Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, 5 Yushan Road, Qingdao 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, No.1 Wenhai Road, Qingdao 266071, China.
Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
Microb Pathog. 2017 Dec;113:152-159. doi: 10.1016/j.micpath.2017.10.043. Epub 2017 Oct 23.
Vibrio ichthyoenteri was an important causative agent of bacterial enteritis in flounder (Paralichthys olivaceus). Outer membrane protein A (OmpA) of Gram-negative pathogen was a major cell surface antigen. In the present study, OmpA of V. ichthyoenteri was recombinantly expressed in Escherichia coli, and the immunogenicity of OmpA was identified by western blotting using flounder anti-rOmpA and anti-V. ichthyoenteri antibodies. The vaccine potential of rOmpA was tested in a flounder model, and a high relative percentage of survival rate was obtained with 73.1% after challenge with V. ichthyoenteri. Meanwhile, the immune response of flounder induced by rOmpA was also investigated, and the results showed that the sIg + lymphocytes in blood, spleen, and pronephros significantly proliferated, and the peak levels occurred at week 4 after immunization. Moreover, rOmpA could induce higher levels of specific serum antibodies than the control group after immunization, and the peak level occurred at week 5 after immunization. Meanwhile, qRT-PCR analysis showed that the expressions of CD4-1, CD8α, IL-1β, IFN-γ, MHCIα and MHCIIα genes were significantly up-regulated after immunization with rOmpA. Taking together, these results demonstrated that rOmpA could evoke highly protective effects against V. ichthyoenteri challenge and induce strong immune response of flounder, which indicated that OmpA was a promising vaccine candidate.
创伤弧菌是牙鲆细菌性肠炎的重要致病菌。革兰氏阴性病原体的外膜蛋白 A (OmpA)是主要的细胞表面抗原。本研究在大肠杆菌中重组表达了创伤弧菌的 OmpA,并使用牙鲆抗 rOmpA 和抗创伤弧菌抗体的 Western blot 鉴定了 OmpA 的免疫原性。在牙鲆模型中测试了 rOmpA 的疫苗潜力,在受到创伤弧菌的挑战后,获得了 73.1%的高相对存活率。同时,还研究了 rOmpA 诱导牙鲆的免疫反应,结果表明,血液、脾脏和前肾中的 sIg+淋巴细胞显著增殖,在免疫后第 4 周达到峰值。此外,rOmpA 可诱导比对照组更高水平的特异性血清抗体,在免疫后第 5 周达到峰值。同时,qRT-PCR 分析表明,rOmpA 免疫后 CD4-1、CD8α、IL-1β、IFN-γ、MHCIα 和 MHCIIα 基因的表达显著上调。总之,这些结果表明 rOmpA 可以引发对创伤弧菌的高度保护作用,并诱导牙鲆强烈的免疫反应,表明 OmpA 是一种很有前途的疫苗候选物。