Departments of Bacteriology, Pasteur Institute of Iran, Tehran, Iran; Departments of Bacteriology, Faculty of Medical sciences, Tarbiat Modares University, Tehran, Iran.
APMIS. 2014 Feb;122(2):115-27. doi: 10.1111/apm.12101. Epub 2013 Jun 12.
The main goal of this study was to develop a vaccination strategy that would enhance the protective response against the recombinant type A flagellin (r-fla-A) of Pseudomonas aeruginosa in the burn wound sepsis model. Inbred mice were immunized with r-fla-A with or without alum adjuvant. The vaccinated mice were burned and challenged with P. aeruginosa. To evaluate the type of induced immune response, sera were analyzed by ELISA for total IgG, IgG1, and IgG2a isotypes. To determine the functional activity of anti r-fla IgG, opsonophagocytic killing and motility inhibition assay was performed. In vivo administration of r-fla-A afforded a remarkable improvement in survival of mice (83.3%) challenged with homologous strain (PAK) in the burn wound infection. The antibodies generated against the r-fla-A achieved 25% survival in immunized mice that had been infected with heterologous strain PAO1. Flagellin also induced high level humoral immune response via high titers of serum IgG1 in the burn and challenged mice. Anti r-fla-A antibody promoted phagocytosis of the PAK strain, and the number of viable bacterial cells decreased over 53.1%; In contrast, low opsonophagocytic killing activity (17.4%) was observed when the antiserum to r-fla-A was treated with the PAO1 strain. The anti r-fla-A antisera was able to inhibit the motility of the homologous strains; however, they did not inhibit the heterologous strains. We concluded that active immunization with recombinant type A-flagellin could protect burn mice against lethal P. aeruginosa challenge via immobilization of the pathogen which promoted the phagocytic activity.
本研究的主要目的是开发一种疫苗接种策略,以增强对铜绿假单胞菌重组 A 型鞭毛蛋白(r-fla-A)的保护反应,这种鞭毛蛋白存在于烧伤创面脓毒症模型中。近交系小鼠用 r-fla-A 加或不加明矾佐剂进行免疫接种。对接种疫苗的小鼠进行烧伤并感染铜绿假单胞菌。为了评估诱导的免疫反应类型,通过 ELISA 分析血清中的总 IgG、IgG1 和 IgG2a 同种型。为了确定抗 r-fla IgG 的功能活性,进行调理吞噬杀伤和运动抑制试验。在烧伤创面感染中,r-fla-A 的体内给药显著提高了同源菌株(PAK)攻击小鼠的存活率(83.3%)。针对 r-fla-A 产生的抗体可使感染异源菌株 PAO1 的免疫小鼠的存活率达到 25%。鞭毛蛋白还通过烧伤和攻毒小鼠血清 IgG1 的高滴度诱导高水平的体液免疫反应。抗 r-fla-A 抗体促进了 PAK 株的吞噬作用,活菌数减少了 53.1%以上;相比之下,当用 r-fla-A 抗血清处理 PAO1 菌株时,观察到调理吞噬杀伤活性较低(17.4%)。抗 r-fla-A 抗血清能够抑制同源株的运动,但不能抑制异源株的运动。我们得出结论,用重组 A 型鞭毛蛋白进行主动免疫接种可以通过固定病原体来保护烧伤小鼠免受致命的铜绿假单胞菌攻击,从而促进吞噬作用。