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石斑鱼(Lutjanus sanguineus)IL-6 基因:鉴定、表达分析及其对哈维弧菌 OmpW DNA 疫苗的佐剂效应。

An IL-6 gene in humphead snapper (Lutjanus sanguineus): Identification, expression analysis and its adjuvant effects on Vibrio harveyi OmpW DNA vaccine.

机构信息

College of Fishery, Guangdong Ocean University, Zhanjiang, PR China; Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, PR China; Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang, PR China; Shenzhen Fisheries Service and Aquatic Product Technology Extension Center, Shenzhen, PR China.

College of Fishery, Guangdong Ocean University, Zhanjiang, PR China; Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, PR China; Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang, PR China; Shenzhen Research Institute of Guangdong Ocean University, Shenzhen, PR China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, PR China; Guangxi Key Lab for Marine Biotechnology, Guangxi Institute of Oceanography, Guangxi Academy of Sciences, Beihai, PR China.

出版信息

Fish Shellfish Immunol. 2019 Dec;95:546-555. doi: 10.1016/j.fsi.2019.11.013. Epub 2019 Nov 5.

Abstract

Interleukin 6 (IL-6) is a pleiotropic cytokine that plays important role in mediating the innate and adaptive immune responses against pathogen infection. In this study, an IL-6 homolog (Ls-IL6) was identified and characterized from humphead snapper, Lutjanus sanguineus. The full-length cDNA of Ls-IL6 was 1066 bp, containing an open reading frame (ORF) of 639 bp encoding 212 amino acids, 5' untranslated region(UTR) of 63 bp and 3' UTR of 605 bp. The predicted Ls-IL6 protein had typical motif of IL-6 family and shared high identities to teleost IL-6s. Ls-IL6 extensively expressed in various tissues, and the highest expression of Ls-IL6 was detected in head kidney, spleen and thymus. In vivo, the transcript levels of Ls-IL6 were significantly up-regulated in response to Vibrio harveyi infection. Moreover, the DNA plasmid containing the OmpW of V. harveyi together with the gene encoding Ls-IL6 were successfully constructed and administered to fish, the protective efficacy of Ls-IL6 was investigated. Compared with the pcDNA-OmpW group, the level of specific antibodies against V. harveyi increased in pcDNA-IL6-OmpW injected group. After V. harveyi infection, the pcDNA-IL6-OmpW vaccinated fish showed higher relative percent survival (76%) than the relative survival of fish immunized with pcDNA-OmpW (60%). These results indicated that Ls-IL6 was involved in immune response against V. harveyi infection and could be applied as a promising adjuvant for DNA vaccines against V. harveyi.

摘要

白细胞介素 6(IL-6)是一种多效细胞因子,在介导针对病原体感染的先天和适应性免疫反应中发挥重要作用。本研究从紫红笛鲷(Lutjanus sanguineus)中鉴定和表征了一种白细胞介素 6 同源物(Ls-IL6)。Ls-IL6 的全长 cDNA 为 1066bp,包含 639bp 的开放阅读框(ORF),编码 212 个氨基酸、5'非翻译区(UTR)63bp 和 3'UTR605bp。预测的 Ls-IL6 蛋白具有典型的 IL-6 家族特征,与鱼类的 IL-6 具有高度同源性。Ls-IL6 在各种组织中广泛表达,在头肾、脾脏和胸腺中表达水平最高。在体内,Ls-IL6 的转录水平在受到哈维弧菌感染后显著上调。此外,成功构建了含有哈维弧菌 OmpW 基因和 Ls-IL6 基因的 DNA 质粒,并将其施用于鱼类,研究了 Ls-IL6 的保护效果。与 pcDNA-OmpW 组相比,pcDNA-IL6-OmpW 注射组针对哈维弧菌的特异性抗体水平升高。感染哈维弧菌后,pcDNA-IL6-OmpW 疫苗接种组的相对存活率(76%)高于 pcDNA-OmpW 免疫组(60%)。这些结果表明,Ls-IL6 参与了对哈维弧菌感染的免疫反应,可作为哈维弧菌 DNA 疫苗的一种有前途的佐剂。

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