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JoVE Journal
Immunology and Infection
建立一个在体外系统来研究细胞内的行为光滑念珠菌在人THP-1巨噬细胞
建立一个在体外系统来研究细胞内的行为光滑念珠菌在人THP-1巨噬细胞
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Immunology and Infection
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JoVE Journal Immunology and Infection
Establishment of an In vitro System to Study Intracellular Behavior of Candida glabrata in Human THP-1 Macrophages

建立一个在体外系统来研究细胞内的行为光滑念珠菌在人THP-1巨噬细胞

Full Text
12,258 Views
13:47 min
December 10, 2013

DOI: 10.3791/50625-v

Maruti Nandan Rai1, Sapan Borah1, Gaurav Bairwa*1, Sriram Balusu*1,2, Neelima Gorityala1, Rupinder Kaur1

1Laboratory of Fungal Pathogenesis,Centre for DNA Fingerprinting and Diagnostics, Andhra Pradesh, India, 2Current location: VIB Department for Molecular Biomedical Research, UGent,Fiers-Schell-Van Montagu Building, Technologiepark 927, B-9052 Ghent (Zwijnaarde), Belgium

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Overview

This article presents a protocol for establishing an in vitro cell culture model to investigate the interaction between the human fungal pathogen Candida glabrata and human macrophages. This model aims to enhance understanding of fungal virulence mechanisms.

Key Study Components

Area of Science

  • Microbiology
  • Immunology
  • Cell Biology

Background

  • Candida glabrata is a facultative intracellular pathogen.
  • Understanding its interaction with macrophages is crucial for insights into fungal infections.
  • THP-1 cell line is commonly used for macrophage differentiation.
  • In vitro models are essential for studying pathogen-host interactions.

Purpose of Study

  • To develop a reliable in vitro model for studying Candida glabrata.
  • To assess the intracellular behavior of the pathogen in human macrophages.
  • To evaluate macrophage responses to Candida infection.

Methods Used

  • THP-1 monocytes are differentiated into macrophages using PMA treatment.
  • Macrophages are infected with Candida glabrata cells.
  • Non-phagocytosed cells are removed through PBS washes.
  • Intracellular cells are recovered for further analysis.

Main Results

  • The protocol allows for the assessment of Candida glabrata virulence in macrophages.
  • It enables the study of cytokine responses in THP-1 macrophages.
  • Insights into the maturation processes of macrophages upon infection are provided.

Conclusions

  • This in vitro model is a valuable tool for studying fungal pathogenesis.
  • It contributes to the understanding of host-pathogen interactions.
  • Future studies can leverage this model to explore therapeutic interventions.

Frequently Asked Questions

What is the significance of studying Candida glabrata?
Candida glabrata is an opportunistic pathogen that poses a significant threat to immunocompromised individuals, making its study crucial for developing effective treatments.
How are THP-1 cells differentiated into macrophages?
THP-1 cells are treated with PMA to induce differentiation into macrophage-like cells.
What techniques are used to analyze intracellular Candida glabrata?
The protocol includes washing non-phagocytosed cells and recovering intracellular cells for further analysis.
What are the expected outcomes of this study?
The study aims to elucidate the virulence mechanisms of Candida glabrata and the immune response of macrophages.
Can this model be used for other pathogens?
Yes, the model can potentially be adapted to study other intracellular pathogens.
What is the role of cytokines in this study?
Cytokines are key indicators of the immune response and are measured to assess macrophage activation during infection.

当前文章概述了一个协议来建立体外细胞培养模型系统来研究一种兼人体细胞内的真菌病原体光滑念珠菌与人体巨噬细胞将是一个有用的工具,以增进我们对真菌毒力机制知识的互动。

该程序的总体目标是建立体外培养模型系统,以研究人类机会性真菌病原体的细胞内行为。来自人单核细胞系 TP one 的巨噬细胞中的犬科实验室大鼠。为了实现这个 THP one,首先用四种酯处理单核细胞,以将它们分化为巨噬细胞。

接下来,THP 1 巨噬细胞在感染后 2 小时被犬 GTA 细胞感染。通过 PBS 垫圈去除非吞噬糖犬 GTA 细胞,并通过感染巨噬细胞的渗透回收细胞内 e 细胞以进行进一步研究。该方案可用于评估巨噬细胞中病毒对犬科 GTA 细胞的评价,以及念珠菌 GTA 感染后宏 FIDS 上 THP 的 faiz 成熟 Ross 和细胞因子反应。

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