Eraghi Vida, Derakhshandeh Abdollah, Hosseini Arsalan, Haghkhah Masoud, Sechi Leonardo A, Motamedi Boroojeni Azar
1Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, 71345-1731 Iran.
2Sezione di Microbiologia e Virologia, Dipartimento di Scienze Biomediche, Università degli Studi di Sassari, 07100 Sassari, Italy.
Gut Pathog. 2019 Jul 9;11:36. doi: 10.1186/s13099-019-0317-6. eCollection 2019.
subsp. (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.
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.
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.
副结核分枝杆菌亚种（MAP）是全球所有反刍动物约翰氏病的病原体。奶牛和绵羊产业的经济问题、公共卫生问题、MAP在环境中的持久性以及缺乏有效的疫苗，都表明有必要研究各种抗原以作为候选疫苗。基于MAP的发病机制，研究生产新的重组蛋白以刺激细胞介导的免疫似乎是有帮助的。本研究描述了一种嵌合融合蛋白的成功表达和纯化，该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域（FAP-P）组成。通过ELISA测定了免疫山羊分离出的外周血单核细胞（PBMC）对重组HBHA-FAP和MAP的所有粗蛋白（PPD）触发的抗原特异性IFN-γ反应。
观察到由重组HBHA-FAP构建的嵌合蛋白和接种PPD的山羊刺激的外周血单核细胞（PBMC）的IFN-γ产生水平显著增加。阳性组（接种PPD）针对重组HBHA-FAP和试验组（接种重组HBHA-FAP）针对PPD的抗原特异性γ干扰素（IFN-γ）分泌，在重组HBHA-FAP和PPD刺激之间也有统计学上不显著的升高，表明我们的嵌合蛋白刺激针对MAP的细胞介导免疫的潜力和特异性。
总体而言这些结果表明重组HBHA-FAP在PBMC培养中引发强烈的IFN-γ产生。因此，应进一步研究本产品作为自然感染动物候选疫苗进行分析其潜力。
