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铜绿假单胞菌重组b型鞭毛蛋白作为候选疫苗的免疫原性:在小鼠烧伤创面脓毒症模型中的保护效果。

Immunogenicity of Pseudomonas aeruginosa recombinant b-type fagellin as a vaccine candidate: Protective efficacy in a murine burn wound sepsis model.

作者信息

Behrouz Bahador, Mahdavi Mehdi, Amirmozafari Nour, Fatemi Mohammad Javad, Irajian Gholamreza, Bahroudi Mahbubeh, Hashemi Farhad B

机构信息

Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

Department of Immunology, Pasteur Institute of Iran, Tehran, Iran.

出版信息

Burns. 2016 May 2. doi: 10.1016/j.burns.2016.03.015.

Abstract

Pseudomonas aeruginosa (PA) is a formidable opportunistic pathogen among patients with burn wound infections. Antimicrobial therapy is often unsuccessful because PA can develop multi-drug resistance; thus, immunotherapy can be a rational alternative. The goal of this study was to evaluate the immunogenicity recombinant type b flagellin (r-b-flagellin) as a potential vaccine against P. aeruginosa in a mouse model for burn wound sepsis. Primary immunization with r-b-flagellin (10μg) followed by two booster shots was sufficient to generate a robust humoral response, which was predominantly a T helper 2 (Th2) type response consisting mainly of subtype IgG1 and low levels of IgG2a. Analysis of the Th1-Th2 response among immunized mice showed an increased production of IL-4, INF-γ and IL-17 by splenocytes upon stimulation by r-b-flagellin. Opsono-phagocytosis assays confirmed the enhanced killing of bacteria by anti r-b-flagellin immune sera. These antibodies were also able to inhibit motility of P. aeruginosa and afforded protection to immunized mice by reducing bacterial load in the site of original infection into the liver of challenged mice. The reduction of systemic bacterial spread resulted in an increase in the survival rate of challenged immunized mice. In conclusion, immunization of mice with r-b-flagellin protein increased the level of humoral and cellular immune response and led to an efficacious protection against P. aeruginosa infection in the burn mouse model.

摘要

铜绿假单胞菌(PA)是烧伤创面感染患者中一种可怕的机会致病菌。抗菌治疗往往不成功,因为PA会产生多重耐药性;因此,免疫治疗可能是一种合理的替代方法。本研究的目的是在烧伤创面脓毒症小鼠模型中评估重组b型鞭毛蛋白(r-b-鞭毛蛋白)作为抗铜绿假单胞菌潜在疫苗的免疫原性。用r-b-鞭毛蛋白(10μg)进行初次免疫,随后进行两次加强注射,足以产生强烈的体液反应,该反应主要是T辅助2(Th2)型反应,主要由IgG1亚型和低水平的IgG2a组成。对免疫小鼠中Th1-Th2反应的分析表明,r-b-鞭毛蛋白刺激后,脾细胞产生的IL-4、INF-γ和IL-17增加。调理吞噬试验证实了抗r-b-鞭毛蛋白免疫血清对细菌的杀伤作用增强。这些抗体还能够抑制铜绿假单胞菌的运动,并通过减少感染小鼠肝脏中原感染部位的细菌载量为免疫小鼠提供保护。全身细菌传播的减少导致受攻击免疫小鼠的存活率增加。总之,用r-b-鞭毛蛋白对小鼠进行免疫可提高体液和细胞免疫反应水平,并在烧伤小鼠模型中有效预防铜绿假单胞菌感染。

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