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Visualization of integrin Mac-1 in vivo

Authors
 Kihong Lim  ;  Young-Min Hyun  ;  Kris Lambert-Emo  ;  David J Topham  ;  Minsoo Kim 
Citation
 Journal of Immunological Methods, Vol.426(-) : 120-127, 2015 
Journal Title
 Journal of Immunological Methods 
ISSN
 0022-1759 
Issue Date
2015
MeSH
Animals ; Bacterial Proteins/genetics ; CD11b Antigen/immunology* ; CD18 Antigens/immunology ; Cell Adhesion ; Cell Movement/immunology ; Extracellular Matrix/immunology ; Flow Cytometry ; Gene Knock-In Techniques ; Intercellular Adhesion Molecule-1/metabolism ; Luminescent Proteins/genetics ; Macrophage-1 Antigen/analysis* ; Macrophage-1 Antigen/immunology* ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microscopy, Fluorescence ; Neutrophils/immunology* ; Phagocytosis/immunology ; Spleen/cytology* ; Spleen/immunology ; Staining and Labeling/methods
Keywords
CD11b ; Imaging ; Integrin ; MAC-1 ; YFP
Abstract
β2 integrins play critical roles in migration of immune cells and in the interaction with other cells, pathogens, and the extracellular matrix. Among the β2 integrins, Mac-1 (Macrophage antigen-1), composed of CD11b and CD18, is mainly expressed in innate immune cells and plays a major role in cell migration and trafficking. In order to image Mac-1-expressing cells both in live cells and mouse, we generated a knock-in (KI) mouse strain expressing CD11b conjugated with monomeric yellow fluorescent protein (mYFP). Expression of CD11b-mYFP protein was confirmed by Western blot and silver staining of CD11b-immunoprecipitates and total cell lysates from the mouse splenocytes. Mac-1-mediated functions of the KI neutrophils were comparable with those in WT cells. The fluorescence intensity of CD11b-mYFP was sufficient to image CD11b expressing cells in live mice using intravital two-photon microscopy. In vitro, dynamic changes in the intracellular localization of CD11b molecules could be measured by epifluorescent microscopy. Finally, CD11b-expressing immune cells from tissue were easily detected by flow cytometry without anti-CD11b antibody staining.
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DOI
10.1016/j.jim.2015.08.012
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Anatomy (해부학교실) > 1. Journal Papers
Yonsei Authors
Hyun, Young-Min(현영민) ORCID logo https://orcid.org/0000-0002-0567-2039
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/157304
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