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Visualization of IL-22-expressing Lymphocytes Using Reporter Mice
Published on: January 25, 2017
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Visualization of IL-22-expressing Lymphocytes Using Reporter Mice
Wei Shen1, Wenqing Li1, Julie A Hixon1
1Laboratory of Molecular Immunoregulation, Cancer and Inflammation Program, Center for Cancer Research, National Cancer Institute, National Institutes of Health.
Journal of Visualized Experiments : Jove
|February 13, 2017
Summary
This study developed a new reporter mouse to track interleukin (IL)-22 gene expression in vivo. The reporter accurately visualizes IL-22-producing cells in tissues, aiding inflammation research.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- Reporter mice are crucial for visualizing gene expression localization in vivo.
- Existing methods for studying interleukin (IL)-22 expression required cell isolation and in vitro stimulation, limiting in vivo accuracy.
- Visualizing IL-22, a cytokine vital for intestinal repair during inflammation, in real-time within living organisms presents a significant challenge.
Purpose of the Study:
- To establish a novel transgenic reporter mouse model for visualizing endogenous IL-22 gene expression.
- To overcome limitations of previous methods by enabling direct in vivo detection of IL-22 producing cells.
- To utilize a fluorescent protein reporter integrated into the IL-22 gene for accurate localization studies.
Main Methods:
- Engineered a bacterial artificial chromosome (BAC) containing the IL-22 gene, incorporating a fluorescent reporter (tdTomato) that is retained intracellularly.
- Introduced the engineered BAC into the mouse genome to create the transgenic reporter line.
- Analyzed reporter gene expression in various tissues (spleen, thymus, lymph nodes, Peyer's patch, intestine) under homeostatic and induced colitis conditions using flow cytometry and tissue section visualization.
Main Results:
- Successfully generated a reporter mouse model with high-fidelity IL-22 expression.
- Observed homeostatic IL-22 reporter expression in key immune tissues.
- Tracked the accumulation of IL-22-producing T cells in mesenteric lymph nodes and intestinal tissues during induced colitis.
Conclusions:
- The developed reporter mouse model provides a robust and simpler alternative to traditional knock-in methods for studying IL-22 dynamics.
- This tool facilitates accurate in vivo visualization of IL-22 producing cells, advancing research in inflammatory diseases and tissue repair.
- The BAC-based strategy offers a reliable method for generating faithful gene expression reporters in mice.
