Visualizing fewer than 10 mouse T cells with an enhanced firefly luciferase in immunocompetent mouse models of cancer

Brian A. Rabinovich, Yang Ye, Tamara Etto, Qing Chen Jie, Hyam I. Levitsky, Willem W. Overwijk, Laurence J N Cooper, Juri Gelovani, Patrick Hwu

Research output: Contribution to journalArticle

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 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

Original languageEnglish (US)
Pages (from-to)14342-14346
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume105
Issue number38
DOIs
StatePublished - Sep 23 2008

Keywords

  • Bioluminescence
  • Immunology
  • Molecular biology

ASJC Scopus subject areas

  • General

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    Rabinovich, B. A., Ye, Y., Etto, T., Jie, Q. C., Levitsky, H. I., Overwijk, W. W., Cooper, L. J. N., Gelovani, J., & Hwu, P. (2008). Visualizing fewer than 10 mouse T cells with an enhanced firefly luciferase in immunocompetent mouse models of cancer. Proceedings of the National Academy of Sciences of the United States of America, 105(38), 14342-14346. https://doi.org/10.1073/pnas.0804105105