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两种类纤维胶凝蛋白在日本沼虾先天免疫防御中的功能

Function of two ficolin-like proteins in innate immune defense of the oriental river prawn, Macrobrachium nipponense.

作者信息

Li Tingting, Wu Lei, Jin Min, Ma Futong, Huang Xin, Ren Qian

机构信息

Jiangsu Key Laboratory for Biodiversity & Biotechnology, Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, PR China.

State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, SOA, Xiamen 361005, PR China.

出版信息

Fish Shellfish Immunol. 2017 Sep;68:488-499. doi: 10.1016/j.fsi.2017.07.059. Epub 2017 Jul 29.

Abstract

Ficolins have crucial functions in recognizing and eliminating pathogens in innate immunity. In this study, we identified two ficolin-like genes from the oriental river prawn, Macrobrachium nipponense. These genes were designated as MnFico1 and MnFico2. MnFico1 cDNA has 1600 bp, whereas MnFico2 has 1486 bp. In addition to a coiled-coil region or a low complexity region, the two ficolins both contained a signal peptide and a fibrinogen-related domain. qRT-PCR results showed that the highest expression level of MnFico1 expression was in the gills, whereas that of MnFico2 was in the heart. The transcripts of MnFico1 and MnFico2 could both respond to bacteria challenge. The transcription of antilipopolysaccharide factors ALFs (MnALF1 and MnALF2) and crustin genes (MnCru4, MnCru5, MnCru6, and MnCru7) was inhibited in the gills of MnFico1 or MnFico2 knockdown prawns at 24 h Vibrio parahaemolyticus challenge. Recombinant proteins of rMnFico1 and rMnFico2 could bind toward diverse bacteria and agglutinate Gram-negative and Gram-positive bacteria with the presence of calcium (Ca). rMnFico1 and rMnFico2 proteins also have lipopolysaccharide and peptidoglycan binding activity. Both recombinant ficolin proteins could help the prawn to facilitate the clearance of V. parahaemolyticus in vivo. Our results suggested that MnFico1 and MnFico2 might serve as pattern recognition receptors in M. nipponense.

摘要

纤维胶凝蛋白在天然免疫中识别和清除病原体方面具有关键作用。在本研究中,我们从日本沼虾(Macrobrachium nipponense)中鉴定出两个类纤维胶凝蛋白基因。这些基因被命名为MnFico1和MnFico2。MnFico1 cDNA有1600 bp,而MnFico2有1486 bp。除了卷曲螺旋区域或低复杂性区域外,这两种纤维胶凝蛋白均含有信号肽和纤维蛋白原相关结构域。qRT-PCR结果显示,MnFico1表达的最高水平在鳃中,而MnFico2的最高水平在心脏中。MnFico1和MnFico2的转录本均可对细菌攻击作出反应。在24小时副溶血性弧菌攻击下,MnFico1或MnFico2敲低的沼虾鳃中,抗脂多糖因子ALFs(MnALF1和MnALF2)和crustin基因(MnCru4、MnCru5、MnCru6和MnCru7)的转录受到抑制。rMnFico1和rMnFico2的重组蛋白可与多种细菌结合,并在有钙(Ca)存在的情况下凝集革兰氏阴性菌和革兰氏阳性菌。rMnFico1和rMnFico2蛋白还具有脂多糖和肽聚糖结合活性。两种重组纤维胶凝蛋白均可帮助沼虾在体内促进副溶血性弧菌的清除。我们的结果表明,MnFico1和MnFico2可能作为日本沼虾中的模式识别受体。

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