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太平洋牡蛎鳃中的造血作用及其在抵御灿烂弧菌二次攻击的免疫反应中的作用。

The hematopoiesis in gill and its role in the immune response of Pacific oyster Crassostrea gigas against secondary challenge with Vibrio splendidus.

作者信息

Li Yiqun, Song Xiaorui, Wang Weilin, Wang Lingling, Yi Qilin, Jiang Shuai, Jia Zhihao, Du Xinyu, Qiu Limei, Song Linsheng

机构信息

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; University of Chinese Academy of Sciences, Beijing 100049, China.

Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian 116023, China.

出版信息

Dev Comp Immunol. 2017 Jun;71:59-69. doi: 10.1016/j.dci.2017.01.024. Epub 2017 Feb 1.

Abstract

Increasing evidences have demonstrated that the invertebrate gill is a predominant tissue participating in the immune response during pathogen challenge. In the present study, the hematopoiesis and immune activities in gill of Pacific oyster Crassostrea gigas were investigated. Stem-like cells with big nuclei and thin cytoplasm were found in the tubules of gill filaments, where DNA synthesis is active and hemocytes production are exuberant. The oysters primarily stimulated by formaldehyde-killed Vibrio splendidus exhibited stronger immune responses and enhanced cell regeneration in gill when they encountered the secondary challenge of live V. splendidus. After the secondary stimulation with V. splendidus, the expression levels of CgClec-4 and CgIFN in the gill of oysters pre-stimulated with formaldehyde-killed V. splendidus were significantly higher (p < 0.05) than that in the oysters pre-stimulated with filter-sterilized (0.22 μm pore size) sea water, while the expression level of CgIL-17 was significantly decreased (p < 0.05). Meanwhile, the protein expression level of hematopoietic transcription factor CgGATA3 and immune-related protein CgEcSOD in gill increased apparently after the secondary challenge with V. splendidus. ROS production was also enhanced (p < 0.05) at 6 h and 24 h after the secondary challenge. The phagocytic rate in gill of oysters pre-stimulated with formaldehyde-killed V. splendidus was significantly increased (p < 0.05) at 6 h after the secondary challenge with live V. splendidus, showing faster response than that pre-stimulated with filter-sterilized sea water. These results collectively showed that the immune parameters in gill were apparently enhanced after secondary challenge with live V. splendidus, indicating that hematopoiesis might participate in immune priming in Pacific oyster C. gigas.

摘要

越来越多的证据表明,无脊椎动物的鳃是病原体攻击期间参与免疫反应的主要组织。在本研究中,对太平洋牡蛎鳃中的造血和免疫活动进行了研究。在鳃丝小管中发现了具有大细胞核和薄细胞质的干细胞样细胞,其中DNA合成活跃,血细胞产生旺盛。用甲醛灭活的灿烂弧菌初次刺激后的牡蛎,在遇到活的灿烂弧菌二次攻击时,表现出更强的免疫反应和鳃中细胞再生增强。用灿烂弧菌二次刺激后,用甲醛灭活的灿烂弧菌预刺激的牡蛎鳃中CgClec-4和CgIFN的表达水平显著高于(p<0.05)用过滤除菌(孔径0.22μm)海水预刺激的牡蛎,而CgIL-17的表达水平显著降低(p<0.05)。同时,用灿烂弧菌二次攻击后,鳃中造血转录因子CgGATA3和免疫相关蛋白CgEcSOD的蛋白表达水平明显增加。二次攻击后6小时和24小时,ROS产生也增强(p<0.05)。用甲醛灭活的灿烂弧菌预刺激的牡蛎鳃在受到活的灿烂弧菌二次攻击后6小时吞噬率显著增加(p<0.05),比用过滤除菌海水预刺激的牡蛎反应更快。这些结果共同表明,用活的灿烂弧菌二次攻击后,鳃中的免疫参数明显增强,表明造血可能参与了太平洋牡蛎的免疫致敏。

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