Negri Donatella, Sestili Paola, Borghi Martina, Ciccolella Maria, Bracci Laura
Department of Infectious Diseases, Istituto Superiore di Sanità, Rome, Italy.
National Centre for the Control and the Evaluation of Medicines, Istituto Superiore di Sanità, Rome, Italy.
Methods Enzymol. 2020;632:457-477. doi: 10.1016/bs.mie.2019.05.058. Epub 2019 Jun 29.
Critical to the advancement of tumor immunotherapy is the reliable identification of responders and the quantification of the tumor-specific immune response elicited by treatments. In this regard, Enzyme-Linked Immunospot assay (ELISpot) is an ideal monitoring technique due to its high sensitivity, ease of execution and cost-effectiveness. Originally developed for the enumeration of B cells secreting antigen-specific antibodies, ELISpot assay has been adapted to detect and quantify cytokine-secreting immune cells present at low frequency in a variety of biological samples, including blood, in response to antigen-specific stimuli. The above-mentioned features emphasize the role of ELISpot as valuable assay for translational research and clinical applications. In the present chapter, we will focus on the use of ELISpot assay for monitoring the tumor-specific effector responses induced by different treatments in preclinical models and will provide some protocols and technical hints for its application.
肿瘤免疫治疗进展的关键在于可靠地识别反应者以及量化治疗引发的肿瘤特异性免疫反应。在这方面,酶联免疫斑点测定法(ELISpot)因其高灵敏度、易于操作和成本效益高而成为一种理想的监测技术。ELISpot测定法最初是为了计数分泌抗原特异性抗体的B细胞而开发的,现已被用于检测和量化在包括血液在内的各种生物样品中对抗原特异性刺激产生反应的低频存在的分泌细胞因子的免疫细胞。上述特点突出了ELISpot作为转化研究和临床应用中有价值测定方法的作用。在本章中,我们将重点介绍ELISpot测定法在临床前模型中监测不同治疗诱导的肿瘤特异性效应反应的应用,并提供一些应用方案和技术提示。