Hop Huynh Tan, Reyes Alisha Wehdnesday Bernardo, Simborio Hannah Leah Tadeja, Arayan Lauren Togonon, Min Won Gi, Lee Hu Jang, Lee Jin Ju, Chang Hong Hee, Kim Suk
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea.
Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
J Microbiol Biotechnol. 2016 Jan;26(1):190-6. doi: 10.4014/jmb.1505.05028.
In this study, the Brucella abortus ohr gene coding for an organic hydroperoxide resistance protein (Ohr) was cloned into a maltose fusion protein expression system (pMAL), inserted into Escherichia coli, and purified, and its immunogenicity was evaluated by western blot analysis using Brucella-positive mouse sera. The purified recombinant Ohr (rOhr) was treated with adjuvant and injected intraperitoneally into BALB/c mice. A protective immune response analysis revealed that rOhr induced a significant increase in both the IgG1 and IgG2a titers, and IgG2a reached a higher level than IgG1 after the second and third immunizations. Additionally, immunization with rOhr induced high production of IFN-γ as well as proinflammatory cytokines such as TNF, MCP-1, IL-12p70, and IL-6, but a lesser amount of IL-10, suggesting that rOhr predominantly elicited a cell-mediated immune response. In addition, immunization with rOhr caused a significantly higher degree of protection against a virulent B. abortus infection compared with a positive control group consisting of mice immunized with maltose-binding protein. These findings showed that B. abortus rOhr was able to induce both humoral and cell-mediated immunity in mice, which suggested that this recombinant protein could be a potential vaccine candidate for animal brucellosis.
在本研究中,编码有机氢过氧化物抗性蛋白(Ohr)的流产布鲁氏菌ohr基因被克隆到麦芽糖融合蛋白表达系统(pMAL)中,插入大肠杆菌并进行纯化,然后使用布鲁氏菌阳性小鼠血清通过蛋白质印迹分析评估其免疫原性。将纯化的重组Ohr(rOhr)用佐剂处理后腹腔注射到BALB/c小鼠体内。保护性免疫反应分析显示,rOhr诱导IgG1和IgG2a滴度显著增加,且在第二次和第三次免疫后IgG2a水平高于IgG1。此外,用rOhr免疫诱导了高水平的IFN-γ以及促炎细胞因子如TNF、MCP-1、IL-12p70和IL-6,但IL-10的量较少,这表明rOhr主要引发细胞介导的免疫反应。此外,与用麦芽糖结合蛋白免疫的小鼠组成的阳性对照组相比,用rOhr免疫对强毒流产布鲁氏菌感染的保护程度显著更高。这些发现表明,流产布鲁氏菌rOhr能够在小鼠体内诱导体液免疫和细胞介导的免疫,这表明这种重组蛋白可能是动物布鲁氏菌病的潜在疫苗候选物。