共查询到20条相似文献，搜索用时 468 毫秒
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Islam MA Cinar MU Uddin MJ Tholen E Tesfaye D Looft C Schellander K 《Veterinary immunology and immunopathology》2012,146(1):62-73
The aim of the present study was to determine the age-related kinetic changes of Toll-like receptors (TLRs) and downstream genes expression, and secretion of cytokine in lipopolysaccharide (LPS) stimulated porcine alveolar macrophages (AM). For this purpose, AMs were isolated from 5-day-old newborn piglets and 120-day-old young pigs. mRNA expression and cytokine measurement was determined by quantitative real-time PCR and ELISA, respectively. First, AMs were incubated for 24 h in the absence or presence of increasing concentrations of LPS. Results showed the up-regulation of TLRs 2, 4, 5 and 9 mRNA from all concentrations of LPS used, as compared to non-stimulated cells, and TLR4 was the highest expression in both ages (P<0.05). Furthermore, quantitative analysis demonstrated increased expression of mRNAs encoding TLRs 2, 4, 5 and 9, LBP, CD14, MD2, MyD88, IRAK4 and TRAF6 in both ages in a time-dependant manner (P<0.05). Overall, LPS inducible mRNA for TLR4, LBP, CD14 and MyD88 had higher expression in newborn piglets compared with those of young pigs (P<0.05). The level of cytokine protein IL6 and TNFα in supernatant fluid significantly varied with time of incubation and age of animals. Their concentration increased immediately at 1 h after LPS stimulation and remained significantly higher up to 48 h in both ages. Production of pro-inflammatory cytokine protein IL6 and TNFα in supernatant was significantly higher in young pigs than those of piglets. This study suggests that differential age-related changes in the expression of TLRs and downstream genes, and pro-inflammatory cytokine could contribute to a different age-related innate immune response during pulmonary infection. Further investigation is warranted to determine the precise effects of LPS on porcine AMs by means of a functional study across a wider age range. 相似文献
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Leptospiral lipopolysaccharide stimulates the expression of toll‐like receptor 2 and cytokines in pig fibroblasts 
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下载免费PDF全文 Yijie Guo Tomokazu Fukuda Kenichiro Donai Kengo Kuroda Mizuki Masuda Shuichi Nakamura Hiroshi Yoneyama Emiko Isogai 《Animal Science Journal》2015,86(2):238-244
Pigs throughout the world are afflicted with leptospirosis, causing serious economic losses and potential hazards to human health. Although it has been known that leptospiral lipopolysaccharide (L‐LPS) is involved in an immunological reaction between an antigen and a host cell, little is known about how the immune system of pigs can respond to L‐LPS. Here, we stimulated pig fibroblasts by L‐LPS and then quantitatively measured gene and protein expression levels of two toll‐like receptors (TLRs), TLR2 and TLR4, by real‐time PCR and Western blotting. As a result, expression of TLR2 was found to be significantly up‐regulated within 24 h after L‐LPS stimulation whereas induction of TLR4 expression was relatively weak. We also revealed that of myeloid differentiation primary response gene 88 (MyD88), interleukin 6 (IL‐6) and IL‐8 gene expressions were markedly up‐regulated by L‐LPS stimulation. These results may suggest that the pig cell can activate TLR2 rather than TLR4 by L‐LPS stimulation, thereby inducing expression of cytokines. 相似文献
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To evaluate effects of treatment with pathogen-associated molecular patterns (PAMPs) on toll-like receptor (TLR), MHC II, B7 and cytokine expression, pig monocytes and monocyte-derived DCs (moDCs) were treated with LPS, CpG, lipoteichoic acid (LTA), poly IC or peptidoglycan (Pep). Monocytes and moDCs treated with LPS, CpG, LTA, poly IC or Pep altered expression of at least one TLR (4, 5 and 9) and up-regulated MHC II and/or B7. The mRNA for IL-4 was not detected after any treatment. Treatment with LPS or LTA tended to up-regulate mRNA for TLR 4, Th-1 (IFN-gamma and IL-12p35) and Th-2 cytokines (IL-10 and IL-13). Poly IC or CpG tended to up-regulate TLR 9 and Th-1 cytokines. Porcine monocytes and moDCs like those of humans and mice responded to microbial PAMPs by altering TLR expression, up-regulating MHC II and B7 and altering cytokine expression toward Th-1 and/or Th-2, which may steer immune response. Hence, porcine moDCs and monocytes are likely able to discriminate between microorganisms using TLRs which determine cytokine expression and immune response bias. 相似文献
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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
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Lipopolysaccharide and cytokines modulate leukotriene (LT)B4 and LTC4 production by porcine endometrial endothelial cells 下载免费PDF全文
下载免费PDF全文 Uterine inflammatory response is mediated by inflammatory mediators including eicosanoids and cytokines produced by immune and endometrial cells. Interactions between lipopolysaccharide (LPS) and cytokines, and leukotrienes (LTs) in endothelium, important for the host defence during the inflammation, are unknown. We studied the effect of LPS, tumour necrosis factor (TNF)‐α, interleukin (IL)‐1β, IL‐4 and IL‐10 on 5‐lipooxygenase (5‐LO), LTA4 hydrolase (LTAH) and LTC4 synthase (LTCS) mRNA and protein expression, LTB4 and LTC4 release from porcine endometrial endothelial cells, and cell viability. For 24 hr, cells were exposed to LPS (10 or 100 ng/ml of medium) and cytokines (each 1 or 10 ng/ml). 5‐LO mRNA/protein expression augmented after incubation with larger doses of LPS, TNF‐α, IL‐4 and IL‐10 and smaller dose of IL‐1β. Larger dose of TNF‐α, smaller doses of LPS and IL‐1β and both doses of IL‐10 increased LTAH mRNA/protein expression. LTAH protein content was up‐regulated by larger dose of LPS, but it was reduced in response to both doses of IL‐4. LTCS mRNA expression was elevated by larger doses of LPS, IL‐4 and IL‐10 or both doses of TNF‐α and IL‐1β. LTCS protein level increased after treatment with both doses of IL‐1β, IL‐4 and IL‐10, smaller dose of LPS and larger dose of TNF‐α. Both doses of LPS and larger doses of TNF‐α and IL‐10 increased LTB4 release. LPS, IL‐1β and IL‐10 at smaller doses, or TNF‐α and IL‐4 at larger doses stimulated LTC4 release. Smaller doses of TNF‐α and IL‐1β or both doses of IL‐4 enhanced the cell viability. This work provides new insight on the participation of LPS, TNF‐α, IL‐1β, IL‐4 and IL‐10 in LTB4 and LTC4 production/release from porcine endometrial endothelial cells, and the effect of above factors on these cells viability. The used cellular model gives the possibility to further establish the interactions between inflammatory mediators. 相似文献
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St Paul M Mallick AI Haq K Orouji S Abdul-Careem MF Sharif S 《Veterinary immunology and immunopathology》2011,144(3-4):228-237
Toll-like receptors (TLRs) are a group of conserved proteins that play an important role in pathogen recognition in addition to the initiation and regulation of innate and adaptive immune responses. To date, several TLRs have been identified in chickens, each recognizing different ligands. TLR stimulation in chickens has been shown to play a role in host-responses to pathogens. However, the mechanisms through which TLRs modulate the chicken immune system have not been well examined. The present study was conducted to characterize the kinetics of responses to TLR4 and TLR21 stimulation in chickens following intramuscular injections of their corresponding ligands, lipopolysaccharide (LPS) and CpG oligodeoxynucleotides (ODNs), respectively. To this end, relative expression of cytokine genes in the spleen was determined at 2, 6, 12 and 24 h after injection of TLR ligands. The results indicated that LPS strongly induced the up-regulation of some immune system genes early on in the response to treatment, including interferon (IFN)-γ, interleukin (IL)-10, and IL-1β. Furthermore, treatment with CpG ODN promoted the up-regulation of major histocompatibility complex (MHC)-II, IFN-γ and IL-10. The response to CpG ODN appeared to be somewhat delayed compared to the response to LPS. Moreover, we found a significant increase in IFN-α gene expression in response to LPS but not CpG ODNs. Future studies may be aimed to further characterize the molecular mechanisms of TLR activation in chickens or to exploit TLR agonists as vaccine adjuvants. 相似文献
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本试验旨在研究亚麻籽油对脂多糖(LPS)刺激仔猪肝脏Toll样受体4(TLR4)和核苷酸结合寡聚化结构域(NOD)信号通路关键基因表达的影响。选取24头断奶仔猪,按体重相近原则随机分为4个组,分别为对照组、LPS组、2.5%亚麻籽油组(2.5%亚麻籽油+LPS)、5.0%亚麻籽油组(5.0%亚麻籽油+LPS),每组6个重复,每个重复1头猪,试验期21 d。试验组注射100μg/kg体重的LPS,对照组注射等量的生理盐水。注射LPS或生理盐水4 h后屠宰仔猪,取肝脏,测定TLR4和NOD信号通路关键基因及相关炎性介质的mRNA表达水平。结果表明:1)LPS刺激显著提高了肝脏肿瘤坏死因子-α(TNF-α)、环氧酶2(COX2)、热休克蛋白70(HSP70)的mRNA相对表达量(P0.05),2.5%亚麻籽油可显著降低COX2、TNF-α的mRNA相对表达量(P0.05),5.0%亚麻籽油可显著降低TNF-α的mRNA相对表达量(P0.05)。2)LPS刺激显著提高了肝脏TLR4、髓样分化因子88(My D88)、白细胞介素-1受体相关激酶1(IRAK1)、NOD1、NOD2、受体互作蛋白2(RIPK2)、核因子-κB(NF-κB)的mRNA相对表达量(P0.05);2.5%亚麻籽油可显著降低NOD1、NOD2的mRNA相对表达量(P0.05),有降低RIPK2 mRNA相对表达量的趋势(0.05≤P0.10);5.0%亚麻籽油可显著降低NOD2的mRNA相对表达量(P0.05)。这表明LPS刺激导致仔猪发生炎症反应,亚麻籽油可能通过抑制NOD信号通路进而缓解肝脏炎症反应。 相似文献
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先分离培养小鼠腹腔巨噬细胞,经差速贴壁法纯化后,随机分为6组：空白对照组、0.5mg/L脂多糖（LPS）组、10-6 mol/L孕酮（P4）组、LPS＋10-5 mol/L P4组、LPS＋10-6 mol/L P4组、LPS＋10-7 mol/L P4组。各组在处理12、24h分别提取上清液,ELISA法测TNF-α和IL-1β的含量;各组在处理24h分别提取细胞总RNA,用RT-PCR法测TLR4、CD14、MD2mRNA的表达。结果显示,处理12、24h,0.5mg/L LPS组TNF-α和IL-1β的含量均极显著高于对照组（P〈0.01）;10-6 mol/L P4组与对照组差异不显著（P〉0.05）;LPS＋10-5 mol/L P4组极显著低于对照组（P〈0.01）;LPS＋10-6 mol/L P4组显著低于对照组（P〈0.05）;而LPS＋10-7 mol/L P4组TNF-α的表达差异不显著（P〉0.05）,IL-1β的表达差异显著（P〈0.05）。说明P4可降低LPS刺激小鼠腹腔巨噬细胞TNF-α和IL-1β的分泌,且呈剂量依赖关系。LPS单独处理,TLR4和CD14mRNA的表达极显著高于对照组（P〈0.01）;10-6 mol/L P4单独处理与对照组无显著差异（P〉0.05）;分别添加1-5、10-6、10-7 mol/L P4组均极显著降低LPS诱导TLR4和CD14mRNA的表达（P〈0.01）,而MD2mRNA的表达差异不显著（P〉0.05）。说明P4可极显著降低LPS刺激小鼠腹腔巨噬细胞TLR4和CD14mRNA表达,但对MD2mRNA表达影响不显著。结果显示,P4能抑制LPS刺激的小鼠腹腔巨噬细胞TNF-α和IL-1β的分泌,此过程与细胞TLR4和CD14表达下降相关,而与MD2的表达无关。 相似文献
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为探究马链球菌马亚种(Streptococcus equi subsp.equi,S.equi)对小鼠骨髓源树突状细胞(bone marrow derived dendritic cells,BMDCs)TLR1、TLR2、TLR6、MyD88、IL-1、IL-6、IL-10、IL-12、TNF-αmRNA表达的影响,用GM-CSF、IL-4细胞因子刺激小鼠骨髓细胞使其诱导分化成未成熟的BMDCs,感染S.equi后16、20、26h,采用SYBR GreenⅠ实时荧光定量PCR检测感染组及未感染组细胞TLRs、MyD88和细胞因子mRNA的表达情况。结果显示,S.equi感染BMDCs后16、20、26h,TLR1、TLR2、TLR6、MyD88mRNA的表达量均显著或极显著的高于未感染组(P0.05或P0.01)。S.equi感染DCs后16、26h,IL-1的分泌量较未感染组没有变化(P0.05),IL-10、TNF-α的分泌量显著或极显著增加(P0.05或P0.01)。IL-6在S.equi感染DCs后16h的分泌量显著增加(P0.05),但感染后26h无显著变化(P0.05)。IL-12在S.equi感染DCs后16h的表达没有变化(P0.05),但感染后26h分泌量有所增加(P0.05)。说明S.equi具有调节DCs TLRs、MyD88和细胞因子表达的能力,从而介导机体的免疫应答反应。 相似文献
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Reasons for performing study: Further knowledge of equine keratinocyte physiology and keratinocyte response to various stimuli is important in developing a better understanding of disease states involving the epidermis. Objectives: To assess the inflammatory cytokine response of cultured equine keratinocytes to various pathogen‐associated molecular pattern molecules (PAMPs) from both Gram‐negative and positive bacteria likely to be present in equine sepsis. Methods: Keratinocytes were isolated from skin of 2 horses and primary cultures performed. Keratinocytes were harvested for RNA extraction after exposure to lipopolysaccharide (LPS), lipoteichoic acid (LTA), peptidoglycan (PGN), bacterial DNA (CpG), flagellin or maintained in medium (controls) for 4 or 24 h. Real time‐quantitative PCR was used to quantify interleukin‐1β (IL‐1β), interleukin‐6 (IL‐6) and CXCL8 mRNA concentrations. Results: Increases (P<0.05) in IL‐1β, IL‐6 and CXCL8 mRNA concentrations were induced by LPS exposure compared to controls. Increased mRNA concentrations of both IL‐6 and CXCL8 were also noted (vs. controls) upon exposure to flagellin. Overall, responses were greater at 4 h. No increases (P>0.05) in cytokine expression by keratinocytes were present after LTA, PGN or CpG exposure. Conclusions: Increased proinflammatory cytokine expression in response to LPS and flagellin indicate that equine keratinocytes have functional TLR4 and TLR5 receptor signalling. However, the lack of keratinocyte stimulation by PGN, LTA or CpG provides no evidence for functional TLR2, TLR9 or NOD receptor signalling. These results suggest that equine keratinocytes are more responsive to PAMPs usually associated with Gram‐negative sepsis and unresponsive to PAMPs most commonly associated with Gram‐positive sepsis. Potential relevance: The increased incidence of injury of epidermal structures in clinical cases of Gram‐negative (vs. Gram‐positive) sepsis in the horse may be due to a lack of functional TLR signalling for Gram‐positive PAMPs in the equine keratinocyte. 相似文献
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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
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为探究Toll样受体(TLRs)介导的信号通路在马链球菌马亚种(S.equi)感染小鼠巨噬细胞RAW264.7中的作用,收集S.equi感染后不同时间点的RAW264.7细胞,提取总RNA并反转录成cDNA,利用实时荧光定量PCR技术检测细胞Toll样受体1、2、6(TLR1、TLR2、TLR6)、接头蛋白骨髓分化蛋白88(MyD88)及细胞因子IL-1、IL-6、IL-10、IL-12、TNF-αmRNA的表达情况。结果显示,S.equi感染RAW264.7细胞后6h时,TLR1、TLR2、TLR6与MyD88mRNA水平均较对照组没有显著差异(P>0.05);感染后12h时,TLR1、TLR2和TLR6mRNA表达量未出现明显上升(P>0.05),而MyD88mRNA水平极显著升高(P<0.01);感染后24h时,TLR1、TLR2和TLR6mRNA表达水平出现极显著升高(P<0.01),MyD88mRNA表达没有显著变化(P>0.05),且IL-10和IL-12mRNA水平与对照组相比极显著升高(P<0.01),IL-1、IL-6和TNF-αmRNA水平均极显著下降(P<0.01)。结果表明,TLRs介导的信号通路参与S.equi感染RAW264.7细胞的免疫应答反应。 相似文献
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《中国畜牧杂志》2019,(4)
本试验旨在研究脂多糖(LPS)刺激后不同时间断奶仔猪肌肉炎症和肌肉蛋白质降解相关基因表达的变化规律。选择42头(7.1±0.9)kg杜×长×大三元杂断奶仔猪,按注射LPS之前(0 h)和注射LPS后1、2、4、8、12、24 h随机分为7个处理,每个处理6头猪。预试14 d后,腹腔注射100μg/kg体重的LPS。按以上时间点将仔猪屠宰,取背最长肌样品待测。结果表明:背最长肌炎性细胞因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6的mRNA表达量在注射LPS 1～2 h后达到峰值;Toll样受体4信号通路关键基因Toll样受体4(TLR4)、骨髓分化因子88(MyD88),核苷酸结合寡聚域信号通路关键基因核苷酸结合寡聚域2(NOD2)、受体互作蛋白激酶2(RIPK2)的mRNA表达量在注射LPS 2～4 h后达到峰值;肌肉蛋白质降解相关基因叉头转录因子-1(FOXO-1)、FOXO-4、肌肉环指蛋白1(MuRF1)、肌萎缩F-box(MAFbx)的mRNA表达量在注射LPS 12 h后达到峰值。可见,LPS刺激诱导肌肉释放大量炎性细胞因子,使TLR4和NOD炎症信号通路关键基因及肌肉蛋白质降解相关基因mRNA显著表达。 相似文献
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Kiku Y Matsuzawa H Ohtsuka H Terasaki N Fukuda S Kon-Nai S Koiwa M Yokomizo Y Sato H Rosol TJ Okada H Yoshino TO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(8):723-726
The effects of chlorpromazine (CPZ), pentoxifylline (PTX) and dexamethasone (DEX) on mRNA expression of lipopolysaccharide (LPS)-induced proinflammatory cytokines were examined in bovine peripheral blood mononuclear cells (PBMCs) in vitro. The expression of inflammatory cytokine mRNAs was analyzed by RT-PCR and Southern blot hybridization in bovine PBMCs. CPZ and DEX decreased the expression of cytokine mRNA (such as interleukin-1 beta and tumor necrosis factor-alpha) after stimulation with LPS in a dose-dependent manner. However, pretreatment with PTX had no inhibitory effect on the mRNA expression of proinflammatory cytokines. These results indicated that pretreatment with CPZ and DEX might be effective to reduce the production of LPS-induced inflammatory cytokines in bovine PBMCs in vitro. 相似文献
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Short-term changes of mRNA expression of various inflammatory factors and milk proteins in mammary tissue during LPS-induced mastitis 总被引：11，自引：0，他引：11
During mammary gland infection, non-specific responses are the predominant ones. The goal of this study was to investigate the mRNA expression of various soluble immune components and of the major milk proteins during the acute phase of mammary inflammation. Five healthy lactating cows were intramammary infused in one quarter with 100 microg Escherichia coli-endotoxin (lipopolysaccharide, LPS) and the contralateral quarter with saline (9 g/l) serving as control. Mammary biopsy samples of both quarters were taken immediately before and at 3, 6, 9 and 12 h after infusion and mRNA expression of various factors was quantified via real-time RT-PCR. Blood samples for determination of leukocyte number were taken simultaneously with the biopsy samples and rectal temperature was measured at 1-h intervals. Rectal temperature increased until 5h (P < 0.05) after LPS administration and remained elevated until 9 h after LPS inoculation. Blood leukocyte number decreased (P < 0.05) from 0 to 3 h from 7.7 +/- 1.1 x 10(9)l(-1) to 5.7 +/- 1.0 x 10(9)l(-1) and thereafter recovered to pre-treatment levels until 12 h after LPS challenge. In LPS-treated quarters, tumor necrosis factor-alpha and cyclooxygenase-2-mRNA expression increased (P < 0.05) to highest values at 3h after LPS challenge. Lactoferrin, lysozyme, inducible nitric oxide synthase increased (P < 0.05) and peaked at 6 h after challenge, and platelet-activating factor acetylhydrolase-mRNA expression tended to increase (P = 0.07). mRNA expression of insulin-like growth factor-I and of alphaS1-casein (CN), alphaS2-CN, beta-CN and beta-lactoglobulin did not change significantly, whereas mRNA expression of 5-lipoxygenase and alpha-lactalbumin decreased (P < 0.05) in both quarters and that of kappa-CN only in the LPS quarter. mRNA expression of some investigated factors (tumor necrosis factor-alpha, lysozyme, 5-lipoxygenase, alpha-lactalbumin) changed in control quarters, however in all respective factors less than in the LPS quarters (P < 0.05). In conclusion, mRNA expression of most inflammatory factors increased within hours, whereas that of most milk proteins remained unchanged. 相似文献
20.
Escherichia coli inoculation of porcine mammary glands affects local mRNA expression of Toll-like receptors and regulatory cytokines 总被引：1，自引：0，他引：1
The expression of mRNAs for the Toll-like receptors (TLRs) TLR2 and TLR4, pro- and anti inflammatory cytokines and their receptors was evaluated in mammary gland biopsy material collected from sows intramammarily inoculated with Escherichia coli strain O127 at parturition. Quantitative real-time RT-PCR analysis showed increased mRNA levels for TLR2, the proinflammatory cytokines interleukin IL-1beta and tumor necrosis factor-alpha TNF-alpha, and the anti-inflammatory cytokine IL-10 in the inoculated mammary glands 24h after inoculation. Increased mRNA levels of the proinflammatory cytokine IL-6 were only observed in the inoculated mammary glands of sows that developed clinical signs of mastitis. In contrast, the expression of the anti-inflammatory cytokine, transforming growth factor-beta 1 (TGF-beta1) mRNA was unaltered, as was mRNA expression for the IL-1 receptor type I (IL-1R1). Furthermore, IL-1beta and IL-10 mRNA expression was higher in the inoculated mammary glands of sows that developed clinical signs of mastitis compared with sows that remained clinically healthy. Notably, sows that developed clinical signs of mastitis had significantly lower pre-inoculation levels of IL-1beta mRNA than sows that remained clinically healthy. These findings suggest that development of coliform mastitis is associated with the level of local expression of regulatory cytokines in response to intramammary E. coli inoculation and infection. 相似文献