Adoptive Transfer of IL-33-Stimulated Macrophages into Bleomycin-Induced Mouse Models to Study Their Effect on Idiopathic Pulmonary Fibrosis <em>In Vivo</em>

Xiaorun Zhai; Jiao Li; Yunjuan Nie

doi:10.3791/64742

Adoptive Transfer of IL-33-Stimulated Macrophages into Bleomycin-Induced Mouse Models to Study Th...

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Immunology and Infection

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JoVE Journal Immunology and Infection

Adoptive Transfer of IL-33-Stimulated Macrophages into Bleomycin-Induced Mouse Models to Study Their Effect on Idiopathic Pulmonary Fibrosis In Vivo

Adoptive Transfer of IL-33-Stimulated Macrophages into Bleomycin-Induced Mouse Models to Study Their Effect on Idiopathic Pulmonary Fibrosis In Vivo

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06:29 min

May 5, 2023

DOI: 10.3791/64742-v

Xiaorun Zhai*¹, Jiao Li*¹, Yunjuan Nie¹

¹Department of Basic Medicine, Wuxi School of Medicine,Jiangnan University

Overview

This protocol describes the isolation of pulmonary interstitial macrophages (IMs) and their adoptive transfer after IL-33 stimulation in a mouse model. This technique facilitates the in vivo study of idiopathic pulmonary fibrosis (IPF).

Key Study Components

Area of Science

Neuroscience
Immunology
Pulmonary Biology

Background

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease.
Macrophages play a crucial role in lung inflammation and fibrosis.
IL-33 is a cytokine involved in immune responses in the lung.
Understanding macrophage function can provide insights into IPF mechanisms.

Purpose of Study

To isolate pulmonary interstitial macrophages (IMs) from mouse lungs.
To investigate the effects of IL-33 on macrophage behavior.
To facilitate in vivo studies of IPF using a mouse model.

Methods Used

Isolation of pulmonary interstitial macrophages (IMs).
Adoptive transfer of IMs after IL-33 stimulation.
Administration of clodronate liposomes to deplete macrophages.
Use of anesthetized mice for nasal administration of treatments.

Main Results

Successful isolation of IMs from mouse lungs.
Demonstrated effects of IL-33 on macrophage function.
Provided a method for studying IPF in vivo.
Highlighted the role of macrophages in lung pathology.

Conclusions

The protocol allows for the study of macrophage dynamics in IPF.
IL-33 stimulation can enhance understanding of macrophage roles.
This method can be applied to further research in pulmonary diseases.

Frequently Asked Questions

What is the significance of isolating pulmonary interstitial macrophages?

Isolating IMs allows researchers to study their specific functions and roles in lung diseases like IPF.

How does IL-33 influence macrophage behavior?

IL-33 is known to activate macrophages, potentially altering their inflammatory responses and functions.

What are clodronate liposomes used for in this protocol?

Clodronate liposomes are used to deplete macrophages in order to study the effects of specific treatments on lung pathology.

Can this method be applied to other lung diseases?

Yes, the protocol can be adapted to study various pulmonary conditions involving macrophage activity.

What animal model is used in this study?

The study utilizes a mouse model to investigate the effects of IL-33 on pulmonary interstitial macrophages.

What is idiopathic pulmonary fibrosis?

IPF is a chronic and progressive lung disease characterized by scarring of lung tissue, leading to respiratory failure.

This protocol describes the isolation of pulmonary interstitial macrophages (IMs) and their adoptive transfer after IL-33 stimulation of the lung alveoli in a mouse model, which can facilitate the in vivo study of idiopathic pulmonary fibrosis (IPF).

The protocol describes the isolation of pulmonary IMS and adoptive transfer after IL-33 determination in a mouse model, which can facilitate the in vivo study of idiopathic pulmonary fibrosis. The technique enables the researchers to explore the function of macrophages, simulated by traditional cytokines in developing IPF. To begin, take out the vial of clodronate liposomes from the refrigerator.

Allow it to warm for 30 minutes at room temperature and invert several times to ensure uniform mixing. Aspirate 60 microliters of clodronate liposomes using a pipette with a sterile suction tip. Administer the control and drug drop-wise into the nasal cavity of the anesthetized mouse.

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