Dakterzada Farida, Mohabati Mobarez Ashraf, Habibi Roudkenar Mehryar, Mohsenifar Afshin
Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Vaccine. 2016 Mar 14;34(12):1472-9. doi: 10.1016/j.vaccine.2016.01.041. Epub 2016 Feb 8.
Flagellin of Pseudomonas aeruginosa is an important vaccine candidate. N-terminal domains are highly conserved in both type a and type b flagellins. The efficacy of gold nanoparticles (AuNPs) conjugated to N-terminal domains of P. aeruginosa flagellin (flagellin(1-161)), as an immunogen in mice, has been assessed. The nanoparticles were conjugated to the recombinant protein through direct interaction of thiol molecules of the cysteines with AuNPs and formation of AuS bond. Flagellin(1-161), AuNP-flagellin(1-161), and flagellin(1-161) emulsified in Freund's adjuvant (FA: complete/incomplete Freund's adjuvant formulation) were administered subcutaneously to BALB/c mice. Mice given AuNP-flagellin(1-161) elicited high titers of anti-flagellin(1-161) antibodies compared with non-immune group and/or mice which received flagellin(1-161) without adjuvant. In whole cell ELISA, these antibodies effectively recognized the native flagellin on the bacteria. Opsonophagocytosis assay demonstrated the functional activity and specificity of anti-flagellin(1-161) antibodies raised by AuNP-flagellin(1-161) against homologous strain. All of the results were comparable with those obtained by use of FA. Taken together, this is the first report of conjugation of AuNPs to flagellin and evaluating its immune response against P. aeruginosa.
铜绿假单胞菌鞭毛蛋白是一种重要的疫苗候选物。a型和b型鞭毛蛋白的N端结构域高度保守。已评估了与铜绿假单胞菌鞭毛蛋白（鞭毛蛋白(1-161)）N端结构域偶联的金纳米颗粒（AuNP）作为小鼠免疫原的效果。纳米颗粒通过半胱氨酸的硫醇分子与AuNP的直接相互作用以及AuS键的形成与重组蛋白偶联。将鞭毛蛋白(1-161)、AuNP-鞭毛蛋白(1-161)以及在弗氏佐剂（FA：完全/不完全弗氏佐剂配方）中乳化的鞭毛蛋白(1-161)皮下注射给BALB/c小鼠。与非免疫组和/或未使用佐剂接受鞭毛蛋白(1-161)的小鼠相比，给予AuNP-鞭毛蛋白(1-161)的小鼠产生了高滴度的抗鞭毛蛋白(1-161)抗体。在全细胞ELISA中，这些抗体有效地识别了细菌上的天然鞭毛蛋白。调理吞噬作用试验证明了由AuNP-鞭毛蛋白(1-161)产生的抗鞭毛蛋白(1-161)抗体对同源菌株的功能活性和特异性。所有结果与使用FA获得的结果相当。综上所述，这是关于AuNP与鞭毛蛋白偶联并评估其对铜绿假单胞菌免疫反应的首次报道。
