title: Sensitive in vivo imaging of T cells using a membrane-bound Gaussia princeps luciferase.
abstract: 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.
authors.affiliation: Department of Radiology, Memorial Sloan Kettering Cancer Center (MSKCC), 1275 York Avenue, New York, New York 10021, USA.
authors.forename: Elmer B
authors.initials: EB
authors.lastname: Santos
authors.forename: Raymond
authors.initials: R
authors.lastname: Yeh
authors.forename: James
authors.initials: J
authors.lastname: Lee
authors.forename: Yan
authors.initials: Y
authors.lastname: Nikhamin
authors.forename: Blesida
authors.initials: B
authors.lastname: Punzalan
authors.forename: Blesserene
authors.initials: B
authors.lastname: Punzalan
authors.forename: Krista
authors.initials: K
authors.lastname: La Perle
authors.forename: Steven M
authors.initials: SM
authors.lastname: Larson
authors.forename: Michel
authors.initials: M
authors.lastname: Sadelain
authors.forename: Renier J
authors.initials: RJ
authors.lastname: Brentjens
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D001517:Coleoptera / Q000201:enzymology; D005434:Flow Cytometry; D006801:Humans; D008156:Luciferases / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology*
chemical_list: D008156:Luciferases