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

被引:0
作者
Baccaglini, L
Hoque, ATMS
Wellner, RB
Goldsmith, CM
Redman, RS
Sankar, V
Kingman, A
Barnhart, KM
Wheeler, CJ
Baum, BJ
机构
[1] Natl Inst Dent & Craniofacial Res, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD 20892 USA
[2] Dept Vet Affairs Med Ctr, Oral Pathol Res Lab, Washington, DC 20422 USA
[3] Natl Inst Dent & Craniofacial Res, Craniofacial Epidemiol & Genet Branch, NIH, Bethesda, MD 20892 USA
[4] Vical Inc, San Diego, CA 92121 USA
关键词
transfection; cationic lipid; salivary gland;
D O I
10.1002/1521-2254(2000)9999:9999<::AID-JGM151>3.0.CO;2-X
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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. Copyright (C) 2000 John Wiley & Sons, Ltd.
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页码:82 / 90
页数:9
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