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Visualization of integrin Mac-1 in vivo
Kihong Lim1, Young-Min Hyun1, Kris Lambert-Emo1
1Department of Microbiology and Immunology, David H. Smith Center for Vaccine Biology and Immunology, University of Rochester, Rochester, NY, USA.
Journal of Immunological Methods
|September 7, 2015
Summary
Researchers developed a novel knock-in mouse model to visualize Mac-1 (CD11b/CD18) expressing immune cells. This tool allows for in vivo and in vitro imaging, aiding the study of immune cell migration and trafficking.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- β2 integrins, particularly Mac-1 (CD11b/CD18), are crucial for innate immune cell migration and interactions.
- Existing methods for studying Mac-1 function in live systems are limited.
Purpose of the Study:
- To generate a genetically modified mouse model for visualizing Mac-1 expressing cells.
- To enable real-time imaging of immune cell dynamics in live animals and in vitro.
Main Methods:
- Generation of a knock-in mouse strain expressing CD11b fused to monomeric yellow fluorescent protein (mYFP).
- Confirmation of CD11b-mYFP expression using Western blot and immunoprecipitation.
- Assessment of Mac-1 function in knock-in neutrophils compared to wild-type cells.
- Intravital two-photon microscopy and epifluorescent microscopy for imaging.
Main Results:
- Successfully generated a CD11b-mYFP knock-in mouse model.
- CD11b-mYFP expression did not impair Mac-1-mediated neutrophil functions.
- Sufficient fluorescence intensity allowed for in vivo imaging of CD11b-expressing cells in live mice.
- Dynamic intracellular CD11b localization was visualized in vitro.
- Immune cells were detected via flow cytometry without antibody staining.
Conclusions:
- The CD11b-mYFP knock-in mouse is a valuable tool for studying Mac-1 biology in live imaging settings.
- This model facilitates the investigation of immune cell migration and trafficking dynamics.
- It offers a non-invasive method for detecting and analyzing CD11b-expressing immune cells.
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