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JoVE Journal
Immunology and Infection
設立インビトロシステムの細胞内挙動を研究するために、カンジダ·グラブラタ人間のTH...
設立インビトロシステムの細胞内挙動を研究するために、カンジダ·グラブラタ人間のTH...
JoVE Journal
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

設立インビトロシステムの細胞内挙動を研究するために、カンジダ·グラブラタ人間のTHP-1マクロファージにおける

Full Text
12,269 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.

現在の記事は、真菌の病原性のメカニズムの知識を進めるために有用なツールとなり、人間のマクロファージとの通性細胞内のヒトの真菌病原体のカンジダ·グラブラタの相互作用を研究するためのin vitroでの細胞培養モデル系を確立するためのプロトコルの概要を説明します。

この手順の全体的な目標は、ヒト日和見真菌病原体の細胞内挙動を研究するためのin vitro培養モデルシステムを確立することです。Canida labは、ヒト単球細胞株TP oneに由来するマクロファージのラッタです。このTHPを達成するために、1つの単球を最初に4つのエステルで処理してマクロファージに分化させます。

次に、THP 1マクロファージは、感染後2時間でcanida GTA細胞に感染します。非食作用のあるcanida GTA細胞はPBSワッシャーによって除去され、細胞内電子細胞は感染したマクロファージの浸透膜によるさらなる研究のために回収されます。このプロトコールは、マクロファージ中のウイルスからcanida GTA細胞、ならびにカンジダGTA感染時のマクロFIDS上のTHPにおけるfaiz成熟ロスおよびサイトカイン応答の両方を評価するために使用することができます。

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