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Issue 1, 2013
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Imaging of genetically engineered T cells by PET using gold nanoparticles complexed to Copper-64

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Abstract

Adoptive transfer of primary T cells genetically modified to have desired specificity can exert an anti-tumor response in some patients. To improve our understanding of their therapeutic potential we have developed a clinically-appealing approach to reveal their in vivo biodistribution using nanoparticles that serve as a radiotracer for imaging by positron emission tomography (PET). T cells electroporated with DNA plasmids from the Sleeping Beauty transposon–transposase system to co-express a chimeric antigen receptor (CAR) specific for CD19 and Firefly luciferase (ffLuc) were propagated on CD19+ K562-derived artificial antigen presenting cells. The approach to generating our clinical-grade CAR+ T cells was adapted for electro-transfer of gold nanoparticles (GNPs) functionalized with 64Cu2+ using the macrocyclic chelator (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, DOTA) and polyethyleneglycol (GNP–64Cu/PEG2000). MicroPET/CT was used to visualize CAR+EGFPffLucHyTK+GNP–64Cu/PEG2000+ T cells and correlated with bioluminescence imaging. These data demonstrate that GNPs conjugated with 64Cu2+ can be prepared as a radiotracer for PET and used to image T cells using an approach that has translational implications.

Graphical abstract: Imaging of genetically engineered T cells by PET using gold nanoparticles complexed to Copper-64

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Supplementary files

Article information


Submitted
23 Apr 2012
Accepted
17 Sep 2012
First published
18 Sep 2012

Integr. Biol., 2013,5, 231-238
Article type
Technical Innovation

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