Name: granulocyte-dependent autoantibody-induced skin blistering
Uploaded: 2012-10-12T13:00:00
Description: Watch this Scientific Journal Video about Granulocyte-dependent Autoantibody-induced Skin Blistering at JoVE.com

作者承认，从德意志研究联合会SI-1281/4-1的和BIOSS从弗赖堡大学医学系（CS）的补助金。

1。致病抗体的制备注意：该抗体的纯化和浓度应在同一天进行，作为抗体是不是要被存储在0.1M甘氨酸pH为2.5-3（洗脱缓冲液）过夜（ON）。 亲和纯化的IgG：使用免疫兔血清25毫升： <ol><li>解冻后的兔血清对在4°C，混合，以1:1 1xPBS（运行缓冲液），并在1260 XG离心10分钟，在20°C。 ，用滤纸过滤步骤可能包括离心后的情况下，血清脂肪。 </li><li>同时，洗G矩...

<ol>
	<li>Sitaru, C., Mihai, S., Otto, C., Chiriac, M. T., Hausser, I., Dotterweich, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+dermal-epidermal+separation+in+mice+by+passive+transfer+of+antibodies+specific+to+type+VII+collagen.">Induction of dermal-epidermal separation in mice by passive transfer of antibodies specific to type VII collagen.</a> J. Clin. Invest. 115, 870-878 (2005).</li><li>Kopecki, Z., Arkell, R. M., Strudwick, X. L., Hirose, M., Ludwig, R. J., Kern, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overexpression+of+the+Flii+gene+increases+dermal-epidermal+blistering+in+an+autoimmune+ColVII+mouse+model+of+epidermolysis+bullosa+acquisita.">Overexpression of the Flii gene increases dermal-epidermal blistering in an autoimmune ColVII mouse model of epidermolysis bullosa acquisita.</a> J. Pathol. 225, 401-413 (2011).</li><li>Kulkarni, S., Sitaru, C., Jakus, Z., Anderson, K. E., Damoulakis, G., Davidson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PI3K&#946;+plays+a+critical+role+in+neutrophil+activation+by+immune+complexes.">PI3K&#946; plays a critical role in neutrophil activation by immune complexes.</a> Sci. Signal. 4, 23-23 (2011).</li><li>Chiriac, M. T., Roesler, J., Sindrilaru, A., Scharffetter-Kochanek, K., Zillikens, D., Sitaru, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NADPH+oxidase+is+required+for+neutrophil-dependent+autoantibody-induced+tissue+damage.">NADPH oxidase is required for neutrophil-dependent autoantibody-induced tissue damage.</a> J. Pathol. 212, 56-65 (2007).</li><li>Mihai, S., Chiriac, M. T., Takahashi, K., Thurman, J. M., Holers, V. M., Zillikens, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+alternative+pathway+of+complement+activation+is+critical+for+blister+induction+in+experimental+epidermolysis+bullosa+acquisita.">The alternative pathway of complement activation is critical for blister induction in experimental epidermolysis bullosa acquisita.</a> J. Immunol. 178, 6514-6521 (2007).</li><li>Sesarman, A., Sitaru, A. G., Olaru, F., Zillikens, D., Sitaru, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+Fc+receptor+deficiency+protects+from+tissue+injury+in+experimental+epidermolysis+bullosa+acquisita.">Neonatal Fc receptor deficiency protects from tissue injury in experimental epidermolysis bullosa acquisita.</a> J. Mol. Med. 86, 951-959 (2008).</li><li>Huang, X. R., Holdsworth, S. R., Tipping, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Th2+responses+induce+humorally+mediated+injury+in+experimental+anti-glomerular+basement+membrane+glomerulonephritis.">Th2 responses induce humorally mediated injury in experimental anti-glomerular basement membrane glomerulonephritis.</a> J. Am. Soc. Nephrol. 8, 1101-1108 (1997).</li><li>Matsumoto, I., Staub, A., Benoist, C., Mathis, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arthritis+provoked+by+linked+T+and+B+cell+recognition+of+a+glycolytic+enzyme.">Arthritis provoked by linked T and B cell recognition of a glycolytic enzyme.</a> Science. 286, 1732-1735 (1999).</li><li>Oswald, E., Sesarman, A., Franzke, C., W&#246;lfle, U., Bruckner-Tuderman, L., Jakob, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+flavonoid+luteolin+inhibits+Fc&#947;-dependent+respiratory+burst+in+granulocytes,+but+not+skin+blistering+in+a+new+model+of+pemphigoid+in+adult+mice.">The flavonoid luteolin inhibits Fc&#947;-dependent respiratory burst in granulocytes, but not skin blistering in a new model of pemphigoid in adult mice.</a> PLoS One. 7, e31066 (2012).</li><li>Liu, Z., Diaz, L. A., Troy, J. L., Taylor, A. F., Emery, D. J., Fairley, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+passive+transfer+model+of+the+organ-specific+autoimmune+disease,+bullous+pemphigoid,+using+antibodies+generated+against+the+hemidesmosomal+antigen,+BP180.">A passive transfer model of the organ-specific autoimmune disease, bullous pemphigoid, using antibodies generated against the hemidesmosomal antigen, BP180.</a> J. Clin. Invest. 92, 2480-2488 (1993).</li><li>Anhalt, G. J., Labib, R. S., Voorhees, J. J., Beals, T. F., Diaz, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+pemphigus+in+neonatal+mice+by+passive+transfer+of+IgG+from+patients+with+the+disease.">Induction of pemphigus in neonatal mice by passive transfer of IgG from patients with the disease.</a> N. Engl. J. Med. 306, 1189-1196 (1982).</li><li>Toyka, K. V., Brachman, D. B., Pestronk, A., Kao, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myasthenia+gravis:+passive+transfer+from+man+to+mouse.">Myasthenia gravis: passive transfer from man to mouse.</a> Science. 190, 397-399 (1975).</li><li>Rose, N. R., Bona, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defining+criteria+for+autoimmune+diseases+(Witebsky's+postulates+revisited.">Defining criteria for autoimmune diseases (Witebsky's postulates revisited.</a> Immunol. Today. 14, 426-430 (1993).</li><li>Sitaru, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+models+of+epidermolysis+bullosa+acquisita.">Experimental models of epidermolysis bullosa acquisita.</a> Exp. Dermatol. 16, 520-531 (2007).</li><li>Sitaru, C., Kromminga, A., Hashimoto, T., Br&#246;cker, E. B., Zillikens, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autoantibodies+to+type+VII+collagen+mediate+Fcgamma-dependent+neutrophil+activation+and+induce+dermal-epidermal+separation+in+cryosections+of+human+skin.">Autoantibodies to type VII collagen mediate Fcgamma-dependent neutrophil activation and induce dermal-epidermal separation in cryosections of human skin.</a> Am. J. Pathol. 161, 301-311 (2002).</li><li>Sesarman, A., Oswald, E., Chiriac, M. T., Csorba, K., Vuta, V., Feldrihan, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+human+pemphigoid+autoantibodies+do+not+trigger+disease+by+the+passive+transfer+into+mice.">Why human pemphigoid autoantibodies do not trigger disease by the passive transfer into mice.</a> Immunol. Lett. , (2012).</li><li>Ishii, N., Recke, A., Mihai, S., Hirose, M., Hashimoto, T., Zillikens, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autoantibody-induced+intestinal+inflammation+and+weight+loss+in+experimental+epidermolysis+bullosa+acquisita.">Autoantibody-induced intestinal inflammation and weight loss in experimental epidermolysis bullosa acquisita.</a> J. Pathol. 224, 234-244 (2011).</li></ol>

被动转移到实验动物的自身抗体是一个重要的方法展示他们的致病性，-12。通过这种方法获得的动物模型，除了是自身免疫13的间接证据，允许传出的调查阶段的致病机理。抗体诱导的粒细胞依赖的皮肤起泡性大疱性表皮松解症acquisita（EBA）的被动转移模型被用于解剖组织损伤的机制，在自身免疫性皮肤病。它的出现，作为一个精美的启发性疾病模型研究的根本，远远超出对胶原?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>重组抗原</td><td></td><td></td><td>的质粒编码的重组形式的小鼠的胶原VII可从相应的作者。 </td></tr><tr><td>免疫兔血清</td><td> <a href="http://www.eurogentec.com/" target="_blank">www.eurogentec.com</a> </td><td></td><td>我们有免疫200μg的抗原，在2周的时间间隔的3倍的新西兰白兔。为了这个目的，我们已经使用了服务Eurogentec SA，比利时。 </td></tr><tr><td> G蛋白琼脂糖</td><td>罗氏应用科学部</td><td> 11243233001 </td><td></td></tr><tr><td> Balb / c小鼠</td><td> Charles River实验室</td><td></td><td></TD&gt; </tr><tr><td> OCT化合物，组织康</td><td>樱花Finetek </td><td> 4583 </td><td> OCT，最佳切削温度</td></tr><tr><td> Cryomold标准</td><td>樱花Finetek </td><td> 4557 </td><td> 25毫米×20毫米×5毫米</td></tr><tr><td> Cryomold中间</td><td>樱花Finetek </td><td> 4566 </td><td> 15毫米×15毫米×5毫米</td></tr><tr><td>统一链接Einbettkassette的</td><td> R. Langenbrinck </td><td> 09-0503 </td><td>组织学处理和嵌入箱</td></tr><tr><td> SPEZIAL-Tatowierfarbe施瓦茨的</td><td> H. Hauptnerund和理查德Herberholz的GMBH＆CO KG </td><td> 71492000 </td><td>纹身贴</td></tr><tr><td> Tatowierzange TZ1 </td><td> EBECO E.贝克尔＆CO GMBH </td><td></td><td>纹身设备</td></tr><tr><td>肝素</td><td>卡尔·罗斯GMBH＆CO。 </td><td> 7692.1 </td><td></td></tr><tr><td>甲醛37％ </td><td>卡尔·罗斯GMBH＆CO。 </td><td> 7386 </td><td></td></tr><tr><td>盐酸氯胺酮</td><td> Sigma-Aldrich公司Chemie GmbH公司</td><td> K2753-1G </td><td></td></tr><tr><td>赛拉嗪盐酸盐</td><td> Sigma-Aldrich公司Chemie GmbH公司</td><td> X1251-1G </td><td></td></tr><tr><td>无菌PBS </td><td> Biochrom </td><td> L182-50 </td><td></td></tr><tr><td>数码相机</td><td>尼康</td><td> COOLPIX 5400数码相机</td><td></td></tr><tr><td>注射器驱动0.45微米过滤装置</td><td>密理博</td><td></td><td></td></tr><tr><td>卡尺</td><td>三丰7309 </td><td> 1667338 </td><td>派睿电子（经销商） </td></tr><tr><td>疾病计分表</td><td></td><td></td><td>例如机箱。中美战略经济对话</td></tr></tbody></table>

granulocyte-dependent autoantibody-induced skin blistering

自身免疫性现象发生在健康人，但，自我宽容失败时，可能会导致自体免疫反应在具体的病理。据Witebsky的假设，诊断为自身免疫性疾病中的标准之一是在实验动物中的疾病的再现由被动转移的自身抗体。大疱性表皮松解症acquisita（EBA），原型器官特异性自身免疫性疾病，皮肤和粘膜，一些实验模型，最近成立的。在动物模型中描述在我们目前的工作中，纯化的IgG抗体对拉伸的200个氨基酸（氨基酸757-967）的胶原纤维VII反复被注入到小鼠体内，再现起泡表型以及直方和免疫病理学特征对人体EBA 1。全面爆发普遍的疾病，通常第一次注射后5-6天，疾病的程度与剂量的管理Çollagen的VII-特异性IgG。在实验EBA组织损伤（水疱形成）根据粒细胞的募集和活化组织结合自身抗体-2，-4。因此，该模型允许粒细胞依赖的炎性途径参与自身免疫性组织损伤的解剖，作为模型只再现传出神经的自体免疫反应的T细胞独立的相位。此外，它的值被强调的一些研究表明自身抗体在体内的泡罩诱发电位和调查-6 EBA 1,3，水疱形成的机制。最后，这种模式将极大地促进新的自身抗体引起的疾病的消炎疗法的发展。总体而言，EBA被动转移动物模型的可访问性和指导性的疾病模型，将有助于研究人员不仅分析的的EBA发病，但回答基本的生物学和临床重要的自身免疫问题。

的被动转移抗原特异性抗体，结果在一个完全成熟的病鼠在临床，病理和免疫病理的水平，类似的人EBA。水疱，痂皮，糜烂和脱发发展的耳朵，猪嘴，爪，腿，背部和眼睛周围的老鼠。第一临床疾病的征兆，很可能会出现在耳朵上和/或头部区域。如果在中冰冻切片的病灶周围皮肤的真皮表皮交界处，检测到特异性的兔IgG和鼠补体C3沉积直接。在皮损部位表皮下疱和炎症浸润的组织学。 <p class="j...

Watch this Scientific Journal Video about Granulocyte-dependent Autoantibody-induced Skin Blistering at JoVE.com

Granulocyte-dependent Autoantibody-induced Skin Blistering

在动物模型中描述在我们目前的工作中，纯化的IgG抗体对拉伸的200个氨基酸（氨基酸757-967）的胶原纤维VII反复被注入到小鼠体内，再现起泡表型以及直方和免疫病理学特征对人体acquisita大疱性表皮松解症（EBA）<sup&gt; 1</sup&gt;。

粒细胞相关的自身抗体引起的皮肤起泡

Autoimmune phenomena occur in healthy individuals, but when self-tolerance fails, the autoimmune response may result in specific pathology. According to ...

Research

JoVE Journal

Immunology and Infection

Cutaneous Leishmaniasis in the Dorsal Skin of Hamsters: a Useful Model for the Screening of Antileishmanial Drugs

在仓鼠的背部皮肤：皮肤利什曼病Antileishmanial药物筛选的一个有用的模型

Traditionally, hamsters are experimentally inoculated in the snout or the footpad. However in these sites an ulcer not always occurs, measurement of ...

科研

免疫与感染

Imaging of HIV-1 Envelope-induced Virological Synapse and Signaling on Synthetic Lipid Bilayers

艾滋病毒的成像1信封引起的病毒学突触信号合成脂双层

Human immunodeficiency virus type 1 (HIV-1) infection occurs most efficiently via cell to cell transmission2,10,11. This cell to cell transfer between ...

Transplantation of Tail Skin to Study Allogeneic CD4 T Cell Responses in Mice

尾部皮肤来研究异基因CD4 + T细胞应答的小鼠移植

The study of T cell responses and their consequences during allo-antigen recognition requires a model that enables one to distinguish between donor and ...

Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture

Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture

从表型分析和人体皮肤淋巴细胞分离<em&gt;离体</em&gt;细胞培养

Human skin has an important barrier function and contains various immune cells that contribute to tissue homeostasis and protection from pathogens. As the ...

Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions

单细胞悬液的细胞荧光分析从不同的鼠标皮肤区域准备

The skin is a barrier organ that interacts with the external environment. Being continuously exposed to potential microbial invasion, the dermis and ...

Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation

从小鼠皮肤使用酶消化和梯度分离提取白细胞浸润的

Dissociating murine skin into a single cell suspension is essential for downstream cellular analysis such as the characterization of infiltrating immune ...

Myeloid Cell Isolation from Mouse Skin and Draining Lymph Node Following Intradermal Immunization with Live Attenuated Plasmodium Sporozoites

Myeloid Cell Isolation from Mouse Skin and Draining Lymph Node Following Intradermal Immunization with Live Attenuated Plasmodium Sporozoites

从老鼠皮肤及引流淋巴结髓细胞分离皮下继免疫减毒活<em&gt;疟原虫</em&gt;孢子

Malaria infection begins when the sporozoite stage of Plasmodium is inoculated into the skin of a mammalian host through a mosquito bite. The highly ...

Vasodilation of Isolated Vessels and the Isolation of the Extracellular Matrix of Tight-skin Mice

隔离容器的血管扩张和细胞外基质的紧皮肤小鼠隔离

The interferon regulatory factor 5 (IRF5) is crucial for cells to determine if they respond in a pro-inflammatory or anti-inflammatory fashion. IRF5's ...

Murine Full-thickness Skin Transplantation

鼠全层皮肤移植

Murine full-thickness skin transplantation is a well-established in vivo model to study alloimmune response and graft rejection. Despite its limited ...

Development of an Economical DNA Delivery System by "Acufection" and its Application to Skin Research

通过“Acufection”和它的皮肤研究中的应用的经济型DNA递送系统的开发

Dysregulation of immune response in skin is associated with numerous human skin disorders. Direct transfer of immune-related genes into skin tissue is a ...