a Applied Biotechnology Research Centre , Baqiyatallah University of Medical Sciences , Tehran , Iran.
b Agriculture Biotechnology Department , National Institute for Genetic Engineering and Biotechnology , Tehran , Iran.
Artif Cells Nanomed Biotechnol. 2018;46(sup3):S1067-S1075. doi: 10.1080/21691401.2018.1529676. Epub 2019 Jan 14.
In the present study, the application of mesoporous silica nanoparticles (MSNPs) loaded with recombinant EspA protein, an immunogen of enterohaemorrhagic E. coli, was investigated in the case of BALB/c mice immunization against the bacterium. MSNPs of 96.9 ± 15.9 nm in diameter were synthesized using template removing method. The immunization of mice was carried out orally and subcutaneously. Significant immune responses to the antigen were observed for the immunized mice when rEspA-loaded MSNPs were administered in both routes in comparison to that of the antigen formulated using a well-known adjuvant, i.e. Freund's. According to the titretitre of serum IL-4, the most potent humoral responses were observed when the mice were immunized subcutaneously with antigen-loaded MSNPs (244, 36 and 14 ng/dL of IL-4 in the serum of mice immunized subcutaneously or orally by antigen-loaded MSNPs, and subcutaneously by Freund's adjuvant formulated-antigen, respectively). However, the difference in serum IgG and serum IgA was not significant in mice subcutaneously immunized with antigen-loaded MSNPs and mice immunized with Freund's adjuvant formulated-antigen. Finally, the immunized mice were challenged orally by enterohaemorrhagic E. coli cells. The amount of bacterial shedding was significantly reduced in faecesfaeces of the animals immunized by antigen-loaded MSNPs in both subcutaneous and oral routes.
在本研究中,研究了负载重组 EspA 蛋白(产志贺样毒素大肠埃希氏菌的免疫原)的介孔硅纳米颗粒(MSNPs)在 BALB/c 小鼠针对该细菌免疫中的应用。采用模板去除法合成了直径为 96.9±15.9nm 的 MSNPs。通过口服和皮下免疫的方式对小鼠进行免疫。与使用知名佐剂弗氏佐剂配制的抗原相比,当以两种途径给予负载 rEspA 的 MSNPs 时,免疫小鼠对抗原表现出显著的免疫反应。根据血清 IL-4 的滴定度,当用负载抗原的 MSNPs 对小鼠进行皮下免疫时,观察到最有效的体液反应(皮下或口服免疫的小鼠血清中 IL-4 的浓度分别为 244、36 和 14ng/dL,皮下免疫用弗氏佐剂配制的抗原)。然而,皮下免疫负载抗原的 MSNPs 的小鼠和皮下免疫弗氏佐剂配制的抗原的小鼠的血清 IgG 和血清 IgA 差异不显著。最后,用产志贺样毒素大肠埃希氏菌细胞对免疫小鼠进行口服攻毒。在皮下和口服两种途径免疫负载抗原的 MSNPs 的动物的粪便中,细菌脱落的量显著减少。