应用果蝇胚胎细胞表达系统分泌表达猪干扰素α的试验

Study of the Secretory expression of pig interferon-alpha in recombinant Drosophila melanogaster S2 cells

干扰素作为一类具有多种生物学功能的细胞因子,已经在人类疾病的防治中得到广泛应用。在兽药行业,大量的临床试验证明了干扰素激活免疫系统快速、广谱的有效性,但干扰素蛋白的活性和使用成本一直是制约其在兽药中应用的重要因素。为了探索人工重组的干扰素在兽医临床上应用的可行性,本文应用果蝇胚胎细胞表达系统筛选出了稳定表达猪干扰素α(rPIFNα)的细胞系,利用Western blot分析了细胞筛选前后的分泌蛋白的表达情况.结果显示,稳定表达细胞系的rPIFNα表达量较高,通过细胞试验证实,分泌表达的rPIFNα具有较强的抗病毒作用。本研究首次利用果蝇胚胎表达系统稳定表达猪干扰素,对干扰素在临床上应用是一个有益的尝试。

As a kind of multifunction cytokines, Interferences （IFN） had been administrated in many diseases of human being. In the study of Veterinary medicine, a great number of clinical tests has showed thePharmacological effects on the activation of the immune cells. However, for the reasons of the biological activity of recombinant proteins or the high-cost of extraction technologies, the administrations of interferon were still not widely used. In this article, we firstly purified recombinant PIFNa using an S2 secretion expression system. The antiviral activity of the rPIFNa was assessed using the VSV-Vero cell model and the result showed that rPIFNa inhibited VSV infection. This study indicates that the pig rPIFNa can be obtained with genetic engineering.