[1]孟庆国,陈 静,黄艳青,等.中华绒螯蟹螺原体类螺旋蛋白SLP25对中华绒螯蟹和RAW264.7细胞免疫反应的研究[J].南京师大学报(自然科学版),2013,36(02):78-83.
　Meng Qingguo,Chen Jing,Huang Yanqing,et al.Immune Responses of Eriocheir Sinensis and RAW264.7 Cells Induced by Spiralin-like Protein SLP25 from Spiroplasma Eriocheiris[J].Journal of Nanjing Normal University(Natural Science Edition),2013,36(02):78-83.
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中华绒螯蟹螺原体类螺旋蛋白SLP25对中华绒螯蟹和RAW264.7细胞免疫反应的研究()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第36卷
期数:
2013年02期
页码:
78-83
栏目:
生命科学
出版日期:
2013-06-30

文章信息/Info

Title:
Immune Responses of Eriocheir Sinensis and RAW264.7 Cells Induced by Spiralin-like Protein SLP25 from Spiroplasma Eriocheiris
文章编号:
1001-4616(2013)02-0078-06
作者:
孟庆国1; 陈 静1; 黄艳青2; 靳明建3; 顾 伟1; 王 文1
1.南京师范大学生命科学学院,江苏省生物多样性与生物技术重点实验室,江苏省水生甲壳动物病害重点实验室,江苏 南京 210023
2.中国水产科学研究院东海水产研究所,农业部海洋与河口渔业资源及生态重点开放实验室,上海 200090
3.江苏省如东县渔业技术推广站,江苏 如东 226400
Author(s):
Meng Qingguo1; Chen Jing1; Huang Yanqing2; Jin Mingjian3; Gu Wei1; Wang Wen1
1.Jiangsu Key Laboratory for Biodiversity and Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, School of Life Sciences,Nanjing Normal University,Nanjing 210023,China
2.Key and Open Laboratory of Marine and Estuary Fisheries,Ministry of Agriculture of China,East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences,Shanghai 200090,China
3.Rudong Station for Fisheries Technology Extension,Rudong 226400,China
关键词:
螺原体; 中华绒螯蟹; 螺旋蛋白; 免疫反应
Keywords:
Spiroplasma; Chinese mitten crab; Spiralin; immune responses
分类号:
Q936
文献标志码:
A
摘要:
中华绒螯蟹螺原体是一种引起中华绒螯蟹颤抖病的新型病原微生物,对中华绒螯蟹养殖业造成了很大的损失.本文研究了螺原体的一个特有蛋白——螺旋蛋白所引起的中华绒螯蟹和小鼠RAW264.7细胞的免疫反应.克隆和原核表达中华绒螯蟹螺原体类螺旋蛋白SLP25后,用以感染中华绒螯蟹和小鼠巨噬细胞RAW264.7,用实时定量PCR和Western Blot的方法检测宿主免疫相关基因或蛋白表达量的变化.类螺旋蛋白SLP25刺激后,中华绒螯蟹的抗脂多糖因子、酚氧化酶原、消极素轻链和Peroxiredoxin 6等基因的mRNA表达量变化显著; RAW264.7细胞内的TNF-α、IL-1β和IL-10等基因的mRNA表达量变化显著,而TGF-β1和CD40的mRNA表达量变化不显著; 同时RAW264.7细胞内P38和ERK蛋白的磷酸化水平变化显著,而JNK蛋白的磷酸化水平变化不显著.通过本文的研究表明类螺旋蛋白SLP25可以引起宿主广泛的免疫变化,进而证明其是重要的毒力相关因子.
Abstract:
Spiroplasma eriocheiris caused mass mortality of Chinese mitten crab Eriocheir sinensis.This paper studied the immune responses of E.sinensis and RAW264.7 cells induced by a unique gene,spiralin-like protein of S.eriocheiris.After cloning and expression the spiralin-like protein in E.coli,the role of this protein in promoting immune responses was characterized through analyzing the interactions between S.eriocheiris and E.sinensis as well as RAW264.7 cells.Real-time PCR and western blot were used for detection of expression levels of immune genes in both hosts.The mRNA expressions of anti-lipopolysaccharide factor,prophenoloxidase,pacifastin light chain and peroxiredoxin 6 in E.sinensis and TNF-α,IL-1β,IL-10 in RAW264.7 cells were induced by spiralin-like protein,whereas TGF-β1 and CD40 in RAW264.7 cells were not.In RAW264.7 cells,the levels of phosphorylated P38 and phosphorylated ERK increased after spiralin-like protein induced,whereas the level of phosphorylated JNK did not change.This study reveals that spiralin-like protein of S.eriocheiris can induce the immune responses of E.sinensis and RAW264.7 cells and the protein may be an important virulence factor of S.eriocheiris.This study gives light to the pathogenesis of the spiroplasmas and provides knowledge of immune reactions of hosts from both invertebrate and vertebrate challenged with spiroplasmas unique protein.

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备注/Memo

备注/Memo:
收稿日期:2012-09-20.
基金项目:国家自然科学基金(31170120)、江苏省自然科学基金(2011104SBZ0122)、江苏省普通高校自然科学基金(2011104TSJ0154)、江苏省水产三项工程项目(PJ2011-65).
通讯联系人:王文,博士,教授,研究方向:水生甲壳动物病害.E-mail:njnuwang@263.net
更新日期/Last Update: 2013-06-30