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JoVE Journal
Immunology and Infection
传染性单核细胞增多症儿童外周血样本中免疫细胞亚群的分离
传染性单核细胞增多症儿童外周血样本中免疫细胞亚群的分离
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Separation of Immune Cell Subpopulations in Peripheral Blood Samples from Children with Infectious Mononucleosis

传染性单核细胞增多症儿童外周血样本中免疫细胞亚群的分离

Full Text
2,941 Views
08:44 min
September 7, 2022

DOI: 10.3791/64212-v

Linlin Zhang1,2, Mengjia Liu1,2, Meng Zhang1,2, Junhong Ai1,2, Jiao Tian1,2, Ran Wang1,2, Zhengde Xie1,2

1Beijing Key Laboratory of Pediatric Respiratory Infectious Diseases, Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, Laboratory of Infection and Virology, Beijing Pediatric Research Institute, Beijing Children's Hospital,Capital Medical University, National Center for Children's Health, 2Research Unit of Critical Infection in Children, 2019RU016,Chinese Academy of Medical Sciences

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article describes a method that combines immunomagnetic beads and fluorescence-activated cell sorting to isolate and analyze specific immune cell subpopulations from peripheral blood mononuclear cells. The method allows for the purification and analysis of magnetic and fluorescently labeled cells.

Key Study Components

Area of Science

  • Immunology
  • Cell Biology
  • Flow Cytometry

Background

  • Peripheral blood mononuclear cells (PBMCs) are crucial for studying immune responses.
  • Understanding immune cell subpopulations can provide insights into various diseases.
  • EBV infection mechanisms can be better characterized through this method.
  • Isolation of specific cell types enhances the ability to study their functions and interactions.

Purpose of Study

  • To isolate and analyze defined immune cell subpopulations.
  • To expand understanding of immune responses in diseases.
  • To demonstrate the procedure for isolating PBMCs effectively.

Methods Used

  • Isolation of PBMCs using centrifugation.
  • Use of CD14 microbeads for monocyte isolation.
  • Fluorescence-activated cell sorting for analysis.
  • Washing and resuspending cells in a prepared buffer.

Main Results

  • Successful isolation of specific immune cell types.
  • Enhanced ability to analyze immune responses in individuals.
  • Demonstration of the method's effectiveness in a laboratory setting.
  • Potential applications in studying EBV infection mechanisms.

Conclusions

  • The method provides a reliable approach for isolating immune cells.
  • It can significantly contribute to research on immune responses.
  • Future studies can leverage this method to explore various diseases.

Frequently Asked Questions

What are PBMCs?
PBMCs, or peripheral blood mononuclear cells, are blood cells that have a round nucleus and include lymphocytes and monocytes.
How does fluorescence-activated cell sorting work?
Fluorescence-activated cell sorting (FACS) uses fluorescent markers to identify and sort specific cell types based on their characteristics.
What is the significance of isolating immune cell subpopulations?
Isolating immune cell subpopulations allows researchers to study their specific functions and roles in immune responses and diseases.
Can this method be applied to other types of cells?
Yes, the method can be adapted to isolate and analyze other cell types beyond immune cells.
What diseases can benefit from this research?
This research can benefit studies on infectious diseases, autoimmune disorders, and cancer.

我们描述了一种结合免疫磁珠和荧光激活细胞分选的方法,以分离和分析外周血单核细胞(单核细胞,CD4 + T细胞,CD8 + T细胞,B细胞和自然杀伤细胞)的已定义免疫细胞亚群。使用这种方法,可以纯化和分析磁性和荧光标记的细胞。

该方法将能够表征IM个体处于相同疾病状态的不同免疫细胞,这将扩展我们对EBV感染机制的理解。演示该程序的将是来自我们实验室的医生张琳琳。首先,将先前分离的PBMC置于1.5毫升微量离心管中,并在室温下以300G旋转10分钟。

弃去上清液,并用80微升制备的缓冲液重新悬浮细胞。接下来,向细胞悬液中加入20微升CD 14微珠,通过移液充分混合,并将管在冰上孵育15分钟。然后使用一毫升缓冲液洗涤PBMC,并在室温下以300G离心10分钟。

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