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首页> 外文期刊>The Journal of Membrane Biology >In Vivo Molecular Imaging and Histological Analysis of Changes Induced by Electric Pulses Used for Plasmid DNA Electrotransfer to the Skin: A Study in a Dorsal Window Chamber in Mice
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In Vivo Molecular Imaging and Histological Analysis of Changes Induced by Electric Pulses Used for Plasmid DNA Electrotransfer to the Skin: A Study in a Dorsal Window Chamber in Mice

机译：用于质粒DNA电转移到皮肤的电脉冲引起的体内分子成像和组织学分析变化：小鼠背窗腔中的一项研究

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摘要

Electropermeabilization/electroporation (EP) is a physical method that by application of electric pulses to cells increases cell membrane permeability and enables the introduction of molecules into the cells. One of the uses of EP in vivo is plasmid DNA electrotransfer to the skin for DNA vaccination. EP of tissues induces reduction of blood flow and, in combination with plasmid DNA, induction of an immune response. One of the EP protocols for plasmid DNA electrotransfer to the skin is a combination of high-voltage (HV) and low-voltage (LV) pulses. However, the effects of this pulse combination on skin-vessel blood flow are not known. Therefore, using intravital microscopy in a dorsal window chamber in mice and fluorescently labeled dextrans, the effects of one HV and eight LV pulses on skin vasculature were investigated. In addition, a detailed histological analysis was performed. Image analysis of fluorescence intensity changes demonstrated that EP induces a transient constriction and increased permeability of blood vessels as well as a “vascular lock.” Histological analysis revealed rounding up of endothelial cells and stacking up of erythrocytes at 1 h after EP. In addition, extravasation of erythrocytes and leukocyte infiltration accompanied by edema were determined up to 24 h after EP. In conclusion, our results show that blood flow modifying effects of EP in skin contribute to the infiltration of immune cells in the exposed area. When combined with plasmid DNA for vaccination, this could enable the initial and prolonged contact of immune cells with encoded therapeutic proteins.
机译：电透化/电穿孔（EP）是一种物理方法，通过向细胞施加电脉冲来增加细胞膜的通透性，并使分子能够引入细胞中。 EP在体内的用途之一是质粒DNA电转移到皮肤上以进行DNA疫苗接种。组织的EP诱导血流量减少，并与质粒DNA结合诱导免疫反应。用于将质粒DNA电转移到皮肤的EP协议之一是高压（HV）和低压（LV）脉冲的组合。但是，这种脉冲组合对皮肤血管血流的影响尚不清楚。因此，在小鼠背窗腔内使用活体显微镜检查和荧光标记的葡聚糖，研究了一个HV和八个LV脉冲对皮肤血管的影响。另外，进行了详细的组织学分析。荧光强度变化的图像分析表明，EP会引起短暂收缩，血管通透性增加以及“血管闭锁”。组织学分析显示，EP后1 h内皮细胞聚集，红细胞堆积。另外，在EP后24小时内确定了红细胞外渗和白细胞浸润并伴有水肿。总之，我们的结果表明，皮肤中EP的血流调节作用有助于暴露区域中免疫细胞的浸润。当与质粒DNA结合用于疫苗接种时，这可以使免疫细胞与编码的治疗性蛋白质初始和长时间接触。

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