Establishment of Viral Infection and Analysis of Host-Virus Interaction in <em>Drosophila Melanogaster</em>

Shuo Yang; Yaya Zhao; Junjing Yu; Zhiqin Fan; Si-tang Gong; Hong Tang; Lei Pan

doi:10.3791/58845

Mise en place d’une Infection virale et analyse de l’Interaction hôte-Virus chez Drosophila M...

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Immunology and Infection

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JoVE Journal Immunology and Infection

Establishment of Viral Infection and Analysis of Host-Virus Interaction in Drosophila Melanogaster

Mise en place d’une Infection virale et analyse de l’Interaction hôte-Virus chez Drosophila Melanogaster

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09:57 min

March 14, 2019

DOI: 10.3791/58845-v

Shuo Yang*^1,2,3, Yaya Zhao*^1,2, Junjing Yu⁴, Zhiqin Fan², Si-tang Gong¹, Hong Tang^1,2, Lei Pan^1,2,3

¹The Joint Center for Infection and Immunity, Guangzhou Institute of Pediatrics,Guangzhou Women and Children's Medical Center, ²CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai,Chinese Academy of Sciences, ³CAS Key Laboratory of Infection and Immunity (CASKLII), Institute of Biophysics,Chinese Academy of Sciences, ⁴Shanghai Institute of Biochemistry and Cell Biology,Chinese Academy of Sciences

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Overview

This protocol describes how to establish viral infection in vivo in Drosophila melanogaster using the nano-injection method and basic techniques to analyze virus-host interaction.

Key Study Components

Area of Science

Neuroscience
Virology
Infectious Disease Research

Background

Understanding virus-host interactions is crucial for developing effective treatments.
Drosophila melanogaster serves as a model organism for studying viral infections.
The nano-injection method allows for precise control over viral dosage.
This technique can be adapted for other microbial pathogens.

Purpose of Study

To establish a reliable method for viral infection in Drosophila.
To investigate the dynamics of virus-host interactions.
To provide a framework for future studies on viral infections.

Methods Used

Grow S2 star cells in complete Schneider's Drosophila Medium.
Prepare Drosophila C Virus (DCV) at a specific multiplicity of infection.
Add virus solution to cell cultures and incubate.
Monitor cell morphology and culture medium for signs of infection.

Main Results

Successful establishment of viral infection in Drosophila.
Clear indicators of infection observed in cell morphology.
Method demonstrated adaptability for other pathogens.
Provides insights into virus-host dynamics.

Conclusions

The nano-injection method is effective for studying viral infections.
Findings contribute to the understanding of virus-host interactions.
This approach can facilitate further research in virology.

Frequently Asked Questions

What is the main focus of this study?

The study focuses on establishing viral infection in Drosophila melanogaster using a nano-injection method.

Why use Drosophila melanogaster for viral studies?

Drosophila serves as a model organism that allows for detailed studies of virus-host interactions.

What are the key steps in the nano-injection method?

Key steps include growing S2 cells, preparing the virus, and monitoring infection indicators.

How does this method improve upon previous techniques?

It allows for precise control of the viral dose and can be adapted for other pathogens.

What are the implications of this research?

The research provides insights into viral infections and can inform future therapeutic strategies.

Ce protocole décrit comment établir une infection virale in vivo chez Drosophila melanogaster en utilisant la méthode de nano-injection et les techniques de base pour analyser l’interaction virus-hôte.

Cette méthode peut aider à répondre aux questions clés dans le domaine de l’interaction virus-hôte sur la façon d’établir efficacement une infection virale chez Drosophila melanogaster. Cette méthode de nanoinjection permet un contrôle précis de la dose d’infection du virus et peut être facilement appliquée aux infections par d’autres agents pathogènes microbiens. Commencez par cultiver une fois 10 à la septième cellule S2 viable par millilitre comme monocouche lâche et semi-adhérente dans un plat de culture cellulaire de 10 centimètres avec 10 millilitres de drosophile moyen complet de Schneider sans dioxyde de carbone supplémentaire à 25 degrés Celsius pendant une heure.

Pendant l’incubation, resuspendez le virus Drosophila C ou DCV fraîchement décongelé à une multiplicité d’infection de 0,01 dans un millilitre de milieu complet et ajoutez une solution virale aux plats de culture cellulaire. Ensuite, retournez la plaque à l’incubateur de 25 degrés Celsius pendant trois à cinq jours. Le virus est prêt pour la collecte lorsque la morphologie cellulaire semble floue et que le milieu de culture est plein de particules noires révélatrices de débris cellulaires.

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