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

被引：166

作者：

Rabinovich, Brian A. ^{[1
]}

Ye, Yang ^{[1
]}

Etto, Tamara ^{[1
]}

Chen, Jie Qing ^{[1
]}

Levitsky, Hyam I. ^{[2
]}

Overwijk, Willem W. ^{[1
]}

Cooper, Laurence J. N. ^{[1
]}

Gelovani, Juri ^{[1
]}

Hwu, Patrick ^{[1
]}

机构：

[1] MD Anderson Canc Ctr, Houston, TX 77054 USA

[2] Johns Hopkins Univ, Sch Med, Sidney Kimmel Comprehens Canc Ctr, Baltimore, MD 21231 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2008年 / 105卷 / 38期

基金：

美国国家卫生研究院;

关键词：

bioluminescence; immunology; molecular biology;

D O I：

10.1073/pnas.0804105105

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：14342 / 14346

页数：5

共 39 条

[21] Phase I study of adoptive T-cell therapy using antigen-specific CD8+ T cells for the treatment of patients with metastatic melanoma [J].