Name: the citrobacter rodentium mouse model: studying pathogen and host contributions to infectious colitis
Uploaded: 2013-02-19T13:00:00
Description: Watch this Scientific Journal Video about The Citrobacter rodentium Mouse Model: Studying Pathogen and Host Contributions to Infectious Colitis at JoVE.com

这项工作是支持的补助金BAV的克罗恩病和结肠炎基金会和加拿大卫生研究院（CIHR）加拿大（CCFC）。 GB CIHR研究生奖学金的资助。 BAV是的儿童肠和肝脏疾病（儿童）基金会主席儿科IBD的研究和加拿大研究主席在儿科胃肠病学。

1。 枸橼酸rodentium接种物的制备和小鼠灌胃<ol><li>准备和高压灭菌卢里亚Bertani肉汤（LB）。 </li><li>获得可行的C. LB琼脂平板上，用无菌接种环或移液器吸头rodentium从冷冻甘油股票和条纹上。孵育，37℃培养过夜。 ，接种3毫升无菌的LB肉汤猎鹰文化的LB平板上的菌落，用接种环或吸液管头管。应做的接种物的制备，使用无菌技术。 </li><li>孵育C. rodentium文化有氧，在37°C在2...

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A., Kalman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TLR+signaling+mediated+by+MyD88+is+required+for+a+protective+innate+immune+response+by+neutrophils+to+Citrobacter+rodentium.">TLR signaling mediated by MyD88 is required for a protective innate immune response by neutrophils to Citrobacter rodentium.</a> Journal of Immunology. 179 (1), 566-577 (2007).</li><li>Bry, L., Brenner, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Critical+role+of+T+cell+dependent+serum+antibody,+but+not+the+gut-associated+lymphoid+tissue,+for+surviving+acute+mucosal+infection+with+Citrobacter+rodentium,+an+attaching+and+effacing+pathogen.">Critical role of T cell dependent serum antibody, but not the gut-associated lymphoid tissue, for surviving acute mucosal infection with Citrobacter rodentium, an attaching and effacing pathogen.</a> Journal of Immunology. 172 (1), 433-441 (2004).</li><li>Simmons, C. P., Clare, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+role+for+B+lymphocytes+and+CD4++T+cells+in+immunity+to+infection+by+the+attaching+and+effacing+pathogen+Citrobacter+rodentium.">Central role for B lymphocytes and CD4+ T cells in immunity to infection by the attaching and effacing pathogen Citrobacter rodentium.</a> Infection &amp; Immunity. 71 (9), 5077-5086 (2003).</li><li>Ahmed, I., Chandrakesan, P., Tawfik, O., Xia, L., Anant, S., Umar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Critical+Roles+of+Notch+and+Wnt/&#946;-Catenin+Pathways+in+the+Regulation+of+Hyperplasia+and/or+Colitis+in+Response+to+Bacterial+Infection.">Critical Roles of Notch and Wnt/&#946;-Catenin Pathways in the Regulation of Hyperplasia and/or Colitis in Response to Bacterial Infection.</a> Infection &amp; Immunity. 80 (9), 3107-3121 (2012).</li></ol>

枸橼酸rodentium感染传染病的小鼠结肠炎的研究提供了一个有价值的模型。这种独特的模式允许两个宿主反应，以及细菌的致病特性的表征。这种模式的成功使用该协议的细节中列出的步骤。 有几个关键的步骤，在此协议时，请记住诱导结肠炎和分析反应。首先，准备一个新鲜的过夜C. rodentium在LB肉汤接种感染小鼠的成功。由于需要的剂量为10 8 -10 9</s...

肠道细菌病原体感染引发胃肠道（GI）的炎症，以及肠道病理学与病理生理学，包括腹泻和肠上皮屏障功能障碍。细菌病原体的殖民他们的主机，以及具体的抵御这种感染的宿主反应，胃肠道的致病机制了解甚少，但最近的进步已经开始在肠道细菌感染的造型，以帮助我们了解这些过程。肠道细菌也被连接到的炎症性肠病（IBDs）。 IBDs克罗恩病（CD）和UC复杂疾病的病因不明的慢性肠道炎症和组织损伤，其特征。许多模型小鼠肠道炎症的存在，从在遗传修饰的菌株，如IL-10 - / - 小鼠的自发炎症的化合物，如葡聚糖硫酸钠（DSS）和d，化学挑战initrobenzene磺酸（DNBS）1。据推测，适应不良的慢性炎症反应在IBD患者在遗传易感个体发展后肠黏膜免疫系统的异常暴露的肠道细菌，因此感染性结肠炎模型的研究也提供了显着的潜力，确定潜在的致病主机肠道细菌的反应。 枸橼酸杆菌rodentium诱导的结肠炎是罕见的模式已感染性结肠炎，以及其特征在于1,3之一，允许对分析的宿主反应，以肠溶性的细菌和潜在的IBD发病机制的进一步理解，一个重要的步骤在开发新的预防和治疗方法。 C. rodentium是革兰氏阴性安装和谦虚（A / E），小鼠的特异性细菌性病原体是密切相关的人体重要病原体肠致病性大肠杆菌大肠杆菌 （EPEC），肠出血性大肠杆菌大肠杆菌 （EHEC）3-8。的家庭的A / E病原体密切附加到根尖盲肠和结肠上皮细胞的宿主细胞的膜，形成一种非侵入性的台座状结构的宿主细胞。口腔挑战C. rodentium 10 8 -10 9生物体产生一个强大的感染性结肠炎模型，其特征在于由结肠的隐窝，单核的免疫细胞浸润和杯状细胞耗竭3,4增生或伸长。初始站点的殖民统治，几个小时后，面临的挑战，是在盲肠修补程序，然后发展到远端结肠感染后2〜3天，3。在免疫功能正常的小鼠品系中，间隙实现的病原体感染后3〜4周1,3,4。然而，许多转基因的菌株， 即基因缺失或基因敲除小鼠（ - / - ），已被发现显示增加sed的感染导致夸张的损坏和/或慢性感染和炎症9-14的易感性。这在这些基因敲除株的感染性结肠炎模型，许多人缺乏与生俱来的信号蛋白，一直是不可或缺的分辨率的肠道感染和炎症的组成部分露出几个宿主蛋白。

<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>卢里亚肉汤</td><td> ABM </td><td> G247 </td><td>添加LB粉25克至1L水。高压灭菌后方可使用。 </td></tr><tr><td>广场底板与电网</td><td>费舍尔</td><td> 08-757-11A </td><td></td></tr><tr><td>猎鹰文化管</td><td>萨尔斯塔特</td><td> 62.515.006 </td><td></td></tr><tr><td>灯泡放倒灌胃针</td><td>精细科学工具</td><td> 18060-20 </td><td></td></tr><tr><td> 1ml注射器</td><td> BD Biosciences公司</td><td> 309659 </td><td></td></tr><tr><td> 4 kDa的FITC-葡聚糖</td><td>西格玛</td><td> FD-4 </td><td></td></tr><tr><td>柠檬酸</td><td>西格玛</td><td> C7129 &lt;/ TD&gt; <td></td></tr><tr><td>柠檬酸钠</td><td>费舍尔</td><td> S279-500 </td><td></td></tr><tr><td>葡萄糖</td><td>费舍尔</td><td> D16.1 </td><td></td></tr><tr><td>柠檬酸盐葡萄糖</td><td></td><td></td><td>的20mM ctiric酸，110 mM柠檬酸钠，5mM的葡萄糖</td></tr><tr><td>黑色96 - 孔板</td><td>费舍尔</td><td> 07-200-762 </td><td></td></tr><tr><td>金属珠（5毫米） </td><td> Qiagen公司</td><td> 69989 </td><td></td></tr><tr><td> 10％的福尔马林</td><td>费舍尔</td><td> 5F93-4 </td><td></td></tr><tr><td> 5毫升小瓶</td><td> DiaMed </td><td> STK3205 </td><td></td></tr><tr><td>苏木</td><td>费舍尔</td><td> H345-23 </td><td></td></tr><tr><td>曙红</td><td>费舍尔</td><td> E511-100 </td><td></td></tr><tr><td>二甲苯</td><td>费舍尔</td><tD&gt; HC700-1GAL </td><td></td></tr><tr><td>吐温20 </td><td>西格玛</td><td> P5927 </td><td></td></tr><tr><td>科普林氏染色缸</td><td> VWR </td><td> 47751-792 </td><td></td></tr><tr><td>柠檬酸钠缓冲液</td><td></td><td></td><td> 10mM柠檬酸钠，0.05％吐温20，pH 6.0的</td></tr><tr><td>巴氏笔</td><td> CEDARLANE </td><td> Mu22 </td><td></td></tr><tr><td>山羊血清</td><td>西格玛</td><td> G902-3 </td><td></td></tr><tr><td>牛血清白蛋白（BSA） </td><td>费舍尔</td><td> BP1600-100 </td><td></td></tr><tr><td>的Triton X-100的</td><td>西格玛</td><td> T8532 </td><td></td></tr><tr><td>叠氮化钠</td><td>西格玛</td><td> SZ002 </td><td></td></tr><tr><td>封闭缓冲液</td><td></td><td></td><td> 2％山羊血清，1％BSA，0.1％Triton X-100，0.05％吐温20，0.05％的氢氧化钠叠氮钠，0.01 M PBS，pH值7.2，混合储存于4°C。 </td></tr><tr><td>抗体稀释液</td><td></td><td></td><td> 0.1％Triton X-100，0.1％BSA，0.05％叠氮钠，0.04％EDTA </td></tr><tr><td>阻断缓冲液和抗体稀释液TIR </td><td></td><td></td><td>与上述相同的配方，但不添加的洗涤剂（TRITON X-100和吐温20） </td></tr><tr><td>延长Gold抗淬灭试剂与DAPI </td><td> Invitrogen公司</td><td> P-36931 </td><td></td></tr><tr><td>盖玻片</td><td>费舍尔</td><td> 12.54SE </td><td></td></tr><tr><td>台式孵化振动筛</td><td> Barnstead实验室线</td><td>最大Q4000 </td><td></td></tr><tr><td>荧光计</td><td> Perkin Elmer公司</td><td> Victor2D </td><td></td></tr><tr><td>冷冻离心机</td><td> Beckman Coulter公司</td><td>微型离心22R</td><td></td></tr><tr><td>汽船</td><td> Black＆Decker公司</td><td></td><td></td></tr><tr><td>荧光显微镜</td><td>蔡司</td><td> Axio上Image.Z1 </td><td></td></tr></tbody></table>

the citrobacter rodentium mouse model: studying pathogen and host contributions to infectious colitis

该协议列出的步骤需要产生一个强大的感染性疾病和肠炎的模型，以及用来描述枸橼酸rodentium感染的小鼠的方法。C. rodentium是一种革兰氏染色阴性，小鼠特异性细菌性病原体，临床上重要的人类病原体肠致病性大肠杆菌是密切相关的大肠杆菌和肠出血性大肠杆菌大肠杆菌 。一旦感染C. rodentium，免疫小鼠患上温和和短暂的体重减轻和腹泻。组织学观察，肠隐窝伸长率，免疫细胞浸润，杯状细胞耗竭。实现间隙感染后3〜4周。肠上皮屏障的完整性，细菌量，以及组织损伤，感染后的不同时间点的测量，让易受感染的小鼠品系的特征。 致病机制s的细菌病原体定植于肠道他们的主机，以及具体的抵御这种感染的宿主反应，却知之甚少。因此，C. rodentium作为一个有价值的工具，以帮助我们了解这些过程模型肠道细菌感染。肠道细菌也被连接到的炎症性肠病（IBDs）。据推测，在肠黏膜免疫系统的异常暴露的肠道细菌基因易感个体适应不良的慢性炎症反应的IBD患者中发展。因此，感染性结肠炎模型的研究提供了重要C.确定潜在的致病性肠道细菌的宿主反应的潜力。 rodentium诱导的结肠炎就是这样一个难得的模式，让肠道细菌的宿主反应的分析，我们进一步了解鸡传染性法氏囊病发病的潜在机制;新的预防和治疗方法的发展至关重要。

在标准的感染实验，小鼠灌胃100μL过夜C.感染者约2.5×10 8 CFU通过rodentium文化。 与 C的C57BL / 6小鼠的感染rodentium结果，在温和和短暂的体重减轻和腹泻。虽然很少发生C57BL / 6小鼠，动物可能生病和需要euthanization。因此，鼠标应该减肥的程度和症状的困扰，如piloerect毛皮和驼背姿势进行监控，以确定不同菌株在何种程度上受到感染。 结果呈现?...

Watch this Scientific Journal Video about The Citrobacter rodentium Mouse Model: Studying Pathogen and Host Contributions to Infectious Colitis at JoVE.com

The Citrobacter rodentium Mouse Model: Studying Pathogen and Host Contributions to Infectious Colitis

枸橼酸rodentium感染提供了一个有价值的模型来研究肠道细菌感染以及宿主的免疫反应和结肠炎的小鼠。该协议概述了屏障的完整性测量，病原体负荷和病理损害，从而为深入鉴定病原体和宿主小鼠的感染性肠炎的贡献。

“<em&gt;枸橼酸rodentium</em&gt;鼠标产品型号：研究感染性结肠炎的病原体和宿主的贡献

This protocol  outlines the steps required to produce a robust model of infectious disease and  colitis, as well as the methods used to characterize ...

Research

JoVE Journal

Immunology and Infection

The Citrobacter rodentium Mouse Model: Studying Pathogen and Host Contributions to Infectious Colitis

Use of an Optical Trap for Study of Host-Pathogen Interactions for Dynamic Live Cell Imaging

动态活细胞成像的宿主 - 病原体相互作用的研究使用的光学陷阱

Dynamic live cell imaging allows direct visualization of real-time interactions between cells of the immune system1, 2; however, the lack of spatial and ...

科研

免疫与感染

Use of the EpiAirway Model for Characterizing Long-term Host-pathogen Interactions

使用的EpiAirway模型，用于描述长期宿主 - 病原体相互作用

Nontypeable Haemophilus influenzae  (NTHi) are human-adapted Gram-negative bacteria that can cause recurrent and chronic infections of the respiratory ...

A Comparative Approach to Characterize the Landscape of Host-Pathogen Protein-Protein Interactions

比较方法来表征景观的宿主 - 病原体蛋白 - 蛋白相互作用

Significant efforts were gathered to generate  large-scale comprehensive protein-protein interaction network maps. This is  instrumental to understand the ...

4D Multimodality Imaging of Citrobacter rodentium Infections in Mice

4D Multimodality Imaging of Citrobacter rodentium Infections in Mice

4D多模态成像<em&gt;柠檬酸rodentium</em在小鼠感染

This protocol outlines the steps required to  longitudinally monitor a bioluminescent bacterial infection using composite 3D diffuse light imaging ...

Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis

使用点击化学测量的影响，病毒感染宿主细胞RNA合成

Many RNA viruses have evolved the ability to inhibit host cell transcription as a means to circumvent cellular defenses. For the study of these viruses, ...

A Mouse Model for Pathogen-induced Chronic Inflammation at Local and Systemic Sites

鼠标型号病原体引起的慢性炎症局部及全身站点

Chronic inflammation is a major driver of pathological tissue damage and a unifying characteristic of many chronic diseases in humans including ...

Experimental Infection with Listeria monocytogenes as a Model for Studying Host Interferon-&#947; Responses

Experimental Infection with Listeria monocytogenes as a Model for Studying Host Interferon-ÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â³ Responses

实验性感染<em&gt;李斯特菌</em&gt;作为研究主机干扰素γ响应模型

L. monocytogenes is a gram-positive bacterium that is a cause of food borne disease in humans. Experimental infection of mice with this pathogen has been ...

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

原发性鸡初囊-细胞培养模型研究传染性囊内病病毒发病机制

Infectious bursal disease virus (IBDV) is a birnavirus of economic importance to the poultry industry. The virus infects B cells, causing morbidity, ...

Using a Bacterial Pathogen to Probe for Cellular and Organismic-level Host Responses

细菌病原体在细胞和有机水平宿主反应探针中的应用

There are a variety of strategies bacterial pathogens employ to survive and proliferate once inside the eukaryotic cell. The so-called 'cytosolic' ...

Evaluation of Host-Pathogen Responses and Vaccine Efficacy in Mice

宿主病原学反应及疫苗效果评价

Vaccines are a 20th century medical marvel. They have dramatically reduced the morbidity and mortality caused by infectious diseases and contributed to a ...