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为研究白介素15(Interleukin-15, IL-15)在松江鲈(Trachidermus fasciatus Heckel)先天免疫中的功能,本研究利用RACE技术克隆得到松江鲈IL-15基因(命名为TfIL-15)的全长cDNA序列,其长度为1140 bp,包括5¢-非编码区(5¢-UTR) 165 bp、开放阅读框(ORF) 522 bp和3¢UTR 453 bp。在5¢UTR区域,存在4个读码框外的AUG翻译起始位点。基因ORF编码173个氨基酸(aa),其中,前59 aa为信号肽序列。成熟肽全长为114 aa,预测分子量为12.975 kDa,理论等电点为5.15。同源比对发现,鱼类IL-15变异程度较高,TfIL-15与其他鱼类IL-15同源性在23%~61%之间。多序列比对和三维结构构建结果显示,TfIL-15具有典型4个α螺旋二级结构,形成二硫键的4个半胱氨酸高度保守。qRT-PCR分析表明,TfIL-15广泛表达于松江鲈各组织中。腹腔注射脂多糖(Lipopolysaccharides, LPS)后,TfIL-15 mRNA在血液、皮肤、肝脏和脾脏中均上调表达。在皮肤和血液中,刺激后2 h表达量迅速上调至最高峰,分别为对照组的74倍和41倍。脾脏和肝脏在刺激后12 h分别达到对照组的3倍和18倍。肝脏中,刺激后96 h,表达量再次上调至对照组的86倍。上述结果表明,TfIL-15可能参与了松江鲈抵抗外界刺激的先天免疫过程。另外,通过构建TfIL-15成熟肽的原核表达载体,成功获得重组蛋白,为进一步研究TfIL-15蛋白的功能奠定了基础。 相似文献
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根据鲤科鱼类同源序列设计并合成了长鳍吻(鳍)(Rhinogobio ventralis)应激因子HSP70、抗体IgM、抑炎性因子IL-10和促炎性因子IL-1b基因特异引物,以长鳍吻(鳍)皮肤组织总RNA为模板,RT-PCR 扩增获得长度分别为412、463、520和217 bp的上述4种免疫因子基因cDNA部分序列.同时通过RT-PCR比较患小瓜虫病长鳍吻1715和对照组长鳍吻(鳍)皮肤和肠道组织中各免疫因子的表达差异,结果显示:在皮肤组织中,IgM和IL-10在感染组鱼体中表达量显著高于对照鱼(P<0.05), HSP 70和 IL-1b的表达则没有差异;在肠道组织中,IgM、IL-10和 IL-1b在感染组鱼体中表达量显著增高,HSP 70的表达量没有差异.本研究首次对长鳍吻(鳍)的免疫因子进行了分析,指出免疫因子IgM和IL-10在鱼体中免疫应答反应中较为灵敏,为养殖过程中长鳍吻(鳍)应激监测方面奠定了理论基础. 相似文献
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白细胞介素-12（IL-12）是一种连接先天性和适应性免疫系统的重要细胞因子,在免疫反应和炎症反应中均发挥着重要作用。本实验以团头鲂为研究对象,克隆了团头鲂IL-12的4个亚基IL-12A、IL-12Ba、IL-12Bb和IL-12Bc的开放阅读框（ORF）序列,长度分别为588、993、939和837 bp。实时荧光定量PCR（RT-qPCR）结果表明,团头鲂IL-12各亚基在检测的10个不同的成鱼健康组织中都有表达,但是在不同组织中的表达模式不同;腹腔注射嗜水气单胞菌后,团头鲂IL-12各亚基在免疫相关组织中显著差异表达;脂多糖（LPS）刺激分离的团头鲂头肾淋巴细胞后,4个IL-12亚基均被快速诱导表达。进一步通过大肠杆菌原核表达系统和变性复性方法得到团头鲂IL-12Bb、IL-12Bc重组蛋白并刺激分离的头肾淋巴细胞,显著上调TNF-α、IL-1β、IFN-γ的表达,表明获得的重组蛋白具有生物学活性。最后对获得的蛋白进行抑菌活性检测,结果表明重组IL-12Bb、IL-12Bc单体蛋白对嗜水气单胞菌、大肠杆菌和金黄色葡萄球菌都有明显的抑制作用。综上,本研究结果表明,团头鲂IL-12在调节免疫应答的过程中发挥着重要作用。 相似文献
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根据鲤科鱼类同源序列设计并合成了长鳍吻(Rhinogobio ventralis)应激因子HSP70、抗体IgM、抑炎性因子IL-10和促炎性因子IL-1b基因特异引物,以长鳍吻皮肤组织总RNA为模板,RT-PCR扩增获得长度分别为412、463、520和217 bp的上述4种免疫因子基因cDNA部分序列。同时通过RT-PCR比较患小瓜虫病长鳍吻和对照组长鳍吻皮肤和肠道组织中各免疫因子的表达差异,结果显示:在皮肤组织中,IgM和IL-10在感染组鱼体中表达量显著高于对照鱼(P<0.05),HSP 70和IL-1b的表达则没有差异;在肠道组织中,IgM、IL-10和IL-1b在感染组鱼体中表达量显著增高,HSP 70的表达量没有差异。本研究首次对长鳍吻的免疫因子进行了分析,指出免疫因子IgM和IL-10在鱼体中免疫应答反应中较为灵敏,为养殖过程中长鳍吻应激监测方面奠定了理论基础。 相似文献
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为了探究鱼类体内过量游离血红蛋白对鱼体的影响，本实验以草鱼为研究对象，通过体内注射血红蛋白模拟体内出血，探究血红蛋白对组织和组织中巨噬细胞的免疫调控作用。体内注射血红蛋白的实验结果显示，血红蛋白的刺激导致头肾和中肾组织中出现明显的血红蛋白沉淀，同时提高了鱼体血清中的抗氧化相关酶活性，促进了头肾组织中炎症因子基因TNF-α、IL-1β和IL-10的mRNA表达水平。普鲁士蓝和间接免疫荧光实验结果显示，头肾巨噬细胞对血红蛋白表现出了吞噬活性。荧光定量PCR检测结果显示，血红蛋白的刺激显著激活了头肾巨噬细胞中CD68、CD86、CSF和MHC-Ⅱ的mRNA表达水平。进一步的细胞因子检测结果显示，血红蛋白刺激12 h后，头肾巨噬细胞中的炎症因子基因(TNF-α、IL-1β和IL-4)、趋化因子基因(CCL20和CCL4)、IFN-α和TLR4的mRNA表达水平被显著上调表达。研究表明，草鱼头肾巨噬细胞对血红蛋白具有吞噬能力，同时血红蛋白也激活了巨噬细胞中多种细胞因子的表达。本研究结果首次阐述了草鱼血红蛋白与巨噬细胞的相互作用关系，丰富了鱼类血液基础免疫学理论，同时也为鱼类健康养殖提供了新的参考... 相似文献
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嗜水气单胞菌感染引起的病害使我国鱼类养殖遭受了严重经济损失，制约了水产养殖业的健康发展。嗜水气单胞菌可产生多种毒力因子，致病过程及影响因素复杂，感染鱼类种类广。鱼类的肠道、脾脏、肝脏和肾脏具有重要的免疫功能，在抵御病原菌入侵中发挥重要作用，其中肠道菌群直接影响着宿主的免疫和炎症反应。本文着重阐述了嗜水气单胞菌感染后鱼类免疫相关器官的组织病理学变化、转录组水平基因的差异表达和通路富集及肠道微生物组成与结构改变，综述了嗜水气单胞菌感染后鱼类病理学和组学水平变化，为研究鱼类免疫应答机制及肠道菌群免疫调节功能提供参考依据。 相似文献
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甘露寡糖对草鱼感染嗜水气单胞菌后细胞因子表达的调节 总被引：1，自引：0，他引：1
在饲料中添加0.2%甘露寡糖(Mannan oligosaccharide,(MOS)),饲养草鱼(Ctenopharyngodon idellus)28 d后,草鱼经腹腔注射3.2×106cell/mL的嗜水气单胞菌(Aeromonas hydrophila,Ah)0.1 mL,在注射嗜水气单胞菌48 h、72 h和96 h后,分别取草鱼头肾、脾脏和肝脏,利用荧光定量PCR分析甘露寡糖对草鱼感染嗜水气单胞菌后各组织中IL-1β和IL-10的表达影响。结果显示,在草鱼头肾中,MOS/I(0.2%甘露寡糖+注射Ah)组IL-1β的表达在注射嗜水气单胞菌后48 h和72 h显著高于CON/I(基础饲料+注射Ah)组;MOS/I组中1L-10的表达在注射嗜水气单胞菌72 h显著高于CON/I组,在96h时显著低于CON/I组。在草鱼脾脏中,MOS/I组IL-1β的表达在注射后72 h和96 h显著低于CON/I组;IL-10的表达在注射嗜水气单胞菌后48 h和96 h显著低于CON/I组。在草鱼肝脏中,MOS/I组IL-1β和IL-10的表达在注射嗜水气单胞菌后48 h显著高于CON/I组,但在注射后72 h时该2种基因的表达显著低于CON/I组。表明甘露寡糖在鱼体感染嗜水气单胞菌时能快速调节头肾、脾脏和肝脏中细胞因子的表达,增强鱼体抵抗细菌性病原的免疫力。 相似文献
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Francesco Buonocore Elisa Randelli Steve Bird Chris J. Secombes Angelo Facchiano Susan Costantini Giuseppe Scapigliati 《Aquaculture (Amsterdam, Netherlands)》2007,270(1-4):512-522
In this paper we investigate the interleukin-10 (IL-10) expression and 3D modelling of the European sea bass (Dicentrarchus labrax L.). IL-10 is a regulatory cytokine that has been intensively studied in mammals and has been found mainly involved in the suppression or deactivation of activated immune responses. The full-length sea bass IL-10 cDNA consists of 1015 bp and is translated in a predicted molecule of 187 amino acids. A multiple alignment of this peptide with other known sequences showed the conservation of fundamental IL-10 family characteristics. Expression analysis by real-time PCR in control fish showed a high basal expression in the head kidney (HK), followed by gut and brain. In vitro treatment of HK leucocytes with LPS showed an intense increase of IL-10 expression at 4 h and a significant decrease at 24 h, with PHA-L no differences were evidenced. A 3D model for sea bass IL-10 was obtained by accurate homology procedures and putative interaction residues with the IL-10 receptor described. The results suggested that sea bass IL-10 could be involved in anti-inflammatory responses, but further experiments are needed to assess its importance in response to pathogenic agents, vaccinations and immunostimulants. 相似文献
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鱼类黏膜相关淋巴组织是抵御病原入侵的第一道防线。其中,鱼类鼻黏膜相关淋巴组织（Nasal-associated lymphoid tissue,NALT）近年来被国内外学者所关注,并被证明是嗅觉器官中抗原识别和启动黏膜免疫应答的重要场所,在细菌、病毒和寄生虫感染后可发挥快速的局部免疫响应。以NALT为靶点,对鱼类开展疫苗鼻内接种可起到良好的免疫保护效果。但目前,对NALT中复杂的免疫细胞与分子网络及其互作机制知之甚少。本文对鱼类嗅觉器官结构与功能、NALT的细胞与分子网络及免疫应答、鼻内接种的应答及免疫保护效果等方面的最新研究进展进行综述,旨在阐明鱼类NALT在黏膜局部发挥的免疫防御机制,以期为新型黏膜疫苗的设计与研发提供参考。 相似文献
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硬骨鱼类免疫球蛋白分类及多样性产生机制研究进展 总被引：4，自引：0，他引：4
鱼类是最先出现免疫球蛋白的低等脊椎动物,对研究免疫球蛋白的起源、进化具有重要的意义。IgM和IgD是最先被人类发现和认识的鱼类免疫球蛋白,随着科技进步和研究深入,陆续在斑马鱼（Danio rerio）、虹鳟（Oncorhynchus mykiss）及鲤鱼（Cyprinus carpio）中发现了IgZ、IgT、IgM-Z等。与哺乳动物的免疫球蛋白一样,不同种鱼,甚至同种鱼不同个体间,免疫球蛋白的形态结构、基因类型、生物学功能等均有差异。为了进一步认识鱼类免疫球蛋白及其免疫应答的机制,本文综述了鱼类免疫球蛋白的分类和多样性机制等。 相似文献
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团头鲂营养需求与健康研究进展 总被引：1，自引：0，他引：1
团头鲂是我国主要的大宗淡水养殖品种之一,其养殖规模在近10年不断扩大。团头鲂的营养物质需求主要以生长和营养缺乏症为评价指标。而营养物质对鱼类健康,如免疫反应和抗病力等影响的研究尚不充分。一般认为,饲料中营养物质搭配合理、品质优良有利于维持鱼类生理健康,并能保护养殖水环境。此外,一些营养物质如维生素,在鱼类的免疫机制中发挥重要作用。未来水产饲料应具有促进水产动物生长和维持健康的双重作用,通过营养调控预防鱼类疾病是保证水产养殖可持续发展的重要策略之一。本研究综述了团头鲂对蛋白质、脂肪、碳水化合物、维生素和矿物质营养需求的研究进展,以及饲料营养元素对团头鲂免疫力及抗病力影响的最新报道,以期为团头鲂高效配合饲料的开发提供科学参考。 相似文献
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Lievens B Frans I Heusdens C Justé A Jonstrup SP Lieffrig F Willems KA 《Journal of fish diseases》2011,34(11):861-875
Fish diseases can be caused by a variety of diverse organisms, including bacteria, fungi, viruses and protozoa, and pose a universal threat to the ornamental fish industry and aquaculture. The lack of rapid, accurate and reliable means by which fish pathogens can be detected and identified has been one of the main limitations in fish pathogen diagnosis and fish disease management and has consequently stimulated the search for alternative diagnostic techniques. Here, we describe a method based on multiplex and broad-range PCR amplification combined with DNA array hybridization for the simultaneous detection and identification of all cyprinid herpesviruses (CyHV-1, CyHV-2 and CyHV-3) and some of the most important fish pathogenic Flavobacterium species, including F. branchiophilum, F. columnare and F. psychrophilum. For virus identification, the DNA polymerase and helicase genes were targeted. For bacterial identification, the ribosomal RNA gene was used. The developed methodology permitted 100% specificity for the identification of the target species. Detection sensitivity was equivalent to 10 viral genomes or less than a picogram of bacterial DNA. The utility and power of the array for sensitive pathogen detection and identification in complex samples such as infected tissue is demonstrated in this study. 相似文献
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高不饱和脂肪酸（HUFAs）在淡水鱼类中的营养作用研究进展 总被引：3，自引：2，他引：1
鱼粉、鱼油资源的短缺促使全球水产养殖业积极寻找其替代原料,而探究鱼粉、鱼油与其替代物之间的差异尤为重要,其中最主要的差异就是高不饱和脂肪酸(highly unsaturated fatty acids,HUFAs)是否存在及其含量多少。HUFAs是一类碳原子数目≥20、双键数≥3的脂肪酸,具有为动物提供能量、构成细胞膜组分、形成高生物活性物质、调控脂质代谢和免疫功能等重要作用,主要存在于鱼油和某些类别的微藻中。淡水鱼类具备自身合成HUFAs的能力,因此,一般认为HUFAs不是淡水鱼类的必需脂肪酸,无需通过饲料提供。但已有研究指出,饲料中添加一定量的HUFAs能够对淡水鱼类产生积极的营养作用,表明淡水鱼类的脂肪酸营养理论尚需进一步完善。本文综述了HUFAs在淡水鱼类生长、脂质代谢、健康免疫、繁殖特性等方面发挥作用的相关研究结果,明确提出淡水鱼类需要摄取一定水平的外源性HUFAs,指出在当前淡水鱼饲料中普遍使用HUFAs相对缺乏的蛋白源和油脂源的背景之下,HUFAs对淡水鱼类的作用应受到关注。最后,本文对今后淡水鱼类HUFAs营养的研究方向,以及新的HUFAs油脂源的开发前景进行了展望。 相似文献
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The use of immunostimulants as an alternative to the drugs, chemicals and antibiotics currently being used to control fish
diseases in fish culture is attracting the attention of many researchers. In this context, many have focused on the use of
medicinal plant products as potential therapeutic measures for modulating the immune response and, specifically, on the use
of herbs to prevent and control fish diseases. Medicinal plants (plant remedies) are a deeply rooted component of the cultural
heritage of many people from diverse cultures and countries and are, as such, closely linked to the maintenance of good health.
The aim of this paper is to review research currently being carried out on the herbs and herbal extracts that have been shown
to modulate the immune system of fish. Special attention is given to the use of Chinese and Indian herbs. 相似文献
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脂肪酸对鱼类免疫系统的影响及调控机制研究进展 总被引：5，自引：3，他引：2
替代脂肪源的开发和利用是解决鱼油短缺问题的必然选择。然而,随着替代水平的提高,鱼体常常表现免疫水平和抗病能力降低。鱼油替代的本质为脂肪酸替代,深入研究脂肪酸与鱼类免疫的关系显得尤为重要。本实验综述了脂肪酸对鱼类免疫性能的影响及调控机制。饱和脂肪酸会降低鱼类免疫力,而适量添加n-3长链多不饱和脂肪酸(LC-PUFA)、共轭亚油酸(CLA)或提高n-3/n-6多不饱和脂肪酸(PUFA)的比例有利于鱼体免疫力发挥;饲料中脂肪酸主要通过细胞膜结构、信号传导、类花生四烯酸、细胞因子和类固醇激素等途径对鱼类免疫进行调控。脂肪酸与鱼类的免疫性能具有高度相关性,而调控机制的研究尚有较大空间。未来研究应该侧重于以下几个方面:脂肪酸对免疫相关转录因子的调控机制;鱼类肠道脂肪酸组成改变与菌群结构和免疫性能之间的相关性;环境因子对鱼体脂肪代谢和免疫力的影响;非脂肪酸成分(矿物质、维生素)对鱼类脂肪酸代谢和免疫过程的调控机制。 相似文献
19.
Megumi Matsumoto Kazuma Hayashi Kyosuke Araki Teruyuki Nakanishi Atsushi Yamamoto 《Journal of fish diseases》2020,43(1):81-89
Mycobacteriosis in cultured fish is a challenge for the aquaculture industry worldwide. Treatment by chemical administration is difficult and no effective vaccine has been developed. Therefore, detection and isolation by early diagnosis are important for prevention of the spread of the disease. In mammals, interferon gamma release assays have been established for detection of tuberculosis; these tests are based on the delayed-type hypersensitivity (DTH) response against culture filtrate protein-10 (CFP-10) and the 6-kDa early secreted antigen target (ESAT-6) of Mycobacterium tuberculosis. On the other hand, little is known about the fish immune response against the ESAT-6 and CFP-10 proteins of mycobacteria, although these responses should find application in the diagnosis of mycobacteriosis in fish. In the present study, we identified ESAT-6 and CFP-10 from Mycobacterium pseudoshottsii and cloned the corresponding genes. Intraperitoneal injection of the corresponding DNA plasmid constructs in ginbuna crucian carp yielded increased expression of the fish interferon-γ1-1-encoding gene (IFN-γ1-1). In contrast, IFN-γ1-1 expression accompanied by DTH response was observed only in the CFP-10-DNA plasmid-injected fish. Furthermore, fish that had been prophylactically injected with CFP-10-DNA plasmid exhibited increased survival of M. pseudoshottsii infection. Taken together, these results suggested that CFP-10 may facilitate diagnosis of mycobacteriosis. 相似文献
20.
B Swain M Samanta M Basu P Panda B R Sahoo N K Maiti B K Mishra A E Eknath 《Aquaculture Research》2012,43(6):897-907
Interleukin‐10 (IL‐10) is a multifunctional cytokine and plays an important role in diseases. In this study, IL‐10 gene was cloned and characterized from catla (Catla catla), which is a highly commercially important fish species in the Indian subcontinent. The result indicated that the full‐length catla IL‐10 (cIL‐10) gene had five exons and four introns with an open reading frame of 540 nucleotides encoding a polypeptide of 179 amino acids. A phylogenetic analysis of cIL‐10 gene sequence showed that cIL‐10 clustered with freshwater carps group as expected. Quantitative real‐time polymerase chain reaction analysis showed that cIL‐10 was expressed in gill, liver, kidney, intestine, skin and heart and its expression profile was up‐regulated in bacterial infection and LPS treatment. A close relationship of high cIL‐10 expression and low pro‐inflammatory cytokine IL‐1β expression was observed in the treated group of fish, which might reveal the role of cIL‐10 as an anti‐inflammatory cytokine. Mechanism of cIL‐10 induction was investigated by blocking nuclear factor (NF)‐κB ‐signalling with BAY 11‐7082 in catla kidney cell culture. Blocking NF‐κB suppressed IL‐10 induction by LPS, and thus it revealed that cIL‐10 was induced through NF‐κB signalling. These data could be helpful to understand the function of IL‐10 in fish in response to vaccinations, probiotics and various diseases. 相似文献