Visualizing fewer than 10 mouse T cells with an enhanced firefly luciferase in immunocompetent mouse models of cancer.
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.
Rabinovich
Brian A
BA
M. D. Anderson Cancer Center, 7455 Fannin Street, Houston, TX 77054, USA. brabinov@mdanderson.org
Ye
Yang
Y
Etto
Tamara
T
Chen
Jie Qing
JQ
Levitsky
Hyam I
HI
Overwijk
Willem W
WW
Cooper
Laurence J N
LJ
Gelovani
Juri
J
Hwu
Patrick
P
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
D010047:Ovalbumin; D049409:Luciferases, Firefly
J Clin Invest. 1996 Jan 15;97(2):515-21
8567975
J Natl Cancer Inst. 1989 Nov 15;81(22):1709-17
2810387
J Virol. 1996 Aug;70(8):5701-5
8764092
Nat Rev Cancer. 2002 Apr;2(4):266-76
12001988
Immunity. 2004 Sep;21(3):315-29
15357943
Mol Imaging. 2006 Apr-Jun;5(2):57-64
16954019
Hum Gene Ther. 2007 Jul;18(7):575-88
17576157
Cancer. 1994 Mar 15;73(6):1731-7
8156501
Blood. 2005 Aug 1;106(3):1113-22
15855275
Am J Med Sci. 1989 Jul;298(1):28-33
2665484
Cancer Res. 2006 Nov 15;66(22):10995-1004
17108138
Mol Ther. 2005 Mar;11(3):435-43
15727940
Int Immunol. 2007 Oct;19(10):1165-73
17698980
Mol Microbiol. 1995 Nov;18(4):593-603
8817482
Nat Med. 2003 Sep;9(9):1144-50
12925844
Cell. 1994 Jan 14;76(1):17-27
8287475
J Immunol. 2001 Aug 15;167(4):2379-87
11490028
Blood. 2007 Oct 15;110(8):2793-802
17638856
J Virol. 1999 Apr;73(4):2886-92
10074136
J Virol. 2004 Nov;78(22):12537-47
15507641
J Immunol. 2007 Sep 1;179(5):3332-41
17709550
Blood. 2006 Dec 1;108(12):3890-7
16926291
Proc Natl Acad Sci U S A. 2004 Oct 5;101 Suppl 2:14639-45
15381769
Blood. 2006 Jul 1;108(1):390-9
16522809
Cancer Res. 2007 Mar 15;67(6):2872-80
17363611
J Clin Oncol. 2006 Nov 1;24(31):5060-9
17075125
J Immunol. 2001 Jun 1;166(11):6500-8
11359800
Blood. 2007 Oct 1;110(7):2620-30
17507664
J Clin Oncol. 1995 Aug;13(8):1939-49
7636534
J Immunol. 2003 Apr 1;170(7):3572-6
12646619
J Clin Invest. 2001 May;107(10):1293-301
11375419
Exp Hematol. 2001 Dec;29(12):1353-60
11750093
J Biomed Opt. 2005 Jul-Aug;10(4):41210
16178634
Science. 2006 Mar 24;311(5768):1780-4
16556847
Am J Med Sci. 1988 Dec;296(6):406-12
3063115
Circulation. 2005 Aug 30;112(9 Suppl):I105-10
16159800
J Clin Invest. 2008 Apr;118(4):1390-7
18357341