Department of Veterinary Medicine, College of Veterinary Medicine, National Pingtung University of Science and Technology, No. 1 Shuefu Road, Neipu, Pingtung 91201, Taiwan, ROC.
Graduate Institute of Animal Vaccine Technology, College of Veterinary Medicine, National Pingtung University of Science and Technology, No. 1 Shuefu Road, Neipu, Pingtung 91201, Taiwan, ROC.
Fish Shellfish Immunol. 2018 Jul;78:35-41. doi: 10.1016/j.fsi.2018.04.024. Epub 2018 Apr 14.
A reverse vaccinology-based survey of potent antigens associated with fish nocardiosis was conducted using the largemouth bass, Micropterus salmoides, with an aim to develop subunit vaccines. The antigens selected from the virulent strain Nocardia seriolae 961113 include the gene products of NGL2579 (GAPDH), NGL5701 (MMP), NGL4377 (OCTase), NGL4486 (ABC transporter), NGL3372 (LLE), NGL3388 (GHf10), NGL6627 (Antigen-85), NGL6696 (Esterase), and NGL6936 (CBP). These antigens were heterologously expressed in E. coli BL21 (DE3) for recombinant protein production. Then fish were vaccinated was these antigens, boosted at 2 weeks, and challenged with N. seriolae at 6 weeks after vaccination. The relative protection survival assay revealed high and significant protection efficacies of 94.45, 50.00, and 44.45 in fish that received the NGL3388 (GHf10), NGL6936 (CBP), and NGL3372 (LLE) vaccines, respectively. There were no apparent relationships or differences in tissue lesions among the administered vaccines. The serum titers against the bacterial preparations were higher for all vaccinated groups than for the control group at 4 weeks after immunization. However, no significant difference in serum titer was found at 6 weeks after immunization. The results of this study demonstrate that subunit vaccines against fish nocardiosis have differential effects, but are highly promising for nocardial prophylaxis.
本研究采用大口黑鲈(Micropterus salmoides)进行反向疫苗学调查,以寻找与鱼类诺卡氏菌病相关的有效抗原,旨在开发亚单位疫苗。从强毒株诺卡氏菌 961113 中筛选出的抗原包括 NGL2579(GAPDH)、NGL5701(MMP)、NGL4377(OCTase)、NGL4486(ABC 转运蛋白)、NGL3372(LLE)、NGL3388(GHf10)、NGL6627(抗原-85)、NGL6696(酯酶)和 NGL6936(CBP)等基因产物。这些抗原在大肠杆菌 BL21(DE3)中异源表达,用于重组蛋白生产。然后将这些抗原对鱼进行免疫接种,在第 2 周加强免疫,在接种后第 6 周用诺卡氏菌进行攻毒。相对保护存活率试验表明,接受 NGL3388(GHf10)、NGL6936(CBP)和 NGL3372(LLE)疫苗的鱼的保护存活率分别为 94.45%、50.00%和 44.45%,具有显著的高保护效果。在给予的疫苗之间,组织病变没有明显的关系或差异。在免疫后 4 周,所有免疫组的血清效价均高于对照组,但在免疫后 6 周时,血清效价没有显著差异。本研究结果表明,针对鱼类诺卡氏菌病的亚单位疫苗具有不同的效果,但对诺卡氏菌病的预防具有很高的应用前景。