Name: accurate and simple measurement of the pro-inflammatory cytokine il-1&beta; using a whole blood stimulation assay
Uploaded: 2011-03-01T18:45:00
Description: Watch this Scientific Journal Video about Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1ÃŽÂ² using a Whole Blood Stimulation Assay at JoVE.com

这项研究是支持的关节炎和肌肉骨骼及皮肤疾病，美国国立卫生研究院国立研究所院内研究计划。

1。收集全血<ol><li>获取从受影响的个人，以及年龄和性别匹配的健康对照最小的10毫升外周血。静脉血应收集到，而不是使用其他抗凝血剂，如EDTA，倒几次，以确保适当的混合与肝素钠肝素钠真空采血管的血液收集管。请注意，10毫升是体积最小的电镀样品需要重复这个特殊的检测全血。 </li><li>样品应尽快处理收集后，以取得最佳效果。但是，肝素血液可以存储在昏暗的灯光下下，长达24小时?...

<ol>
	<li>Martinon, F., Mayor, A., Tschopp, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+inflammasomes:+guardians+of+the+body.">The inflammasomes: guardians of the body.</a> Annu Rev Immunol. 27, 229-265 (2009).</li><li>Thurm, C. W., Halsey, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cytokine+production+using+whole+blood.">Measurement of cytokine production using whole blood.</a> Curr Protoc Immunol. 66, 7-18 .</li><li>Mariathasan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cryopyrin+activates+the+inflammasome+in+response+to+toxins+and+ATP.">Cryopyrin activates the inflammasome in response to toxins and ATP.</a> Nature. 440, 228-232 (2006).</li><li>Gabay, C., Lamacchia, C., Palmer, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-1+pathways+in+inflammation+and+human+diseases.">IL-1 pathways in inflammation and human diseases.</a> Nat Rev Rheumatol. 4, 232-241 (2010).</li><li>Franchi, L., Elgenbrod, T., Mu&#241;oz-Planillo, R., Nu&#241;ez, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+inflammasome:+a+caspase-1-activation+platform+that+regulates+immune+responses+and+disease+pathogenesis.">The inflammasome: a caspase-1-activation platform that regulates immune responses and disease pathogenesis.</a> Nat Immunol. 3, 241-247 (2009).</li><li>Sims, J. E., Smith, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+IL-1+family:+regulators+of+immunity.">The IL-1 family: regulators of immunity.</a> Nat Rev Immunol. 2, 89-102 (2010).</li><li>Haug, R., Jo&#248;, G. B., Westvik, A. B., Kierulf, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemokine+production+and+pattern+recognition+receptor+(PRR)+expression+in+whole+blood+stimulated+with+pathogen-associated+molecular+patterns+(PAMPs.">Chemokine production and pattern recognition receptor (PRR) expression in whole blood stimulated with pathogen-associated molecular patterns (PAMPs.</a> Cytokine. 32, 304-315 (2005).</li><li>Netea, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+requirement+for+the+activation+of+the+inflammasome+for+processing+and+release+of+IL-1bin+monocytes+and+macrophages.">Differential requirement for the activation of the inflammasome for processing and release of IL-1bin monocytes and macrophages.</a> Blood. 113, 2324-2335 (2009).</li><li>Latz, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+inflammasomes:+mechanisms+of+activation+and+function.">The inflammasomes: mechanisms of activation and function.</a> Curr Opin Immnol. 22, 28-33 (2010).</li><li>Masters, S. L., Simon, A., Aksentijevich, I., Kastner, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Horror+autoinflammaticus:+the+molecular+pathophysiology+of+autoinflammtory+disease.">Horror autoinflammaticus: the molecular pathophysiology of autoinflammtory disease.</a> Annu Rev Immunol. 27, 621-668 (2009).</li></ol>

整个血液检测是一个简单的方法，更加紧密地与其他的技术，将要求细胞的分离和广泛的来样加工和操纵相比，模仿生理条件。这种方法利用全血中的白细胞计数，因此，提供了一种基于细胞的数量要准确规范样品与AO Cellometer视觉。 必须指出，这个实验中，全血应卷入肝素钠收集管其他抗凝血剂已知钙螯合剂，从而影响检测结果。血清自由的媒体也应该使用，因为这可能会?...

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<td>Sodium Heparin Blood Collection Tubes</td>
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<td>Bio-Plex Pro human cytokine x-plex assay</td>
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accurate and simple measurement of the pro-inflammatory cytokine il-1&beta; using a whole blood stimulation assay

从免疫细胞可溶性介质的分泌导致的炎症过程，导致各种表现在皮肤，关节和其他组织以及改变细胞因子的动态平衡。先天免疫系统中起着至关重要的作用在识别病原体和其他内源性危险刺激。先天免疫细胞释放的主要因子之一，是白细胞介素（IL）-1。因此，我们利用全血刺激试验，以衡量炎性细胞因子的分泌和专门的促炎症细胞因子IL -1β1，2，3 。 与先天免疫系统，导致autoinflammatory综合征的遗传功能障碍的患者表现夸张成熟后，单纯用LPS刺激IL -1β的释放。为了评估先天免疫与炎症相关的病症的患者组成部分，我们使用一个特定的免疫检测病原体相关分子模式（PAMPS），如革兰阴性细菌内毒素，细胞免疫反应，脂多糖（LPS ）。这些PAMPS上的先天免疫系统4，5，6，7细胞中发现的病原体识别受体（PRRS），确认。原始信号，脂多糖，结合次要的信号，ATP，是必要的炎性激活，一个多蛋白复合物，处理亲IL -1β，其成熟， 生物活性，4，5，6，8 ，9， 10。 全血检测需要最少的样品操作，以评估需要劳力密集的隔离和特定的细胞群的培养等方法相比，细胞因子的产生。此方法不同于其他刺激全血化验的RPMI媒体的比例与稀释的样品，而不是我们执行一个白细胞计数直接从稀释的全血，因此，刺激的白血细胞在培养2。这个特殊的全血检测结果表明了一种新技术，有助于阐明病人同伙提出autoinflammatory pathophysiologies。

Watch this Scientific Journal Video about Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1ÃŽÂ² using a Whole Blood Stimulation Assay at JoVE.com

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1ÃƒÅ½Ã‚Â² using a Whole Blood Stimulation Assay

我们描述了一个简单的免疫autoinflammatory表型的患者，来衡量生产促炎性细胞因子，如IL -1β生产。通过激活细胞与病原体相关分子模式，特别是与脂多糖，全血培养，细胞因子的分泌，可以方便地评估全血上清。

使用全血刺激试验的促炎性细胞因子IL -1β的精确和简单的测量

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

Research

JoVE Journal

Medicine

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

在单绒毛膜胎盘血管吻合用有色染料的精确和简单的评价

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia ...

科研

医学

Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System

利用经颅磁刺激，以研究人类的神经肌肉系统

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade.  While ...

Epidural Intracranial Pressure Measurement in Rats Using a Fiber-optic Pressure Transducer

硬膜外颅内压测量中使用的光纤压力传感器的大鼠

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and ...

Rapid Point-of-Care Assay of Enoxaparin Anticoagulant Efficacy in Whole Blood

快速点护理依诺肝素的抗凝血剂功效的测定全血中

There is the need for a clinical assay to determine the extent to which a patient's blood is effectively anticoagulated by the ...

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

脑Microslices的兴奋性刺激作为<em&gt;在体外</em&gt;中风模型

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

Fast and Accurate Exhaled Breath Ammonia Measurement

快速，准确的呼出气测量氨

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that ...

Real-time Cytotoxicity Assays in Human Whole Blood

实时的细胞毒性分析在人全血

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

一个简单的设备，迅速准备小鼠肠的全坐骑

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

使用Saccadometry与深部脑刺激来研究正常和病理脑功能

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...

Long-term Blood Pressure Measurement in Freely Moving Mice Using Telemetry

自由活动小鼠使用遥测长期血压测量

During the development of new vasoactive agents, arterial blood pressure monitoring is crucial for evaluating the efficacy of the new proposed drugs. ...