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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Production of Genetically Engineered Biotinylated Interleukin-2 and Its Application in a Rapid Nonradioactive Assay for T-Cell Activation
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Production of Genetically Engineered Biotinylated Interleukin-2 and Its Application in a Rapid Nonradioactive Assay for T-Cell Activation

机译：基因工程生物素化白介素2的生产及其在T细胞活化快速非放射性分析中的应用

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The development of reliable assay systems that can measure lymphocyte activation in vitro has been a major goal of immunodiagnostics. Traditionally, tritiated thymidine incorporation has been used to monitor T-cell activation. Other methods include enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunospot assay, and colorimetric assays. We have established a lymphocyte activation assay that utilizes fluorescein isothiocyanate (FITC)-streptavidin bound to recombinant biotinylated human interleukin-2 (IL-2). Utilizing recombinant DNA technology, a unique monobiotinylated human IL-2 has been created and isolated using the Promega PinPoint vector system. ELISA has been used to demonstrate streptavidin binding and recognition by a human IL-2-specific antibody. IL-2 function has been demonstrated using a murine IL-2-dependent T-cell line, CTLL-2, responsive to human IL-2. Recombinant biotinylated human IL-2 conjugated to streptavidin-FITC or streptavidin-horseradish peroxidase has been used to monitor T-cell activation in the presence of antigen as well as mitogen. The sensitivity and convenience of this method make this lymphocyte activation assay an attractive alternative to tritiated thymidine incorporation as a method for monitoring T-cell activation. In addition, the availability of a recombinant biotinylated human IL-2 will permit the production of a uniform product suitable for diagnostic and clinical application.
机译：可以测量体外淋巴细胞活化的可靠测定系统的开发一直是免疫诊断的主要目标。传统上，tri化胸腺嘧啶核苷的掺入已用于监测T细胞活化。其他方法包括酶联免疫吸附测定（ELISA），酶联免疫斑点测定和比色测定。我们已经建立了利用异硫氰酸荧光素（FITC）-链霉亲和素与重组生物素化人白介素2（IL-2）结合的淋巴细胞活化测定法。利用重组DNA技术，使用Promega PinPoint载体系统创建并分离了独特的生物素化人IL-2。 ELISA已用于证明链霉亲和素与人IL-2特异性抗体的结合和识别。已经使用对人IL-2有反应的鼠IL-2依赖性T细胞系CTLL-2证明了IL-2的功能。与抗生蛋白链菌素-FITC或抗生蛋白链菌素-辣根过氧化物酶偶联的重组生物素化人IL-2已用于监测抗原和促细胞分裂剂存在下的T细胞活化。这种方法的灵敏性和便利性使该淋巴细胞活化测定法成为tri化胸苷掺入的一种有吸引力的替代方法，作为监测T细胞活化的一种方法。另外，重组生物素化的人IL-2的可用性将允许产生适用于诊断和临床应用的均匀产品。

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