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Identification, functional characterization and the potential role of variable lymphocyte receptor EsVLRA from Eriocheir sinensis in response to secondary challenge after Vibrio parahaemolyticus vaccine.
Fish & Shellfish Immunology ( IF 4.7 ) Pub Date : 2020-01-07 , DOI: 10.1016/j.fsi.2020.01.011
Hourong Liu 1 , Chengwen Song 2 , Junhao Ning 1 , Yuan Liu 3 , Zhaoxia Cui 4
Affiliation  

Variable lymphocyte receptors (VLRs) play an important role via their antigen-special reorganization in jawless vertebrates (agnathans) adaptive immune response. In the present study, the open reading frame (ORF) of Eriocheir sinensis VLRA (designated as EsVLRA) was identified. EsVLRA comprised a 799-amino-acid polypeptide with one LRR_NT domain, thirteen LRR domains and one LRR_CT domain, which showed a high domain consistency of the VLR genes in lamprey (Petromyzon marinus). The transcript of EsVLRA was detected in all examined tissues with the highest level detected in hepatopancreas. Notably, the expression of EsVLRA in hepatopancreas, gonads, gill and intestine of male crabs was significantly higher than that in females. The recombinant EsVLRA exhibited strong bacteria-binding activity rather than antibacterial activity, suggesting its crucial role in immune recognition. Furthermore, 6 h earlier response and a significantly higher peak of EsVLRA mRNA expression was observed after challenge with live Vibrio parahaemolyticus (240.6-fold, P < 0.01, crabs receive secondary challenge after V. parahaemolyticus vaccine to the carbs only receive twice PBS injection, N = 6), compared with those only received first injection with formalin-inactivated V. parahaemolyticus (39.7-fold, P < 0.01, challenge 6 h to vaccination 12 h). The findings of this study together demonstrated that EsVLRA plays an important role in the immune system of E. sinensis, serving as a pattern recognition receptor and involving in the immune priming.

中文翻译:

副溶血弧菌疫苗后继发攻击的中华绒螯蟹可变淋巴细胞受体EsVLRA的鉴定,功能表征和潜在作用。

可变淋巴细胞受体(VLR)通过在无颚脊椎动物(agnathans)适应性免疫反应中的抗原特异性重组而发挥重要作用。在本研究中,确定了中华绒螯蟹VLRA(称为EsVLRA)的开放阅读框(ORF)。EsVLRA包含具有一个LRR_NT结构域,十三个LRR结构域和一个LRR_CT结构域的799个氨基酸多肽,其在七lamp鳗(Petromyzon marinus)中显示了VLR基因的高结构域一致性。在所有检查过的组织中均检测到EsVLRA的转录本,在肝胰腺中检测到的水平最高。值得注意的是,EsVLRA在雄性蟹的肝胰腺,性腺,g和肠中的表达明显高于雌性。重组EsVLRA表现出很强的细菌结合活性,而不是抗菌活性,提示其在免疫识别中的关键作用。此外,用活副溶血弧菌攻击后,观察到反应较早6 h,EsVLRA mRNA表达明显升高(240.6倍,P <0.01,螃蟹接受副溶血弧菌疫苗的二次攻击后,只接受两次PBS注射, N = 6),与仅接受福尔马林灭活副溶血性弧菌首次注射者相比(39.7倍,P <0.01,在接种后12小时攻击6小时)。这项研究的结果共同证明,EsVLRA在中华绒螯蟹的免疫系统中起着重要的作用,它是一种模式识别受体,并参与了免疫引发。副溶血弧菌活菌攻击后6 h较早响应,观察到EsVLRA mRNA表达明显更高的峰(240.6倍,P <0.01,螃蟹接受副溶血弧菌对碳水化合物的疫苗继发攻击,仅注射两次PBS,N = 6),与仅接受福尔马林灭活副溶血性弧菌首次注射者相比(39.7倍,P <0.01,接种6小时后接种疫苗12小时)。这项研究的结果共同证明,EsVLRA在中华绒螯蟹的免疫系统中起着重要的作用,它是一种模式识别受体,并参与了免疫引发。副溶血弧菌活菌攻击后6 h较早响应,观察到EsVLRA mRNA表达明显更高的峰(240.6倍,P <0.01,螃蟹接受副溶血弧菌对碳水化合物的疫苗继发攻击,仅注射两次PBS,N = 6),与仅接受福尔马林灭活副溶血性弧菌首次注射者相比(39.7倍,P <0.01,接种6小时后接种疫苗12小时)。这项研究的结果共同证明,EsVLRA在中华绒螯蟹的免疫系统中起着重要的作用,它是一种模式识别受体,并参与了免疫引发。与仅接受首次福尔马林灭活副溶血性弧菌注射的患者相比(39.7倍,P <0.01,对疫苗接种12 h攻击6 h)。这项研究的结果共同证明,EsVLRA在中华绒螯蟹的免疫系统中起着重要的作用,它是一种模式识别受体,并参与了免疫引发。与仅接受首次福尔马林灭活副溶血性弧菌注射的患者相比(39.7倍,P <0.01,对疫苗接种12 h攻击6 h)。这项研究的结果共同证明,EsVLRA在中华绒螯蟹的免疫系统中起着重要的作用,它是一种模式识别受体,并参与了免疫引发。
更新日期:2020-01-07
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