Name: visualization of inflammatory caspases induced proximity in human monocyte-derived macrophages
Uploaded: 2022-04-06T12:45:00
Description: Watch this Scientific Journal Video about Visualization of Inflammatory Caspases Induced Proximity in Human Monocyte-Derived Macrophages at JoVE.com

我们感谢LBH实验室过去和现在的成员，他们为这项技术的发展做出了贡献。该实验室由NIH / NIDDK T32DK060445（BEB），NIH / NIDDK F32DK121479（BEB），NIH / NIGMS R01GM121389（LBH）提供支持。图2是使用Biorender软件绘制的。

该协议遵循贝勒医学院人类研究伦理委员会关于操纵人类样本的指导方针。血液样本的处理遵循人体样本的机构安全指南。血液样本在区域血库获得，在那里用柠檬酸磷酸葡萄糖（CPD）溶液收集。然而，用其他抗凝剂如肝素钠，锂肝素或EDTA收集的血液也可用于该方案28，29。
1.人单核细胞的分离和分化成巨噬细胞
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该协议描述了从从人血液样本中分离的单核细胞中获取巨噬细胞的工作流程，以及一种在不影响细胞活力和行为的情况下将炎症性半胱天冬酶BiFC报告基因有效地引入人MDM的方法。
该协议利用BiFC技术35 用分裂荧光蛋白Venus的非荧光片段标记半胱天冬酶募集结构域（CARD）处的炎症半胱天冬酶。炎症性半胱天冬酶编码由大（p20）和小（p10）亚基组成的催化结构域...

半胱天冬酶是半胱氨酸天冬氨酸蛋白酶的一个家族，可以分为引发子半胱天冬酶和刽子手半胱天冬酶。刽子手半挂酶包括半胱天冬酶-3、-6和-7。它们以二聚体的形式天然存在于细胞中，并被引发剂半胱天冬酶切割以执行细胞凋亡1。引发剂半胱天冬酶包括人半胱天冬酶-1、-2、-4、-5、-8、-9、-10和-12。它们被发现为无活性酶原（前半胱天冬酶），通过接近诱导的二聚化激活，并通过自体蛋白水解裂解2，3稳定。炎症性半胱天冬酶是引发子半胱天冬酶2 的一个子集，包括人类中的半胱天冬酶-1、-4、-5 和 -12，以及小鼠4、5 中的半胱天冬酶-1、-11 和 -12。它们不是凋亡作用，而是在炎症中起核心作用。它们介导蛋白水解处理和促白细胞介素（IL）-1β和亲IL-186，7的分泌，这是响应致病性入侵者释放的第一批细胞因子8，9。半叶酶-1在其激活平台募集时被激活;称为炎症小体的大分子量蛋白质复合物（图1A）10。半胱天冬酶-4、-5 和 -11 的二聚化通过非规范的炎性小体途径11，12 独立于这些平台发生。
典型炎性粒体是胞质多聚体蛋白复合物，由炎性粒体传感器蛋白、适应蛋白ASC（含有CARD的凋亡相关斑点样蛋白）和效应蛋白半胱天冬酶-110组成。研究最充分的规范炎小体是含有pyrin结构域（NLRP），NLRP1和NLRP3的NOD样受体家族，含有CARD（NLRC），NLRC4的NLR家族以及黑色素瘤2（AIM2）中不存在的。它们各自包含一个pyrin域，一个CARD或两个域。CARD域介导含CARD的半链酶与其上游激活剂之间的相互作用。因此，由N端pyrin结构域（PYD）和C端CARD基序13，14组成的支架分子ASC是将半胱天冬酶-1募集到NLRP110，NLRP315和AIM216 炎性小体所必需的。
每个炎症小体都以其独特的传感器蛋白命名，该传感器蛋白可识别不同的促炎刺激（图1B）。该途径的激活剂被称为规范刺激。炎症小体充当微生物成分和组织应激的传感器，并通过激活炎症半胱天冬酶17来组装以触发强大的炎症反应。炎症小体组装启动半胱天冬酶-1活化以介导其主要底物pro-IL-1β和pro-IL-18的成熟和分泌。此过程通过两步机制进行。首先，启动刺激通过激活NF-κB途径上调某些炎症小体蛋白和pro-IL-1β的表达。其次，细胞内（规范）刺激诱导炎症小体组装和前卡星酶-16，7的募集。
半胱天冬酶-4和半胱天冬酶-5是小鼠半胱天冬酶-11 11的人类同源物。它们通过细胞内脂多糖（LPS）以炎症小体独立的方式被激活，LPS是在革兰氏阴性细菌18，19，20的外膜中发现的分子，以及细胞外血红素，红细胞溶血21的产物。已经提出LPS直接与这些蛋白质的CARD基序结合并诱导其寡聚化20。半胱天冬酶-4或半胱天冬酶-5的活化通过裂解形成孔隙蛋白gasdermin D（GSDMD）18，19诱导称为焦磷酸化的炎症形式的细胞死亡来促进IL-1β的释放。此外，由半胱天冬酶-4和GSDMD介导的热融性死亡引起的钾离子外排诱导NLRP3炎症小体的活化和随后的半胱天冬酶-122，23的活化。因此，半胱天冬酶-4、-5 和 -11 被认为是 LPS 的细胞内传感器，它们能够诱导焦磷酸化和半胱天冬酶-1 激活以响应特定的刺激11，24。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/63162/63162fig01.jpg"> 图1：炎症半胱天冬酶和半胱天冬酶 - 双分子荧光互补（BiFC）测定。 （A）显示半胱天冬酶-BiFC系统的图表，其中与金星的每个非荧光片段（Venus-C或Venus-N）相连的两个半胱天冬酶-1蛋白结构域（C1-pro）被招募到NLRP3激活平台，迫使金星重新折叠并发出荧光。该复合物在显微镜下显示为绿点，并用作炎症性半胱天冬酶诱导的接近的读数，这是引发半胱天冬酶激活的第一步。（B）显示炎症小体成分和炎症性半胱天冬酶的结构域组织的示意图。 <a href="https://www.jove.com/files/ftp_upload/63162/63162fig01large.jpg" target="_blank">请点击此处查看此图的大图。</a>
测量特异性引发剂半胱天冬酶活化是困难的，并且通过成像方法没有多少方法可以做到这一点。半胱天冬酶双分子荧光互补（BiFC）可用于直接在活细胞中可视化炎症半胱天冬酶活化（图1A）25。该技术最近已被改编用于人单核细胞来源的巨噬细胞（MDM）21。半胱天冬酶BiFC测量炎症性半胱天冬酶活化的第一步，诱导接近以促进二聚化。使用编码含CARD的半胱天冬酶pro结构域的质粒的表达，这些质粒融合到光稳定黄色荧光蛋白Venus（Venus-C [VC]）和Venus-N [VN]）的非荧光片段中。当两个半胱天冬酶前跨域被招募到它们的激活平台或经历诱导的接近时，金星的两半被靠近并被迫重新折叠和发出荧光（见 图1A，B）。这提供了特定炎症性半胱天冬酶激活的实时读数。
人类MDM大量表达炎症小体基因和模式识别受体，识别危险信号和病原体产物。这为炎症半胱天冬酶途径的询问提供了理想的细胞类型。此外，它们可以来自外周血，甚至来自患者样本，以评估特定疾病状态下的炎症性半胱天冬酶激活。该协议描述了如何使用核感染，一种基于电穿孔的转染方法将BiFC半胱天冬酶报告基因引入MDM，如何处理细胞以诱导炎症性半胱天冬酶活化，以及如何使用显微镜方法可视化活性半胱天冬酶复合物。此外，该方法可以适于确定这些复合物的分子组成，亚细胞定位，动力学和这些高度有序结构的大小25，26，27。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>48 well tissue culture2:34 plates</td>
			<td>Genesee Scientific</td>
			<td>25-108</td>
			<td></td>
		</tr>
		<tr>
			<td>10 cm Tissue Culture Dishes</td>
			<td>VWR</td>
			<td>25382-166</td>
			<td></td>
		</tr>
		<tr>
			<td>2 Mercaptoethanol 1000x</td>
			<td>Thermo Fisher Scientific</td>
			<td>21985023</td>
			<td></td>
		</tr>
		<tr>
			<td>8 well chambered coverglass with 1.5 HP coverglass</td>
			<td>Cellvis</td>
			<td>c8-1.5H-N</td>
			<td></td>
		</tr>
		<tr>
			<td>AutoMACS columns</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-021-101</td>
			<td>For automated separation using AutoMACS Pro Separator only</td>
		</tr>
		<tr>
			<td>AutoMACS Pro Separator</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-092-545</td>
			<td>For automated separation using AutoMACS Pro Separator only</td>
		</tr>
		<tr>
			<td>AutoMACS Pro Washing Solution</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-092-987</td>
			<td>For automated separation using AutoMACS Pro Separator only</td>
		</tr>
		<tr>
			<td>AutoMACS Rinsing Solution</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-091-222</td>
			<td>For automated separation using AutoMACS Pro Separator only</td>
		</tr>
		<tr>
			<td>AutoMacs running buffer</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-091-221</td>
			<td>For manual or automated separation using QuadroMACS or AutoMACS pro Separator</td>
		</tr>
		<tr>
			<td>Axio Observer Z1 motorized inverted microscope equipped with a CSU-X1A 5000 spinning disk unit</td>
			<td>Zeiss</td>
			<td></td>
			<td>Any confocal microscope equipped with a laser module fitted with laser lines of 568 nm (RFP) and 488 or 512 nm (GFP or YFP) wavelengths can be used</td>
		</tr>
		<tr>
			<td>AxioObserver A1, Research Grade Inverted Microscope</td>
			<td>Zeiss</td>
			<td></td>
			<td>Any epifluorescence microscope with fluorescence filters capable of exciting 568 nm (RFP) and 488 or 512 nm (GFP or YFP) wavelengths can be used</td>
		</tr>
		<tr>
			<td>CD14+ MICROBEADS</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-050-201</td>
			<td>For manual or automated separation using QuadroMACS or AutoMACS pro Separator</td>
		</tr>
		<tr>
			<td>DPBS without calcium chloride and magnesium chloride</td>
			<td>Sigma</td>
			<td>D8537-6x500ML</td>
			<td></td>
		</tr>
		<tr>
			<td>DsRed mito plasmid</td>
			<td>Clontech</td>
			<td>632421</td>
			<td>Similar plasmids that can be used as fluorescent reporters can be found on Addgene</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Thermo Fisher Scientific</td>
			<td>10437028</td>
			<td></td>
		</tr>
		<tr>
			<td>Ficoll-Paque PLUS 6 x 100 mL</td>
			<td>Sigma</td>
			<td>GE17-1440-02</td>
			<td></td>
		</tr>
		<tr>
			<td>GlutaMAX Supplement (100x)</td>
			<td>Thermo Fisher Scientific</td>
			<td>35050079</td>
			<td></td>
		</tr>
		<tr>
			<td>GM-CSF</td>
			<td>Thermo Fisher Scientific</td>
			<td>PHC2011</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemin BioXtra, from Porcine, &#8805;96.0% (HPLC)</td>
			<td>Sigma</td>
			<td>51280-1G</td>
			<td></td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>Thermo Fisher Scientific</td>
			<td>15630106</td>
			<td></td>
		</tr>
		<tr>
			<td>Inflammatory caspase BiFC plasmids</td>
			<td></td>
			<td></td>
			<td>Available by request from LBH lab</td>
		</tr>
		<tr>
			<td>LPS-EB Ultrapure</td>
			<td>Invivogen</td>
			<td>TLRL-3PELPS</td>
			<td></td>
		</tr>
		<tr>
			<td>LS Columns</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-042-401</td>
			<td>For manual separation using QuadroMACS Separator only</td>
		</tr>
		<tr>
			<td>MACS 15 mL Tube Rack</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-091-052</td>
			<td>For manual separation using QuadroMACS Separator only</td>
		</tr>
		<tr>
			<td>MACS MultiStand</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-042-303</td>
			<td>For manual separation using QuadroMACS Separator only</td>
		</tr>
		<tr>
			<td>mCherry plasmid</td>
			<td>Yungpeng Wang Lab</td>
			<td></td>
			<td>Similar plasmids that can be used as fluorescent reporters can be found on Addgene</td>
		</tr>
		<tr>
			<td>Neon Transfection System</td>
			<td>Thermo Fisher Scientific</td>
			<td>MPK5000</td>
			<td>Includes Neon electroporation device, pipette and pipette station</td>
		</tr>
		<tr>
			<td>Neon Transfection System 10 &#181;L Kit</td>
			<td>Thermo Fisher Scientific</td>
			<td>MPK1096</td>
			<td>Includes resuspension buffer R, resuspension buffer T, electrolytic buffer E, 96 x 10 &#181;L Neon tips and Neon electroporation tubes</td>
		</tr>
		<tr>
			<td>Nigericin sodium salt, ready made solution</td>
			<td>Sigma</td>
			<td>SML1779-1ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin (10,000 U/mL)</td>
			<td>Thermo Fisher Scientific</td>
			<td>15140122</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly-D-Lysine Hydrobromide</td>
			<td>Sigma</td>
			<td>P7280-5mg</td>
			<td></td>
		</tr>
		<tr>
			<td>QuadroMACS Separator</td>
			<td>Miltenyi (Biotec)</td>
			<td>130-090-976</td>
			<td>For manual separation using QuadroMACS Separator only</td>
		</tr>
		<tr>
			<td>qVD-OPh</td>
			<td>Fisher (ApexBio)</td>
			<td>50-101-3172</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI 1640 Medium</td>
			<td>Thermo Fisher Scientific</td>
			<td>11875119</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin-EDTA (0.25%), phenol red</td>
			<td>Thermo Fisher Scientific</td>
			<td>25200072</td>
			<td></td>
		</tr>
		<tr>
			<td>UltraPure 0.5 M EDTA, pH 8.0</td>
			<td>Thermo Fisher Scientific</td>
			<td>15575020</td>
			<td></td>
		</tr>
		<tr>
			<td>Zeiss Zen 2.6 (blue edition) software</td>
			<td>Zeiss</td>
			<td></td>
			<td>Any software used to operate the confocal microscope of choice</td>
		</tr>
	</tbody>
</table>

visualization of inflammatory caspases induced proximity in human monocyte-derived macrophages

炎症性半胱天冬酶包括半胱天冬酶-1、-4、-5、-11 和 -12，属于引发者半胱天冬酶的亚组。半胱天冬酶-1是确保炎症信号传导正确调节所必需的，并且在募集到炎症小体后通过接近诱导的二聚化激活。半胱天冬酶-1在单核细胞谱系中含量丰富，并诱导促炎细胞因子白细胞介素（IL）-1β和IL-18向活性分泌分子成熟。其他炎症性半胱天冬酶，半胱天冬酶-4和-5（及其小鼠同源半胱天冬酶-11）通过诱导焦磷酸化来促进IL-1β的释放。半胱天冬酶双分子荧光互补（BiFC）是一种用于测量炎症性半胱天冬酶诱导的接近度作为半胱天冬酶活化的读数的工具。半胱天冬酶-1，-4或-5原结构域，其中包含与炎症小体结合的区域，与黄色荧光蛋白Venus（Venus-N [VN]或Venus-C [VC]）的非荧光片段融合，当半胱天冬酶发生诱导接近时，这些片段与荧光金星复合物结合以重整荧光金星复合物。该协议描述了如何使用细胞核感染将这些报告基因引入原代人单核细胞来源的巨噬细胞（MDM），处理细胞以诱导炎症性半胱天冬酶活化，并使用荧光和共聚焦显微镜测量半胱天冬酶活化。这种方法的优点是它可用于鉴定活细胞中炎症性半胱天冬酶活化复合物的组分，要求和定位。然而，需要考虑仔细的控制，以避免损害细胞活力和行为。该技术是一种强大的工具，用于分析炎症小体水平的动态半胱天冬酶相互作用，以及询问活MDM和来自人类血液样本的单核细胞中的炎症信号级联。

图2所示的方案概述了如何获得，转染和成像人MDM.在选定的CD14 +单核细胞与GM-CSF孵育7天后，细胞形态在分化期间发生变化（图3A），从球形悬浮细胞到旋转和完全附着（第3天和第4天）， 最后，在完全分化时（第7天）进行更多的扩散细胞。然后将完全分化的细胞从平板中分离出来，并用半胱天冬酶BiFC对（VC和VN）以及报告质粒（例如，dsRedmito，一种编?...

Watch this Scientific Journal Video about Visualization of Inflammatory Caspases Induced Proximity in Human Monocyte-Derived Macrophages at JoVE.com

Visualization of Inflammatory Caspases Induced Proximity in Human Monocyte-Derived Macrophages

该协议描述了从人血液样本中获取单核细胞来源巨噬细胞（MDM）的工作流程，一种在不损害细胞活力和行为的情况下将炎症半胱天冬酶双分子荧光互补（BiFC）报告基因有效引入人MDM的简单方法，以及一种基于成像的方法来测量活细胞中的炎症半胱天冬酶活化。

炎症性半月酶诱导的人单核细胞来源巨噬细胞邻近性的可视化

Inflammatory caspases include caspase-1, -4, -5, -11, and -12 and belong to the subgroup of initiator caspases. Caspase-1 is required to ensure correct ...

该协议具有重要意义，因为它可用于在特定疾病状态下的活细胞中分子水平上可视化和询问炎症半胱天冬酶途径。该技术的主要优点是它可用于鉴定原代免疫细胞中炎症半胱天冬酶活化复合物的成分，要求和定位。通过微小的优化，该方法可以适应其他原代细胞和通过寡聚化激活的一系列蛋白质，并且其适用性不仅限于微观方法。首先，在10厘米培养皿上从完全分化的巨噬细胞中吸取培养基，并用温热的血清游离RPMI-1640培养基洗涤细胞单层，完全除去所有培养基。通过每10厘米培养皿中加入两毫升0.25%温胰蛋白酶- EDTA溶液来收获细胞。用P-1000微量移液管轻轻地将胰蛋白酶-EDTA上下移液到整个培养皿上，然后将悬浮液转移到含有5毫升温完全培养基的15毫升锥形管中。在明场显微镜下检查细胞脱落，并在需要时再次分离。吸出培养基并将细胞重悬于10毫升预热至37摄氏度的1X无菌PBS中。然后取20微升等分试样，使用血细胞计数器确定细胞数。每次转染10至5个细胞，取1至2次，并将其放入15毫升管中。将最终体积为15毫升，预热的1X无菌PBS。吸出PBS，并以250×g离心一次，持续一分钟。使用P-200微量移液管从细胞沉淀中除去任何残留的PBS。每次转染服用1.5毫升无菌微管，并在烟罩中加入适当的报告质粒和半胱天冬酶BiFC质粒。将移液器站、设备、吸头、电穿孔管和移液器放在无菌层流罩中。连接并打开核体测量设备。在显示的启动屏幕中输入事务参数。按“电压”，输入 1000，然后按“完成”将其设置为 1000 伏。按“宽度”，输入 40，然后按“完成”将脉冲设置为 40 毫秒。最后，按脉冲，输入 2，然后按完成将电脉冲数设置为 2。取其中一个电穿孔管，并在室温下用三毫升电解缓冲液E填充。将电穿孔管插入移液器站上的移液器支架中，确保管子侧面的电极朝内，并且在插入管子时听到咔嗒声。取细胞沉淀，向第五个细胞中加入10微升预热的重悬R缓冲液，每次1至2次10次，并与P-20微量移液管轻轻混合。向每个转染管中加入10微升细胞悬浮液，并与P-20微量移液管轻轻混合。通过按下按钮到第二个停止点将吸头插入移液器，确保夹具完全拾取吸头中活塞的安装杆。接下来，按下移液器上的按钮到第一个停止点，然后浸入第一个含有细胞或质粒DNA混合物的管中以吸出样品。将带有样品的移液器非常小心地插入移液器支架中，确保移液器卡住并正确放置。在触摸屏上按“开始”，然后等待电脉冲发出。从工作站中缓慢取出移液器，然后通过缓慢按下按钮到第一个停止处，立即将横切的细胞悬浮液与预热的无抗生素培养基一起加入相应的孔中。用横切细胞轻轻摇动板，并在加湿的组织培养箱中孵育一到三个小时，然后向每个孔中加入200微升预热的培养基。将培养皿放入培养箱中，等待至少24小时的基因表达。第二天，使用落射荧光显微镜检查细胞活力和转染效率。用P-1000微量移液管小心地从细胞中取出培养基，并在孔的侧面加入500微升的刺激溶液。要运行未经处理的控制孔，请添加没有刺激的成像介质。要使用落射荧光或共聚焦显微镜观察细胞，请按照说明打开显微镜和荧光光源。选择10倍或20倍物镜，并将培养皿放在显微镜载物台上。使用目镜在568纳米滤光片下找到细胞。并专注于表达报告基因红细胞的细胞。计算视野中的所有红细胞。在同一视野中，更改为 488 或 512 过滤器，计算同样为绿色的红色单元格的数量。计数至少100个至少三个字段的红色单元格。计算每个视野中金星阳性横切细胞的百分比，并平均每个治疗孔的结果百分比，以获得标准偏差。要使用落射荧光或共聚焦显微镜对细胞进行成像，请在计算机屏幕上可视化由相机获取的细胞的实时图像。使用操纵杆控制和对焦轮微调单元的焦点和位置。设置512纳米或488纳米和568纳米激光器的百分比激光功率和曝光时间，以使图像中的信号看起来良好而不会饱和。打开实时捕获并检查生成的图像，确保在两个通道的显示直方图中，每个楼层都可以看到不同的峰值。在可视化细胞的实时图像时，拍摄一个场的多个代表性图像，该场包含一个或多个细胞，这些细胞表达板中每个孔的mCherry或dsRedmito报告基因。与GM-CSF一起孵育七天的选定的CD-14阳性单核细胞在分化期间显示出细胞形态的变化，从球形悬浮细胞到旋转和完全附着，最后，在完全分化时到更多的扩散细胞。用半胱天冬酶BiFC对VC和VN以及报告质粒dsRedmito横切分化完全分化的细胞，用于标记横切细胞。未经处理的细胞没有金星荧光。在尼日利亚红素处理的细胞中，半胱天冬酶-1 BiFC以单一的点状物出现，具有ASC斑点的典型形状。Venus阳性MDM的定量显示，在LPS加nigericin治疗组中，半胱天冬酶-1 BiFC的百分比最高。在半胱天冬酶-1、4 和 5 亲 BiFC 对的质粒滴定中，金星阳性细胞的比例最高分别来自 400、500 和 1，000 纳克横切质粒。在横断术后24小时用细胞场的20倍物镜获得的共聚焦图像表明，未经处理的横切细胞是有活力的，因为线粒体是完整的。用血红素治疗后，半胱天冬酶-1复合物表现为类似于尼日利亚霉素诱导的单个绿色点。请记住，在分化、转染和治疗过程中避免苛刻的条件和对细胞的过度操作，因为它会导致自发活化和高背景水平的半胱天冬酶活化。

Research

JoVE Journal

Immunology and Infection

An In vitro Co-infection Model to Study Plasmodium falciparum-HIV-1 Interactions in Human Primary Monocyte-derived Immune Cells

An In vitro Co-infection Model to Study Plasmodium falciparum-HIV-1 Interactions in Human Primary Monocyte-derived Immune Cells

一个<em&gt;在体外</em&gt;共感染模型研究<em&gt;恶性疟原虫</em人原发性单核细胞源性的免疫细胞-HIV-1相互作用

Plasmodium falciparum, the causative agent  of the deadliest form of malaria, and human immunodeficiency virus type-1 (HIV-1) are among the most important ...

科研

免疫与感染

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1&#946; in Human Monocyte-derived Dendritic Cells

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1ÃƒÅ½Ã‚Â² in Human Monocyte-derived Dendritic Cells

激活和NLRP3炎性活性的人单核细胞来源的树突状细胞用IL-1β测量

Inflammatory processes resulting from the secretion of Interleukin (IL)-1 family cytokines by immune cells lead to local or systemic inflammation, tissue ...

Culture of Macrophage Colony-stimulating Factor Differentiated Human Monocyte-derived Macrophages

巨噬细胞集落刺激因子分化的人单核细胞衍生的巨噬细胞的培养

A protocol is presented for cell culture of macrophage colony-stimulating factor (M-CSF) differentiated human monocyte-derived macrophages. For initiation ...

Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols

通过成像流式细胞仪人单核细胞来源的树突状细胞的特征：两个单核细胞分离方法的比较

Dendritic cells (DCs) are antigen presenting cells of the immune system that play a crucial role in lymphocyte responses, host defense mechanisms, and ...

Generation of Human Monocyte-derived Dendritic Cells from Whole Blood

从全血人单核细胞来源的树突状细胞的生成

Dendritic cells (DCs) recognize foreign structures of different pathogens, such as viruses, bacteria, and fungi, via a variety of pattern recognition ...

Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry

通过流式细胞术分析来自中枢神经系统的小胶质细胞和单核细胞衍生的巨噬细胞

Numerous studies have demonstrated the role of immune cells, in particular macrophages, in central nervous system (CNS) pathologies. There are two main ...

Quantification of Monocyte Chemotactic Activity In Vivo and Characterization of Blood Monocyte Derived Macrophages

单核细胞化学策略活性在Vivo的定量和血液单核细胞衍生巨噬细胞的特征

Tissue homeostasis and repair are critically dependent on the recruitment of monocyte-derived macrophages. Both under- and over-recruitment of ...

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase (G6PI)-Induced RA Mice

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase G6PI-Induced RA Mice

葡萄糖-6-磷酸异构酶（G6PI）诱导RA小鼠中iNKT细胞的采用免疫治疗

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T ...

In Vitro Stimulation and Visualization of Extracellular Trap Release in Differentiated Human Monocyte-derived Macrophages

分化人类单细胞衍生巨噬细胞中细胞外陷阱释放的体外刺激与可视化

The release of extracellular traps (ETs) by neutrophils has been identified as a contributing factor to the development of diseases related to chronic ...

Determination of Vaccine Immunogenicity Using Bovine Monocyte-Derived Dendritic Cells

使用牛单核细胞来源的树突状细胞测定疫苗免疫原性

Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs) within the immune system. They patrol the organism looking for pathogens and ...