Mirzaei Maryam, Xu Yan, Elias Cynthia B, Prakash Satya
Biomedical Technology and Cell Therapy Research Laboratory, Department of Biomedical Engineering, and Artificial Cells and Organs Research Centre, Faculty of Medicine, McGill University, 3775 University Street, Montreal, PQ, Canada H3A 2B4.
J Biomed Biotechnol. 2009;2009:637942. doi: 10.1155/2009/637942. Epub 2008 Oct 15.
Human interleukin-7 (hIL-7) is a cytokine secreted by the stromal cells of the red marrow. It is important for proliferation during certain stages of B-cell maturation and for T and NK cell survival, development, and homeostasis. It is a critical growth factor for enhancement and recovery of the immune T-cell. Because of its strong immunomodulatory effects, hIL-7 may become a valuable supplementary agent for immunotherapeutical treatments in patients with HIV infection or immunodeficiency. Human IL-7 has previously been produced in various protein expression systems. In this paper, we present an alternative expression system, in Spodoptera frugiperda cells, for the production of hIL-7 using nonlytic vector systems. This system allows generation of correctly translated and accurately processed heterologous proteins as soluble recombinant proteins. Here we report plasmid construction, transfection, and consequent expression of hIL-7 using this nonlytic insect cell expression system. The levels of secreted hIL-7 in a small scale experiment reached a level of 1.7 microg x 1(-1) under serum-free cell culture conditions.
人白细胞介素-7（hIL-7）是由红骨髓基质细胞分泌的一种细胞因子。它在B细胞成熟的某些阶段对增殖以及T细胞和NK细胞的存活、发育和稳态维持很重要。它是增强和恢复免疫T细胞的关键生长因子。由于其强大的免疫调节作用，hIL-7可能成为HIV感染或免疫缺陷患者免疫治疗的有价值的辅助药物。人IL-7以前已在各种蛋白质表达系统中产生。在本文中，我们提出了一种在草地贪夜蛾细胞中使用非裂解载体系统生产hIL-7的替代表达系统。该系统能够产生正确翻译和精确加工的异源蛋白，作为可溶性重组蛋白。在此我们报告使用这种非裂解昆虫细胞表达系统进行hIL-7的质粒构建、转染及后续表达。在无血清细胞培养条件下，小规模实验中分泌的hIL-7水平达到了1.7μg×1(-1)。
