Related Experiment Video
Updated: Nov 21, 2025
08:25
Flow Cytometric Characterization of Murine B Cell Development
Published on: January 22, 2021
17.6K
Evaluating cytokine production by flow cytometry using brefeldin A in mice
Stephen B Kovacs1,2,3, Changhoon Oh4, Youssef Aachoui4
1Department of Immunology, Duke University, Durham, NC 27710, USA.
STAR Protocols
|January 18, 2021
Summary
This study introduces a new method using brefeldin A injection for in situ cytokine analysis in mice during bacterial infections. This technique overcomes limitations of traditional reporter mice for studying immune responses.
Area of Science:
- Immunology
- Microbiology
- Cellular Biology
Background:
- Understanding in situ cytokine production is crucial for analyzing immune responses.
- Existing cytokine reporter mouse models have limitations in flexibility and availability.
- Developing alternative methods is necessary for comprehensive immunologic studies.
Purpose of the Study:
- To present a novel protocol for analyzing in situ cytokine production in mice.
- To overcome the limitations associated with traditional cytokine reporter mouse models.
- To enable flow cytometric analysis of cytokine production during bacterial infections.
Main Methods:
- Injection of brefeldin A into mice.
- Flow cytometric analysis of cytokine production.
- Evaluation of interferon-gamma (IFN-γ) production during Burkholderia thailandensis infection.
Main Results:
- The brefeldin A injection method facilitates in situ cytokine analysis.
- This protocol allows for the assessment of cytokine production during active bacterial infection.
- The method is adaptable for various cytokines and mouse models.
Conclusions:
- The brefeldin A injection protocol provides a viable alternative for studying in situ cytokine production.
- This method enhances the ability to investigate immune responses in mouse models.
- The protocol is applicable to a range of cytokines and infection models.
