共查询到19条相似文献，搜索用时 208 毫秒
1.
根据鸡β-干扰素(ChIFN-β)基因保守序列设计引物建立了检测鸡IFN-βmRNA表达水平的荧光定量RT-PCR(RRT-PCR)方法,并对H5N1亚型禽流感病毒(AIV)和新城疫病毒强毒株(vNDV)感染后3、6、12、24、30 h的鸡胚成纤维细胞(CEF)中IFN-βmRNA表达水平进行了检测。结果显示,建立的RRT-PCR特异性好,对鸡IFN-βmRNA的扩增效率为94.18%,线性范围为10-8~10-3,相关系数为0.992,最低能检出48拷贝/反应。AIV在感染CEF后6、12 h显著抑制IFN-βmRNA的表达,24 h开始诱导IFN-βmRNA表达,30 h时IFN-βmRNA水平显著升高;vNDV在感染CEF后的24 h内显著抑制IFN-βmRNA的表达,30 h时则显著诱导IFN-βmRNA表达。本试验结果为进一步研究H5N1和NDV与机体的相互作用提供了有价值的信息。 相似文献
2.
为研究病毒与机体的相互作用,本研究参考GenBank中鸡Toll样受体21 (ChTLR21)的基因序列设计实时定量PCR特异性引物,以鸡核糖体蛋白L4 (RPL4)为内参基因,建立检测ChTLR21 mRNA相对转录水平的实时定量PCR方法,分析ChTLR21在禽传染性喉气管炎病毒(ILTV)感染的鸡胚成纤维细胞(CEF)中和感染SPF雏鸡免疫器官脾脏、法氏囊和胸腺组织中的转录水平.结果显示:ILTV感染CEF后2h、4h、8h和18h时间点ChTLR21 mRNA转录水平分别为未感染对照细胞的1 540.53、0.98、1.19和3.70倍.但仅在ILTV感染2h时引起ChTLR21转录水平升高,与对照组相比差异显著(P＜0.05).ILTV感染SPF雏鸡后6h、24 h和30 h脾脏中ChTLR21 mRNA转录水平分别为未感染对照的56.34、59.85和3.61倍;法氏囊中分别为0.03、25.98和3.08倍;胸腺中分别为2.52、50和7.32倍.在感染初期,脾脏中ChTLR21转录量显著升高,随后有所降低,但均高于未感染对照(p＜0.05);法氏囊中仅在感染6h时呈显著抑制(p＜0.01);胸腺中呈波动性转录水平升高,但与对照组无统计学差异(p＞0.05).本研究证明ChTLR21参与了鸡体对ILTV感染的应答,并在体内外感染模型中呈现不同的表达规律. 相似文献
3.
根据鸡β-干扰素（ChIFN-β）基因保守序列设计引物建立了检测鸡IFN-β mRNA表达水平的荧光定量RT-PCR (RRT-PCR)方法,并对H5N1亚型禽流感病毒（AIV）和新城疫病毒强毒株（vNDV）感染后3、6、12、24、30 h的鸡胚成纤维细胞（CEF）中IFN-β mRNA表达水平进行了检测。结果显示,建立的RRT-PCR特异性好,对鸡IFN-β mRNA的扩增效率为94.18%,线性范围为10-8～10-3,相关系数为0.992,最低能检出48拷贝/反应。AIV在感染CEF后6、12 h显著抑制IFN-β mRNA的表达,24 h开始诱导IFN-β mRNA表达,30 h时IFN-β mRNA水平显著升高;vNDV在感染CEF后的24 h内显著抑制IFN-β mRNA的表达,30 h时则显著诱导 IFN-β mRNA表达。本试验结果为进一步研究H5N1和NDV与机体的相互作用提供了有价值的信息。 相似文献
4.
雏鸡不同组织TLR1、TLR2、TLR4、TLR5和TLR15 mRNA转录水平相对定量研究 总被引：1，自引：0，他引：1
参考GenBank登录的ChTLRs基因序列设计实时定量PCR特异性引物,建立检测鸡Toll样受体(ChTLR)mRNA相对转录水平的实时定量PCR方法,分析ChTLR1、ChTLR2、ChTLR4、ChTLR5和ChTLR15在雏鸡不同器官组织中的转录水平。结果显示5种ChTLRs在脾脏、法氏囊、胸腺和各段肠道组织中均有转录。其中,ChTLR1 mRNA在法氏囊、肾脏和盲肠组织中转录水平较高;ChTLR2 mRNA在脾脏、法氏囊和肝脏等组织中转录水平较高,在肾脏、肺脏和皮肤未检测到转录;ChTLR4 mRNA在所检测组织中转录水平差异较小,在脾脏、十二指肠和胸腺转录水平较高;ChTLR5 mRNA在肾脏、脾脏和空肠中的转录水平较高;ChTLR15 mRNA在法氏囊中转录水平最高,其次为脾脏和盲肠。本研究建立了检测ChTLRs mRNA在不同器官组织中表达水平的实时定量PCR方法,ChTLRs mRNA在雏鸡各器官组织中转录水平差异较大,可能与雏鸡各器官组织对病原的识别和抵抗能力有关。 相似文献
5.
《中国家禽》2019,(11)
为探究TLR4基因在三黄鸡不同器官中的表达规律,试验通过运用real-time PCR方法检测TLR4基因在100日龄三黄鸡公、母鸡不同器官中的表达情况。结果显示:三黄鸡母鸡的法氏囊、肌肉、肺、心、肝、下丘脑中TLR4的表达水平最高,脾脏、十二指肠中TLR4的表达水平较高,腺胃、胸腺、卵巢、回肠、肌胃、垂体、肾、盲肠、空肠和大脑最低。三黄鸡公鸡的心、胸腺、下丘脑、肺、法氏囊、垂体、肌肉、肝、脾中TLR4的表达水平显著高于其他器官。而公、母鸡对比,公鸡胸腺中TLR4的表达水平显著低于母鸡,但是法氏囊中TLR4的表达水平显著高于母鸡。研究结果为TLR4的深入研究提供了理论基础,同时为三黄鸡的抗病育种提供新思路。 相似文献
6.
从日本大耳白兔的脾脏组织克隆出Toll样受体基因2(RTLR2)和Toll样受体基因4(RTLR4),并对其在兔的17种组织中的表达分布进行了检测。RTLR2和RTLR4核苷酸序列与GenBank中登载的穴兔(Oryctolagus cuniculus)TLR2序列(NM_001082781)、TLR4序列(NM_001082781)的相似性分别为99%和100%;推导氨基酸序列的同源性比对发现,与人、马、野猪、猫、猩猩、牛、绵羊和鼠等8种动物相比,RTLR2与马的同源性最高,为80%,与鼠最低,为61%,而RTLR4与这几种动物的同源性在62%~75%之间;兔的胸腺、盲肠、睾丸等15种组织中均检测到TLR2和TLR4 mRNA的转录产物,但骨骼中未发现TLR2和TLR4,TLR4在皮肤中也未见表达。 相似文献
7.
为探讨产肠毒素大肠杆菌（ETEC） K88感染仔猪发生炎症反应的分子机制,试验用ETEC K88灌服断奶仔猪,ELISA法检测攻毒后仔猪血清中白细胞介素8（IL-8）含量,实时荧光定量PCR方法检测淋巴结中Toll样受体2（TLR2）、Toll样受体4（TLR4）及其信号通路相关基因（髓样分化因子88（MyD88）、Toll相互作用蛋白（Tollip）、B细胞淋巴瘤因子3（Bcl3））的mRNA相对表达水平。结果发现,仔猪攻毒ETEC K88后6和24 h血清IL-8含量和淋巴结TLR2/4的表达水平均极显著或显著高于对照组（P<0.01;P<0.05）,且感染后24 h显著低于感染后6 h（P<0.05）;仔猪感染ETEC K88后24 h淋巴结中MyD88、Tollip和Bcl3的表达水平均极显著高于对照组（P<0.01）,但是感染后6 h时与对照组相比均无显著差异（P>0.05）。综上所述,ETEC K88感染仔猪可能是通过TLR2/4-MyD88信号通路产生炎症因子IL-8,促使仔猪出现炎症反应,且该炎症反应可能受Tollip和Bcl3蛋白的调控而被减弱。 相似文献
8.
为研究传染性法氏囊病病毒(IBDV)感染对鸡细胞模式识别受体(PRRs)及天然免疫抗病毒基因转录的影响,本实验分别采用IBDV弱毒感染DT40细胞和强毒感染SPF鸡,以荧光定量RT-PCR(q RT-PCR)检测感染细胞和法氏囊组织中PRRs(TLR2、TLR3、TLR4、TLR7、MDA5和LGP2)及天然免疫抗病毒基因(IPS-1、IRF3、PKR、OAS、Mx)的m RNA转录变化。在IBDV感染DT40细胞的2 h到24 h内,ch MDA5、ch TLR3、ch LGP2、IRF3、PKR、OAS、Mx的转录水平显著上升,分别为324、22、61、29、16、225 000和18 700倍。当采用si RNA分别敲除DT40细胞中ch MDA5、ch TLR3和ch LGP2时,IPS-1、IRF3、PKR、OAS和Mx在IBDV感染后的2 h到24 h差异变化不显著。在IBDV感染SPF鸡法氏囊组织细胞中,ch MDA5和ch TLR3的表达显著降低,而ch LGP2表达无显著差异,IRF3、PKR、OAS和Mx的转录水平显著上升。TLR4和TLR7在IBDV的体外和体内感染试验中均未检测到,而IPS-1在IBDV的体外和体内感染试验中均变化不显著。本实验结果表明,ch MDA5、ch TLR3和ch LGP2在识别IBDV的感染中起着关键作用,IBDV感染严重抑制了PRRS ch MDA5和ch TLR3的转录,并且提示其抑制作用在体内与体外可能存在不同的分子机制。 相似文献
9.
试验旨在从免疫遗传学的角度初步探讨藏鸡（TC）和隐性白羽鸡（RWC）对柔嫩艾美耳球虫（Eimeria tenella）易感性差异的分子机制,分别用1×105个柔嫩艾美耳球虫孢子化卵囊感染对球虫具有抗性的藏鸡和易感的隐性白羽鸡。应用实时荧光定量PCR检测藏鸡和隐性白羽鸡感染前0 d和感染后第2、4、6和8天脾脏、盲肠、胸腺、法氏囊中γ-干扰素（IFN-γ）、白细胞介素-2（IL-2）、IL-16、Toll样受体（TLR3）和TLR15免疫相关基因的转录水平变化。结果显示,藏鸡脾脏IFN-γ、IL-2、IL-16及TLR3、TLR15免疫相关基因转录水平于感染后第4和8天明显上调,隐性白羽鸡则无明显变化。藏鸡盲肠IFN-γ转录水平在感染后第2天显著上调（P<0.05）,TLR3在感染后第4天起显著上调（P<0.05）,其余免疫相关基因变化幅度不大;隐性白羽鸡盲肠IFN-γ转录水平在感染后第8天显著上调（P<0.05）,IL-2在感染后第2天起显著上调（P<0.05）,IL-16在感染后第6天起显著上调（P<0.05）,TLR3在感染后第2和8天显著上调（P<0.05）,TLR15变化幅度不大。各免疫相关基因在2个品种鸡胸腺和法氏囊中均出现上调或下调,但除藏鸡法氏囊TLR3和TLR15转录水平变化幅度相对较大外,其余免疫相关基因与感染前相比变化幅度不大。以上结果显示,球虫感染主要导致藏鸡和隐性白羽鸡脾脏和盲肠中的各免疫相关基因出现显著变化,表明宿主的遗传背景在一定程度上可影响球虫感染的免疫应答。 相似文献
10.
根据GenBank中马Toll样受体基因序列设计特异性引物,建立检测马Toll样受体（TLRs）mRNA转录水平的SYBR GreenⅠ实时荧光定量PCR方法,检测TLR4、TLR2、TLR1和TLR6在蒙古马不同组织器官中的转录水平。4种TLRs在心脏、肝脏、脾脏、肺脏、肾脏、胃、十二指肠、空肠、盲肠和骨髓中均有转录。其中,TLR4mRNA除在空肠和肝脏外,在其他组织器官中的表达水平均高于TLR2、TLR1、TLR6。免疫器官中,TLR4、TLR1mRNA在骨髓中表达量高于脾脏,而TLR2、TLR6mRNA在脾脏中表达量高于骨髓。各肠段,TLR4、TLR2、TLR1、TLR6mRNA表达水平之间在空肠的差异不是很大,而在十二指肠和盲肠中差异很大。结果表明,TLRs mRNA在马各组织器官转录水平差异较大,可能与其对病原体的识别和抵抗能力有关。 相似文献
11.
Zhang YW Davis EG Blecha F Wilkerson MJ 《Veterinary immunology and immunopathology》2008,124(3-4):209-219
Innate immunity relies on a series of germline-encoded pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), to detect conserved microbial components. TLR9 is typically expressed intracellularly in immune cells such as dendritic cells and recognizes unmethylated bacterial or viral cytosine-phosphate-guanine DNA (CpG-DNA). To investigate innate immune responses through TLR9 signaling pathway in horses, we cloned and characterized equine TLR9. Protein sequence analysis shows that equine TLR9 has a typically conserved cytosolic Toll/interleukin-1 receptor (TIR) domain, three leucine-rich repeat (LRR) motifs, with greater than 82% identity to human, monkey, bovine, canine, feline, porcine and ovine orthologs. Equine TLR9 mRNA expression was characterized for spleen, lymph node, and peripheral blood leukocyte samples. Flow cytometric analysis of equine TLR9 expression using a cross-reactive TLR9 mAb identified high constitutive expression of equine TLR9 in PMNs, CD4(+) and CD8(+) T-lymphocytes as well as other leukocytes; similar to human TLR9 expression. The conservation of equine TLR9 and high expression profile in leukocytes suggests that equine TLR9 is a frequent target for unmethylated CpG-DNA, an essential mechanism for the activation of innate immunity. 相似文献
12.
13.
Advanced molecular immunoassay system for immunobiotic lactic acid bacteria using a transfectant of Toll-like receptor 2 总被引：1，自引：1，他引：1
Masanori TOHNO Takeshi SHIMOSATO Yasushi KAWAI Hisashi ASO Shuji IKEGAMI Naoki TAKETOMO Tadao SAITO Haruki KITAZAWA 《Animal Science Journal》2007,78(2):195-205
Toll‐like receptor 2 (TLR2) is a receptor for a variety of microbial components, and it also mediates activation signals in the cell relating to the innate immune system. In order to evaluate the precise molecular immunoregulation by various strains of lactic acid bacteria (LAB) via TLR2, the swine TLR2 (sTLR2)‐expressing transfectant was constructed using human embryonic kidney (HEK) 293 cells. It is demonstrated that intact immunobiotic LAB can induce immune responses through TLR2, and that different nuclear factor‐κB (NF‐κB) activities of various strains can be accurately detected by sTLR2‐expressing HEK293 cells. Furthermore, cellular activation of NF‐κB via TLR2 is reflected in enhanced binding and uptake of LAB. The sTLR2‐expressing HEK293 cells were also useful for characterizing the expression pattern of type I helper T (Th1) and type II helper T (Th2) cytokines by the stimulation of immunobiotic LAB. These results suggest that sTLR2‐expressing HEK293 cells may be useful in certain molecular immunoassay systems for producing new physiologically functional foods with intestinal immunomodulatory abilities, such as the maintenance of Th1/Th2 polarization. 相似文献
14.
In Drosophila, Toll signaling cascade, which resembles the mammalian Toll-like receptor (TLR)/IL-1R signaling pathways and regulates the expression of anti-microbial peptide genes, mainly relies on peptidoglycan recognition proteins (PGRPs) for the detection of bacterial pathogens. To explore the effect of zebrafish peptidoglycan recognition protein 6 (zfPGRP6) on Toll-like receptor signaling pathway, RNA interference (siRNA) and real time quantitative PCR (RQ-PCR) methods were used to identify differentially expressed genes regulated by zfPGRP6. The target genes included TLR2, TLR3, TLR5, TLR7, TLR8, IL1R, Sterile-alpha and Armadillo motif containing protein (SARM), myeloid differentiation factor 88 (MyD88) and nuclear factor (NF)-kappa B2 (p100/p52). The results of RQ-PCR showed that RNAi-mediated suppression of zfPGRP6 significantly down-regulated the expression of TLR2, TLR5, IL1R, SARM, MyD88 and p100/p52. The expression of beta-defensin-1 was also down-regulated in those embryos silenced by zfPGRP6. In challenge experiments to determine the anti-bacterial response to Gram-negative bacteria, RNAi knock-down of zfPGRP6 markedly increased susceptibility to Flavobacterium columnare. 相似文献
15.
Toll‐like receptor and related cytokine mRNA expression in bovine corpora lutea during the oestrous cycle and pregnancy 
下载免费PDF全文
下载免费PDF全文 JE Gadsby AM Tyson Nipper HA Faircloth M D'Annibale‐Tolhurst J Chang PW Farin IM Sheldon DH Poole 《Reproduction in domestic animals》2017,52(3):495-504
Improving our understanding of the mechanisms controlling the corpus luteum (CL) and its role in regulating the reproductive cycle should lead to improvements in the sustainability of today's global animal industry. The corpus luteum (CL) is a transient endocrine organ composed of a heterogeneous mixture steroidogenic, endothelial and immune cells, and it is becoming clear that immune mechanisms play a key role in CL regulation especially in luteolysis. Toll‐like receptors (TLR) mediate innate immune mechanisms via the production of pro‐inflammatory cytokines, especially within various tissues, although the role of TLR within CL remains unknown. Thus, the objectives of this study were to characterize TLR mRNA expression in the CL during the oestrous cycle and in pregnancy (day 30–50), and to examine the role of TLR signalling in luteal cells. Corpora lutea were collected at various stages of the cycle and pregnancy and analysed for TLR and cytokine mRNA expression. In addition, luteal cells were cultured with the TLR4 ligand (lipopolysaccharide, LPS) for 24 h to evaluate the role of TLR4 in regulating luteal function. Toll‐like receptors 1, 2, 4, 6, tumour necrosis factor alpha (TNF), interferon gamma (IFN‐G), and interleukin (IL)‐12, mRNA expressions were greatest in regressing CL compared with earlier stages (p < .05), whereas no change was observed for IL‐6 mRNA expression. Cytokine mRNA expression in cultured luteal cells was not altered by LPS. Based on these data, one or more of the TLRs found within the CL may play a role in luteolysis, perhaps via pro‐inflammatory cytokine mRNA expression. 相似文献
16.
Development of immune assay system for both CpG and non-CpG DNA from lactic acid bacteria using a transfectant of swine Toll-like receptor 9 总被引：2，自引：1，他引：2
Takeshi SHIMOSATO Haruki KITAZAWA Masanori TOHNO Shinichiro KATOH Yasushi KAWAI Tadao SAITO 《Animal Science Journal》2004,75(4):377-382
Bacterial DNA is expected to be a potent immune stimulating agent to Toll‐like receptor (TLR) 9 expressed cells such as macrophages, monocytes, B lymphocytes, NK cells and dendritic cells. In the present study, we constructed a transfectant of swine TLR9 with mammalian cells. We demonstrated that the transfectant, recognizing both CpG and non‐CpG oligonucleotides from lactic acid bacteria, induced NF‐κB activation by gene reporter assay. These findings indicate that the swine TLR9 transfectant will be highly useful for the screening of immunostimulatory DNA from lactic acid bacteria. 相似文献
17.
将40只雌性ICR小鼠,受孕后随机分为试验组和对照组。雌鼠产后10-11d,试验组经第4对乳头灌注LPS,对照组灌注生理盐水,分别于灌注后不同时间采集样本,组织学分析乳腺病理变化;分析对各组乳腺组织中TLR4和TNF-α mRNA表达变化。组织学结果显示,灌注1．5h后乳腺组织中炎性细胞增多,6、12h乳腺腺泡内有大量的炎性细胞浸润,腺泡结构崩解;6h TLR4 mRNA表达极显著高于对照组（P〈0．01）;4个试验组中TNF-αmRNA表达极显著高于对照组（P〈0．01）。试验结果表明LPS能够增强TLR4和TNF-α mRNA表达。 相似文献
18.
Differential expression of Toll-like receptor mRNA in White Leghorn and indigenous chicken of India 总被引：1，自引：0，他引：1
Kannaki T. Ramasamy Maddula R. Reddy Dhanutha N. Raveendranathan Shanmugam Murugesan Rudra N. Chatterjee Rajkumar Ullengala Santosh Haunshi 《Veterinary research communications》2010,34(7):633-639
In the present experiment, the expression profile of Toll-like receptor mRNA in indigenous and pure line chickens was studied.
The expression of TLR3, TLR4, TLR5 and TLR7 were quantified in heterophils of Aseel, Kadaknath, Naked neck, Dwarf and White
Leghorn lines by Quantitative Real-time PCR. White Leghorns expressed significantly (P < 0.01) higher levels of TLR3 mRNA
compared to other lines. TLR4 and TLR5 mRNA were significantly highly expressed in Kadaknath line. Among the TLRs investigated
TLR5 was more expressed in all lines studied. TLR7 was highly expressed in indigenous chicken Aseel and Kadaknath than other
lines. Dwarf chicken expressed significantly (P < 0.01) lower levels of all TLRs investigated. On the basis of the present
study we conclude that the differential expression of TLR mRNA in the heterophils of indigenous and other chicken breeds might
contribute to their variable disease resistance/susceptibility. 相似文献
19.
为了探究猪Toll样受体（Toll-like receptor 5,TLR5） 基因表达水平与F18大肠杆菌抗性的关系,试验通过不同血清型产肠毒素大肠杆菌（F18ab和F18ac）侵染猪小肠上皮细胞（IPEC-J2）,同时通过脂多糖（LPS）分别诱导IPEC-J2细胞4和8 h,利用实时荧光定量PCR检测TLR5基因表达水平变化,并利用Western blotting进行蛋白表达分析。结果显示,不同血清型大肠杆菌（F18ab和F18ac）菌体侵染IPEC-J2细胞后,TLR5基因表达水平均极显著上调（P<0.01）;LPS诱导IPEC-J2细胞4和8 h后,TLR5基因表达水平均极显著上调（P<0.01）,且在LPS诱导IPEC-J2细胞8 h后,TLR5基因表达水平明显高于诱导4 h。与对照组相比,细胞中TLR5蛋白的表达水平极显著上调（P<0.01）,与LPS诱导及F18大肠杆菌菌体刺激IPEC-J2细胞后mRNA表达水平结果相一致。本研究在细胞水平上分析了TLR5表达水平和F18大肠杆菌侵染的相关性,进一步证实猪TLR5基因的表达水平在细胞抵抗F18大肠杆菌的侵染过程中发挥了重要的调控作用,为今后关于TLR5基因功能及其在大肠杆菌腹泻遗传育种应用的研究奠定基础。 相似文献