Jiang Jie, Zheng Zonglin, Wang Kaiyu, Wang Jun, He Yang, Wang Erlong, Chen Defang, Ouyang Ping, Geng Yi, Huang Xiaoli
Department of Basic Veterinary, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang District Huimin Road No. 211, Chengdu 611130, China.
Department of Aquaculture, Rongchang Campus, Southwest University, Chongqing 402460, China.
Int J Mol Sci. 2015 Nov 25;16(12):28001-13. doi: 10.3390/ijms161226082.
Channel catfish (Ictalurus punctatus) is an important agricultural fish that has been plagued by Streptococcus iniae (S. iniae) infections in recent years, some of them severe. C5a peptidase is an important virulent factor of S. iniae. In this study, the subunit vaccine containing the truncated part of C5a peptidase (pSCPI) was mixed with aluminum hydroxide gel (AH), propolis adjuvant (PA), and Freund's Incomplete Adjuvant (FIA). The immunogenicity of the pSCPI was detected by Western-blot in vitro. The relative percent survival (RPS), lysozyme activity, antibody titers, and the expression of the related immune genes were monitored in vivo to evaluate the immune effects of the three different adjuvants. The results showed that pSCPI exerted moderate immune protection (RPS = 46.43%), whereas each of the three adjuvants improved the immune protection of pSCPI. The immunoprotection of pSCPI + AH, pSCPI + PA, and pSCPI + FIA was characterized by RPS values of 67.86%, 75.00% and, 85.71%, respectively. Further, each of the three different adjuvanted pSCPIs stimulated higher levels of lysozyme activity and antibody titers than the unadjuvanted pSCPI and/or PBS buffer. In addition, pSCPI + FIA and pSCPI + PA induced expression of the related immune genes under investigation, which was substantially higher than the levels stimulated by PBS. pSCPI + AH significantly stimulated the induction of MHC II β, CD4-L2, and IFN-γ, while it induced slightly higher production of TNF-α and even led to a decrease in the levels of IL-1β, MHC I α, and CD8 α. Therefore, we conclude that compared with the other two adjuvants, FIA combined with pSCPI is a more promising candidate adjuvant against S. iniae in channel catfish.
斑点叉尾鮰（Ictalurus punctatus）是一种重要的养殖鱼类，近年来一直受到海豚链球菌（S. iniae）感染的困扰，其中一些感染较为严重。C5a肽酶是海豚链球菌的一种重要毒力因子。在本研究中，将含有截短型C5a肽酶部分的亚单位疫苗（pSCPI）与氢氧化铝凝胶（AH）、蜂胶佐剂（PA）和弗氏不完全佐剂（FIA）混合。通过Western-blot在体外检测pSCPI的免疫原性。在体内监测相对存活率（RPS）、溶菌酶活性、抗体效价以及相关免疫基因的表达，以评估这三种不同佐剂的免疫效果。结果表明，pSCPI发挥了中等程度的免疫保护作用（RPS = 46.43%），而这三种佐剂均增强了pSCPI的免疫保护作用。pSCPI + AH、pSCPI + PA和pSCPI + FIA的免疫保护作用的RPS值分别为67.86%、75.00%和85.71%。此外，三种不同佐剂增强的pSCPIs刺激产生的溶菌酶活性和抗体效价均高于未加佐剂的pSCPI和/或PBS缓冲液。另外，pSCPI + FIA和pSCPI + PA诱导了所研究的相关免疫基因的表达，其水平显著高于PBS刺激的水平。pSCPI + AH显著刺激了MHC II β、CD4-L2和IFN-γ的诱导，同时它诱导产生的TNF-α水平略高，甚至导致IL-1β、MHC I α和CD8 α水平降低。因此，我们得出结论，与其他两种佐剂相比，FIA与pSCPI联合使用是斑点叉尾鮰抗海豚链球菌更有前景的候选佐剂。
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