Name: transplantation of tail skin to study allogeneic cd4 t cell responses in mice
Uploaded: 2014-07-25T13:00:00
Description: Watch this Scientific Journal Video about Transplantation of Tail Skin to Study Allogeneic CD4 T Cell Responses in Mice at JoVE.com

This work was supported by SNF grants PPOOA-_119204 and PPOOP3_144918 to S.W.R. We thank E. Palmer and B.T.H. Hausmann for mice and technical expertise.

在这个视频发布和协议，所有动物的程序按照批准由州局巴塞尔城市州，瑞士动物的协议进行的。执行在无菌条件下尽可能的所有程序。 外科1。准备<ol><li>高压灭菌所有的手术器械，使用前纱布。 </li><li>热保暖垫，并组织手术器械上的表 （表材料/方法 ）。 </li><li>打开手指带绷带。应用凡士林的伤口垫用棉签（确保纱布完全覆盖）。 </li><li>称取?...

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皮肤移植是一种广泛使用的方法为研究抑制和容忍依赖于T细胞。自成立以来的皮肤移植模型中，几个修改和变化都得到了应用。在描述的过程中，使用麻醉气体（异氟烷）进行IA BM12尾部皮肤移植。使用气体麻醉的降低执行和小鼠恢复，从而减少了应力对移植小鼠的时间。该程序使用组织粘合剂来固定尾部皮肤，这容易卷曲超过耳或背皮肤组织。此外，它弄平切割和修整的边缘，因为这降...

固体器官，如皮肤，心脏和肾脏移植是目前在全世界的医疗实践1的标准程序。成功移植的器官可以通过激活受体的免疫系统，其中确认捐助的主要组织相容性抗原被拒绝。因此移植的患者需要使用免疫抑制药物治疗2。异体皮移植在小鼠体内建立了梅达沃和他的同事于1955年，是识别后世誉为主要组织相容性复合体（MHC）I类和II目标分子的帮助。从那以后，皮肤移植模型得到了不断的修改和调整，研究T细胞亚群的作用和抑制移植排斥2-4化学和生物干预的相关性。从耳朵和躯干的皮肤都比较困难的准备，而且更容易受到缺氧和坏死较尾皮5;然而，移植过程是类似的。在另外的尾部皮肤移植物的监测是很容易由于皮肤的特征头发纹理。 本文提供的详细步骤MHC II类错配尾部皮肤移植，允许的CD4 + T细胞介导的移植排斥反应和耐受小鼠不同方面的研究。在MHC II类分子IA b将天然三点突变（称为IA BM12）6-9是足以引起排斥反应的皮肤移植物在C57BL / 6小鼠8。在IA BM12分子激活CD4 + T细胞与各种αβT细胞受体（TCR）从C57BL / 6小鼠链，其中Vα2Vβ8-TCR特异性T细胞，以产生一个TCR转基因小鼠10识别。 Vα2Vβ8-TCR特异性T细胞的过继转移已被用于建立一个排斥模型在免疫缺陷C57BL / 6的Rag2 - / -移植与IA BM12皮肤老鼠。 捐赠者和接受者之间的基因差异影响移植接受和拒绝的结果。有不同类型的移植：自体移植是从接收者个人自身移植; syngrafts和移植物分别从基因上相同和基因无关个体移植。不同的异体器官移植的接受已经证明了患者和小鼠模型11,3,4化学和生物干预。在一个基本的方法，抗-CD3抗体处理的C57BL / 6小鼠表现出IA BM12尾部皮肤（未发表数据），生存期延长。 CD4 +和CD8 + T细胞移植受体小鼠前的枯竭导致接受MHC I类和II不匹配的移植（修订版12）。有趣的是，抑制皮肤移植取决于CD4 +的存在</sup&gt; T细胞（版本12）。在此模型中，通过阻断共刺激分子的抗体或抑制与调节性T细胞靶向不同的免疫细胞之间的特异性相互作用可能诱导耐受（未发表数据）。事实上，无论阻断CD40和CD28分子导致了长期的皮肤移植耐受13,14。 相比于其它器官的移植尾部皮肤移植容易进行并易于监测。此外，尾部皮肤移植很容易准备，而且不太容易受到缺血较其他皮肤组织。而相比之下，注射麻醉剂，移植过程中使用麻醉气体（异氟烷）的缩短两者的程序和收件人的恢复时间。尾部皮肤移植，这可能会导致不完全的伤口愈合，移植物排斥的卷曲，可以防止通过应用组织粘合剂。此外，IA BM12尾部皮肤移植模型的专门激活的CD4 <sup&gt; + T细胞（相同遗传背景）两个免疫和免疫缺陷小鼠促进结果的解释。 本协议描述了一个可靠，重现性好，易于监控的小鼠模型，允许化学和生物干预。该模型的目的是调查排斥和耐受诱导的尾部皮肤移植手术。

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			<td style="width:231px;">Comments/Description</td>
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			<td>Mundipharma</td>
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			<td height="21" style="height:21px;">Cotton swab</td>
			<td style="width:183px;">Carl Roth GmbH</td>
			<td style="width:119px;">31025060</td>
			<td></td>
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			<td height="21" style="height:21px;width:299px;">Dafalgan , UPSA</td>
			<td>Bristol Myers Squibb SA</td>
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			<td></td>
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			<td height="21" style="height:21px;">Hansaplast Finger Strips</td>
			<td>Beierdorf AG</td>
			<td style="width:119px;">REF.76861</td>
			<td></td>
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			<td height="21" style="height:21px;">Histoacryl Tissue adhesive</td>
			<td style="width:183px;">Braun</td>
			<td style="width:119px;">REF.1050052</td>
			<td></td>
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			<td height="21" style="height:21px;">Leukotape classic , 2cm x 10 m</td>
			<td>BSN Medical SAS</td>
			<td>REF.02204-00</td>
			<td></td>
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			<td height="21" style="height:21px;">PBS, Phosphate-Buffered Saline, pH 7.4</td>
			<td style="width:183px;">Invitrogen</td>
			<td style="width:119px;">10010015</td>
			<td>GIBCO</td>
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			<td height="21" style="height:21px;">Sterile gauze, 5 x 5cm, 8ply</td>
			<td>MaiMed GmbH</td>
			<td>21010</td>
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			<td height="21" style="height:21px;">Surgical instruments&#160;</td>
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			<td>Narrow pattern forceps,&#160;</td>
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			<td style="width:119px;">14095-11</td>
			<td>Fine iris scissor curved,</td>
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			<td height="21" style="height:21px;width:299px;"></td>
			<td style="width:183px;"></td>
			<td style="width:119px;">14094-11</td>
			<td>Fine iris scissor&#160;</td>
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			<td height="21" style="height:21px;width:299px;"></td>
			<td style="width:183px;"></td>
			<td style="width:119px;">14010-15</td>
			<td>Mayo scissors,</td>
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			<td height="21" style="height:21px;width:299px;"></td>
			<td style="width:183px;"></td>
			<td style="width:119px;">14080-11</td>
			<td>Artery scissors ball tip 11.5 cm&#160;</td>
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			<td height="21" style="height:21px;width:299px;"></td>
			<td style="width:183px;"></td>
			<td style="width:119px;">11021-14</td>
			<td>Tissue forceps</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Surgical Blade No.20</td>
			<td style="width:183px;">Swann-Morton LTD</td>
			<td style="width:119px;">3006</td>
			<td>Carbon Steel</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Surgical blade Handles</td>
			<td style="width:183px;">Swann-Morton LTD</td>
			<td style="width:119px;"></td>
			<td></td>
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			<td height="21" style="height:21px;">Syringe, 1ml</td>
			<td style="width:183px;">ARTSANA</td>
			<td style="width:119px;"></td>
			<td>disposable</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Temgesic , Buprenorphine</td>
			<td style="width:183px;">ESSEX Chemie AG</td>
			<td style="width:119px;"></td>
			<td>0.3mg/ml</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Tissue Culture dishes 10 cm, 60.1 cm2</td>
			<td style="width:183px;">TPP</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:299px;">Vaseline</td>
			<td style="width:183px;">Vifor SA</td>
			<td style="width:119px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:299px;">Warm pad</td>
			<td style="width:183px;">Solis</td>
			<td style="width:119px;"></td>
			<td>Type 223</td>
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transplantation of tail skin to study allogeneic cd4 t cell responses in mice

The study of T cell responses and their consequences during allo-antigen recognition requires a model that enables one to distinguish between donor and host T cells, to easily monitor the graft, and to adapt the system in order to answer different immunological questions. Medawar and colleagues established allogeneic tail-skin transplantation in mice in 1955. Since then, the skin transplantation model has been continuously modified and adapted to answer specific questions. The use of tail-skin renders this model easy to score for graft rejection, requires neither extensive preparation nor deep anesthesia, is applicable to animals of all genetic background, discourages ischemic necrosis, and permits chemical and biological intervention. 
In general, both CD4+ and CD8+ allogeneic T cells are responsible for the rejection of allografts since they recognize mismatched major histocompatibility antigens from different mouse strains. Several models have been described for activating allogeneic T cells in skin-transplanted mice. The identification of major histocompatibility complex (MHC) class I and II molecules in different mouse strains including C57BL/6 mice was an important step toward understanding and studying T cell-mediated alloresponses. In the tail-skin transplantation model described here, a three-point mutation (I-Abm12) in the antigen-presenting groove of the MHC-class II (I-Ab) molecule is sufficient to induce strong allogeneic CD4+ T cell activation in C57BL/6 mice. Skin grafts from I-Abm12 mice on C57BL/6 mice are rejected within 12-15 days, while syngeneic grafts are accepted for up to 100 days. The absence of T cells (CD3-/- and Rag2-/- mice) allows skin graft acceptance up to 100 days, which can be overcome by transferring 2 x 104 wild type or transgenic T cells. Adoptively transferred T cells proliferate and produce IFN-&#947; in I-Abm12-transplanted Rag2-/- mice.

在第一种方法中的C57BL / 6小鼠移植IA BM12同种异体移植物和IA b syngrafts。拆除绷带后，移植物伤口愈合的迹象明显没有在C57BL / 6小鼠（ 图1A）的接触带的闭合。拆除绷带后，CD4 + T细胞介导的炎症导致的坏死区域的外观（红点）和内移植（ 图1B-C，实心）后13天C57BL / 6小鼠抑制IA BM12移植，而syngrafts被容忍长达100天（ 图1B-C，空心符...

Watch this Scientific Journal Video about Transplantation of Tail Skin to Study Allogeneic CD4 T Cell Responses in Mice at JoVE.com

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

Tail-skin transplantation is a powerful model for studying T cell-dependent rejection and tolerance induction during allogeneic immune responses in mice. The advantages of this protocol are minor invasive surgery, and ease of monitoring with no need to sacrifice the recipient mouse.

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 ...

Research

JoVE Journal

Immunology and Infection

Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant

Multivirus特异性T细胞的生成，预防/治疗异基因造血干细胞移植后的病毒感染

Viral infections cause morbidity and mortality in allogeneic hematopoietic stem cell transplant (HSCT) recipients. We and others have successfully ...

科研

免疫与感染

Accelerated Type 1 Diabetes Induction in Mice by Adoptive Transfer of Diabetogenic CD4+ T Cells

加速1型糖尿病感应在小鼠体内的过继转移致糖尿病的CD4 + T细胞

The nonobese diabetic (NOD) mouse spontaneously develops autoimmune diabetes after 12 weeks of age and is the most extensively studied animal model of ...

Adenoviral Transduction of Naive CD4 T Cells to Study Treg Differentiation

腺病毒转导朴素的CD4 T细胞调节性T细胞分化研究

Regulatory T cells (Tregs) are essential to provide immune tolerance to self as well as to certain foreign antigens. Tregs can be generated from naive CD4 ...

Isolation and Th17 Differentiation of Na&iuml;ve CD4 T Lymphocytes

Isolation and Th17 Differentiation of Na&#239;ve CD4 T Lymphocytes

隔离和幼稚的CD4 T细胞的Th17细胞分化

Th17 cells are a distinct subset of T cells that have been found to produce interleukin 17 (IL-17), and differ in function from the other T cell subsets ...

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

诱导小鼠肠道炎症由效应CD4的过继转移<sup&gt; +</sup&gt; CD45RB<sup&gt;高</sup&gt; T细胞免疫缺陷小鼠

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages ...

Mouse Na&#239;ve CD4+ T Cell Isolation and In vitro Differentiation into T Cell Subsets

Mouse Na&#239;ve CD4+ T Cell Isolation and In vitro Differentiation into T Cell Subsets

鼠标朴素CD4<sup&gt; +</sup&gt; T细胞分离和<em&gt;在体外</em&gt;分化为T细胞亚群

Antigen inexperienced (na&#239;ve) CD4+ T cells undergo expansion and differentiation to effector subsets at the time of T cell receptor (TCR) recognition ...

Imaging CD4 T Cell Interstitial Migration in the Inflamed Dermis

成像的CD4 T细胞间质迁移发炎真皮

The ability of CD4 T cells to carry out effector functions is dependent upon the rapid and efficient migration of these cells in inflamed peripheral ...

In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients

In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients

体外和活体评估移植受体的 T、B 和髓细胞抑制活性和体液反应

The main concern in transplantation is to achieve specific tolerance through induction of regulatory cells. The understanding of tolerance mechanisms ...

A Suction Blister Protocol to Study Human T-cell Recall Responses In Vivo

A Suction Blister Protocol to Study Human T-cell Recall Responses In Vivo

一种研究人体 T 细胞召回反应的吸水泡协议

Cutaneous antigen-recall models allow for studies of human memory responses in vivo. When combined with skin suction blister (SB) induction, this model ...

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice

感染 Ifnar1 小鼠 Immunoprivileged 器官寨卡病毒特异 T 细胞反应的评价

The Zika virus (ZIKV) can induce inflammation in immunoprivileged organs (e.g., the brain and testis), leading to the Guillain-Barr&#233; syndrome and ...