溃疡性结肠炎患者肠道菌群变化与细胞因子、TLRs分子表达的相关性研究

目的 探讨溃疡性结肠炎患者肠道菌群变化与细胞因子、TOLL样受体(Toll-like receptors,TLRs)分子表达的相关性.方法 将2015年6月—2016年12月在山东省医学科学院第三附属医院确诊并接受治疗的溃疡性结肠炎患者78例作为试验组,同时选择未患溃疡性结肠炎的80例健康者作为对照组.分别对试验组和对照组进行肠道菌群检测,肠黏膜TLR2、TLR4、TLR5、TLR9分子表达检测和外周血IL-4、IL-6、IL-17、IL-23、TNF-α等炎性细胞因子表达检测,分析炎性细胞因子和TLRs表达与肠道菌群变化的关系.结果 试验组双歧杆菌、乳杆菌含量明显低于对照组(P均<0.05),拟杆菌、肠杆菌、肠球菌、梭杆菌含量明显高于对照组(P均<0.05);试验组肠黏膜组织中TLR2、TLR4、TLR5、TLR9表达明显高于对照组(P均<0.05);试验组外周血IL-4表达低于对照组,IL-6、IL-17、IL-23、TNF-α等炎性细胞因子表达高于对照组(P均<0.05).Pearson相关性分析显示,TLR2、TLR4、TLR5、TLR9表达与拟杆菌、肠杆菌、肠球菌含量呈正相关,与双歧杆菌、乳杆菌含量呈负相关;与IL-6、IL-17、IL-23、TNF-α表达呈正相关,与IL-4表达呈负相关.结论 溃疡性结肠炎患者正常肠道菌群平衡被打破,促炎因子表达增加,抑炎因子表达减少,TLRs分子表达增加.肠道菌群紊乱可能通过增强TLRs分子表达来促进促炎因子的分泌,介导肠黏膜炎性反应.     

Toll样受体家族基因多态性与结核易感性研究进展

结核病是由结核分枝杆菌引起的慢性传染性疾病。固有免疫在对结核分枝杆菌的应答中起到重要的作用,其中Toll样受体（TLRs）是固有免疫中一种重要的模式识别受体,它是激活固有免疫的一个开关,在识别病原体相关成分的过程中举足轻重。TLR1、TLR2、TLR4、TLR9在结核分枝杆菌感染过程中对菌体相关成分进行识别从而促进固有免疫应答,其中TLR1基因的单核苷酸多态性位点rs4833095、rs5743618、rs3923647,TLR2基因的rs57473708、rs3804099位点和TLR9基因的rs352139、rs5743836等位点的变异在某些人群中与结核易感性密切相关,而TLR3、TLR6、TLR7、TLR8、TLR10基因多态性与结核易感性存在一定关系。TLRs功能的正常发挥保证了机体对结核分枝杆菌正常免疫反应。TLRs基因的多样性使不同个体应对相同的病原体产生不同的反应。对TLRs单核苷酸多态性位点与结核易感性关系的研究,可以预测某些人群的结核病易感倾向,提供药物新靶点。     

Toll样受体与输卵管疾病

Toll样受体(TLRs)是机体通过直接感知病原体做出防御反应的第一个天然免疫受体,为机体防御微生物入侵的第一道防线。近年来的研究发现,TLRs在生殖道表达,且与输卵管妊娠、输卵管慢性炎症密切相关,尤其是Toll样受体2(TLR2)和Toll样受体4(TLR4)。本文就TLRs在输卵管妊娠、输卵管炎的相关研究进展进行综述,旨在探讨其参与疾病发生、发展的机制,为后续的研究提供一些参考。     

Toll样受体单核苷酸多态性与疾病易感性的关系

基因多态性与机体对疾病的遗传易感性相关。Toll样受体(TLRs)在识别病原微生物、启动炎性应答中起重要作用。越来越多数据表明,TLRs单核苷酸多态性(SNPs)与炎性应答损伤及感染性疾病的遗传易感性相关,如TLR4基因Asp299Gly和Thr399Ile SNPs与多种感染性疾病正相关,而在动脉粥样硬化及其相关疾病中则起保护作用[1]。笔者就TLRs SNPs与感染性疾病的遗传易感性进行综述。1 Toll样受体与免疫应答近来认为,机体针对病原微生物等病原相关分子模式的入侵所产生的先天及特异性免疫由模式识别受体(PRRs)启动。TLRs是主要的PRRs,能调节…     

Toll样受体基因表达水平在儿童时期的变化及其意义

【目的】观测单个核细胞Toll样受体(Toll-like receptor,TLR)2、3、4的基因表达水平在儿童时期的变化,探讨其在天然免疫发育中的意义。【方法】收集0~15岁健康儿童的外周血,共259份标本,分离单个核细胞,采用RT-荧光定量PCR法测定TLR2、3、4 mRNA。【结果】婴幼儿TLRs基因表达水平较高,其后有所下降,男童至青春期达到第二个高峰。性别对TLRs mRNA水平具有影响,表现为学龄前期儿童TLR2、3、4 mRNA水平女童高于男童,而9岁以后TLR2、3、4 mRNA水平女童普遍低于男童。【结论】TLRs基因表达水平存在年龄和性别相关的变化。年幼儿TLRs的高表达表明在婴幼儿时期,天然免疫系统对外界病原体的识别就已经相当完善。学龄前期及青春期前后儿童,确实存在性别有关的TLRs的表达差异,可能与其性激素的水平有关。     

Toll样受体与常见病原菌感染之间关系的研究进展

Toll样受体(Toll-like receptors, TLRs)是近年来发现的先天性免疫系统中的细胞跨膜受体及病原模式识别受体之一,迄今为止,已在哺乳动物中发现了11种TLRs分子,其中TLR2和TLR4的作用尤为突出.笔者就TLRs的分子结构特点、信号转导方式及其在革兰阳性菌、革兰阴性菌、病毒、真菌及结核菌等病原菌所致感染中的作用做一综述.     

Toll样受体7与哮喘的关系

人类Toll样受体（toll-like receptors,TLRs）是近年来发现的一类信号传导跨膜受体,可以识别、结合病原体相关分子模式,启动免疫反应,是人体先天免疫和获得性免疫的桥梁.TLR7作为TLRs家族中重要的成员,在生理、病理中均起着重要的作用,维持它的正常功能对人体的健康非常重要,如Mφller-Larsen等发现TLR7和TLR8两个受体在支气管哮喘中有着重要的作用机制.本文着重对TLR7在生理和病理中的作用并与哮喘的关系,参考国内国外的文献最新进展作一综述.     

支气管哮喘合并感染性肺炎患儿中性粒细胞CD64及Toll样受体的表达及临床意义

目的研究支气管哮喘合并感染性肺炎患儿外周血中性粒细胞CD64及Toll样受体(TLRs)表达的变化及作用机制。方法选取2015年1月-2018年12月天津市北辰区中医医院儿科支气管哮喘合并感染性肺炎患儿118例(感染组),并根据病原体种类,将感染组分为细菌性肺炎组(56例)、病毒性肺炎组(24例)、支原体肺炎组(20例)及衣原体肺炎组(18例);同期选取儿科支气管哮喘非感染性肺炎患儿30例(非感染组)以及健康体检儿童30例(健康组)。入院次日清晨或体检当天采空腹外周静脉血,应用流式细胞术检测外周血中性粒细胞CD64及Toll样受体TLR2、TLR4、TLR7及TLR9水平,电化学发光法检测降钙素原(PCT),并对中性粒细胞CD64指数与TLR2、TLR4、TLR7及TLR9水平进行相关性分析。结果 3组中性粒细胞CD64指数及PCT比较差异有统计学意义(P<0. 05),感染组(4. 82±1. 55,5. 19±5. 34)>非感染组(2. 46±0. 99,0. 35±0. 13)>健康组(0. 99±0. 29,0. 23±0. 09);感染组中,细菌性肺炎组CD64指数及PCT水平高于病毒性肺炎组、支原体肺炎组及衣原体肺炎组,病毒性肺炎组CD64指数高于支原体肺炎组及衣原体肺炎组(P<0. 05); 3组中性粒细胞TLR2、TLR4、TLR7及TLR9表达差异有统计学意义(P<0. 05),感染组TLRs水平高于非感染组及健康组,非感染组TLRs水平高于健康组;CD64指数与TLR2、TLR4、TLR7呈正相关(r=0. 421、0. 355、0. 458,均P<0. 05)。结论感染性肺炎与儿童支气管哮喘发病密切相关,感染时中性粒细胞CD64及TLR2、TLR4、TLR7及TLR9表达增加,TLRs固有免疫调节在感染性肺炎患儿支气管哮喘发病机制中有着重要作用。     

血液透析巨细胞病毒感染炎症反应状态和 TLRs/MYD88信号通路水平

目的探究血液透析(HD)巨细胞病毒(CMV)感染炎症反应状态和Toll样受体/髓样分化因子88(TLRs/MYD88)信号通路水平。方法选择2017年12月-2019年12月青海省人民医院住院治疗的HD患者150例,根据患者是否合并CMV感染将其分为HD合并CMV组(n=37)和HD组(n=113),比较两组患者T淋巴细胞亚群(CD_3~+、CD_4~+、CD_8~+、CD_4~+/CD_8~+)水平、炎性因子[白细胞介素(IL)-2、IL-6、IL-10、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)、中性粒细胞明胶酶相关脂质运载蛋白(NAGL)]水平以及TLRs/MYD88信号通路(TLR2、TLR4、MYD88 mRNA)表达水平。结果 HD合并CMV组患者CD_4~+、CD_4~+/CD_8~+水平低于HD组、CD_8~+水平高于HD组(均P0.05),CD_3~+水平与HD组比较,无统计学差异;HD合并CMV组患者IL-2、IL-6、TNF-α、IFN-γ、NAGL水平高于HD组、IL-10水平低于HD组(均P0.05);HD合并CMV组患者TLR2、TLR4、MYD88 mRNA表达水平高于HD组(P0.05)。结论 CMV感染可导致HD患者T淋巴细胞异常,加重患者炎性反应,其机制可能为上调TLR2、TLR4、MYD88表达,从而激活TLRs/MYD88信号通路。     

膀胱癌尿路感染患者免疫和炎症因子的表达及临床诊断价值

目的 探讨尿路感染对膀胱癌患者Toll样受体(TLRs)信号通路和炎症、免疫功能的影响。方法 选择2017年4月-2020年4月唐山市工人医院收治的105例膀胱癌术后尿路感染患者为研究组，另择同期收治的55例未发生尿路感染膀胱癌患者及50名健康体检者纳入非感染组及对照组，分析各组TLR4、TLR9基因相对表达量；测定CD_3⁺、CD_4+、CD_4⁺、CD_8+、CD_8⁺ T淋巴细胞及免疫球蛋白M(IgM)、IgG、IgA表达水平；测定C-反应蛋白(CRP)、降钙素原(PCT)、白细胞介素(IL)-1、IL-6炎症因子水平。结果 105例感染患者共分离病原菌112株，其中革兰阴性菌71株(63.39%)。感染组及非感染组TLR4、TLR9表达量高于对照组，感染组高于非感染组(P<0.05)。感染组患者CD_3+ T淋巴细胞及免疫球蛋白M(IgM)、IgG、IgA表达水平；测定C-反应蛋白(CRP)、降钙素原(PCT)、白细胞介素(IL)-1、IL-6炎症因子水平。结果 105例感染患者共分离病原菌112株，其中革兰阴性菌71株(63.39%)。感染组及非感染组TLR4、TLR9表达量高于对照组，感染组高于非感染组(P<0.05)。感染组患者CD_3⁺、CD_4+、CD_4⁺、CD_4+、CD_4⁺/CD_8+/CD_8⁺、IgM、IgG、IgA免疫功能指标低于非感染组及对照组，非感染组IgM、IgA水平低于对照组(P<0.05)。感染组患者CRP、PCT、IL-1、IL-6炎症因子水平高于非感染组及对照组，非感染组高于对照组(P<0.05)。TLR4、TLR9、CD_4+、IgM、IgG、IgA免疫功能指标低于非感染组及对照组，非感染组IgM、IgA水平低于对照组(P<0.05)。感染组患者CRP、PCT、IL-1、IL-6炎症因子水平高于非感染组及对照组，非感染组高于对照组(P<0.05)。TLR4、TLR9、CD_4⁺/CD_8+/CD_8⁺、IgM、IgG、IgA、CRP、PCT、IL-1、IL-6指标诊断膀胱癌术后尿路感染的曲线下面积均>60.0%。结论 膀胱癌术后尿路感染患者存在TLRs信号通路活性的增强，免疫、炎症因子表达失调，这些指标水平的变化可作为诊断尿路感染及评估病情的参考指标。     

Ionizing radiation affects the expression of Toll-like receptors 2 and 4 in human monocytic cells through c-Jun N-terminal kinase activation

Pattern recognition receptors recognize pathogen-associated molecular patterns. Among these, Toll-like receptors (TLRs) have well-characterized roles in antibacterial and antiviral immunity. In the present study, the effects of ionizing radiation on the expression of TLRs and cellular responses to ligands were investigated in THP1 monocytes (human monocytic leukemia cells) and THP1-derived macrophage cells (macrophage-like cells), which are induced by culturing in the presence of phorbol 12-myristate 13-acetate. TLR2 and TLR4 expression was detected in THP1 and macrophage-like cells. X-irradiation caused increased expression of these TLRs in THP1 and decreased expression in macrophage-like cells. Responses to FSL-1 (TLR2 ligand) and lipopolysaccharide (LPS, TLR4 ligand) were estimated by determining the induction of tumor necrosis factor-α (TNF-α). After FSL-1 or LPS stimulation, TNF-α induction was greater in X-irradiated THP1 monocytes than in non-irradiated cells. However, although TNF-α expression was not affected by X-irradiation in macrophage-like cells, the expression of LPS-inducible interferon-β was lower following X-irradiation of macrophage-like cells. To clarify the mechanisms of TLR2 and TLR4 regulation by X-irradiation, expression of mitogen-activated protein kinase was investigated. These experiments showed that c-Jun N-terminal kinase (JNK) mediated increases in TLR expression in X-irradiated THP1 monocytes and decreases in TLR expression in X-irradiated macrophage-like cells. This study demonstrates that ionizing radiation modulates ligand-responsive TLR expression through the JNK pathway, depending on differentiation state.     

Toll-like receptors and the significance for clinical medicine

The recently-discovered class of toll-like receptors (TLRs) play an essential role in the complex defence system against microorganisms. TLRs are the first to detect potential pathogens, initiate immune responses and form the crucial link between the innate and acquired immune systems. TLRs also play an important role in the pathophysiology of infectious diseases, inflammatory diseases such as Crohn's disease and atherosclerosis, and possibly play a role in autoimmune diseases. Common polymorphisms in TLR genes are associated with predisposition to severe infections. Drugs that target the TLRs offer new opportunities for the development of therapeutics against a wide variety of diseases such as sepsis syndrome, asthma, inflammatory-bowel diseases and cancer. The first drug that works by modulating the TLR response has already been registered.     

TLRs信号通路和炎症因子的表达与反复呼吸道感染的关系分析

目的 研究Toll样受体(TLRs)信号通路和炎症因子的表达与反复呼吸道感染的关系,为临床治疗提供监测靶点。方法 将汕头市龙湖区第二人民医院儿科2019年1-12月门诊收治的150例反复呼吸道感染患儿纳入研究,记作病例组,另取同期于本院进行体检的健康儿童150例作为对照组。比较病变组和对照组儿童TLRs信号通路和炎症相关因子水平,并作相关性分析。此外,对比病变组和对照组儿童氧化应激反应相关指标水平。结果 病例组患儿Toll样受体2(TLR2)及TLR4水平显著高于对照组,差异有统计学意义(t=69.981,101.427,P＜0.001)。病例组患儿血清白细胞介素-4(IL-4)、白细胞介素-10(IL-10)及肿瘤坏死因子-α(TNF-α)水平显著高于对照组(t=78.453,63.388,116.697,P<0.001),干扰素γ(INF-γ)显著低于对照组(t=70.489,P<0.001)。Pearson相关性分析显示:反复呼吸道感染患儿TLR2、TLR4和血清IL-4、IL-10、TNF-α水平均呈正相关(P<0.05或<0.01),与INF-γ水平呈负相关(P<0.001)。病变组超氧化物歧化酶(SOD)、丙二醛(MDA)及过氧化氢酶(CAT)水平与对照组相比,差异均有统计学意义(t=28.633,57.277,50.103,P＜0.001)。结论 TLRs信号通路和炎症因子均可能参与反复呼吸道感染的发生、发展过程,且反复呼吸道感染患者存在明显的氧化应激状态异常。     

婴儿肝病综合征患儿外周血有核细胞Toll样受体的表达

目的:探讨婴儿肝病综合征患儿外周血有核细胞Toll样受体(TLR s)的表达。方法:婴儿肝病综合征患儿25例,正常对照组12例。采用RT-PCR法检测外周血有核细胞TLR 4、TLR 2 mRNA的变化,采用ELISA法检测血清TNFα、IL-6水平,同时检测内毒素水平,并分析TLR 4、TLR 2 mRNA水平与血清总胆红素(TB)、直接胆红素(DB)的关系。结果:婴儿肝病综合征患儿TLR 4、TLR 2 mRNA的表达及TNFα、IL-6水平明显高于正常对照组,差异有极显著性意义(P<0.01)。两组内毒素水平无明显差异(P>0.05)。TLR 4、TLR 2 mRNA的表达与血清TB、DB无相关关系(P>0.05)。结论:婴儿肝病综合征患儿体内TLR s的表达增强,提示TLR s在婴儿肝病综合征发病机理中起作用。     

Abnormal expression of TLRs may play a role in lower embryo quality of women with polycystic ovary syndrome

Toll-like receptors (TLRs) localize in mammalian ovary, including granulosa cells, cumulus cells, and theca cells. Previous studies demonstrated that TLRs may be important for the cumulus-oocyte complex expansion and fertilization. There is no evidence to indicate that the deletion of TLRs will induce infertility; however, the abnormal expression of TLRs may decrease oocyte quality and fertility rate. In the present study, we investigated the effects of polycystic ovary syndrome (PCOS) on the expression of TLRs in cumulus cells by using western-blot and quantitative real-time PCR (qRT-PCR) analyses. We found that the expression of TLR4 and 9 in cumulus cells was influenced significantly by PCOS. We also observed that overweight/obesity changed the expression of TLR2 and 5 in cumulus cells of PCOS subjects. In addition, we found that the rate of available embryos of women with PCOS was slightly lower. These results indicate that the abnormal expression of TLRs in cumulus may be a reason for the lower embryo quality of women with PCOS.

Abbreviations: ART: assisted reproductive technology BMI: body mass index COC: cumulus-cell-oocyte complex PCOS: polycystic ovary syndrome qRT-PCR: quantitative real-time PCR TLRs: Toll-like receptors     

脐血单个核细胞TLR及SOCS mRNA的表达及意义

目的:观察LPS刺激新生儿脐血单个核细胞(MNC)TLR4、TLR2及SOCS1、SOCS3mRNA的表达变化,探讨新生儿感染时机体防御反应机制。方法:分离脐血MNC,LPS(1μg/m l)培养0,1,3,6,12 h后收集细胞及上清液,RT-PCR方法测定TLR及SOCSmRNA表达情况,ELISA检测上清的TNF-α水平。结果:脐血MNC在LPS刺激1,3,6,12 h后,TLR 4、TLR 2、SOCS 1、SOCS 3 mRNA及TNF-α水平均增高,与0 h组比较,差异有极显著或显著性意义(P<0.01或P<0.05)。TLR 4 mRNA在1h表达最高,TLR 2 mRNA在3 h表达最高,SOCS 1、SOCS 3在1 h表达最强,TNF-α水平在3 h增加尤为明显。结论:TLR参与了LPS诱导炎症因子的产生,SOCS可能是TLR信号通路的负反馈调节因子。     

Acute exposure to ethanol affects Toll-like receptor signaling and subsequent responses: an overview of recent studies.

Ethanol suppresses innate resistance to a variety of microbes, and findings of studies from both our laboratory and other laboratories indicate suppression of responses is mediated through two Toll-like receptors (TLRs): TLR3 and TLR4. In this article, we review recent findings from studies in our laboratory, indicating that ethanol also suppresses responses mediated through other TLRs. Considering the importance of TLR-mediated responses in innate immunity, this supports the possibility that suppression of these responses may constitute a major mechanism by which ethanol suppresses innate immunity. In addition, ethanol-induced changes in cellular signaling and in patterns of gene expression induced through TLR3 were examined in mouse peritoneal macrophages, and these results are reviewed in this article. Signaling through TLR3 was inhibited, and results of DNA microarray analysis supported the notion that inhibition of an interferon-related amplification loop might be responsible for suppression of gene expression for several effector molecules of innate immunity and inflammation not previously known to be altered by ethanol. Thus, ethanol alters responses through most or all mouse TLRs, and this suppresses expression of a wide range of innate immune mediators.     

Role of TLRs in Brucella mediated murine DC activation in vitro and clearance of pulmonary infection in vivo

Brucellosis is worldwide zoonoses affecting 500,000 people annually with no approved human vaccines available. Live attenuated Brucella abortus vaccine strain RB51 protects cattle through CD4 and CD8 T-cell mediated responses. However, limited information is known regarding how Brucella stimulate innate immunity. Although the most critical toll like receptors (TLRs) involved in the recognition of Brucella are TLR2, TLR4 and TLR9, it is important to identify the essential TLRs that induce DC activation/function in response to Brucella, to be able to upregulate both vaccine strain RB51-mediated protection, and clearance of pathogenic strain 2308. Furthermore, in spite of the importance of aerosol transmission of Brucella, no published studies have addressed the role of TLRs in the clearance of strain 2308 or strain RB51 from intranasally infected mice. Therefore, we used a (a) bone marrow derived dendritic cell model in TLRKO and control mice to assess the differential role of pathogenic and vaccine strains to induce DC activation and function in vitro, and (b) respiratory model in TLRKO and control mice to assess the critical roles for TLRs in clearance of strains in vivo. In support of the essential TLRs in clearance and protection, we performed challenge experiments to identify if these critical TLRs (as agonists) could enhance vaccine induced protection against pathogenic strain 2308 in a respiratory model. We determined: vaccine strain RB51 induced significant (p≤0.05) DC activation vs. strain 2308 which was not dependent on a specific TLR; strain RB51 induced TNF-α production was TLR2 and TLR9 dependent, and IL-12 production was TLR2 and TLR4 dependent; TLR4 and TLR2 were critical for clearance of vaccine and pathogenic Brucella strains respectively; and TLR2 (p<0.05), TLR4 (p<0.05) and TLR9 (p=0.075) agonists enhanced vaccine strain RB51-mediated protection against respiratory challenge with strain 2308 in the lung.     

Pathogen sensors and chemokine receptors in dendritic cell subsets

Pathogen sensors such as Toll-like receptors (TLRs) detect microorganism- or host-derived conserved molecular structures, including lipids or nucleic acids and provoke activation of Ag presenting cells such as dendritic cells (DCs). Several synthetic TLR ligands, especially oligonucleotides, are being developed as promising vaccines for infectious diseases, cancers or allergies. DCs are heterogeneous and consist of various subsets, each of which expresses a subset-specific repertoire of TLRs and responds to the TLR signaling in a subset-specific manner. Furthermore, each DC subset expresses a set of chemokine receptors that regulate its function and behavior. Here I review the functions of two DC subsets and how chemokine receptors function in these subsets. One is the plasmacytoid DC (pDC), which expresses nucleic acid sensing receptors TLR7 and TLR9 and secretes large amounts of type I interferons in response to TLR7/9 signaling. The other is splenic CD8α⁺ conventional DC (cDC). This DC subset expresses lipid sensors, TLR2 and TLR4, and nucleic acid sensors, TLR3, TLR9 and TLR13 and is specialized for antigen crosspresentation. Several chemokine receptors are differentially expressed on these DC subsets. The homologues of these murine DC subsets are also found in humans. Understanding how these DC subsets function and respond to TLR ligands and chemokines should be important for development of effective vaccines.