8) PFU). Nonetheless, cationic liposome-mediated gene transfer to salivary glands may be useful for potential therapeutic applications.

摘要

背景

此前我们已经表明,尽管重组腺病毒介导的基因转移到唾液腺上皮细胞的反应是短暂的,并会引发强烈的宿主免疫反应,但这种基因转移很容易发生。本研究的目的是评估使用阳离子脂质体在体外和体内介导基因转移至大鼠唾液细胞的可行性。

方法

最初,为了进行体外转染,我们使用了两种与编码人生长激素(hGH)的质粒复合的阳离子脂质体制剂(GAP-DLRIE/DOPE和DOSPA/DOPE),hGH作为报告基因。此后,使用GAP-DLRIE/DOPE将质粒体内转移至大鼠唾液腺,并测量唾液、血清和腺体提取物中的hGH水平。

结果

在最佳条件下,始终观察到大鼠下颌下腺(SMG)的转染。用编码β-半乳糖苷酶的质粒转染的细胞中约95%为腺泡细胞。使用编码hGH cDNA并与GAP-DLRIE/DOPE复合的质粒,在转染后的前48小时内获得了最大的hGH表达。在腺体提取物和唾液中检测到hGH,偶尔在血清中也能检测到。未始终观察到明显的全身或局部腺体病理变化。

结论

GAP-DLRIE/DOPE介导的基因转移后获得的报告基因产物hGH的水平(低于0.5%)比腺病毒载体(10(8) PFU)介导的水平低得多。尽管如此,阳离子脂质体介导的基因转移至唾液腺可能对潜在的治疗应用有用。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验