We developed a new approach to bioluminescent T cell imaging using a membrane-anchored form of the Gaussia luciferase (GLuc) enzyme, termed extGLuc, which we could stably express in both mouse and human primary T cells. In vitro, extGLuc+ cells emitted significantly higher bioluminescent signal when compared to cells expressing GLuc, Renilla luciferase (RLuc) or membrane-anchored RLuc (extRLuc). In vivo, mouse extGLuc+ T cells showed higher bioluminescent signal when compared to GLuc+ and RLuc+ T cells. Application of this imaging approach to human T cells genetically modified to express tumor-specific chimeric antigen receptors (CARs) enabled us to show in vivo CAR-mediated T cell accumulation in tumor, T cell persistence over time and concomitant imaging of T cells and tumor cells modified to express firefly luciferase. This sensitive imaging technology has application to many in vivo cell-based studies in a wide array of mouse models.
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This work was supported by US National Institutes of Health grants CA95152, CA059350, CA08748, CA086438, CA096945, CA094060 and CA083084, The Alliance for Cancer Gene Therapy, the Damon Runyon Clinical Investigator Award (R.J.B.), The Annual Terry Fox Run for Cancer Research organized by the Canada Club of New York, Kate's Team, W.H. Goodwin and A. Goodwin and the Commonwealth Cancer Foundation for Research and the Experimental Therapeutics Center of MSKCC, the Geoffrey Beene Cancer Foundation, the Chronic Lymphocytic Leukemia Foundation, the Ludwig Center for Cancer Immunotherapy and the Bocina Cancer Research Fund.
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Santos, E., Yeh, R., Lee, J. et al. Sensitive in vivo imaging of T cells using a membrane-bound Gaussia princeps luciferase. Nat Med 15, 338–344 (2009). https://doi.org/10.1038/nm.1930
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