Luo Deyan, Ni Bing, Li Peng, Shi Wei, Zhang Songle, Han Yue, Mao Liwei, He Yangdong, Wu Yuzhang, Wang Xiliang
State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing, China.
Infect Immun. 2006 May;74(5):2734-41. doi: 10.1128/IAI.74.5.2734-2741.2006.
This study was designed to evaluate the immunogenicity and the protective efficacy of a divalent fusion DNA vaccine encoding both the Brucella abortus L7/L12 protein (ribosomal protein) and Omp16 protein (outer membrane lipoprotein), designated pcDNA3.1-L7/L12-Omp16. Intramuscular injection of this divalent DNA vaccine into BALB/c mice elicited markedly both humoral and cellular immune responses. The specific antibodies exhibited a dominance of immunoglobulin G2a (IgG2a) over IgG1. In addition, the dual-gene DNA vaccine elicited a strong T-cell proliferative response and induced a large amount of gamma interferon-producing T cells upon restimulation in vitro with recombinant fusion protein L7/L12-Omp16, suggesting the induction of a typical T-helper-1-dominated immune response in vivo. This divalent DNA vaccine could also induce a significant level of protection against challenge with the virulent strain B. abortus 544 in BALB/c mice. Furthermore, the protection level induced by the divalent DNA vaccine was significantly higher than that induced by the univalent DNA vaccines pcDNA3.1-L7/L12 or pcDNA3.1-Omp16. Taken together, the results of this study verify for the first time that the Omp16 gene can be a candidate target for a DNA vaccine against brucellosis. Additionally, a divalent genetic vaccine based on the L7/L12 and Omp16 genes can elicit a stronger cellular immune response and better immunoprotection than the relevant univalent vaccines can.
本研究旨在评估一种编码流产布鲁氏菌L7/L12蛋白(核糖体蛋白)和Omp16蛋白(外膜脂蛋白)的二价融合DNA疫苗(命名为pcDNA3.1-L7/L12-Omp16)的免疫原性和保护效力。将这种二价DNA疫苗肌肉注射到BALB/c小鼠体内可显著激发体液免疫和细胞免疫反应。特异性抗体表现为免疫球蛋白G2a(IgG2a)比IgG1占优势。此外,双基因DNA疫苗引发了强烈的T细胞增殖反应,在用重组融合蛋白L7/L12-Omp16体外再次刺激时诱导产生了大量产生γ干扰素的T细胞,这表明在体内诱导了典型的以辅助性T细胞1为主导的免疫反应。这种二价DNA疫苗还能在BALB/c小鼠中诱导出显著水平的针对强毒株流产布鲁氏菌544攻击的保护作用。此外,二价DNA疫苗诱导的保护水平显著高于单价DNA疫苗pcDNA3.1-L7/L12或pcDNA3.1-Omp16诱导的保护水平。综上所述,本研究结果首次证实Omp16基因可作为抗布鲁氏菌病DNA疫苗的候选靶点。此外,基于L7/L12和Omp16基因的二价基因疫苗比相关单价疫苗能引发更强的细胞免疫反应和更好的免疫保护作用。