A Robotic Microscope System to Examine T Cell Receptor Acuity Against Tumor Neoantigens: A New Tool for Cancer Immunotherapy Research
A Robotic Microscope System to Examine T Cell Receptor Acuity Against Tumor Neoantigens: A New Tool for Cancer Immunotherapy Research
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DOI：
10.1109/lra.2019.2894466
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发表时间：
2019-04-01
影响因子：
5.2
通讯作者：
Asada, H. Harry
中科院分区：
文献类型：
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作者：
Ong, Lee-Ling Sharon;Zhu, Hai;Asada, H. Harry
在免疫监视过程中，细胞毒性 T 淋巴细胞 (CTL) 可以通过识别与排列在癌细胞表面的主要组织相容性复合体分子 (pMHC) 结合的肿瘤特异性肽（新抗原）来选择性地识别和破坏肿瘤细胞。 CTL 使用相同的机制来破坏显示病原体特异性 pMHC 的病毒感染细胞，同时留下表达正常自身 pMHC 的完整健康细胞。我们推出了一款机器人显微镜，使科学家能够以高通量进行高灵敏度和选择性的 T 细胞-pMHC 研究。我们的系统操纵涂有特定 pMHC 的微米珠，将它们呈现给 T 细胞，并产生检测 T 细胞与新抗原的敏锐度所需的皮牛顿级分子间力。我们的系统集成了光镊、精密纳微米级以及两种放大倍数的落射/透射照明方案。我们创建了一个坐标参考系统，根据细胞引起的每个焦平面的特征强度变化，在大空间内对半透明 T 细胞进行三维定位。我们的系统执行自动化实验，通过测量下游细胞反应来检测特定 pMHC 的 T 细胞敏锐度水平。可以选择性地回收高敏锐度 T 细胞用于单细胞分析。我们的新方法和工具将对癌症免疫治疗和免疫学研究产生重大影响。
During immune surveillance, cytotoxic T lymphocytes (CTL) can selectively identify and destroy tumor cells by recognizing tumor-specific peptides (neoantigens), bound to major histocompatibility complex molecules (pMHC) arrayed on cancer cell surfaces. CTL use the same machinery to destroy virally infected cells displaying pathogen-specific pMHC, while leaving intact healthy cells expressing normal self-pMHC. We present a robotic microscope that allows scientists to conduct highly sensitive and selective T cell-pMHC studies with high throughput. Our system manipulates micro-meter beads coated with particular pMHC, presents them to T cells and generates piconewton level intermolecular forces required to detect T cell acuity with a neoantigen. Our system integrates optical tweezers, precision nanomicro stages, and episcopic/diascopic illumination schemes at two magnifications. We create a coordinate referencing system to locate translucent T cells in three-dimensional, over a large space, based on the characteristic intensity change at each focal plane caused by the cells. Our system performs automated experiments to detect the level of T cell acuity with specific pMHCs, by measuring the downstream cellular responses. High acuity T cells can be selectively recovered for single cell analysis. Our new methodology and tool will have a significant impact on cancer immunotherapy and immunology research.