Bidmon Nicole, Attig Sebastian, Rae Richard, Schröder Helene, Omokoko Tana A, Simon Petra, Kuhn Andreas N, Kreiter Sebastian, Sahin Ugur, Gouttefangeas Cécile, van der Burg Sjoerd H, Britten Cedrik M
Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
Translational Oncology, University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany; University Medical Center, Johannes Gutenberg University, 55131 Mainz, Germany;
J Immunol. 2015 Jun 15;194(12):6177-89. doi: 10.4049/jimmunol.1400958. Epub 2015 May 8.
The systematic assessment of the human immune system bears huge potential to guide rational development of novel immunotherapies and clinical decision making. Multiple assays to monitor the quantity, phenotype, and function of Ag-specific T cells are commonly used to unravel patients' immune signatures in various disease settings and during therapeutic interventions. When compared with tests measuring soluble analytes, cellular immune assays have a higher variation, which is a major technical factor limiting their broad adoption in clinical immunology. The key solution may arise from continuous control of assay performance using TCR-engineered reference samples. We developed a simple, stable, robust, and scalable technology to generate reference samples that contain defined numbers of functional Ag-specific T cells. First, we show that RNA-engineered lymphocytes, equipped with selected TCRs, can repetitively deliver functional readouts of a controlled size across multiple assay platforms. We further describe a concept for the application of TCR-engineered reference samples to keep assay performance within or across institutions under tight control. Finally, we provide evidence that these novel control reagents can sensitively detect assay variation resulting from typical sources of error, such as low cell quality, loss of reagent stability, suboptimal hardware settings, or inaccurate gating.
对人类免疫系统进行系统评估，在指导新型免疫疗法的合理研发及临床决策方面具有巨大潜力。多种用于监测抗原特异性T细胞数量、表型和功能的检测方法，常用于揭示患者在各种疾病背景及治疗干预期间的免疫特征。与检测可溶性分析物的检测方法相比，细胞免疫检测的变异性更高，这是限制其在临床免疫学中广泛应用的一个主要技术因素。关键解决方案可能来自于使用TCR工程化参考样本对检测性能进行持续控制。我们开发了一种简单、稳定、可靠且可扩展的技术，以生成包含特定数量功能性抗原特异性T细胞的参考样本。首先，我们证明配备选定TCR的RNA工程化淋巴细胞能够在多个检测平台上重复提供大小可控的功能性读数。我们进一步描述了应用TCR工程化参考样本的概念，以严格控制机构内部或不同机构之间的检测性能。最后，我们提供证据表明，这些新型对照试剂能够灵敏地检测出由典型误差源导致的检测变异性，如细胞质量低、试剂稳定性丧失、硬件设置欠佳或门控不准确等。
