Lehmann Alexander, Megyesi Zoltan, Przybyla Anna, Lehmann Paul V
Research and Development Department, CTL, Shaker Heights, OH 44122, USA.
Department of Cancer Immunology, Chair of Medical Biotechnology, Poznan University of Medical Sciences, 61-701 Poznan, Poland.
Cells. 2018 Jan 3;7(1):3. doi: 10.3390/cells7010003.
ELISPOT assays enable the detection of the frequency of antigen-specific T cells in the blood by measuring the secretion of cytokines, or combinations of cytokines, in response to antigenic challenges of a defined population of PBMC. As such, these assays are suited to establish the magnitude and quality of T cell immunity in infectious, allergic, autoimmune and transplant settings, as well as for measurements of anti-tumor immunity. The simplicity, robustness, cost-effectiveness and scalability of ELISPOT renders it suitable for regulated immune monitoring. In response to the regulatory requirements of clinical and pre-clinical immune monitoring trials, tamper-proof audit trails have been introduced to all steps of ELISPOT analysis: from capturing the raw images of assay wells and counting of spots, to all subsequent quality control steps involved in count verification. A major shortcoming of ELISPOT and other related cellular assays is presently the lack of audit trails for the wet laboratory part of the assay, in particular, the assurance that no pipetting errors have occurred during the plating of antigens and cells. Here, we introduce a dye-based reagent tracking platform that fills this gap, thereby increasing the transparency and documentation of ELISPOT test results.
酶联免疫斑点（ELISPOT）检测法通过测量细胞因子或细胞因子组合在应对特定群体外周血单核细胞（PBMC）抗原刺激时的分泌情况，能够检测血液中抗原特异性T细胞的频率。因此，这些检测方法适用于确定感染、过敏、自身免疫和移植环境中T细胞免疫的强度和质量，以及用于抗肿瘤免疫的测量。ELISPOT检测法的简单性、稳健性、成本效益和可扩展性使其适用于规范的免疫监测。为响应临床和临床前免疫监测试验的监管要求，在ELISPOT分析的所有步骤中都引入了防篡改审计追踪：从捕获检测孔的原始图像和斑点计数，到计数验证中涉及的所有后续质量控制步骤。目前，ELISPOT检测法及其他相关细胞检测法的一个主要缺点是检测的湿实验室部分缺乏审计追踪，特别是无法确保在抗原和细胞铺板过程中没有发生移液错误。在此，我们引入了一种基于染料的试剂追踪平台来填补这一空白，从而提高ELISPOT检测结果的透明度和记录。
