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大肚子海马(Hippocampus abdominalis)中DC-SIGN样分子的鉴定与分子特征分析及其在宿主免疫中的潜在相关性推断

Identification and molecular profiling of DC-SIGN-like from big belly seahorse (Hippocampus abdominalis) inferring its potential relevancy in host immunity.

作者信息

Jo Eunyoung, Elvitigala Don Anushka Sandaruwan, Wan Qiang, Oh Minyoung, Oh Chulhong, Lee Jehee

机构信息

Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea; Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology (KIOST), Jeju Special Self-Governing Province, 63349, Republic of Korea.

Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea; Department of Chemistry, University of Colombo, Colombo 03, 00300, Sri Lanka.

出版信息

Dev Comp Immunol. 2017 Dec;77:270-279. doi: 10.1016/j.dci.2017.08.017. Epub 2017 Sep 1.

Abstract

Dendritic-cell-specific ICAM-3-grabbing non-integrin (DC-SIGN) is a C-type lectin that functions as a pattern recognition receptor by recognizing pathogen-associated molecular patterns (PAMPs). It is also involved in various events of the dendritic cell (DC) life cycle, such as DC migration, antigen capture and presentation, and T cell priming. In this study, a DC-SIGN-like gene from the big belly seahorse Hippocampus abdominalis (designated as ShDCS-like) was identified and molecularly characterized. The putative, complete ORF was found to be 1368 bp in length, encoding a protein of 462 amino acids with a molecular mass of 52.6 kDa and a theoretical isoelectric point of 8.26. The deduced amino acid sequence contains a single carbohydrate recognition domain (CRD), in which six conserved cysteine residues and two Ca-binding site motifs (QPN, WND) were identified. Based on pairwise sequence analysis, ShDCS-like exhibits the highest amino acid identity (94.6%) and similarity (97.4%) with DC-SIGN-like counterpart from tiger tail seahorse Hippocampus comes. Quantitative real-time PCR revealed that ShDCS-like mRNA is transcribed universally in all tissues examined, but with abundance in kidney and gill tissues. The basal mRNA expression of ShDCS-like was modulated in blood cell, kidney, gill and liver tissues in response to the stimulation of healthy fish with lipopolysaccharides (LPS), Edwardsiella tarda, or Streptococcus iniae. Moreover, recombinant ShDCS-like-CRD domain exhibited detectable agglutination activity against different bacteria. Collectively, these results suggest that ShDCS-like may potentially involve in immune function in big belly seahorses.

摘要

树突状细胞特异性细胞间黏附分子3抓取非整合素(DC-SIGN)是一种C型凝集素,作为模式识别受体通过识别病原体相关分子模式(PAMP)发挥作用。它还参与树突状细胞(DC)生命周期的各种事件,如DC迁移、抗原捕获与呈递以及T细胞激活。在本研究中,鉴定并对来自大腹海马(Hippocampus abdominalis)的一个类DC-SIGN基因(命名为ShDCS-like)进行了分子特征分析。发现推定的完整开放阅读框长度为1368 bp,编码一个462个氨基酸的蛋白质,分子量为52.6 kDa,理论等电点为8.26。推导的氨基酸序列包含一个单一的碳水化合物识别结构域(CRD),其中鉴定出六个保守的半胱氨酸残基和两个钙结合位点基序(QPN、WND)。基于成对序列分析,ShDCS-like与来自虎尾海马(Hippocampus comes)的类DC-SIGN对应物具有最高的氨基酸同一性(94.6%)和相似性(97.4%)。定量实时PCR显示,ShDCS-like mRNA在所有检测组织中均有转录,但在肾脏和鳃组织中丰度较高。在用脂多糖(LPS)、迟缓爱德华氏菌或海豚链球菌刺激健康鱼类后,血细胞、肾脏、鳃和肝脏组织中ShDCS-like的基础mRNA表达受到调节。此外,重组的ShDCS-like-CRD结构域对不同细菌表现出可检测的凝集活性。总体而言,这些结果表明ShDCS-like可能潜在参与大腹海马的免疫功能。

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