Name: skin tattooing as a novel approach for dna vaccine delivery
Uploaded: 2012-10-18T13:00:00
Description: Watch this Scientific Journal Video about Skin Tattooing As A Novel Approach For DNA Vaccine Delivery at JoVE.com

我们想感谢所有成员的香港实验室和邓焰博士为的显微镜核心，办公室协同科学，NYUMC他们的援助和技术支持。这项工作是支持的试点补助金从纽约大学艾滋病研究中心（CFAR，美国国立卫生研究院授予AI027742）。

1。质粒DNA纯化<ol><li>变换到DH5α 大肠杆菌感受态细胞的真核质粒DNA编码EGFP（PCX-EGFP）。空PCX载体也可以使用作为阴性对照。 </li><li>文化和收获DH5α 大肠杆菌细胞和纯化的质粒DNA根据到的Qiagen公司EndoFree的质粒纯化手册“。 </li><li>筛选pDNA的溶液通过0.22μm的PVDF无菌过滤器，并将其储存在-20℃直到使用。 </li></ol> 2。纹身系统准备<ol><li>手持式单元和控制踏板连接到?...

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	<li>Bins, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rapid+and+potent+DNA+vaccination+strategy+defined+by+in+vivo+monitoring+of+antigen+expression.">A rapid and potent DNA vaccination strategy defined by in vivo monitoring of antigen expression.</a> Nat Med. 11, 899-904 (2005).</li><li>Pokorna, D., Rubio, I., Muller, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA-vaccination+via+tattooing+induces+stronger+humoral+and+cellular+immune+responses+than+intramuscular+delivery+supported+by+molecular+adjuvants.">DNA-vaccination via tattooing induces stronger humoral and cellular immune responses than intramuscular delivery supported by molecular adjuvants.</a> Genet. Vaccines Ther. 6, 4 (2008).</li><li>van den Berg, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+intradermal+vaccination+by+DNA+tattooing+in+human+skin.">Optimization of intradermal vaccination by DNA tattooing in human skin.</a> Hum. Gene Ther. 20, 181-189 (2009).</li><li>Liu, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA+vaccines:+a+review.">DNA vaccines: a review.</a> J. Intern. Med. 253, 402-410 (2003).</li><li>Koide, Y., Nagata, T., Yoshida, A., Uchijima, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA+vaccines.">DNA vaccines.</a> Jpn. J. Pharmacol. 83, 167-174 (2000).</li><li>Peachman, K. K., Rao, M., Alving, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunization+with+DNA+through+the+skin.">Immunization with DNA through the skin.</a> Methods. 31, 232-242 (2003).</li><li>Verstrepen, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+HIV-1+specific+T-cell+responses+by+short-interval+DNA+tattooing+as+compared+to+intramuscular+immunization+in+non-human+primates.">Improved HIV-1 specific T-cell responses by short-interval DNA tattooing as compared to intramuscular immunization in non-human primates.</a> Vaccine. 26, 3346-3351 (2008).</li><li>Potthoff, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunogenicity+and+efficacy+of+intradermal+tattoo+immunization+with+adenoviral+vector+vaccines.">Immunogenicity and efficacy of intradermal tattoo immunization with adenoviral vector vaccines.</a> Vaccine. 27, 2768-2774 (2009).</li><li>Quaak, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA+tattoo+vaccination:+effect+on+plasmid+purity+and+transfection+efficiency+of+different+topoisoforms.">DNA tattoo vaccination: effect on plasmid purity and transfection efficiency of different topoisoforms.</a> J. Control Release. 139, 153-159 (2009).</li><li>Pokorna, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vaccination+with+human+papillomavirus+type+16-derived+peptides+using+a+tattoo+device.">Vaccination with human papillomavirus type 16-derived peptides using a tattoo device.</a> Vaccine. 27, 3519-3529 (2009).</li></ol>

DNA疫苗比传统的疫苗接种策略，被认为是更安全，因为它不要求操作，或暴露疫苗，活的或减毒的病原体。然而，DNA疫苗接种的结果，在很大程度上依赖于配送路线。皮肤是丰富的抗原呈递细胞，如朗格汉斯细胞和树突状细胞1，从而在一个理想的站点用于免疫的免疫原性和易于访问5,6。其结果是，皮内接种策略是用于DNA疫苗的最流行的选择之一。在这个视频中，DNA纹身是一个简?...

<table border="1"><tbody><tr><td>的试剂/材料名称</td><td>公司</td><td>目录编号</td><td>评论</td></tr><tr><td> PCX-EGFP质粒DNA </td><td> Clontech公司</td><td></td><td></td></tr><tr><td> STEALTH旋转纹身系统</td><td>全球纹身器材</td><td> STEALTH-L </td><td></td></tr><tr><td>纹身针</td><td>全球纹身器材</td><td> 1207RSB </td><td></td></tr><tr><td> EndoFree质粒试剂盒</td><td> Qiagen公司</td><td> 12362 </td><td></td></tr><tr><td> 0.22微米PVDF无菌过滤器</td><td>密理博</td><td> SLGV013SL </td><td></td></tr><tr><td>电发剪</td><td>市售</td><td></td><td></td></tr><tr><td>一次性使用的安全剃须刀</td><td>市售</td><td></td><td></td></tr><tr><td>银磺胺嘧啶霜</td><td>沃森</td><td> NDC 0591-0810-55 </td><td></td></tr><tr><td>氯胺酮HCL </td><td></td><td> NDC 0856-2012-01 </td><td></td></tr><tr><td> Zylazine无菌溶液</td><td></td><td> NADA 139-236 </td><td></td></tr></tbody></table>

skin tattooing as a novel approach for dna vaccine delivery

基于核酸的疫苗接种是一个主题的日益增长的兴趣，特别是质粒DNA（pDNA的）编码免疫学上重要的抗原。工程质粒DNA疫苗给药后，它被转录和翻译成免疫原的蛋白质，可以从免疫系统引起的反应。提供DNA疫苗的方法很多被调查，但每次送货路线都有自己的优点和缺陷。皮肤纹身是一种新的技术是安全的，符合成本效益，方便的。此外，穿刺针造成，也可以作为一种有效的佐剂。在这里，我们a）表现出增强型绿色荧光蛋白的质粒DNA编码的皮内交付（PCX-EGFP）在小鼠模​​型中，使用纹身的移动设备和b）在皮肤细胞中，使用共聚焦显微镜，确认有效的EGFP表达。

在小鼠皮肤细胞中可以观察到在488nm和发射峰在509 nm处的激发峰的EGFP的表达。从1.875μg剂量的DNA，含有约3×10 17个拷贝的质粒，我们通常观察到10-20的EGFP信号在1厘米2纹身面积。这种相对较低的转染细胞的数量是先前的研究3的结果相一致。 EGFP表达（ 图1）提供的证据表明，EGFP质粒被输送到动物的皮肤细胞，使用的DNA纹身技术。 <p class="jove_content" fo:keep-togethe...

Watch this Scientific Journal Video about Skin Tattooing As A Novel Approach For DNA Vaccine Delivery at JoVE.com

Skin Tattooing As A Novel Approach For DNA Vaccine Delivery

皮肤纹身是一个强大的和安全的方式来传递DNA疫苗皮内。在这里，一个DNA质粒EGFP交付纹身实验室小鼠的皮肤，和EGFP在皮肤细胞中的表达，然后通过激光共聚焦显微镜检查。

皮肤纹身作为一种新的方法DNA疫苗交货

Nucleic acid-based vaccination is a topic of growing interest, especially plasmid DNA (pDNA) encoding immunologically important antigens. After the ...

Research

JoVE Journal

Bioengineering

Cell Squeezing as a Robust, Microfluidic Intracellular Delivery Platform

电池挤压作为一种强大的，微流控细胞内传送平台

Rapid mechanical deformation of cells has emerged as a promising, vector-free method for intracellular delivery of macromolecules and nanomaterials. This ...

科研

生物工程

An Improved Method for the Preparation of Type I Collagen From Skin

对于I型胶原蛋白的制备皮肤方法的改进

Soluble type 1 collagen (COL1) is used extensively as an adhesive substrate for cell cultures and as a cellular scaffold for regenerative applications. ...

Characterization of Complex Systems Using the Design of Experiments Approach: Transient Protein Expression in Tobacco as a Case Study

烟草中瞬时表达的蛋白作为一个案例研究：复杂系统采用的实验方法设计的表征

Plants provide multiple benefits for the production of biopharmaceuticals including low costs, scalability, and safety. Transient expression offers the ...

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

一个完整的皮肤缺损模型，以评估生物材料的血管化<em&gt;体内</em

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to ...

Microscale Vortex-assisted Electroporator for Sequential Molecular Delivery

微型涡辅助电穿孔的顺序传递分子

Electroporation has received increasing attention in the past years, because it is a very powerful technique for physically introducing non-permeant ...

Fabrication of a Functionalized Magnetic Bacterial Nanocellulose with Iron Oxide Nanoparticles

一机能磁性纳米纤维素的细菌与氧化铁纳米粒子的制备

In this study, bacterial nanocellulose (BNC) produced by the bacteria Gluconacetobacter xylinus is synthesized and impregnated in situ with iron oxide ...

Development of a Multicellular Three-dimensional Organotypic Model of the Human Intestinal Mucosa Grown Under Microgravity

生长在微重力人体肠道黏膜的多细胞三维器官模型的建立

Because cells growing in a three-dimensional (3-D) environment have the potential to bridge many gaps of cell cultivation in 2-D environments (e.g., ...

Mesenchymal Stromal Cell Culture and Delivery in Autologous Conditions: A Smart Approach for Orthopedic Applications

间质干细胞培养和交付自体条件：用于骨科应用程序的智能方法

Human Mesenchymal Stromal Cells (hMSCs) are cultured in vitro with different media. Limits on their use in clinical settings, however, mainly depend on ...

DNA Nanotubes as a Versatile Tool to Study Semiflexible Polymers

DNA 纳米管作为研究半聚合物的万能工具

Mechanical properties of complex, polymer-based soft matter, such as cells or biopolymer networks, can be understood in neither the classical frame of ...

Bioprinting of Cartilage and Skin Tissue Analogs Utilizing a Novel Passive Mixing Unit Technique for Bioink Precellularization

Bioink Precellularization 无源混合装置打印软骨及皮肤组织类似物的应用

Bioprinting is a powerful technique for the rapid and reproducible fabrication of constructs for tissue engineering applications. In this study, both ...