Sensitive in vivo imaging of T cells using a membrane-bound Gaussia princeps luciferase.
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.
Santos
Elmer B
EB
Department of Radiology, Memorial Sloan Kettering Cancer Center (MSKCC), 1275 York Avenue, New York, New York 10021, USA.
Yeh
Raymond
R
Lee
James
J
Nikhamin
Yan
Y
Punzalan
Blesida
B
Punzalan
Blesserene
B
La Perle
Krista
K
Larson
Steven M
SM
Sadelain
Michel
M
Brentjens
Renier J
RJ
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*
D008156:Luciferases