title: Visualizing fewer than 10 mouse T cells with an enhanced firefly luciferase in immunocompetent mouse models of cancer.
abstract: Antigen specific T cell migration to sites of infection or cancer is critical for an effective immune response. In mouse models of cancer, the number of lymphocytes reaching the tumor is typically only a few hundred, yet technology capable of imaging these cells using bioluminescence has yet to be achieved. A combination of codon optimization, removal of cryptic splice sites and retroviral modification was used to engineer an enhanced firefly luciferase (ffLuc) vector. Compared with ffLuc, T cells expressing our construct generated >100 times more light, permitting detection of as few as three cells implanted s.c. while maintaining long term coexpression of a reporter gene (Thy1.1). Expression of enhanced ffLuc in mouse T cells permitted the tracking of <3 x 10(4) adoptively transferred T cells infiltrating sites of vaccination and preestablished tumors. Penetration of light through deep tissues, including the liver and spleen, was also observed. Finally, we were able to enumerate infiltrating mouse lymphocytes constituting <0.3% of total tumor cellularity, representing a significant improvement over standard methods of quantitation including flow cytometry.
authors.affiliation: M. D. Anderson Cancer Center, 7455 Fannin Street, Houston, TX 77054, USA. brabinov@mdanderson.org
authors.forename: Brian A
authors.initials: BA
authors.lastname: Rabinovich
authors.forename: Yang
authors.initials: Y
authors.lastname: Ye
authors.forename: Tamara
authors.initials: T
authors.lastname: Etto
authors.forename: Jie Qing
authors.initials: JQ
authors.lastname: Chen
authors.forename: Hyam I
authors.initials: HI
authors.lastname: Levitsky
authors.forename: Willem W
authors.initials: WW
authors.lastname: Overwijk
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
authors.forename: Juri
authors.initials: J
authors.lastname: Gelovani
authors.forename: Patrick
authors.initials: P
authors.lastname: Hwu
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology; D004195:Disease Models, Animal; D015971:Gene Expression Regulation, Enzymologic; D005822:Genetic Vectors / Q000235:genetics; D007121:Immunocompetence; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements; D051379:Mice; D008810:Mice, Inbred C57BL; D009369:Neoplasms / Q000276:immunology*; D010047:Ovalbumin / Q000276:immunology; D015202:Protein Engineering; D012190:Retroviridae / Q000235:genetics; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: D010047:Ovalbumin; D049409:Luciferases, Firefly
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