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

Estabelecimento de um In vitro Sistema para estudar o comportamento intracelular de ...

JoVE Journal
Immunology and Infection
Author Produced

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Immunology and Infection

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

Estabelecimento de um In vitro Sistema para estudar o comportamento intracelular de Candida glabrata Em humanos THP-1 macrófagos

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

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

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.

O presente artigo descreve um protocolo para estabelecer um sistema modelo de cultura de células in vitro para estudar a interação de um patógeno humano intracelular facultativo de fungos Candida glabrata com macrófagos humanos, que será uma ferramenta útil para aumentar o nosso conhecimento dos mecanismos de virulência de fungos.

O objetivo geral deste procedimento é estabelecer um sistema modelo de cultura in vitro para estudar o comportamento intracelular de um patógeno fúngico oportunista humano. Canida lab ratta em macrófagos derivados da linha celular monocítica humana TP um. Para conseguir isso, os monócitos THP um são primeiro tratados com quatro ésteres para diferenciá-los em macrófagos.

Em seguida, os macrófagos THP um são infectados com células GTA canida duas horas após a infecção. As células GTA da canida não fagocitadas são removidas por arruelas de PBS e as células e intracelulares são recuperadas para estudos posteriores por osmos de macrófagos infectados. Este protocolo pode ser usado para avaliar tanto as células GTA virais para canida em macrófagos, quanto a maturação de Ross faiz e a resposta de citocinas em THP em macro FIDS após infecção por candida GTA.

View the full transcript and gain access to thousands of scientific videos