Name: identification and characterization of immunogenic rna species in hdm allergens that modulate eosinophilic lung inflammation
Uploaded: 2020-05-30T15:00:00
Description: Watch this Scientific Journal Video about Identification and Characterization of Immunogenic RNA Species in HDM Allergens that Modulate Eosinophilic Lung Inflammation at JoVE.com

我们感谢卡拉·戈雷纳女士在流动细胞学方面提供技术援助。L.S. 得到中国奖学金理事会和湖南省研究生创新基金会 （CX201713068） 的支持。H.H.A. 由沙特阿拉伯萨卡卡朱夫大学应用医学学院临床实验室科学系提供支持。X.D.L. 由 UT 健康圣安东尼奥医学院创业基金和马克斯和明奈沃尔克基金支持。

这里描述的实验程序得到了德克萨斯大学圣安东尼奥分校机构动物护理和使用委员会的批准。
1. 点印，以显示 HDM 总 RNA 中存在 dsRNA 结构
<ol><li>与过敏原、昆虫和非昆虫过敏原的总RNA分离<ol>
<li>将活捉或商业获得的 HDM、昆虫或非昆虫动物放入 50 mL 管中，然后快速用液体-N2 冻结。然后在-70°C储存，以进行后续总RNA分离。 注：在这个实验中，HDM，昆虫和非昆虫动物?...

<ol>
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目前的协议描述了如何评估过敏原相关微生物RNA的免疫刺激特性及其对过敏性哮喘小鼠模型中嗜酸性肺炎症发展的影响。虽然长dsRNA被称为许多病毒的复制中间体，可以有力地激活哺乳动物细胞中的干扰素反应，它们在HDM过敏原中的存在一直未知，直到我们最近的工作15。本手稿中介绍的RNA点印迹、RT-qPCR和FACS分析的组合，可能为解剖先天成分（如环境过敏原中的 dsRNA 物种）提供...

根据Strachan1最初提出的卫生假说，幼儿接触环境微生物因素，如内毒素可以防止过敏性疾病2，3,的发展。在微生物感染期间，例如病毒感染，异核酸（RNA/DNA）的先天免疫检测触发宿主防御反应4,4，5，6。,6然而，免疫原核酸的存在和流行，如长双链RNA（dsRNA）物种在家庭尘虫（HDM）或其他昆虫过敏原仍然未知。该协议旨在确定HDM或昆虫和非昆虫过敏原是否含有长dsRNA物种，可以激活保护性免疫反应，以抵消过敏性哮喘小鼠模型中严重嗜酸性肺炎症的发展。在这里，我们提供三种简单而快速的方法来评估HDM总RNA的结构决定因素，这些决定因素是调节过敏原诱发的嗜酸性肺炎症所需的。
粘膜免疫系统是人体最大的免疫器官，是抵御微生物感染和过敏侮辱的第,一道防线。长dsRNA，许多病毒的复制中间体，被称为病原体相关的分子模式（PAMP），通过像3（TLR3）这样的收费受体（TLR3）有效刺激先天反应，以诱导干扰素刺激基因（ISG）9、10、11、12、13、14,10,11,12,13的表达。我们最近表明，HDM总RNA含有dsRNA结构，当通过心内注射在HDM提取物引起的过敏性哮喘的murine模型中施用时，可调节 ISG 的表达，减少严重的嗜酸性肺炎。肺炎症的严重程度是通过分析支气管（BAL）和肺组织的免疫细胞类型，通过流细胞学16，17，18，19，2016,17,18,19确定肺炎症的严重程度。
该协议包括三种检测：1）使用小鼠单克隆抗体J2快速检测具有RNA点印迹的dsRNA结构，该抗体以与序列无关的方式与dsRNA（±40bp）结合;2） 使用RT-qPCR测量 ISG 的诱导，快速评估小鼠肺部免疫刺激RNA的体内效应;3） 使用流量细胞学分析，在HDM引起的肺部炎症背景下准确定量BAL和肺的嗜酸性。
上述检测不仅可用于研究过敏性肺病，还可用于研究呼吸道细菌和病毒感染。例如，dsRNA特异性J2抗体也可用于其他应用，如免疫仿彩色谱、免疫组织化学、酶链接免疫吸附剂测定（ELISA）和免疫染色21、22、23。21,22,23此外，BAL流体收集下游的几种应用可用于使用ELISA量化可溶性物质，如细胞因子和化疗素，以及气道细胞的转录分析（如道道巨噬细胞）。虽然文献中提供了多种评估肺病的协议，但大多数协议通常侧重于目标验证。此处描述的程序可用于识别环境过敏原中对调节过敏性疾病发展非常重要的成分。

<table>
	<tbody>
		<tr>
			<td>0.40 &#181;m Falcon Cell Strainer</td>
			<td>Thermo Fisher Scientific</td>
			<td>08-771-1</td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL syringes</td>
			<td>Henke Sass Wolf</td>
			<td>5010.200V0</td>
			<td></td>
		</tr>
		<tr>
			<td>15 mL Tube</td>
			<td>TH.Geyer</td>
			<td>7696702</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL Tube</td>
			<td>TH.Geyer</td>
			<td>7696705</td>
			<td></td>
		</tr>
		<tr>
			<td>70% ethanol</td>
			<td>Decon Labs</td>
			<td>2701</td>
			<td></td>
		</tr>
		<tr>
			<td>Absolute Counting Beads</td>
			<td>Life Technologies Europe B.V.</td>
			<td>C36950</td>
			<td></td>
		</tr>
		<tr>
			<td>ACK-RBC lysing buffer</td>
			<td>Lonza</td>
			<td>10-548E</td>
			<td></td>
		</tr>
		<tr>
			<td>Amersham Hybond-N+ Membrane</td>
			<td>GE Healthcare</td>
			<td>RPN203B</td>
			<td></td>
		</tr>
		<tr>
			<td>Ant</td>
			<td>San Antonio</td>
			<td>Note: Locally collected</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody dilution buffer</td>
			<td></td>
			<td>(see Table 5 for recipe)</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Mouse CD11b V450 Rat (clone M1/70)</td>
			<td>BD Bioscience</td>
			<td>560456</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Mouse CD11c PE-Cy7 (clone N418)</td>
			<td>BioLegend</td>
			<td>117317</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Mouse CD19 Alexa Flour 647 (clone 1D3)</td>
			<td>eBioscience</td>
			<td>15-0193-81</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Mouse CD3e APC (clone 145-2C11)</td>
			<td>Invitrogen</td>
			<td>15-0031-81</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Mouse CD45 APC-Cy7 (clone: 30-F11)</td>
			<td>BioLegend</td>
			<td>103130</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Mouse Fixable Viabillity Dye eFluor 506</td>
			<td>Invitrogen</td>
			<td>65-0866-14</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Mouse IgG (H+L), AP Conjugate</td>
			<td>Promega</td>
			<td>S3721</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Mouse Ly-6G FITC (clone RB6-8C5)</td>
			<td>Invitrogen</td>
			<td>11-5931-82</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Mouse MHC II APC-eFluor 780 (clone M5/114.15.2)</td>
			<td>eBioscience</td>
			<td>47-5321-80</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Mouse Siglec-F PE (clone E50-2440)</td>
			<td>BD Pharmingen</td>
			<td>552126</td>
			<td>1 to 200 dilution</td>
		</tr>
		<tr>
			<td>BCIP/NBT substrate</td>
			<td>Thermo Fisher Scientific</td>
			<td>PI34042</td>
			<td></td>
		</tr>
		<tr>
			<td>Blocking Buffer</td>
			<td></td>
			<td>(see Table 5 for recipe)</td>
			<td></td>
		</tr>
		<tr>
			<td>Cannual, 20G X 1.5&#8221;</td>
			<td>CADENCE SCIENCE</td>
			<td>9920</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Thermo Fisher Scientific</td>
			<td>75004030</td>
			<td></td>
		</tr>
		<tr>
			<td>CFX384 Touch Real-Time PCR Detection System</td>
			<td>Bio-Rad Laboratories</td>
			<td>1855485</td>
			<td></td>
		</tr>
		<tr>
			<td>Chloroform</td>
			<td>Thermo Fisher Scientific</td>
			<td>C298-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Cockroach</td>
			<td>Greer Laboratories</td>
			<td>B26</td>
			<td></td>
		</tr>
		<tr>
			<td>Counting beads</td>
			<td>Thermo Fisher Scientific</td>
			<td>01-1234-42</td>
			<td></td>
		</tr>
		<tr>
			<td>D. farinae</td>
			<td>Greer Laboratories</td>
			<td>B81</td>
			<td></td>
		</tr>
		<tr>
			<td>D. pteronyssinus</td>
			<td>Greer Laboratories</td>
			<td>B82</td>
			<td></td>
		</tr>
		<tr>
			<td>Denville Cell Culture Plates with lid, 96 well cell culture plate</td>
			<td>Thomas Scientific</td>
			<td>1156F03</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital Dry Bath - Four Blocks</td>
			<td>Universal Medical, Inc.</td>
			<td>BSH1004</td>
			<td></td>
		</tr>
		<tr>
			<td>Earthworm</td>
			<td>San Antonio</td>
			<td>Note: Locally collected</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylenediaminetetraacetic acid (EDTA)</td>
			<td>Sigma-Aldrich</td>
			<td>E6511</td>
			<td></td>
		</tr>
		<tr>
			<td>FACS buffer</td>
			<td></td>
			<td>(see recipe in Table 5)</td>
			<td></td>
		</tr>
		<tr>
			<td>Falcon Round-Bottom Polypropylene Tubes, 5 mL</td>
			<td>STEMCELLTM TECHNOLOGIES</td>
			<td>38056</td>
			<td></td>
		</tr>
		<tr>
			<td>Flow cytometer (BD FACS Celesta)</td>
			<td>BD Biosciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fly</td>
			<td>Greer Laboratories</td>
			<td>B8</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>Roboz Surgical Instrument</td>
			<td>RS-5135</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemocytometer</td>
			<td>Hausser Scientific</td>
			<td>3110</td>
			<td></td>
		</tr>
		<tr>
			<td>HT-DNA</td>
			<td>Sigma</td>
			<td>D6898</td>
			<td></td>
		</tr>
		<tr>
			<td>In Vivo MAb anti-mouse CD16/CD32 (clone: 2.4G2)</td>
			<td>Bio X Cell</td>
			<td>BE0307</td>
			<td></td>
		</tr>
		<tr>
			<td>iScript cDNA Synthesis Kit</td>
			<td>Bio-Rad Laboratories</td>
			<td>1708891</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Abbott Labs</td>
			<td>sc-363629Rx</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropanol</td>
			<td>Thermo Fisher Scientific</td>
			<td>BP2618500</td>
			<td></td>
		</tr>
		<tr>
			<td>J2 anti-dsRNA monoclonal antibody</td>
			<td>SCICONS</td>
			<td>10010200</td>
			<td></td>
		</tr>
		<tr>
			<td>Lung digestion solution</td>
			<td></td>
			<td>(see recipe in Table 5)</td>
			<td></td>
		</tr>
		<tr>
			<td>Lysing Matrix D</td>
			<td>MP Biomedicals</td>
			<td>116913050-CF</td>
			<td></td>
		</tr>
		<tr>
			<td>Lysing Matrix D, 2 mL tube</td>
			<td>MP Biomedicals</td>
			<td>SKU:116913100</td>
			<td></td>
		</tr>
		<tr>
			<td>Mice (female, 8-12 weeks old, C57BL/6J)</td>
			<td>Jackson Laboratory</td>
			<td>#000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Microcentrifuge tube 1.5 mL</td>
			<td>Sigma-Aldrich</td>
			<td>30120.094</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Olympus</td>
			<td>CK30</td>
			<td></td>
		</tr>
		<tr>
			<td>Mini-BeadBeater</td>
			<td>Homogenizers</td>
			<td>SKU:BS:607</td>
			<td></td>
		</tr>
		<tr>
			<td>Mini-Beadbeater-16</td>
			<td>Biospec</td>
			<td>607</td>
			<td></td>
		</tr>
		<tr>
			<td>Mosquito</td>
			<td>Greer Laboratories</td>
			<td>B55</td>
			<td></td>
		</tr>
		<tr>
			<td>NanoDrop 2000C</td>
			<td>Thermo Scientific Spectophotometer Medex Supply</td>
			<td>TSCND2000C</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle, 21 G x 1 1/2 in</td>
			<td>BD Biosciences</td>
			<td>305167</td>
			<td></td>
		</tr>
		<tr>
			<td>Non-fat milk</td>
			<td>Bio-Rad Laboratories</td>
			<td>1706404</td>
			<td></td>
		</tr>
		<tr>
			<td>Nylon string</td>
			<td>Dynarex</td>
			<td>3243</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate-buffered Saline (PBS)</td>
			<td>Lonza</td>
			<td>BE17-516F</td>
			<td></td>
		</tr>
		<tr>
			<td>RNase III</td>
			<td>Thermo Fisher Scientific</td>
			<td>AM2290</td>
			<td></td>
		</tr>
		<tr>
			<td>RNase T1</td>
			<td>Thermo Fisher Scientific</td>
			<td>AM2283</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Roboz Surgical Instrument</td>
			<td>RS-6802</td>
			<td></td>
		</tr>
		<tr>
			<td>Shaker or Small laboratory mixer</td>
			<td>Boekel Scientific</td>
			<td>201100</td>
			<td></td>
		</tr>
		<tr>
			<td>SPHERO AccuCount Fluorescent</td>
			<td>Spherotech</td>
			<td>ACFP-70-5</td>
			<td>1 to 10 dilution</td>
		</tr>
		<tr>
			<td>Spider</td>
			<td>San Antonio</td>
			<td>Note: Locally collected</td>
			<td></td>
		</tr>
		<tr>
			<td>TBS</td>
			<td></td>
			<td>(see recipe in Table 5)</td>
			<td></td>
		</tr>
		<tr>
			<td>TBS-T</td>
			<td></td>
			<td>(see recipe in Table 5)</td>
			<td></td>
		</tr>
		<tr>
			<td>Total cell medium</td>
			<td></td>
			<td>(see recipe in Table 5)</td>
			<td></td>
		</tr>
		<tr>
			<td>TRIzol Reagent</td>
			<td>Thermo Fisher Scientific</td>
			<td>15596018</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween 20</td>
			<td>Sigma-Aldrich</td>
			<td>P9416</td>
			<td></td>
		</tr>
		<tr>
			<td>UV Stratalinker 2400 UV</td>
			<td>LabX</td>
			<td>20447</td>
			<td></td>
		</tr>
		<tr>
			<td>Wasp</td>
			<td>San Antonio</td>
			<td>Note: Locally collected</td>
			<td></td>
		</tr>
	</tbody>
</table>

identification and characterization of immunogenic rna species in hdm allergens that modulate eosinophilic lung inflammation

环境过敏原，如家庭尘虫（HDM）通常以复杂形式，既含有过敏性蛋白质，驱动异常2型反应，又含有诱导先天免疫反应的微生物物质。这些过敏原相关的微生物成分在调节过敏性哮喘等2型炎症疾病的发展方面起着重要作用。然而，基本机制基本上仍未确定。此处提出的协议确定了过敏原相关免疫刺激RNA的结构特征和体内活性。具体来说，在HDM引起的过敏性哮喘小鼠模型中，对常见过敏原检查是否存在双链RNA（dsRNA）物种，这些物种可以刺激肺的 IFN 反应，并抑制严重肺嗜酸性病的发展。在这里，我们包括以下三种检测方法：点印显示从包括HDM物种在内的过敏原中分离出的DNA中的dsRNA结构;RT-qPCR测量HDM RNA在小鼠肺部干扰素刺激基因（ISG）表达（ISG）中的活动;FACS分析，分别确定HDM RNA对BAL和肺嗜酸性生物数量的影响。

使用 dsRNA 特异性小鼠单克隆抗体 J2（±40bp）对HDM、昆虫和非昆虫小动物中长 dsRNA 结构的存在进行了点印检查。RNase III 用于将 dsRNA 消化成 12~15 bp dsRNA 片段，J2 检测不到这些片段（图 1）。
RT-qPCR分析了HDM总RNA以剂量依赖的方式刺激小鼠肺部先天免疫反应的能力（图2，上部）。RNase III 治疗废除了 HDM 总 RNA 的免疫刺激活性，表明 HDM ?...

Watch this Scientific Journal Video about Identification and Characterization of Immunogenic RNA Species in HDM Allergens that Modulate Eosinophilic Lung Inflammation at JoVE.com

Identification and Characterization of Immunogenic RNA Species in HDM Allergens that Modulate Eosinophilic Lung Inflammation

环境过敏原，如家庭尘虫（HDM），通常含有微生物物质，激活先天免疫反应，以调节过敏性炎症。此处提出的协议演示了 HDM 过敏原中 dsRNA 物种的识别，并表征了其在调节嗜酸性肺炎症时免疫原性活动。

HDM过敏原中免疫原RNA物种的识别和特征，可调节嗜酸性肺炎

Environmental allergens such as house dust mites (HDM) are often in complex forms containing both allergic proteins that drive aberrant type 2 responses ...

该协议旨在解决一个非常重要和未解决的问题，微生物物质存在和环境过敏原如何可以调节过敏性炎症的发展。具体来说，这里介绍的方法可以帮助回答家庭尘虫中的双链RNA物种是否在体内具有免疫原性，并能够调节嗜酸性肺炎症。这些技术的主要优点是简单、高效、可重复性高，可以使用常用的工具和设备执行。这些方法还可用于评估由呼吸道病毒感染引起的肺部疾病。李女士，我实验室的研究生，将帮助演示这个程序。首先从过敏原、昆虫和非昆虫过敏原中隔离总RNA。将适当数量的试样转移到含有1.4毫米陶瓷珠的两毫升管中，并在液氮容器中冷冻约10分钟。在每个管中加入一毫升的硫化铀基RNA分离试剂。然后用高能细胞干扰器以最高速度打破昆虫和非昆虫小动物45秒。在冰上冷却样品，然后重复此过程两次。将溶液转移到新的 1.5 毫升管中。在每个管中加入200微升氯仿，并旋转样品。在4摄氏度下以14，000次g离心14分钟。然后将上水相转移到含有500微升异丙醇的新管中。混合样品。然后在摄氏4度下以14，000次g离心14分钟。小心吸上一道。然后用 500 微升 75%乙醇清洗 RNA 颗粒，并在 4 摄氏度下以 7，500 次 g 离心，10 分钟。小心地去除所有液体。空气干燥颗粒，用20至50微升无RNase水溶解RNA。为了检测总RNA中的双链RNA结构，将每微升200纳米RNA样本的两微升检测到带正素的尼龙膜上。在UV交联器中，将样品以1200微焦耳和100微焦耳将样品交联到膜上。重复发现和交联两次，这将导致每印1.2微克RNA。在室温下摇动时，在 TBS-T 中与 5% 牛奶进行非特异性结合一小时。然后去除阻断溶液，在TBS-T中以1至1000%的稀释剂加入抗双链RNA J2抗体。在摄氏四度下孵育膜过夜。第二天，用TBS-T清洗膜三次，每次洗涤五分钟。加入二次抗体，在室温下孵育膜一小时。然后用 TBS-T 重复洗涤。加入基板，孵育膜5至15分钟，直到所需信号可见。用双蒸馏水冲洗膜，停止反应。要收集肺样本，请将肺放在纸巾上，并切除每个肺叶的一小块。将每块放入含有珠子的两毫升管中。要收集支气管，用70%乙醇对安乐死小鼠进行消毒，并使用剪刀将皮肤从腹部的上部区域切到颈部。用钳子轻轻拉开唾液腺和胸腺肌肉，露出气管。然后在它下面放置一根尼龙绳。在气管下切开大约两毫升，刚好足以插入一个管，并打结气管和气管周围的字符串。将注射器连接到管的端，然后用新鲜的 PBS-EDTA 将其加载。然后将溶液的一毫升注射并吸入肺部。将注射器从管子上分离，然后将溶液喷射到 15 毫升管中。使用新鲜的 PBS-EDTA 将此过程重复到冰中，然后将熔岩池池。在4摄氏度下以500倍g离心管7分钟。然后记录体积，然后将上流液转移到两个1.5毫升管，而不会干扰颗粒。重新暂停颗粒后，将 150 微升转移到 96 井板中，并在 500 倍 g 和 4 摄氏度下离心板 7 分钟。然后快速反转纸巾上的盘子，取出上一除。在2.4G2阻断抗体的情况下，用抗体在FACS缓冲液中染色细胞，并在室温下在黑暗中孵育板30分钟。染色后，将板离心，在4摄氏度下以500倍g的分粒7分钟。将板倒置在纸巾上，去除染色溶液。添加 100 微升的 FACS 缓冲液洗涤细胞，然后重复离心。将样品重新填充在 150 微升的 FACS 缓冲液中，然后将它们传输到包含另外 350 微升缓冲液的 FACS 管中。在每个样品中加入25微升的计数珠，然后进行流式细胞测量分析。使用双链、RNA特异性小鼠单克隆抗体，用点印来检查家庭尘虫、昆虫和非昆虫小动物中长双链RNA结构的存在。RNase III 用于将双链 RNA 消化成 12 到 15 个基对片段，J2 无法检测到这些片段。RT-qPCR证明了家尘虫总RNA刺激小鼠肺部先天免疫反应的能力。RNase III 治疗废除了家庭尘虫总 RNA 的免疫刺激活性，表明双链 RNA 结构对肺部的先天免疫活动至关重要。通过FACS分析，评价了家尘虫总RNA对严重二型肺炎症发育的抑制作用。嗜酸性肺炎是由尘虫提取物引起的，这些提取物使用或没有RNase III进行治疗。长双链RNA物种的降解导致严重的二型肺炎症，这反映在支气管和肺部的嗜酸性果蝇数量增加。值得注意的是，经过尘虫总RNA处理的组的嗜酸性蛋白患者数量与使用原尘虫提取物处理的组相当，该组内源性地含有长双链RNA物种。尝试此协议时，在注射和回忆 BAL 液体时要非常小心，因为在此阶段对肺部的任何损害都可能会改变最终结果。除此程序外，还可以执行其他方法（如 ELISA）来分析 BAL 流体中存在的非细胞溶性物质。开发后，该技术为研究人员探索环境过敏原中其他微生物物质铺平了道路，例如可调节过敏性肺炎症发育的非双链RNA因子。

Research

JoVE Journal

Immunology and Infection

A 1.5 Hour Procedure for Identification of Enterococcus Species Directly from Blood Cultures

A 1.5 Hour Procedure for Identification of Enterococcus Species Directly from Blood Cultures

一个1.5小时的程序，鉴定肠球菌直接从血培养的物种

Enterococci are a common cause of bacteremia with E. faecalis being the predominant species followed by E. faecium.  Because resistance to ampicillin and ...

科研

免疫与感染

Using Eggs from Schistosoma mansoni as an In vivo Model of Helminth-induced Lung Inflammation

Using Eggs from Schistosoma mansoni as an In vivo Model of Helminth-induced Lung Inflammation

从使用鸡蛋<em&gt;血吸虫</em&gt;作为<em&gt;在体内</em&gt;蠕虫引起的肺部炎症模型

Schistosoma parasites are blood flukes that infect an estimated 200 million people worldwide 1. In chronic infection with Schistosoma, the severe ...

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

总蛋白提取和二维凝胶电泳方法<em&gt;鼻疽</em&gt;物种

The investigation of  the intracellular protein levels of bacterial species is of importance to  understanding the pathogenic mechanisms of diseases ...

Nucleocapsid Annealing-Mediated Electrophoresis (NAME) Assay Allows the Rapid Identification of HIV-1 Nucleocapsid Inhibitors

Nucleocapsid Annealing-Mediated Electrophoresis NAME Assay Allows the Rapid Identification of HIV-1 Nucleocapsid Inhibitors

核衣壳退火介导电泳（NAME）分析允许HIV-1核衣壳抑制剂的快速鉴定

RNA or DNA folded in stable tridimensional folding are interesting targets in the development of antitumor or antiviral drugs. In the case of HIV-1, viral ...

Loop-mediated Isothermal Amplification (LAMP) Assays for the Species-specific Detection of Eimeria that Infect Chickens

Loop-mediated Isothermal Amplification LAMP Assays for the Species-specific Detection of Eimeria that Infect Chickens

环介导等温扩增（LAMP）法进行的物种特异性检测<em&gt;艾美耳球虫</em&gt;可感染鸡

Eimeria species parasites, protozoa which cause the enteric disease coccidiosis, pose a serious threat to the production and welfare of chickens. In the ...

Isolation, Characterization and Functional Examination of the Gingival Immune Cell Network

分离，鉴定和牙龈免疫细胞网络的功能性考试

Immune cell networks in tissues play a vital role in mediating local immunity and maintaining tissue homeostasis, yet little is known of the resident ...

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

高通量设置的建立筛选小分子能调节C-二GMP信令<em&gt;绿脓杆菌</em

Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. ...

Functional Characterization of Regulatory Macrophages That Inhibit Graft-reactive Immunity

抑制接枝反应性免疫的调节性巨噬细胞的功能表征

Macrophage accumulation in transplanted organs has long been recognized as a feature of allograft rejection1. Immunogenic monocytes infiltrate the ...

Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation

受肥胖相关炎症组织的巨噬细胞的分离、鉴定和纯化

Obesity promotes a chronic inflammatory state that is largely mediated by tissue-resident macrophages as well as monocyte-derived macrophages. ...

Precision-cut Mouse Lung Slices to Visualize Live Pulmonary Dendritic Cells

精密切割的小鼠肺切片可视化现场肺树突状细胞

Inhalation of allergens and pathogens elicits multiple changes in a variety of immune cell types in the lung. Flow cytometry is a powerful technique for ...