Femtosecond laser: a new intradermal DNA delivery method for efficient, long-term gene expression and genetic immunization
Femtosecond laser: a new intradermal DNA delivery method for efficient, long-term gene expression and genetic immunization
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DOI：
10.1096/fj.06-7528com
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发表时间：
2007-11-01
期刊：
影响因子：
4.8
通讯作者：
Galun, Eithan
中科院分区：
文献类型：
--
作者：
Zeira, Evelyne;Manevitch, Alexandra;Galun, Eithan
飞秒激光束基因转导（SG-LBGT）系统被描述为皮内（i.d.）利用飞秒激光脉冲透化细胞在小鼠中进行非病毒基因递送。使用这种方法，获得了持续> 7个月的显著基因表达和有效的皮肤转导。在BALB/C小鼠中测试了这种新的DNA基因转移方法增强基因疫苗接种的能力。只有一个身份证。注射含有乙肝B病毒（HBV）表面抗原（HBsAg）的质粒（10 μ g），然后进行激光脉冲，诱导持续> 210天的高滴度的HBsAg特异性抗体，并增加IgG 1、IgG 2 a、IFN γ和IL-4的水平，表明Th 1和Th 2细胞的活化。此外，使用SG-LBGT接种然后用pHBV攻击的小鼠显示出对病毒攻击的保护增加，如通过HBV DNA水平降低所检测到的，这表明对HBV感染的复制细胞的有效Th 1效应。在用表达HBsAg的同基因肿瘤攻击的接种小鼠中诱导肿瘤生长迟缓。在大多数测试的参数中，质粒给药后激光应用比单独质粒给药显著更有效和延长。未检测到组织损伤，可能未发生质粒整合到宿主基因组DNA中。我们认为，LBGT方法是一种有效和安全的技术，在体内基因表达和疫苗接种，并强调其潜在的治疗应用i.d.非病毒基因递送。
A femtosecond laser beam gene transduction (SG-LBGT) system is described as a novel and efficient method of intradermal (i.d.)non viralgene delivery in mice by permeabilizing cells utilizing femtosecond laser pulses. Using this approach, significant gene expression and efficient dermal transduction lasting for > 7 months were obtained. The ability of this new DNA gene transfer method to enhance genetic vaccination was tested in BALB/C mice. A single i.d. injection of a plasmid (10 mu g) containing the hepatitis B virus (HBV) surface antigen (HBsAg), followed by pulses of laser, induced high titers of HBsAg-specific antibodies lasting for > 210 days and increased levels of IgG1, IgG2a, IFN gamma, and IL-4, indicating the activation of both Th1 and Th2 cells. Moreover, mice vaccinated using the SG-LBGT followed by challenge with pHBV showed increased protection against viral challenge, as detected by decreased levels of HBV DNA, suggesting an efficient Th1 effect against HBV-infected replicating cells. Tumor growth retardation was induced in vaccinated mice challenged with an HBsAg-expressing syngeneic tumor. In most of the parameters tested, administration of plasmid followed by laser application was significantly more effective and prolonged than that of plasmid alone. Tissue damage was not detected and integration of the plasmid into the host genomic DNA probably did not occur. We suggest that the LBGT method is an efficient and safe technology for in vivo gene expression and vaccination and emphasizes its potential therapeutic applications for i.d. nonviral gene delivery.