PMID- 26342259
OWN - NLM
STAT- MEDLINE
DCOM- 20160323
LR  - 20181113
IS  - 1872-7905 (Electronic)
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 426
DP  - 2015 Nov
TI  - Visualization of integrin Mac-1 in vivo.
PG  - 120-7
LID - S0022-1759(15)30039-9 [pii]
LID - 10.1016/j.jim.2015.08.012 [doi]
AB  - beta2 integrins play critical roles in migration of immune cells and in the 
      interaction with other cells, pathogens, and the extracellular matrix. Among the 
      beta2 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.
CI  - Copyright (c) 2015 Elsevier B.V. All rights reserved.
FAU - Lim, Kihong
AU  - Lim K
AD  - Department of Microbiology and Immunology, David H. Smith Center for Vaccine 
      Biology and Immunology, University of Rochester, Rochester, NY, USA. Electronic 
      address: Kihong_Lim@urmc.rochester.edu.
FAU - Hyun, Young-Min
AU  - Hyun YM
AD  - Department of Microbiology and Immunology, David H. Smith Center for Vaccine 
      Biology and Immunology, University of Rochester, Rochester, NY, USA.
FAU - Lambert-Emo, Kris
AU  - Lambert-Emo K
AD  - Department of Microbiology and Immunology, David H. Smith Center for Vaccine 
      Biology and Immunology, University of Rochester, Rochester, NY, USA.
FAU - Topham, David J
AU  - Topham DJ
AD  - Department of Microbiology and Immunology, David H. Smith Center for Vaccine 
      Biology and Immunology, University of Rochester, Rochester, NY, USA.
FAU - Kim, Minsoo
AU  - Kim M
AD  - Department of Microbiology and Immunology, David H. Smith Center for Vaccine 
      Biology and Immunology, University of Rochester, Rochester, NY, USA. Electronic 
      address: minsoo_kim@urmc.rochester.edu.
LA  - eng
GR  - P01 AI102851/AI/NIAID NIH HHS/United States
GR  - P01 HL018208/HL/NHLBI NIH HHS/United States
GR  - HL018208/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20150902
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Bacterial Proteins)
RN  - 0 (CD11b Antigen)
RN  - 0 (CD18 Antigens)
RN  - 0 (Luminescent Proteins)
RN  - 0 (Macrophage-1 Antigen)
RN  - 0 (yellow fluorescent protein, Bacteria)
RN  - 126547-89-5 (Intercellular Adhesion Molecule-1)
SB  - IM
MH  - Animals
MH  - Bacterial Proteins/genetics
MH  - CD11b Antigen/*immunology
MH  - CD18 Antigens/immunology
MH  - Cell Adhesion
MH  - Cell Movement/immunology
MH  - Extracellular Matrix/immunology
MH  - Flow Cytometry
MH  - Gene Knock-In Techniques
MH  - Intercellular Adhesion Molecule-1/metabolism
MH  - Luminescent Proteins/genetics
MH  - Macrophage-1 Antigen/*analysis/*immunology
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Transgenic
MH  - Microscopy, Fluorescence
MH  - Neutrophils/*immunology
MH  - Phagocytosis/immunology
MH  - Spleen/*cytology/immunology
MH  - Staining and Labeling/methods
PMC - PMC4651804
MID - NIHMS721555
OTO - NOTNLM
OT  - CD11b
OT  - Imaging
OT  - Integrin
OT  - MAC-1
OT  - YFP
EDAT- 2015/09/08 06:00
MHDA- 2016/03/24 06:00
PMCR- 2016/11/01
CRDT- 2015/09/07 06:00
PHST- 2015/05/21 00:00 [received]
PHST- 2015/08/19 00:00 [revised]
PHST- 2015/08/25 00:00 [accepted]
PHST- 2015/09/07 06:00 [entrez]
PHST- 2015/09/08 06:00 [pubmed]
PHST- 2016/03/24 06:00 [medline]
PHST- 2016/11/01 00:00 [pmc-release]
AID - S0022-1759(15)30039-9 [pii]
AID - 10.1016/j.jim.2015.08.012 [doi]
PST - ppublish
SO  - J Immunol Methods. 2015 Nov;426:120-7. doi: 10.1016/j.jim.2015.08.012. Epub 2015 
      Sep 2.