Ling Xiao-Dong, Dong Wei-Tao, Zhang Yong, Hu Jun-Jie, Zhang Wang-Dong, Wu Jin-Tang, Liu Ji-Xing, Zhao Xing-Xu
Department of Clinical Veterinary, College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, China.
Product R&D, Lanzhou Weitesen Biological Technology Co. Ltd., Lanzhou, China.
Arch Insect Biochem Physiol. 2019 Apr;100(4):e21539. doi: 10.1002/arch.21539. Epub 2019 Feb 21.
Silkworm (Bombyx mori) larvae are widely used to express exogenous proteins. Moreover, some silkworm pupal proteins can be used as drug-loading materials for selfexpressed oral tolerance drugs. However, several proteins expressed in silkworm pupae cause severe allergic reactions in humans and animals. Interestingly, some baculovirus vectors have been shown to alter the host gene and its expression in insect cells, but this has not been confirmed in silkworm. Here, we analyzed the effects of infection with an empty B. mori baculovirus (BmNPV) vector on silkworm pupal protein expression. Using a proteomics approach, the allergens thiol peroxiredoxin (Jafrac1), 27-kDa glycoprotein (p27k), arginine kinase, and paramyosin as well as 32 additional differentially expressed proteins were identified. Downregulation of the messenger RNA expression of the four known allergens was observed after BmNPV infection; subsequent changes in protein expression were confirmed by the western blot analysis using polyclonal antibodies prepared with recombinant proteins of the four allergens. Collectively, these data indicate that the four known allergens of silkworm pupae can be reduced by infection ith an empty BmNPV vector to increase the safety of silkworm pupa-based exogenous protein expression and drug delivery of oral pharmaceuticals. In addition, the four recombinant allergen proteins may contribute to the diagnosis of allergic diseases of silkworm pupa.
家蚕(Bombyx mori)幼虫被广泛用于表达外源蛋白。此外,一些家蚕蛹蛋白可作为自表达口服耐受药物的载药材料。然而,在家蚕蛹中表达的几种蛋白会在人和动物中引起严重的过敏反应。有趣的是,一些杆状病毒载体已被证明可改变昆虫细胞中的宿主基因及其表达,但在家蚕中尚未得到证实。在此,我们分析了用空的家蚕杆状病毒(BmNPV)载体感染对家蚕蛹蛋白表达的影响。采用蛋白质组学方法,鉴定出过敏原硫醇过氧化物酶(Jafrac1)、27 kDa糖蛋白(p27k)、精氨酸激酶和副肌球蛋白以及另外32种差异表达蛋白。BmNPV感染后观察到四种已知过敏原的信使核糖核酸表达下调;随后使用由四种过敏原的重组蛋白制备的多克隆抗体通过蛋白质印迹分析证实了蛋白质表达的变化。总体而言,这些数据表明,用空的BmNPV载体感染可降低家蚕蛹的四种已知过敏原,以提高基于家蚕蛹的外源蛋白表达和口服药物递送的安全性。此外,这四种重组过敏原蛋白可能有助于家蚕蛹过敏性疾病的诊断。