Recombinant carcinoembryonic antigen as a reporter gene for molecular imaging.

European Journal of Nuclear Medicine and Molecular Imaging
Vania E KenanovaAnna M Wu

Abstract

Reporter genes can provide a way of noninvasively assessing gene activity in vivo. However, current reporter gene strategies may be limited by the immunogenicity of foreign reporter proteins, endogenous expression, or unwanted biological activity. We have developed a reporter gene based on carcinoembryonic antigen (CEA), a human protein with limited normal tissue expression. To construct a CEA reporter gene for PET, a CEA minigene (N-A3) was fused to the extracellular and transmembrane domains of the human Fc gamma RIIb receptor. The NA3-Fc gamma RIIb recombinant gene, driven by a CMV promoter, was transfected in Jurkat (human T cell leukemia) cells. Expression was analyzed by flow cytometry, immunohistochemistry (IHC), and microPET imaging. Flow cytometry identified Jurkat clones stably expressing NA3-Fc gamma RIIb at low, medium, and high levels. High and medium NA3-Fc gamma RIIb expression could also be detected by Western blot. Reporter gene positive and negative Jurkat cells were used to establish xenografts in athymic mice. IHC showed staining of the tumor with high reporter gene expression; medium and low N-A3 expression was not detected. MicroPET imaging, using an anti-CEA (124)I-labeled single-chain Fv-Fc antibody frag...Continue Reading

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Citations

Jul 16, 2010·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·Bhaswati BaratAnna M Wu
Feb 8, 2014·European Journal of Nuclear Medicine and Molecular Imaging·Yingfeng TuBaozhong Shen
Jul 30, 2019·Immunological Reviews·David AkhavanChristine E Brown
Dec 17, 2014·Physics in Medicine and Biology·Katherine L WalkerGregory S Mitchell
Apr 25, 2020·Frontiers in Physiology·Madeleine Iafrate, Gilbert O Fruhwirth
Apr 4, 2020·Molecular Therapy : the Journal of the American Society of Gene Therapy·Candice Ashmore-HarrisGilbert O Fruhwirth
Sep 23, 2021·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·Anna M Wu, Neeta Pandit-Taskar
Apr 2, 2021·Journal of Nuclear Medicine : Official Publication, Society of Nuclear Medicine·Mathew VealBart Cornelissen

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