Im Young Bin, Jung Myunghwan, Shin Min-Kyoung, Kim Suk, Yoo Han Sang
Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul, 08826, South Korea.
College of Veterinary Medicine, Gyeongsang National University, Jinju, 52828, South Korea.
Vet Res. 2016 Feb 11;47:30. doi: 10.1186/s13567-016-0311-7.
Brucellosis is a clinically and economically important disease. Therefore, eradication programs of the disease have been implemented in several countries. One hurdle in these programs is the detection of infected animals at the early stage. Although the protein antigens as diagnostic antigens have recently received attention, the exact mechanisms at the beginning of immune responses are not yet known. Therefore, genes encoding five B. abortus cellular proteins were cloned and the expressed recombinant proteins were purified. The expression of several cytokine genes (IL-1β, IL-4, IL-6, IL-12p40, IFN-γ, TNF-α, and iNOS) was analyzed in bovine peripheral blood mononuclear cells (bPBMC) after stimulation with the recombinant proteins. Three apoptosis-related genes, Bax, Bcl-2, and TLR4, were also included in the analysis to find out the adverse effects of the proteins to the cells. Each protein induced different patterns of cytokine expression depending on the stimulation time and antigen dose. Expression of IL-6, IL-12p40, and IFN-γ was induced with all of the proteins while IL-1β, IL-4, TNF-α, and iNOS gene expression was not. Expression of apoptosis-related genes was not altered except TLR4. These results suggest that the cellular antigens of B. abortus induce both humoral and cellular immunity via the production of IL-6, IL-12p40, and IFN-γ in bPBMC without exerting any adverse effects on the cells.
布鲁氏菌病是一种在临床和经济方面都很重要的疾病。因此,多个国家已实施了该疾病的根除计划。这些计划中的一个障碍是在早期阶段检测出受感染的动物。尽管蛋白质抗原作为诊断抗原最近受到了关注,但免疫反应开始时的确切机制尚不清楚。因此,克隆了编码五种流产布鲁氏菌细胞蛋白的基因,并对表达的重组蛋白进行了纯化。在用重组蛋白刺激牛外周血单核细胞(bPBMC)后,分析了几种细胞因子基因(IL-1β、IL-4、IL-6、IL-12p40、IFN-γ、TNF-α和iNOS)的表达。分析中还包括三个凋亡相关基因Bax、Bcl-2和TLR4,以了解这些蛋白质对细胞的不利影响。每种蛋白质根据刺激时间和抗原剂量诱导出不同的细胞因子表达模式。所有蛋白质均诱导了IL-6、IL-12p40和IFN-γ的表达,而IL-1β、IL-4、TNF-α和iNOS基因的表达未被诱导。除TLR4外,凋亡相关基因的表达未发生改变。这些结果表明,流产布鲁氏菌的细胞抗原通过在bPBMC中产生IL-6、IL-12p40和IFN-γ诱导体液免疫和细胞免疫,且对细胞没有任何不利影响。