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In vivo detection of macrophage recruitment in hind-limb ischemia using a targeted near-infrared fluorophore

Macrophages are an essential component of the immune system and have protective and pathogenic functions in various diseases. Imaging of macrophages in vivo could furnish new tools to advance evaluation of disease and therapies. Critical limb ischemia is a disease in which macrophages have considera...

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Published in:PloS one 2014-07, Vol.9 (7), p.e103721-e103721
Main Authors: Yoo, Jung Sun, Das, Raj Kumar, Jow, Zhi Yen, Chang, Young-Tae
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description Macrophages are an essential component of the immune system and have protective and pathogenic functions in various diseases. Imaging of macrophages in vivo could furnish new tools to advance evaluation of disease and therapies. Critical limb ischemia is a disease in which macrophages have considerable pathogenic roles, and are potential targets for cell-based immunotherapy. We sought to develop a new near-infrared fluorescence (NIRF) imaging probe to target macrophages specifically in vivo in various pathological states, including hind-limb ischemia. We rapidly screened the photostable cyanine-based NIRF library against different blood cell lines. The identified monocyte/macrophage-selective hit was tested in vitro in live-cell labeling assay. Non-invasive NIRF imaging was performed with murine models of paw inflammation by lipopolysaccharide challenge and hind-limb ischemia with femoral artery ligation. in vivo macrophage targeting was further evaluated using intravital microscopy with Csf1r-EGFP transgenic mice and immunofluorescent staining with macrophage-specific markers. We discovered MF800, a Macrophage-specific near-infrared Fluorophore, which showed selective live-cell imaging performance in a panel of cell lines and primary human blood samples. MF800 outperforms the clinically-available NIRF contrast agent ICG for in vivo specificity in paw inflammation and hind-limb ischemia models. We observed a marked overlap of MF800-labeled cells and EGFP-expressing macrophages in intravital imaging of Csf1r-EGFP transgenic mice. In the histologic analysis, MF800-positive cells also expressed the macrophage markers CD68 and CD169. NIRF imaging showcased the potential of using MF800 to understand macrophage behavior in vivo, characterize macrophage-associated diseases, and may help in assessing therapeutic responses in the clinic.
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subjects Animal models
Animals
Atherosclerosis
Biology and Life Sciences
Biotechnology
Blood
Blood cells
Cell Line, Tumor
Cell lines
Consortia
Disease
Disease Models, Animal
FDA approval
Femoral artery
Femur
Fluorescence
Fluorescent Antibody Technique
Fluorescent Dyes - chemical synthesis
Fluorescent Dyes - chemistry
Fluorescent Dyes - metabolism
Gene expression
Genetic engineering
Granulocytes
Homeostasis
Humans
I.R. radiation
Immune system
Immunotherapy
Indoles - chemical synthesis
Indoles - chemistry
Indoles - metabolism
Inflammation
Inflammation - diagnostic imaging
Infrared imaging
Ionizing radiation
Ischemia
Ischemia - diagnostic imaging
Ischemia - metabolism
Ischemia - pathology
Laboratories
Leg
Lipopolysaccharides
Macrophages
Macrophages - chemistry
Macrophages - cytology
Macrophages - metabolism
Markers
Medical imaging
Medical screening
Mice
Mice, Inbred C57BL
Mice, Transgenic
Microscopy
Microscopy, Fluorescence
Monocytes
NMR
Nuclear magnetic resonance
Quantum dots
Radiography
Research and Analysis Methods
Spectroscopy, Near-Infrared
Target recognition
Transgenic mice
title In vivo detection of macrophage recruitment in hind-limb ischemia using a targeted near-infrared fluorophore
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