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JoVE Journal
Immunology and Infection
量激活的主人微血管内皮细胞在静态条件下测量的检测,以中性粒细胞粘附
量激活的主人微血管内皮细胞在静态条件下测量的检测,以中性粒细胞粘附
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions

量激活的主人微血管内皮细胞在静态条件下测量的检测,以中性粒细胞粘附

Full Text
18,004 Views
11:22 min
August 23, 2013

DOI: 10.3791/50677-v

Kevin Wilhelmsen1, Katherine Farrar1,2, Judith Hellman1

1Department of Anesthesia and Perioperative Care,University of California, San Francisco, 2Graduate Program in Biomedical Sciences,University of California, San Francisco

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Overview

This study presents a neutrophil binding assay designed to quantify the adherence of primary human neutrophils to activated endothelial cells under static conditions. The assay is crucial for understanding the inflammatory response during infections.

Key Study Components

Area of Science

  • Neuroscience
  • Immunology
  • Cell Biology

Background

  • Neutrophils play a vital role in the host's inflammatory response.
  • Endothelial cells become activated during inflammation, facilitating neutrophil adhesion.
  • Understanding this process is essential for developing therapeutic strategies.
  • The assay allows for the quantification of neutrophil-endothelial interactions.

Purpose of Study

  • To determine the extent of microvascular endothelial cell activation.
  • To quantify neutrophil adherence under various inflammatory conditions.
  • To enhance understanding of the inflammatory response mechanisms.

Methods Used

  • Isolation of neutrophils from healthy human volunteers.
  • Labeling neutrophils with calcium am.
  • Adhering neutrophils to treated endothelial cell monolayers.
  • Measuring adherence using fluorescence spectro geometry.

Main Results

  • Quantitative data on neutrophil adherence to endothelial cells.
  • Assessment of the effects of different inflammatory stimuli.
  • Comparison of pre-wash and post-wash measurements.
  • Insights into the dynamics of neutrophil-endothelial interactions.

Conclusions

  • The assay effectively quantifies neutrophil adherence.
  • Findings contribute to the understanding of inflammatory responses.
  • Potential implications for therapeutic interventions in inflammatory diseases.

Frequently Asked Questions

What is the purpose of the neutrophil binding assay?
The assay quantifies the adherence of neutrophils to activated endothelial cells, helping to understand inflammatory responses.
How are neutrophils isolated for the assay?
Neutrophils are isolated from healthy human volunteers for use in the assay.
What measurement technique is used in this study?
Fluorescence spectro geometry is used to measure neutrophil adherence.
What conditions are tested in the assay?
The assay tests neutrophil adherence under various inflammatory stimuli.
What are the implications of this research?
The findings may inform therapeutic strategies for managing inflammatory diseases.

中性粒细胞活化的内皮细胞在感染部位是坚持宿主的炎症反应的一个组成部分。本报告中所述的是中性粒细胞的结合测定法,它允许对

该程序的总体目标是通过定量静态条件下在体外粘附在内皮细胞上的中性粒细胞的相对数量来确定微血管内皮细胞炎症激活的程度。这是通过首先在第二步中用所需的实验刺激处理健康的融合单层微血管内皮细胞来实现的。分离来自健康人类志愿者的中性粒细胞,然后用钙 am 标记。

接下来,将标记的中性粒细胞添加到内皮细胞中,并使其粘附 20 分钟,在此期间通过荧光光谱几何获得预洗测量。在最后一步中,洗去非贴壁的中性粒细胞,然后进行洗涤后测量。最终,可以确定在每种处理条件下钙标记的中性粒细胞对内皮细胞单层的百分比和相对粘附性。

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78期 免疫学 细胞生物学 感染 分子生物学 医学 生物医学工程 生物物理 内皮 血管 中性粒细胞 炎症 炎症介质 中性粒细胞 白细胞粘附 血管内皮细胞 检测

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