Evaluation of Two Internalizing Carcinoembryonic Antigen Reporter Genes for Molecular ImagingReport as inadecuate




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Molecular Imaging and Biology

, Volume 13, Issue 3, pp 526–535

First Online: 14 July 2010

Abstract

PurposeThe objective of this article is to develop internalizing positron emission tomography PET reporter genes for tracking genetically modified T cells in vivo.

ProceduresThe transmembrane and cytoplasmic domains of the human transferrin receptor TfR and CD5 were each fused to the carcinoembryonic CEA minigene N-A3 and expressed in Jurkat T cells. Internalization was evaluated by confocal microscopy or by intracellular uptake of I-labeled anti-CEA scFv-Fc. Reporter gene-transfected Jurkat xenografts in mice were analyzed by immunohistochemistry IHC and imaged by PET using I- or Cu-scFv-Fc as tracers.

ResultsSurface expression of TR1–99-NA3 was lower than that of NA3-CD5. Both reporter genes were internalized following binding of the anti-CEA antibody fragment. IHC of tumors showed strong staining of NA3-CD5, whereas TR1–99-NA3 stained weakly. Specific targeting of TR1–99-NA3 or NA3-CD5 was shown by PET in xenografted mice.

ConclusionsThe in vivo imaging studies suggest a potential application of the internalizing form of CEA N-A3 as a PET reporter gene.

Key wordsAntibody fragment CEA reporter gene Internalization Jurkat PET Category and significance: An original article describing the design, generation, expression, and both in vitro and in vivo evaluation of two versions of an internalizing reporter gene. The significance of this approach lies in utilizing reporter protein internalization as a means of accumulation of the tracer in the target cells.

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Author: Bhaswati Barat - Vania E. Kenanova - Tove Olafsen - Anna M. Wu

Source: https://link.springer.com/







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