Janetzki Sylvia, Britten Cedrik M
ZellNet Consulting, Inc., Fort Lee, NJ, USA.
Methods Mol Biol. 2012;792:25-36. doi: 10.1007/978-1-61779-325-7_2.
During more than 25 years of application in immunological sciences, ELISPOT has been established as a routine, robust, versatile, and reliable assay. From basic research to clinical immune monitoring, ELISPOT is being used to address the quantification and (to a lesser extent) functional characterization of immune cells secreting different molecules in the context of health and disease, immune intervention, and therapy in humans and other species [Kalyuzhny (Ed.) (2005) Handbook of Elispot: methods and protocols, Vol. 302, Humana Press Inc., Totowa, NJ]. Over the last decade, ELISPOT assays have been increasingly implemented as an immune-monitoring tool in clinical trials [Schmittel et al. J Immunother 23:289-295, 2000; Whiteside Immunol Invest 29:149-162, 2000; Nagata et al. Ann N Y Acad Sci 1037:10-15, 2004; Cox et al. (2005) Cellular immune assays for evaluation of vaccine efficacy in developing countries., In Manual of Clinical Immunology Laboratory (Rose, N. R., Hamilton, R. G., and Detrick, B., Eds.), p 301, ASM Press, Washington, DC; Cox et al. Methods 38:274-282, 2006]. While the principles of the original protocol have changed little since its first introduction [Czerkinsky J Immunol Methods 110:29-36, 1988], individual laboratories have adapted assay procedures based on experimental needs, availability of reagents and equipment, obtained recommendations, and gained experience, leading to a wide disparity of applied ELISPOT protocols with inevitable consequences. This chapter addresses the resulting challenges for ELISPOT use in clinical trial settings, and discusses the influence of harmonization strategies as a tool for overcoming these challenges. Furthermore, harmonization is discussed in the context of assay standardization and validation strategies.
在免疫科学领域应用的25年多时间里，酶联免疫斑点技术（ELISPOT）已成为一种常规、稳健、多功能且可靠的检测方法。从基础研究到临床免疫监测，ELISPOT被用于在健康与疾病、免疫干预以及人类和其他物种的治疗背景下，对分泌不同分子的免疫细胞进行定量分析以及（在较小程度上）功能表征[卡柳日尼（主编）（2005年）《ELISPOT手册：方法与方案》，第302卷，人类出版社，新泽西州托托瓦]。在过去十年中，ELISPOT检测越来越多地被用作临床试验中的免疫监测工具[施米特尔等人，《免疫治疗杂志》23:289 - 295，2000年；怀特赛德，《免疫研究》29:149 - 162，2000年；永田等人，《纽约科学院学报》1037:10 - 15，2004年；考克斯等人（2005年），《发展中国家疫苗效力评估的细胞免疫检测》，载于《临床免疫实验室手册》（罗斯，N.R.、汉密尔顿，R.G.和德特里克，B.主编），第301页，美国微生物学会出版社，华盛顿特区；考克斯等人，《方法》38:274 - 282，2006年]。虽然自最初引入以来，原始方案的原理变化不大[切尔金斯基，《免疫方法杂志》110:29 - 36，1988年]，但各个实验室根据实验需求、试剂和设备的可用性、获得的建议以及积累的经验对检测程序进行了调整，导致应用的ELISPOT方案差异很大，不可避免地产生了一些后果。本章阐述了在临床试验环境中使用ELISPOT所面临的这些挑战，并讨论了作为克服这些挑战的工具的协调策略的影响。此外，还在检测标准化和验证策略的背景下讨论了协调问题。
