Details

Autor(en) / Beteiligte

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

Titel

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

Ist Teil von

• Proceedings of the National Academy of Sciences - PNAS, 2008-09, Vol.105 (38), p.14342-14346

Ort / Verlag

United States: National Academy of Sciences

Erscheinungsjahr

2008

Quelle

MEDLINE

Beschreibungen/Notizen

• 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 10⁴ 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.