Document Detail


Direct observation to chemokine receptor 5 on T-lymphocyte cell surface using fluorescent metal nanoprobes.
MedLine Citation:
PMID:  20705055     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Chemokine receptor 5 (CCR5) is a cell surface protein required for HIV-1 infection. It is important to detect the amount and observe the spatial distribution of the CCR5 receptors on the cell surfaces. In this report, we describes the metal nanoparticles which were specially designed as molecular fluorescent probes for imaging of CCR5 receptors on the T-lymphocytic PM1 cell surfaces. These CCR5 monoclonal antibodies (mAbs) metal complexes were prepared by labeling mAbs with Alexa Fluor 680 followed by covalent binding the labeled mAbs on the 20 nm silver nanoparticles. Compared with the labeled mAbs without metal, the mAb-metal complexes were found to display enhanced emission intensity and shortened lifetime due to interactions between fluorophores and metal. The mAb-metal complexes were incubated with the PM1 cell lines. The confocal fluorescent intensity and lifetime cell images were recorded on single cells. It was observed that the mAb-metal complexes could be clearly distinguished from the cellular autofluorescence. By analyzing a pool of cell images, we observed that most CCR5 receptors appeared as clusters on the cell surfaces. The fluorophore-metal complexes developed in this report are generally useful for detection of cell surface receptors and provide a new class of probe to study the interaction between the CCR5 receptors with viral gp120 during HIV infections.
Authors:
Jian Zhang; Yi Fu; Ge Li; Kazimierz Nowaczyk; Richard Y Zhao; Joseph R Lakowicz
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2010-08-10
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  400     ISSN:  1090-2104     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-09-13     Completed Date:  2010-10-11     Revised Date:  2011-09-26    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  111-6     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier Inc. All rights reserved.
Affiliation:
Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, MD 21201, USA. jian@cfs.bioment.umaryland.edu
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal / immunology
Cell Line
Cell Membrane / chemistry,  immunology*
HIV Envelope Protein gp120 / immunology
Humans
Metal Nanoparticles / chemistry*
Molecular Probes
Receptors, CCR5 / analysis*,  immunology
Spectrometry, Fluorescence / methods*
T-Lymphocytes / chemistry,  immunology*,  virology
Grant Support
ID/Acronym/Agency:
CA134386/CA/NCI NIH HHS; EB006521/EB/NIBIB NIH HHS; EB009509/EB/NIBIB NIH HHS; HG-002655/HG/NHGRI NIH HHS; HG005090/HG/NHGRI NIH HHS; R01 EB006521-03/EB/NIBIB NIH HHS; R01 HG002655-08/HG/NHGRI NIH HHS; R21 CA134386-02/CA/NCI NIH HHS; R21 EB009509-01A2/EB/NIBIB NIH HHS; R21 EB009509-02/EB/NIBIB NIH HHS; R21 HG005090-03/HG/NHGRI NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/HIV Envelope Protein gp120; 0/Molecular Probes; 0/Receptors, CCR5
Comments/Corrections

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