期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 105, 期 38, 页码 14342-14346出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0804105105
关键词
bioluminescence; immunology; molecular biology
资金
- National Institutes of Health [RO1 CA116206, CA120956, R21 CA129390]
- Anderson Cancer Center [CA16672]
- Netherlands Organisation for Scientific Research Veni [916.046.014]
- Anderson Cancer Center Specialized Programs of Research Excellence [P50 CA093459]
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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据