Background Mycobacterium avium subsp. paratuberculosis (MAP) is a causative agent of Johne's disease in all ruminants worldwide. Economic problems in dairy cattle and sheep industries, public health concern, persistence of MAP in the environment and lack of effective vaccines mentioned necessity of research about various antigens to introduce as vaccine candidates. Based on MAP pathogenesis, it seems that research about the production of new recombinant proteins to stimulate cell-mediated immunity is helpful. This study describes successful expression and purification of a chimeric fusion protein which consists of Heparin-Binding Hemagglutinin Adhesin (HBHA) and high antigenic region of Fibronectin Attachment Protein (FAP-P). Triggered antigen-specific IFN-γ response of isolated PBMCs from immunized goats to rHBHA-FAP and all crude proteins of MAP (PPD), was measured by ELISA. Results Significant increases were observed in the IFN-γ production level of peripheral blood mononuclear cells (PBMCs) stimulated by constructed chimeric protein from rHBHA-FAP and PPD vaccinated goats. Antigen-specific gamma interferon (IFN-γ) secretion in positive group (immunized by PPD) against rHBHA-FAP and test group (immunized by rHBHA-FAP) against PPD, also statistically insignificant rises between stimulation with rHBHA-FAP and PPD, suggested the potential and specificity of our chimeric protein to stimulate cell mediated immunity against MAP. Conclusions Collectively, these results demonstrate that rHBHA-FAP elicits a strong IFN-γ production in PBMC culture. Therefore, further studies of the present product as a candidate vaccine in naturally infected animals should be conducted, to analyze its potential.

中文翻译：

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鸟分枝杆菌亚种的肝素结合血凝素粘附素和纤连蛋白附着蛋白 (rHBHA-FAP) 的重组融合蛋白。副结核病在外周血单核细胞培养中引起强烈的 γ 干扰素反应。

背景鸟分枝杆菌亚种。副结核病 (MAP) 是全球所有反刍动物中约内氏病的病原体。奶牛和绵羊行业的经济问题、公共卫生问题、环境中 MAP 的持续存在以及缺乏有效疫苗都提到了研究各种抗原作为候选疫苗的必要性。基于 MAP 发病机制，关于产生新的重组蛋白以刺激细胞介导免疫的研究似乎是有帮助的。本研究描述了嵌合融合蛋白的成功表达和纯化，该融合蛋白由肝素结合血凝素粘附素 (HBHA) 和纤连蛋白附着蛋白的高抗原区 (FAP-P) 组成。通过ELISA测量来自免疫山羊的分离的PBMC对rHBHA-FAP和MAP的所有粗蛋白（PPD）的触发的抗原特异性IFN-γ反应。结果 rHBHA-FAP 和 PPD 免疫山羊构建的嵌合蛋白刺激外周血单个核细胞 (PBMC) 的 IFN-γ 产生水平显着增加。阳性组（通过 PPD 免疫）针对 rHBHA-FAP 和测试组（通过 rHBHA-FAP 免疫）针对 PPD 的抗原特异性 γ 干扰素 (IFN-γ) 分泌，在 rHBHA-FAP 和 PPD 刺激之间也有统计学意义的上升，提示我们的嵌合蛋白刺激细胞介导的针对 MAP 的免疫的潜力和特异性。结论 总的来说，这些结果表明 rHBHA-FAP 在 PBMC 培养物中引发了强烈的 IFN-γ 产生。所以，