Name: assessment of lymphocyte migration in an ex vivo transmigration system
Uploaded: 2019-09-20T14:30:00
Description: Watch this Scientific Journal Video about Assessment of Lymphocyte Migration in an Ex Vivo Transmigration System at JoVE.com

这项工作由美国肺脏协会（K.J.W.）、授予T.A.W.和K.J.W.的纪念尤金·肯尼基金、犹他大学为K.J.W.提供的慷慨启动支持以及退伍军人事务部授予T.A.W.（VA I01BX0003635）的资助。T.A.W. 是退伍军人事务部研究职业科学家奖 （IK6 BX003781） 的获得者。作者希望感谢丽莎·丘多梅尔卡女士的编辑协助。作者感谢UNMC流式细胞学核心在收集为本手稿生成的流式细胞学数据方面给予的支持。

这里描述的所有方法都经过内布拉斯加大学医学中心（UNMC）和犹他大学动物护理和使用委员会的审查和批准。
1. 试剂的建立和制备
<ol><li>
准备完整的RPMI（罗斯威尔公园纪念研究所）媒体。
	<ol>
<li>将10 mL的热灭活胎儿牛血清 （FBS） 添加到 90 mL 的 RPMI 中。</li>
		<li>将1mL的100x青霉素-链霉素-谷氨酰胺添加到100 mL的10%FBS RPMI中。</li>
	</ol>
</li>
	<li>
准备 ILC2 扩?...

<ol>
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在这里，我们提出了一个成熟的方法，以评估在外体迁移系统中淋巴细胞引起的化学素迁移。该协议中有几个关键步骤，第一个步骤是验证实验中免疫细胞上正确的化学素受体的表达。在我们的手中，我们选择CCR4是因为文献中强调了CCR4在过敏性炎症中T2帮助T细胞的重要性。Ovalbumin引起的炎症先前被证明至少受到两个CCR4拮抗剂24，25的限制;然而，这是之...

这种原始的迁移方法是由斯蒂芬·博伊登于1962年在《实验医学杂志1》上提出的。如果没有博伊登室的发展，我们对化学和化学动力学的了解是不可能实现的。在1977年发现第一个化学药物之前，前体迁移系统被用来了解血清因子，这些血清因子可以阻止巨噬细胞的细胞运动，同时放大中性粒细胞1、2的细胞活力。关于免疫细胞迁移，已经发展了大量的知识，到目前为止，已经发现了47个化学素与19个相应的受体3，4。此外，这些化学药物通路的多种抑制剂/增强剂已发展为治疗目的5，6，7，8。其中许多化合物已经在类似的迁移室中进行了测试，以了解化合物与免疫细胞对给定化学素9的反应之间的直接相互作用。
迁移，或二足动物，进入发炎的组织是一个必要的过程，以健康的炎症反应清除感染10，11。博伊登室、迁移系统或跨井装置一般由由多孔膜1、12隔开的两个腔室组成。底部腔通常装有含有感兴趣的化学素的介质，而白细胞则放置在顶部腔室中。可以根据感兴趣的细胞的大小选择膜中的孔隙大小。对于这个项目，我们选择了3μm多孔膜，因为淋巴细胞的大小是7-20μm，这取决于细胞发育的阶段。这种孔径大小确保这些细胞不会被动地从毛孔中掉落，而是主动迁移以响应化学素梯度。
该协议的主要优点是其成本效益。体内迁移是困难的，因为它需要动物处理和手术方面的广泛培训，并且通常涉及研究人员并不总是可用的高能显微镜。在体内成像之前，可以对被认为能增强或抑制迁移的化合物进行具有成本效益的筛选。由于迁移系统受到严格控制，细胞可以先进行处理，然后添加到转井装置中，反之亦然，可先使用化学抑制剂处理化学药物抑制剂，然后将细胞添加到转井装置中。最后，内皮细胞和/或基底膜蛋白可以在转井实验前1-2天添加到转井插入的底部，以了解这些屏障细胞在化学动力学中的参与程度。同样，这些对系统的操作提供了一种强有力的手段，用于在更复杂的体内研究之前确定有关给定化合物有效性的重要信息。
利用移外室系统是评估淋巴细胞在各种体内和体外条件下的流动性的有效方法12，13，14。在这里，我们描述了一种优化的方法，用于评估外体淋巴细胞对迁移室中化学素的反应。在此示例实验中，CD4+ T 细胞和组 2 先天淋巴细胞 （ILC2） 在 OVA 过敏原暴露后从雄性小鼠和雌性 BALB/c 小鼠中分离。生成一个假设，CCR4+ CD45+系-（LIN-） ILC2 来自过敏原挑战小鼠将比 CCR4 + CD4+ T 帮助细胞更有效地迁移到 CCL17 和 CCL22。CCL17和CCL22是一种化学素，通常由M2（过敏）表型的树突细胞和巨噬细胞等细胞产生，在过敏15、16中。CCL17和CCL22可被视为过敏性炎症的生物标志物，因为它们在气道恶化16、17、18期间很容易在肺部检测到。重要的是，CCR4的表达与未经处理的对照组相比是升高的，从从室内尘虫处理动物中分离出的生物信息数据中所揭示，同样，ILC2来自使用IL-33处理的幼化动物的ILC2（过敏原促进先天细胞因子） 上调节 CCR419，20.此外，根据免疫基因组项目数据库（www.immgen.org）中的ILC2数据，CCR4 mRNA在这些先天免疫细胞中高度表达。迄今为止，关于将ILC2贩运到组织方面鲜为人知，但ILC2和CD4+T细胞很可能使用类似的化学素和受体作为化学和化学动力学，因为它们表达类似的转录因子和受体。因此，我们比较了ILC2和CD4+T淋巴细胞的CCL17与CCL22的响应能力，这些细胞来自男性和女性，OVA挑战动物。

<table>
	<tbody>
		<tr>
			<td>0.4% Trypan Blue</td>
			<td>Sigma-Aldrich</td>
			<td>15250061</td>
		</tr>
		<tr>
			<td>1 mL syringe</td>
			<td>BD Bioscience</td>
			<td>329424</td>
			<td>U-100 Syringes Micro-Fine 28G 1/2&#34; 1cc</td>
		</tr>
		<tr>
			<td>100x Penicillin-Streptomycin, L-Glutamine</td>
			<td>Gibco</td>
			<td>10378-016</td>
			<td>Dilute to 1x in RPMI media</td>
		</tr>
		<tr>
			<td>15 mL conical tubes</td>
			<td>Olympus Plastics</td>
			<td>28-101</td>
			<td>polypropylene tubes</td>
		</tr>
		<tr>
			<td>3 um transwell inserts</td>
			<td>Genesee Scientific</td>
			<td>25-288</td>
			<td>24-well plate containing 12 transwell inserts</td>
		</tr>
		<tr>
			<td>3x stabilizing fixative</td>
			<td>BD Pharmigen</td>
			<td>338036</td>
			<td>Prepare 1x solution according to manufacturers protocol</td>
		</tr>
		<tr>
			<td>5 mL polystyrene tubes</td>
			<td>STEM Cell Technologies</td>
			<td>38007</td>
		</tr>
		<tr>
			<td>50 mL conical tubes</td>
			<td>Olympus Plastics</td>
			<td>28-106</td>
			<td>polypropylene tubes</td>
		</tr>
		<tr>
			<td>8-chamber easy separation magnet</td>
			<td>STEM Cell Technologies</td>
			<td>18103</td>
		</tr>
		<tr>
			<td>ACK Lysing Buffer</td>
			<td>Life Technologies Corporation</td>
			<td>A1049201</td>
		</tr>
		<tr>
			<td>Advanced cell strainer, 40 um</td>
			<td>Genesee Scientific</td>
			<td>25-375</td>
			<td>nylon mesh, 40 micron strainers</td>
		</tr>
		<tr>
			<td>Aluminum Hydroxide, Reagent Grade</td>
			<td>Sigma-Aldrich</td>
			<td>239186-25G</td>
			<td>20 mg/mL</td>
		</tr>
		<tr>
			<td>anti- mouse CCR4; APC-conjugated</td>
			<td>Biolegend</td>
			<td>131211</td>
			<td>0.5 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse CD11b</td>
			<td>BD Pharmigen</td>
			<td>557396</td>
			<td>0.5 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse CD11c; PE eFluor 610</td>
			<td>Thermo-Fischer Scientific</td>
			<td>61-0114-82</td>
			<td>0.25 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse CD16/32, Fc block</td>
			<td>BD Pharmigen</td>
			<td>553141</td>
			<td>0.5 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse CD19; APC-eFluor 780 conjugated</td>
			<td>Thermo-Fischer Scientific</td>
			<td>47-0193-82</td>
			<td>0.5 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse CD3; PE Cy 7-conjugated</td>
			<td>BD Pharmigen</td>
			<td>552774</td>
			<td>0.25 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse CD45; PE conjugated</td>
			<td>BD Pharmigen</td>
			<td>56087</td>
			<td>0.5 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse ICOS (CD278)</td>
			<td>BD Pharmigen</td>
			<td>564070</td>
			<td>0.5 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse NK1.1 (CD161); FITC-conjugated</td>
			<td>BD Pharmigen</td>
			<td>553164</td>
			<td>0.25 ug/test</td>
		</tr>
		<tr>
			<td>anti-mouse ST2 (IL-33R); PerCP Cy5.5 conjugated</td>
			<td>Biolegend</td>
			<td>145311</td>
			<td>0.5 ug/test</td>
		</tr>
		<tr>
			<td>Automated Cell Counter</td>
			<td>BIORAD</td>
			<td>1450102</td>
		</tr>
		<tr>
			<td>Automated Dissociator</td>
			<td>MACS Miltenyi Biotec</td>
			<td>130-093-235</td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin, Lyophilized Powder</td>
			<td>Sigma-Aldrich</td>
			<td>A2153-10G</td>
			<td>0.5% in serum-free RPMI</td>
		</tr>
		<tr>
			<td>Cell Counter Clides</td>
			<td>BIORAD</td>
			<td>1450015</td>
		</tr>
		<tr>
			<td>Chicken Egg Ovalbumin, Grade V</td>
			<td>Sigma-Aldrich</td>
			<td>A5503-10G</td>
			<td>500 ug/mL</td>
		</tr>
		<tr>
			<td>Collagenase, Type 1, Filtered</td>
			<td>Worthington Biochemical Corporation</td>
			<td>CLSS-1, purchase as 5 X 50 mg vials (LS004216)</td>
			<td>25 U/mL in RPMI</td>
		</tr>
		<tr>
			<td>compensation beads</td>
			<td>Affymetrix</td>
			<td>01-1111-41</td>
			<td>1 drop per contol tube</td>
		</tr>
		<tr>
			<td>Dissociation Tubes</td>
			<td>MACS Miltenyi Biotec</td>
			<td>130-096-335</td>
		</tr>
		<tr>
			<td>FACS Buffer</td>
			<td>BD Pharmigen</td>
			<td>554657</td>
			<td>1x PBS + 2% FBS, w/ sodium azide; stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Heat Inactivated-FBS</td>
			<td>Genesee Scientific</td>
			<td>25-525H</td>
			<td>10% in complete RPMI &#38; ILC2 Expansion Media</td>
		</tr>
		<tr>
			<td>mouse CCL17</td>
			<td>GenScript</td>
			<td>Z02954-20</td>
			<td>50 ng/mL</td>
		</tr>
		<tr>
			<td>mouse CCL22</td>
			<td>GenScript</td>
			<td>Z02856-20</td>
			<td>50 ng/mL</td>
		</tr>
		<tr>
			<td>mouse CD4+ T cell enrichment kit</td>
			<td>STEM Cell Technologies</td>
			<td>19852</td>
		</tr>
		<tr>
			<td>mouse IL-2</td>
			<td>GenScript</td>
			<td>Z02764-20</td>
			<td>20 ng/mL</td>
		</tr>
		<tr>
			<td>mouse ILC2 enrichment kit</td>
			<td>STEM Cell Technologies</td>
			<td>19842</td>
		</tr>
		<tr>
			<td>mouse recombinant IL-33</td>
			<td>STEM Cell Technologies</td>
			<td>78044</td>
			<td>20 ng/mL</td>
		</tr>
		<tr>
			<td>RPMI</td>
			<td>Life Technologies Corporation</td>
			<td>22400071</td>
		</tr>
		<tr>
			<td>Separation Buffer</td>
			<td>STEM Cell Technologies</td>
			<td>20144</td>
			<td>1 X PBS + 2% FBS; stored at 4C</td>
		</tr>
		<tr>
			<td>small animal nebulizer and chamber</td>
			<td>Data Sciences International</td>
		</tr>
		<tr>
			<td>sterile saline</td>
			<td>Baxter</td>
			<td>2F7124; NDC 0338-0048-04</td>
			<td>0.9% Sodium Chloride</td>
		</tr>
	</tbody>
</table>

assessment of lymphocyte migration in an ex vivo transmigration system

在这里，我们提出了一个有效的方法，可以执行与基本的实验室技能和材料，以评估淋巴细胞化学动力学运动在外体迁移系统。组2先天淋巴细胞（ILC2）和CD4+T助剂细胞从脾脏和肺部分离出，对受挑战的BALB/c小鼠。我们比较地确认了CCR4在CD4+T细胞和ILC2上的表达。CCL17 和 CCL22 是 CCR4 的已知配体;因此，使用这种活体迁移方法，我们检查了CCL17- CCL22诱导的CCR4+ 淋巴细胞的运动。为了建立化学梯度，CCL17和CCL22被放置在迁移系统的底部腔室。分离的淋巴细胞然后被添加到顶室，并在48小时期间淋巴细胞通过3μm孔主动迁移到底部腔室的化学素。这是一个有效的系统，以确定淋巴细胞的化学动力学，但可以理解的是，不模仿在体内器官微环境中发现的复杂性。这是通过增加正在研究的器官和淋巴细胞的原位成像来克服的方法的一个限制。相反，这种方法的优点是，入门级技术人员可以以比现场成像更具成本效益的速度执行。当治疗化合物可用于增强迁移时，如肿瘤渗透细胞毒性免疫细胞或抑制迁移，也许在免疫病理学备受关注的自身免疫性疾病中，此方法可用作筛选工具。一般来说，如果感兴趣的化学基因在统计上高于介质控制水平生成化学动力学，则该方法是有效的。在这种情况下，也可以确定给定化合物的抑制/增强程度。

CD4+ T 细胞和 ILC2 上的 CCR4 表达。
为了进行活体迁移实验的成功，必须确定淋巴细胞是否对CCL17和CCL22通过CCR4作出反应;因此，我们通过流动细胞测定测定了CD4+T细胞和ILC2的CCR4表达。虽然众所周知，OVA 特异性 CD4+帮助器 T 细胞表示 CCR4，但对 ILC2 上 CCR4 的表达知之甚少。图1显示了来自雄性和雌性、OVA挑战BALB/c小鼠的CCR4表达的?...

Watch this Scientific Journal Video about Assessment of Lymphocyte Migration in an Ex Vivo Transmigration System at JoVE.com

Assessment of Lymphocyte Migration in an Ex Vivo Transmigration System

在此协议中，淋巴细胞被放置在迁移系统的顶室中，由多孔膜与底部腔室隔开。化学因子被添加到底部腔室，从而诱导沿化学基质梯度的活性迁移。48小时后，淋巴细胞在两个腔室计数，以定量迁移。

外迁移系统中淋巴细胞迁移的评估

Herein, we present an efficient method that can be executed with basic laboratory skills and materials to assess lymphocyte chemokinetic movement in an ex ...

淋巴细胞迁移是任何免疫反应的一个重要特征。因此，这个协议是重要的，因为它允许研究人员评估淋巴细胞在一个明确定义的体外系统的流动性。该技术的主要优点是，可以评估各种化疗素，以确定其诱导迁移在最近定义的细胞，如组两个先天淋巴细胞，或 ILC2 的有效性。第二个优点是，在与动物进行更昂贵的实验之前，可以用这种方法测试抑制剂或生物疗法，以阻断特定的化疗途径。首先从安乐死的雌性小鼠中排出肺和肺，每组使用两到三只动物。根据组织类型和动物性别，将切除的组织放在单独的分离管中。将肺组织放在500微升的肺分离培养子中，放在分离管中，将管放在自动组织分离器中，并使用肺方案将其分离。重复这些步骤总共两次。要分离脾脏组织，请将其放在分离器中 500 微升的完整 RPMI 介质中，然后使用脾脏协议。从现在起，使用无菌技术在生物安全柜中工作，通过 40 微米细胞滤株过滤每个分离组织，并收集到 50 毫升锥形管中。用五毫升的额外肺分离介质冲洗含有肺均质的分离管，用五毫升完整的 RPMI 中含有脾质的管，并通过用于相应组织的过滤器过滤其内容。为了进一步分离肺组织，在37摄氏度的培养箱中孵育肺均质15至30分钟，并吸收5%的二氧化碳。然后向肺和脾均质和离心机添加五毫升完整的 RPMI。丢弃上流剂后，将同性动物群中含有血细胞和肺细胞的颗粒组合成一个50毫升的圆锥管。在每个管中加入10毫升完整的RPMI，然后重新暂停。使用自动单元格计数器确定总单元格计数。在分离缓冲液中，将男性和女性细胞悬浮液调整为每毫升1亿个细胞，然后加入5毫升聚苯乙烯管。在将两个先天淋巴细胞从手稿中描述的2/3细胞中分离后，使用剩余的细胞进行CD4阳性T细胞分离程序。要开始CD4阳性T细胞分离，将大鼠血清加入CD4 T细胞富集悬浮液。然后将分离鸡尾酒加入细胞样本，在室温下孵育10分钟。涡流快速球体30秒，并添加到细胞样品中，实现每毫升75微升的稀释。轻轻混合，在室温下孵育2 1~2分钟。在样品顶部，用约两毫升的分离缓冲液，最多三毫升，将五毫升管放入八室易分离磁铁中，并在室温下孵育五分钟。然后将磁铁向前倾斜，将每个细胞悬浮液移液器插入一个干净的五毫升管中。在每个管和离心机中加入1.5毫升无血清RPMI。从上一提液中倒出，在无血清RPMI中以每毫升1000万细胞重新浇注CD4阳性T细胞颗粒。要开始协议的这一部分，请确定 ILC2 和 CD4 阳性 T 细胞实验所需的 Transwell 插入件数，如手稿中所述。用移液器尖端和戴手套的手，轻轻地将三微米跨井刀片从 24 井板的中间行移动到同一 24 井板的顶部和底部行。在24孔板的中间行中，用CCL17加入500微升迁移介质，将CCL17添加到1/3的井中。将 500 微升迁移介质与 CCL22 添加到另外 1/3 的油井中。最后，在井的最后1/3中加入500微升不含化疗的RPMI。将板盖清楚地贴上底部井中放置适当迁移介质的名称。然后，将Transwell插入回包含各种处理的井中。轻轻将 100 微升 CD4 T 细胞或 ILC2 添加到每个刀片的顶部井中，确保不要在 Transwells 上下混合或移液细胞悬浮液。将板盖清楚地贴上每个井中的单元格类型标签，并写下添加单元格的日期和时间。从从板中卸下 Transwell 刀片开始重复此过程，直到所有电池都已放置在带介质的 Transwell 刀片中。一旦感兴趣的细胞被分离，最重要的步骤是跟踪放置在特定孔中的化疗和细胞类型，轻轻地将细胞添加到顶部腔室，最后避免在48小时的孵化过程中与迁移板进行不必要的接触。将板轻轻放在37摄氏度的培养箱中，用5%的二氧化碳孵化48小时，确保在孵化期间尽量减少与板的接触。轻轻地从培养箱中取出板后，将中间行的所有 Transwell 刀片拆下到空井中，然后位于上方或下方。从 Transwell 的顶部和底部孔收集细胞插入与 CCL17、CCL22 或介质、具有细胞类型和复制编号的上或下标记的管中。用 500 微升 1x PBS 冲洗底井，收集到相应的管中，对顶部井进行相同的操作。在室温下以378倍g离心管5分钟。使用移液器轻轻去除所有上流液，然后用 50 微升 1x PBS 重新暂停 T 细胞和 ILC2 颗粒。去除10微升的细胞悬浮液后，加入90微升0.4%的蓝板。计算自动单元格计数器上的单元格。对于每个样本，记录每毫升的生存能力和细胞计数百分比，并确定顶部和底部腔室中每次治疗的细胞总数。当通过流式细胞测定对CD4阳性T细胞和LC2进行CCR4表达评估时，男性和女性宿主之间没有差异。然而，与CD4阳性T细胞相比，CCR4在每细胞的表达高于CD4阳性T细胞。当Transwell仪器的底部腔室充满未经处理的细胞培养基、含有CCL22的介质或含有CCL17的介质时，在介质控制条件下迁移的细胞不到14%。作为对CCL22的回应，这两种细胞类型，无论它们来自雄性还是雌性宿主，都响应了CCL22。此外，CCL22在男性CLC2中诱发的迁移比CCL17大。另一方面，CCL17诱导女性CD4 T细胞和LC2与介质相比大量迁移。CCL17治疗与雄性CD4阳性T细胞或男性LC2的介质无不同。在次优条件下，当带CD4细胞的板在室温下保持24小时，然后进入孵化器时，没有向CCL22和CCL17迁移。此外，底部腔室中的细胞的生存能力明显较低，表明使用本协议中描述的正确温度和条件实现最佳结果的重要性。一旦迁移程序完成，与介质控制井相比，人们应该看到大量迁移到感兴趣的化学。如果没有看到显著的迁移，可以假设程序不能正常工作，或者淋巴细胞对有关化疗没有反应。当淋巴细胞经过广泛的体内治疗时，这种技术可以广泛用于理解淋巴细胞迁移。

Research

JoVE Journal

Immunology and Infection

Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro

Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro

迁移人T淋巴细胞分离，培养和分析<em在体外</em

The migration of T lymphocytes involves the adhesive interaction of cell surface integrins with ligands expressed on other cells or with extracellular ...

科研

免疫与感染

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

白细胞的归位和迁移的非侵入性的成像在体内</em

Two-photon (2P) microscopy is a high resolution imaging technique that has been broadly adapted by biologists.  The value of 2P microscopy is that it ...

Quantitative Assessment of Human Neutrophil Migration Across a Cultured Bladder Epithelium

人中性粒细胞迁移的跨越培养膀胱上皮定量评价

The recruitment of immune cells from the periphery to the site of inflammation is an essential step in the innate immune response at any mucosal surface. ...

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

评估的HIV-1感染的CD4的先天传感<sup&gt; +</sup&gt;性T细胞浆细胞样树突细胞使用<em&gt;体外</em&gt;共培养体系。

HIV-1 innate sensing requires direct contact of infected CD4+ T cells with plasmacytoid dendritic cells (pDCs). In order to study this process, the ...

An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices

An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices

一个<em&gt;体外</em&gt;急性脑片的少突胶质细胞定向T细胞攻击模式

Death of oligodendrocytes accompanied by destruction of neurons and axons are typical histopathological findings in cortical and subcortical grey matter ...

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 ...

Isolation and Ex Vivo Culture of V&#948;1+CD4+&#947;&#948; T Cells, an Extrathymic &#945;&#946;T-cell Progenitor

Isolation and Ex Vivo Culture of VÃƒÅ½Ã‚Â´1+CD4+ÃƒÅ½Ã‚Â³ÃƒÅ½Ã‚Â´ T Cells, an Extrathymic ÃƒÅ½Ã‚Â±ÃƒÅ½Ã‚Â²T-cell Progenitor

分离和<em&gt;离体</emVδ1的&gt;企业文化<sup&gt; +</sup&gt; CD4<sup&gt; +</sup&gt;γδT细胞，胸腺外的αβT细胞祖

The thymus, the primary organ for the generation of &#945;&#946; T cells and backbone of the adaptive immune system in vertebrates, has long been ...

Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10

粒子结合Bet v 1的变应原PM10的流式细胞分析

Flow cytometry is a method widely used to quantify suspended solids such as cells or bacteria in a size range from 0.5 to several tens of micrometers in ...

Murine Lymphocyte Labeling by 64Cu-Antibody Receptor Targeting for In Vivo Cell Trafficking by PET/CT

Murine Lymphocyte Labeling by 64Cu-Antibody Receptor Targeting for In Vivo  Cell Trafficking by PET/CT

小鼠淋巴细胞通过标签<sup&gt; 64</sup&gt;的Cu-抗体受体靶向用于<em&gt;体内</em&gt;由PET / CT细胞运输

This protocol illustrates the production of 64Cu and the chelator conjugation/radiolabeling of a monoclonal antibody (mAb) followed by murine lymphocyte ...