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大黄鱼(Larimichthys crocea)白细胞介素-11 的分子和免疫特性。

Molecular and immunological characterizations of interleukin-11 in large yellow croaker (Larimichthys crocea).

机构信息

Fisheries College, Jimei University, Xiamen, 361021, PR China; Pilot National Laboratory for Marine Science and Technology, Qingdao, PR China.

Fisheries College, Jimei University, Xiamen, 361021, PR China; Pilot National Laboratory for Marine Science and Technology, Qingdao, PR China; State Key Laboratory of Large Yellow Croaker Breeding, Ningde Fufa Fisheries Company Limited, Fujian, Ningde, 352103, PR China.

出版信息

Fish Shellfish Immunol. 2020 May;100:9-17. doi: 10.1016/j.fsi.2020.02.065. Epub 2020 Mar 1.

Abstract

Interleukin (IL)-11 is a multifunctional cytokine that exerts a series of important immunomodulatory effects and exists in many tissues and cells. A 1106-bp nucleotide sequence representing the complete cDNA of IL-11 was obtained from large yellow croaker (Larimichthys crocea), containing an open reading frame (ORF) of 603 bp encoding for 200 amino acids (aa). The predicted LcIL-11 protein included a 12aa signal peptide and a conserved IL-11 domain. The polypeptide sequence identities between LcIL-11 and its counterparts in mammals and other fish are from 84% to 92% with known fish IL-11a and 22%-27% with fish IL-11b. LcIL-11 mRNA existed in most tissues with the most predominant expression in the gill. After immune challenge, the expression levels of LcIL-11 were induced largely in vivo and in vitro, with the peak-value of 32 times as much as the control in the liver at 24 h after Vibrio parahaemolyticus injection (p < 0.05) and the greatest value of 13.9 times as much as the control in LCK cells at 12 h after poly I:C stimulation (p < 0.05). Furthermore, the overexpression vector pcDNA3.1-LcIL-11 was constructed and transfected to LCK cells. Our results showed that the transcriptional expression levels of tumor necrosis factor (TNF)-α and myxovirus resistant protein (Mx) significantly up-regulated in LCK cells after LcIL-11 overexpression (p < 0.05). However, no significant changes of IL-1β, janus kinase (JAK)2 and signal transducers and activators of transcription (STAT)5 was detected. Our finding indicated that LcIL-11 might enhance TNF-α and antiviral protein Mx expression in large yellow croaker.

摘要

白细胞介素 (IL)-11 是一种多功能细胞因子,具有一系列重要的免疫调节作用,存在于许多组织和细胞中。从大黄鱼 (Larimichthys crocea) 中获得了代表 IL-11 全长 cDNA 的 1106bp 核苷酸序列,其中包含一个编码 200 个氨基酸 (aa) 的 603bp 开放阅读框 (ORF)。预测的 LcIL-11 蛋白包含一个 12aa 的信号肽和一个保守的 IL-11 结构域。LcIL-11 与哺乳动物及其它鱼类的同源物的多肽序列同一性为 84%-92%,与已知的鱼类 IL-11a 为 22%-27%。LcIL-11 mRNA 存在于大多数组织中,在鳃中表达最丰富。在免疫挑战后,LcIL-11 的表达水平在体内和体外均被诱导,在注射副溶血弧菌 24 小时后肝脏中的表达水平达到对照的 32 倍(p<0.05),在 poly I:C 刺激 12 小时后 LCK 细胞中的表达水平达到对照的 13.9 倍(p<0.05)。此外,构建了 pcDNA3.1-LcIL-11 过表达载体并转染至 LCK 细胞。结果表明,LcIL-11 过表达后,LCK 细胞中肿瘤坏死因子 (TNF)-α 和抗粘病毒蛋白 (Mx) 的转录表达水平显著上调(p<0.05)。然而,IL-1β、Janus 激酶 (JAK)2 和信号转导和转录激活因子 (STAT)5 的表达没有明显变化。研究结果表明,LcIL-11 可能增强大黄鱼 TNF-α 和抗病毒蛋白 Mx 的表达。

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