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包含汉坦病毒核衣壳蛋白和糖蛋白Gn的融合蛋白的免疫学特性

Immunological properties of a fusion protein containing nucleocapsid protein and glycoprotein Gn of Hantaan virus.

作者信息

Luo W, Zhang F, Yan Y, Wu X, Liu Y, Bai W, Zhao Q, Wang H, Xu Z

机构信息

Department of Microbiology, Fourth Military Medical University, 17 West Changle Rd, Xi'an 710032, PR China.

出版信息

Acta Virol. 2008;52(4):243-9.

Abstract

Escherichia coli and the baculovirus Bac-to-Bac system were used to express chimeric gene GnS0.7 consisting of glycoprotein Gn gene and the 0.7 kb fragment of S genome segment carrying nucleoprotein (N) gene of Hantaan virus (HTNV). The expressed fusion protein GnN0.7 was recognized by monoclonal antibodies (MAbs) to HTNV Gn glycoprotein and N protein, respectively. Its molecular mass as determined by Western blot analysis corresponded to the predicted value. The mice immunized with insect Spodoptera frugiperda (Sf9) cells transfected with recombinant baculovirus carrying GnS0.7 gene, produced serum antibodies with titer up to 3,200 as assayed by immunofluorescence. Moreover, immunized mice showed positive proliferation index for splenocytes stimulated with HTNV Gn and N, respectively. These results indicated that insect Sf9 cells infected with the recombinant baculovirus expressed a fully biologically active fusion protein that elicited not only humoral but also cellular immune response in mice. Hence, this protein may be used as a genetically engineered subunit HTNV vaccine representing efficacious and safe alternative to traditional vaccines.

摘要

利用大肠杆菌和杆状病毒杆状病毒表达系统来表达嵌合基因GnS0.7,该基因由汉坦病毒(HTNV)糖蛋白Gn基因和携带核蛋白(N)基因的S基因组片段的0.7 kb片段组成。表达的融合蛋白GnN0.7分别被针对HTNV Gn糖蛋白和N蛋白的单克隆抗体(MAb)识别。通过蛋白质印迹分析测定的其分子量与预测值相符。用携带GnS0.7基因的重组杆状病毒转染的昆虫草地贪夜蛾(Sf9)细胞免疫的小鼠,通过免疫荧光测定产生的血清抗体效价高达3200。此外,免疫小鼠对分别用HTNV Gn和N刺激的脾细胞显示出阳性增殖指数。这些结果表明,感染重组杆状病毒的昆虫Sf9细胞表达了一种完全具有生物活性的融合蛋白,该蛋白在小鼠中不仅引发了体液免疫反应,还引发了细胞免疫反应。因此,这种蛋白可作为一种基因工程亚单位HTNV疫苗,是传统疫苗有效且安全的替代品。

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