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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Visualizing Fewer Than 10 Mouse T Cells With An Enhanced Firefly Luciferase In Immunocompetent Mouse Models Of Cancer
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Visualizing Fewer Than 10 Mouse T Cells With An Enhanced Firefly Luciferase In Immunocompetent Mouse Models Of Cancer

机译：在癌症的免疫能力强的小鼠模型中，用增强的萤火虫萤光素酶可视化少于10个小鼠T细胞

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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 (Thy 1.1). Expression of enhanced ffLuc in mouse T cells permitted the tracking of < 3 × 10~4 adoptively transferred T cells infiltrating sites of vaccination and preestab-lished 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.
机译：抗原特异性T细胞向感染或癌症部位的迁移对于有效的免疫反应至关重要。在癌症的小鼠模型中，到达肿瘤的淋巴细胞数量通常只有几百个，但是尚未能够实现使用生物发光成像这些细胞的技术。密码子优化，去除隐蔽剪接位点和逆转录病毒修饰的组合被用于设计增强的萤火虫荧光素酶（ffLuc）载体。与ffLuc相比，表达我们的构建体的T细胞产生的光多100倍以上，从而可以检测到多达3个植入s.c的细胞。同时保持报告基因的长期共表达（Thy 1.1）。在小鼠T细胞中表达增强的ffLuc，可以追踪＜3×10〜4个过继转移的T细胞渗入疫苗和预先确定的肿瘤的浸润部位。还观察到光穿透包括肝脏和脾脏在内的深层组织。最后，我们能够枚举构成总肿瘤细胞<0.3％的浸润小鼠淋巴细胞，这比包括流式细胞术在内的标准定量方法有了显着改进。

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