Direct observation to chemokine receptor 5 on T-lymphocyte cell surface using fluorescent metal nanoprobes.
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.
Zhang
Jian
J
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
Fu
Yi
Y
Li
Ge
G
Nowaczyk
Kazimierz
K
Zhao
Richard Y
RY
Lakowicz
Joseph R
JR
D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D015335:Molecular Probes; D019713:Receptors, CCR5 / Q000032:analysis* / Q000276:immunology; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology* / Q000821:virology
D000911:Antibodies, Monoclonal; D015699:HIV Envelope Protein gp120; D015335:Molecular Probes; D019713:Receptors, CCR5