Establishment of an <em>In vitro</em> System to Study Intracellular Behavior of <em>Candida glabrata</em> in Human THP-1 Macrophages

Maruti Nandan Rai; Sapan Borah; Gaurav Bairwa; Sriram Balusu; Neelima Gorityala; Rupinder Kaur

doi:10.3791/50625

Establecimiento de un In vitro Sistema para estudiar el comportamiento intracelular de

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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

Establecimiento de un In vitro Sistema para estudiar el comportamiento intracelular de Candida glabrata En humanos THP-1 macrófagos

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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.

El presente artículo describe un protocolo para establecer un sistema modelo de cultivo celular in vitro para estudiar la interacción de un hongo patógeno intracelular facultativo humano Candida glabrata con macrófagos humanos que será una herramienta útil para avanzar en el conocimiento de los mecanismos de virulencia de hongos.

El objetivo general de este procedimiento es establecer un sistema modelo de cultivo in vitro para estudiar el comportamiento intracelular de un patógeno fúngico oportunista humano. Canida lab ratta en macrófagos derivados de la línea celular monocítica humana TP one. Para lograr esta THP, primero se tratan los monocitos con cuatro ésteres para diferenciarlos en macrófagos.

A continuación, los macrófagos THP uno se infectan con células GTA canida dos horas después de la infección. Las células GTA de la cánida no fagocitosa se eliminan mediante lavadores PBS y las células e intracelulares se recuperan para estudios posteriores mediante ósmosis de macrófagos infectados. Este protocolo se puede utilizar para evaluar tanto el virus de las células GTA de canida en macrófagos, como la maduración de faiz Ross y la respuesta de citocinas en THP en macro FIDS tras la infección por GTA por candida.

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