Gholami M, Chirani A S, Razavi S, Falak R, Irajian G
Department of Microbiology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Department of Medical Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Lett Appl Microbiol. 2017 Nov;65(5):439-445. doi: 10.1111/lam.12796. Epub 2017 Oct 3.
Interference with bacterial adhesion is a new means to prevent or treat bacterial infections. In this experimental study we evaluated the immunogenic properties of a chimeric protein composed of PilQ and disulphide turn region of PilA from Pseudomonas aeruginosa in mice as an anti-adhesion based vaccine. First of all, a chimeric bivalent protein composed of PilQ and PilA was constructed and following subcutaneous immunization with merely the purified protein or in its admixed form with alum, the immunogenicity of the chimeric antigen was assessed in BALB/c mice. Then, the characteristics of the developed antibodies were studied by ELISA. Furthermore, the immunoreactivity of the purified recombinant protein was confirmed by immunoblotting. Alum as a common adjuvant boosted immunogenicity of the construct, resulting significantly greater anti-pili IgG titre. Mice antibody response consisted of IgG1, IgG2a, IgG2b and IgG3 subtypes with predominance of IgG1 subclass. The developed antibodies were capable to inhibit motility of PAO1 strain. In conclusion, our primary results revealed that the designed recombinant protein is a protective construct and may be used as a potential candidate for prophylactic purposes against P. aeruginosa infection.
In this study we examined the potential of integrated PilQ/PilA (QA) antigen as a vaccine candidate against Pseudomonas aeruginosa. Nowadays, anti-adhesion based vaccines are considered as new means to prevent or treat bacterial infections. Our study revealed that chimeric protein PilQ and disulphide turn region of PilA triggers production of specific antibodies. This humoral immune responses augmented when QA was administered in combination with an adjuvant. The results demonstrated efficacy of the designed recombinant chimeric antigen as an effective candidate in prevention of P. aeruginosa infection.
干扰细菌黏附是预防或治疗细菌感染的一种新方法。在本实验研究中，我们评估了由铜绿假单胞菌的PilQ和PilA的二硫键转折区组成的嵌合蛋白在小鼠体内作为基于抗黏附的疫苗的免疫原性。首先，构建了由PilQ和PilA组成的嵌合二价蛋白，然后仅用纯化蛋白或其与明矾的混合形式进行皮下免疫，在BALB/c小鼠中评估嵌合抗原的免疫原性。然后，通过ELISA研究产生的抗体的特性。此外，通过免疫印迹证实了纯化重组蛋白的免疫反应性。明矾作为一种常见佐剂增强了构建体的免疫原性，导致抗菌毛IgG滴度显著更高。小鼠抗体反应由IgG1、IgG2a、IgG2b和IgG3亚型组成，以IgG1亚类为主。产生的抗体能够抑制PAO1菌株的运动性。总之，我们的初步结果表明，设计的重组蛋白是一种保护性构建体，可作为预防铜绿假单胞菌感染的潜在候选物。
在本研究中，我们研究了整合的PilQ/PilA（QA）抗原作为抗铜绿假单胞菌疫苗候选物的潜力。如今，基于抗黏附的疫苗被认为是预防或治疗细菌感染的新方法。我们的研究表明，嵌合蛋白PilQ和PilA的二硫键转折区可触发特异性抗体的产生。当QA与佐剂联合使用时，这种体液免疫反应会增强。结果证明了设计的重组嵌合抗原作为预防铜绿假单胞菌感染的有效候选物的有效性。
