Name: intralymphatic immunotherapy and vaccination in mice
Uploaded: 2014-02-02T14:00:00
Description: Watch this Scientific Journal Video about Intralymphatic Immunotherapy and Vaccination in Mice at JoVE.com

作者感谢的实验帮助发展中国家从虹膜埃德曼，芭芭拉·冯·博伊斯特，朱莉娅玛丽亚·马丁内斯·戈麦斯小鼠淋巴免疫的方法。还要感谢玛吉阿拉斯和尼古拉Cesarovic您让我们使用他们的手术剧场这个视频制作。

1。小鼠麻醉<ol><li>在缓冲液混合氯胺酮（分离性麻醉剂）和甲苯噻嗪（镇静，镇痛）准备麻醉剂。氯胺酮和甲苯噻嗪最终溶液中的浓度是12.5和2毫克/毫升。 </li><li>使用注射器用25-30号针头通过腹膜内给药注射麻醉​​剂将小鼠。使用0.1 ml/10克的机身重量。 </li><li>为了防止干燥出的角膜应用眼用软膏对小鼠的眼睛。 </li><li>确保鼠标充分捏它的脚或脚趾与镊子麻醉。如果鼠标不与反射的捏反应，?...

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A., 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=Enhancing+DNA+vaccination+by+sequential+injection+of+lymph+nodes+with+plasmid+vectors+and+peptides.">Enhancing DNA vaccination by sequential injection of lymph nodes with plasmid vectors and peptides.</a> Vaccine. 27, 2603-2615 (2009).</li><li>Kreiter, 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=Intranodal+vaccination+with+naked+antigen-encoding+RNA+elicits+potent+prophylactic+and+therapeutic+antitumoral+immunity.">Intranodal vaccination with naked antigen-encoding RNA elicits potent prophylactic and therapeutic antitumoral immunity.</a> Cancer Res. 70, 9031-9040 (2010).</li><li>Johansen, P., 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=Antigen+kinetics+determines+immune+reactivity.">Antigen kinetics determines immune reactivity.</a> Proc. 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淋巴内免疫接种和免疫治疗已被证明是合适的两个抗体应答和T细胞应答的刺激。这表现这个视频文章中，接种疫苗的淋巴管内的程序是一个快速简便的方法刺激强烈的免疫反应的小鼠。一个训练有素的外科医生可以在3-4分钟执行程序。这次会议也可两名外科医生，其中一个通常做麻醉和缝合，第二个外科医生在做切口和注射之间共享。该过程的效率，更取决于运营商以外的免疫方法，尤其是当外...

疫苗通常是皮下或肌内注射对免疫应答的刺激。这个程序的成功需要疫苗的有效引流淋巴结其中抗原呈递细胞可以与淋巴细胞相互作用用于生成的T-和B-细胞应答。此外，强度和诱导免疫反应的类型也取决于这种相互作用的密度或频率以及微环境本身，细胞因子，特别是内容。作为注入到外周组织的疫苗只有一小部分到达淋巴结，小鼠和人类通过直接注射该疫苗进入淋巴结的疫苗，该网站被产生的免疫反应，已被执行。在男人中，程序是通过超声波，一个程序也可以用于可视化和诊断在淋巴系统成像剂的给药指导。在小鼠体内，是侵入性检查。在这里，一个小的（5-10毫米）incision被制成在麻醉的动物1的腹股沟区域，淋巴结进行定位和固定钳，和10-20微升的疫苗的一定体积的视觉控制下注入; 10微升用于第一注射和在年轻小鼠用小淋巴结，而20微升可以注入更旧的淋巴结或已催芽的小鼠，其具有较大的淋巴结。切口可闭合使用外科缝合线的单个针迹。通过该方法，将小鼠已接种的质粒DNA 2,3，信使RNA 4，1,3,5,6肽，蛋白质7-10，颗粒11，细菌12以及辅助剂7,13，和免疫大幅改善对所有类型的疫苗反应已​​被观察到。接种淋巴方法特别适合在常规疫苗接种产生的免疫力不足或可用疫苗的量是有限的或版本的情况下Ÿ昂贵。在人类，免疫的淋巴管内的方法已被应用到过敏患者14,15或癌症患者16-21。虽然目前的看法是，在淋巴管内的方法比其它可注射的方法，如肌内和皮下注射更具侵入性的，疼痛感觉不大于静脉穿刺15后更高。预期淋巴内注射疫苗将成为一种替代或补充的预防和治疗尤其是疫苗的其它方法。本文详细阐述了如何淋巴管内接种疫苗的过程中小鼠进行。描述的所有程序，批准了苏黎世和州兽医局，并根据瑞士联邦指导方针以及用于科学目的的动物的保护指令执行。

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	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">Ketamine (Ketasol-100)</td>
			<td style="width:135px;">Graeub AG, Switzerland</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Anesthetics</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:233px;">Xylazine (Rompun)</td>
			<td style="width:135px;">Bayer, Germany</td>
			<td style="width:119px;"></td>
			<td>Anesthetics</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Viscotears Eye-Gel</td>
			<td style="width:135px;">Novartis, Switzerland</td>
			<td style="width:119px;"></td>
			<td>To keep eyes from drying out during anesthesia.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">BD Micro-Fine 0.5 ml</td>
			<td style="width:135px;">BD Medical, France</td>
			<td style="width:119px;"></td>
			<td>29 G Insulin syringes with permanently attached needles</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">6-0 Dermalon Monofilament nylon</td>
			<td style="width:135px;">Covidien, MA, USA</td>
			<td style="width:119px;"></td>
			<td>For sutures (0.7 metric, 18G, 45 cm, Blue)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">Curved forceps, 4.5 inch</td>
			<td style="width:135px;">Polymed, Switzerland</td>
			<td style="width:119px;"></td>
			<td>For incision and holding of lymph node</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">Straight surgical scissors, 4.5 inch</td>
			<td style="width:135px;">Polymed, Switzerland</td>
			<td style="width:119px;"></td>
			<td>For incision</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">Needle holder, 5.5 inch</td>
			<td style="width:135px;">Polymed, Switzerland</td>
			<td style="width:119px;"></td>
			<td>To close incision with suture</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:233px;"></td>
			<td style="width:135px;"></td>
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			<td></td>
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intralymphatic immunotherapy and vaccination in mice

疫苗一般是皮下或肌内注射对免疫应答的刺激。这样做的成功需要疫苗的有效引流淋巴结，其中抗原呈递细胞可以与淋巴细胞相互作用的一代想要的免疫反应。的强度和诱导免疫反应的类型也取决于相互作用的密度或频率以及微环境，细胞因子，特别是内容。作为外周注射疫苗仅一分钟级分到达淋巴结，小鼠和人类的疫苗通过直接注射疫苗被执行到腹股沟淋巴结， 即淋巴内注射。在男人中，程序是由超声引导。在小鼠中，小（5-10毫米）切口，在麻醉的动物的腹股沟区域，淋巴结进行定位和固定钳，和10-20微升的疫苗的一定体积的视觉控制下喷射。切口闭合使用手术缝合线单针。小鼠接种的质粒DNA，RNA，肽，蛋白质，颗粒，细菌以及佐剂，并观察到对所有类型的疫苗的免疫应答的强改善。接种淋巴方法特别适合在常规疫苗接种产生的免疫力不足或可用的疫苗量有限的情况下。

在小鼠淋巴内注射的方法，尽管在手术的性质，是直线前进和比较快的。一个训练有素的人能在3-4分钟执行程序。是与一针封闭切口通常在两天内愈合（ 图1） 淋巴内注射疫苗或免疫接种用的mRNA被执行时，质粒DNA，肽，蛋白质，病毒和细菌。 图2示出了免疫与蛋白质磷脂酶A2（PLA2），主要蜜蜂毒液变应原后的抗体产生。 CBA小鼠进行免疫在0天，14，和2...

Watch this Scientific Journal Video about Intralymphatic Immunotherapy and Vaccination in Mice at JoVE.com

Intralymphatic Immunotherapy and Vaccination in Mice

淋巴免疫治疗和疫苗接种小鼠

预防性和治疗性疫苗往往不能刺激强烈的免疫反应，由于疫苗的周向引流淋巴结细胞和免疫细胞的参与因而不佳。通过直接注射疫苗到淋巴结的，所谓的淋巴内注射，疫苗的有效性，可以大大地改善和疫苗的剂量可以减少。

Vaccines are typically injected subcutaneously or intramuscularly for stimulation of immune responses. The success of this requires efficient drainage of ...

Research

JoVE Journal

Immunology and Infection

Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model

实验转移和CTL过继转移免疫小鼠模型

Experimental metastasis mouse model is a simple and yet physiologically relevant metastasis model. The tumor cells are injected intravenously (i.v) into ...

科研

免疫与感染

Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection

协议RBD - SARS疫苗的重组蛋白的制备，动物疫苗接种和中检测

Based on their safety profile and ability to induce potent immune responses against infections, subunit vaccines have been used as candidates for a wide ...

Preparation of Tumor Antigen-loaded Mature Dendritic Cells for Immunotherapy

制备肿瘤抗原负载成熟树突状细胞免疫治疗

While clinical studies have established that antigen-loaded DC vaccines are safe and promising therapy for tumors 1, their clinical efficacy remains to be ...

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses

高通量筛选RNA病毒的广谱化学抑制剂

RNA viruses are responsible for major human diseases such as flu, bronchitis, dengue, Hepatitis C or measles. They also represent an emerging threat ...

Measuring Local Anaphylaxis in Mice

小鼠量度本地过敏反应

Allergic responses are the result of the activation of mast cells and basophils, and the subsequent release of vasoactive and proinflammatory mediators. ...

Generation of CAR T Cells for Adoptive Therapy in the Context of Glioblastoma Standard of Care

CAR性T细胞过继治疗的护理标准胶质母细胞瘤的上下文生成

Adoptive T cell immunotherapy offers a promising strategy for specifically targeting and eliminating malignant gliomas. T cells can be engineered ex vivo ...

In Vivo Assay for Detection of Antigen-specific T-cell Cytolytic Function Using a Vaccination Model

In Vivo Assay for Detection of Antigen-specific T-cell Cytolytic Function Using a Vaccination Model

在体内应用免疫模型检测抗原特异性 T 细胞细胞功能的试验

Current methodologies for antigen-specific killing are limited to in vitro use or utilized in infectious disease models. However, there is not a protocol ...

Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

Monoclonal antibodies (mAbs) are powerful tools useful for both fundamental research and in biomedicine. Their high specificity is indispensable when the ...

Stereotactic Adoptive Transfer of Cytotoxic Immune Cells in Murine Models of Orthotopic Human Glioblastoma Multiforme Xenografts

立体定向移植人胶质瘤多形性移植小鼠细胞毒性免疫细胞模型的研究

Glioblastoma multiforme (GBM), the most frequent and aggressive primary brain cancer in adults, is generally associated with a poor prognosis, and scarce ...

Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination

优化干扰素伽隆弹性体检测在几内亚猪模型接种疫苗后 t 细胞反应

The guinea pig has played a pivotal role as a relevant small animal model in the development of vaccines for infectious diseases such as tuberculosis, ...