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新PAIR-T:使用CRISPR工程的Jurkat Reporter系统对新抗原-TCR对进行功能映射

Koji Nagaoka¹, Yukari Kobayashi¹, Kazuhiro Kakimi¹

¹Department of Immunology, Kindai University Faculty of Medicine, Sakai 590-0197, Osaka, Japan.

|November 26, 2025

概括

这项研究引入了NeoPAIR-T,这是一种用于识别功能性新抗原-T细胞受体 (TCR) 配对的新试验. 这种方法通过有效选针对疫苗和TCR-T细胞应用的瘤反应型TCR来简化个性化癌症免疫疗法.

科学领域:

免疫学免疫学免疫学
在瘤学瘤学.
生物技术是生物技术.

背景情况:

针对新抗原的个性化免疫疗法显示出希望,但在识别真正的新抗原和相关的T细胞受体 (TCRs) 方面面临挑战.
对新抗原的计算预测通常是不确定的,大多数瘤透淋巴细胞是旁观者,需要功能验证.
目前的方法需要广泛的合成,并且在选新抗原-TCR对中缺乏效率.

研究的目的:

开发一种功能性试验,NeoPAIR-T (使用记者T细胞进行Neoantigen-TCR配对试验),用于有效查和识别功能性新抗原-TCR对.
克服针对个性化癌症免疫治疗的新抗原预测和TCR反应性评估的局限性.
为疫苗和TCR-T细胞疗法确定新抗原-TCR对提供简化方法.

主要方法:

开发了NeoPAIR-T,一种共同培养试验,使用工程TCR-T记者细胞 (Jurkat-derived with luciferase/eGFP双记者) 和自身抗原呈现细胞 (APCs) 感染了编码预测的新抗原的协同小基因 (TMGs).
集成的TCRα-knock-out与有针对性的TCRβ-knock-in,以防止TCR错配并使多个记者T细胞克隆的并行测试成为可能.
应用全外体和RNA测序到肺癌样本,以预测新抗原,组装成TMG,并使用单细胞RNA/TCR测序来识别TCR克隆类型,用于记者T细胞工程.

主要成果:

在肺癌样本中,NeoPAIR-T成功识别了两个功能性的新抗原-TCR对.
该试验通过平行测试8种TCR克隆类型与表达TMG的APC对抗,证明了高效率.
功能对通过测定得到验证,显示出高亲和力 (EC50: 10^-9.2 到 10^-6.7 M).

结论:

新PAIR-T是一种有效和精简的功能测试,用于识别新抗原-TCR对.
这种测试克服了新抗原和TCR识别的关键挑战,促进了个性化癌症免疫疗法.
新PAIR-T对新型癌症疫苗和TCR-T细胞疗法的开发有重大影响.

关键词:

这就是CRISPR/Cas9的作用.这种T细胞受体是T细胞受体.功能查功能查新抗原是一种新抗原.个性化免疫疗法是个性化的免疫疗法.一个单细胞分析.坦德姆迷你发动机双重迷你发动机

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