Lin Y, Robb R J, Gray J E, Simon P
E. I. Du Pont and Co., Glenolden Laboratory, PA 19036.
J Immunol. 1988 Dec 1;141(11):3847-51.
A polylysine extended gibbon IL-2 (IL-2-L) was constructed by the addition of a lysine-rich oligopeptide, Gly3-(Lys-Lys-Asp)3-Leu-Glu to the C terminus of gibbon IL-2 by using rDNA technology. The bioactivity of the hybrid molecule was comparable to that of normal human rIL-2 in assays measuring T cell proliferation, and in generation of lymphokine-activated killer cells from PBL. Furthermore, the addition of the lysine-rich segment made the molecule more amenable to the chemical derivatization of amino groups. After biotinylation, IL-2-L retained a greater proportion of its bioactivity than normal rIL-2. In addition, biotinylated IL-2-L was capable of simultaneously binding to cell surface IL-2R and streptavidin. Thus, the addition of the lysine-rich oligopeptide facilitated the generation of an active form of biotinylated IL-2 which acts as a bridge between IL-2R-positive cells and avidin-coupled reagents and affinity supports. Such a selective means of labeling and binding IL-2-responsive cells may have great practical utility for IL-2-based immunotherapy.
通过重组DNA技术，在长臂猿IL-2的C末端添加富含赖氨酸的寡肽Gly3-(Lys-Lys-Asp)3-Leu-Glu，构建了一种多聚赖氨酸延伸的长臂猿IL-2（IL-2-L）。在检测T细胞增殖以及从外周血淋巴细胞（PBL）产生淋巴因子激活的杀伤细胞的实验中，该杂合分子的生物活性与正常人重组IL-2相当。此外，富含赖氨酸片段的添加使该分子更易于进行氨基的化学衍生化。生物素化后，IL-2-L保留的生物活性比例比正常重组IL-2更高。此外，生物素化的IL-2-L能够同时结合细胞表面的IL-2受体和链霉亲和素。因此，富含赖氨酸寡肽的添加促进了生物素化IL-2活性形式的产生，该活性形式可作为IL-2R阳性细胞与抗生物素蛋白偶联试剂及亲和支持物之间的桥梁。这种标记和结合IL-2反应性细胞的选择性方法可能对基于IL-2的免疫治疗具有很大的实际应用价值。
