TY - JOUR
T1 - Visualizing fewer than 10 mouse T cells with an enhanced firefly luciferase in immunocompetent mouse models of cancer
AU - Rabinovich, Brian A.
AU - Ye, Yang
AU - Etto, Tamara
AU - Jie, Qing Chen
AU - Levitsky, Hyam I.
AU - Overwijk, Willem W.
AU - Cooper, Laurence J N
AU - Gelovani, Juri
AU - Hwu, Patrick
PY - 2008/9/23
Y1 - 2008/9/23
N2 - 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
AB - 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
KW - Bioluminescence
KW - Immunology
KW - Molecular biology
UR - http://www.scopus.com/inward/record.url?scp=55749089404&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=55749089404&partnerID=8YFLogxK
U2 - 10.1073/pnas.0804105105
DO - 10.1073/pnas.0804105105
M3 - Article
C2 - 18794521
AN - SCOPUS:55749089404
SN - 0027-8424
VL - 105
SP - 14342
EP - 14346
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 38
ER -