Toll样受体研究进展

Toll-like receptors(TLRs)是存在于机体的一类重要的模式识别受体,在机体天然免疫中起着重要的作用.TLRs还是连接天然免疫和适应性免疫的桥梁.目前TLR1-9研究的较清楚和详尽.不同TLRs识别的病原体相关分子模式各不相同.TLRs介导的机体信号传导途径分为MyD88依赖性和非MyD88依赖性.多种生物TLRs已被克隆和表达,通过进化分析,脊椎动物的TLRs可被分为6大基因家族,识别相似PAMP的TLRs聚为一簇.TLRs编码序列和功能是高度保守的.     

Toll样受体研究进展

免疫系统识别"非我"和"自我"的过程是依赖于不同的受体来完成的,作为先天性免疫系统的重要组成部分及连接获得性免疫与先天性免疫的"桥梁",Toll样受体(Toll-like receptors,TLRs)是生物的一种模式识别受体(pattern recognition receptor,PRR),它主要通过识别病原相关分子模式(pathogen-associated molecularpatterns,PAMPs)来启动免疫反应。已发现TLRs在炎症、细胞信号转导、细胞凋亡、肿瘤等发生过程中扮演重要角色。随着分子细胞生物学的发展,有关TLRs的研究必将更加深入,同时也会进一步拓展对机体免疫机制的认识。     

鸡Toll样受体研究进展

Toll样受体(TLRs)是一类典型的模式识别受体(PRR),是先天性免疫系统中的细胞跨膜受体以及病原模式识别(PAMP)受体之一。在鸡体中发现的TLRs(chTLRs)已经超过10种,其在进化中趋于保守,能识别保守的微生物成分。TLRs对病原相关分子模式的识别不仅在先天性免疫中有重要作用,还能够有效地启动获得性免疫。论文对部分TLRs在鸡体中的分布及参与激活获得性免疫途径的研究进行综述。     

Toll样受体信号通路在两栖类中的研究进展

两栖类处于脊椎动物由水生向陆生进化的过渡阶段,进化地位重要。近年来因疾病的爆发、生境破碎化、环境污染、紫外辐射增加、人为过度捕捉和生物入侵等诸多因素引起的全球范围内两栖类种群数量锐减已引起人们的广泛关注。Toll样受体(Toll-like receptor,TLR)家族是一类从线虫到哺乳动物普遍存在的高度保守的模式识别受体(PRRs),可以识别侵入机体病原体的病原相关分子模式(PAMPs),是两栖类先天性免疫防御系统的重要组成部分。本文结合近些年国内外对两栖类TLRs的研究对两栖类TLRs结构特征、进化特点、在两栖类发育早期的差异表达特征和两栖类TLRs信号通路中相关分子的表达研究进行概述。目前对于两栖类TLRs的研究发现两栖类TLRs兼具鱼类和哺乳类TLRs的特征,并在从鱼类到两栖类到哺乳类的进化过程中,TLRs家族部分成员如TLR2的配体识别功能可能发生了一定程度的特化;在两栖类早期发育阶段,获得性免疫不够完善,以TLRs为主的先天免疫防御系统在抵御病原体的侵袭方面发挥了重要作用;在特定病原菌和病毒的胁迫下,可能激活了TLRs下游不同的途径参与两栖类对病原体的免疫应答反应。     

禽Toll样受体研究进展

Toll样受体家族（Toll-like receptors,TLRs）作为一种膜表面分子,是模式识别受体（pattern recognition receptors,PRRs）的主要组分,在脊椎动物和无脊椎动物中都可识别入侵的病原体相关分子模式（pathogen associated molecular patterns,PAMPs）。通过对鸡和斑胸草雀的基因组进行比对分析,发现禽中存在10种Toll样受体,其中TLR2 a、b、3、4、5和7已经证明和哺乳动物同源;有些经过基因倍增生成两个基因,TLR1La和TLR1Lb,TLR2 a和2 b;禽TLR21可能与鱼类和两栖动物的同源;禽TLR15是禽类特有的一类受体。各种禽Toll样受体在抵御各种微生物的感染过程中发挥各自重要的作用,本文就禽Toll样受体的研究进展进行简要综述。     

The response of HEK293 cells transfected with bovine TLR2 to established pathogen-associated molecular patterns and to bacteria causing mastitis in cattle

Toll-like receptors (TLRs) are key sensors of pathogen-associated molecular patterns (PAMPs). Their role in immunity is difficult to examine in species of veterinary interest, due to restricted access to the knockout technology and TLR-specific antibodies. An alternative approach is to generate cell lines transfected with various TLRs and to examine the recognition of PAMPs or relevant bacteria. In this report, we examined whether recognition of various PAMPs and mastitis-causing bacteria is achieved by transfection of recombinant bovine TLR2 (boTLR2). Therefore, human embryonic kidney (HEK) 293 cells were transfected by whole boTLR2. A clonal analysis of stably transfected cells disclosed variable recognition of several putative TLR2 agonists although expressing similar amounts of the transgene and endogenous TLR6. One clone (clone 25) reacted by copious interleukin-8 (IL-8) production to several stimulants of TLR2 such as di-palmitoylated cysteyl-seryl-lysyl-lysyl-lysyl-lysine (Pam2), a biochemical preparation of lipoteichoic acid from Staphylococcus aureus, a commercial preparation of peptidoglycan from S. aureus, and heat-killed Listeria monocytogenes (HKLM). TLR2-dependent induction of IL-8 release was stronger in medium containing human serum albumin than in medium containing fetal calf serum. Clone 25 cells responded to high concentrations of S. aureus and to Escherichia coli causing mastitis, but not to Streptococcus uberis and to Streptococcus agalactiae which also cause mastitis. Stimulation by S. aureus was relatively weak when compared (i) with stimulation of the same cells by HKLM and PAMPs derived from S. aureus, (ii) with a clone stably transfected with TLR4 and MD-2 and stimulated by E. coli causing mastitis, and (iii) with interferon-gamma-costimulated bovine macrophages stimulated by S. aureus and S. agalactiae. Thus, clone 25 is suitable for studying the interaction of putative TLR2 agonists with bovine TLR2-transfected cells, provides a cell to search for TLR2-specific antibodies, and is a tool for studying the interaction of TLR2 with bacteria causing disease, e.g. mastitis, in cattle.     

Pattern recognition receptors in companion and farm animals - the key to unlocking the door to animal disease?

The innate immune system is essential for host defence and is responsible for early detection of potentially pathogenic microorganisms. Upon recognition of microbes by innate immune cells, such as macrophages and dendritic cells, diverse signalling pathways are activated that combine to define inflammatory responses that direct sterilisation of the threat and/or orchestrate development of the adaptive immune response. Innate immune signalling must be carefully controlled and regulation comes in part from interactions between activating and inhibiting signalling receptors. In recent years, an increasing number of pattern recognition receptors (PRRs), including C-type lectin receptors and Toll-like receptors (TLRs), has been described that participate in innate recognition of microbes, especially through the so called pathogen-associated molecular patterns (PAMPs). Recent studies demonstrate strong interactions between signalling through these receptors. Whereas useful models to study these receptors in great detail in the murine and human system are now emerging, relatively little is known regarding these receptors in companion and farm animals. In this review, current knowledge regarding these receptors in species of veterinary relevance is summarised.     

Toll样受体9介导的免疫激活研究进展

在细菌和DNA病毒的基因组中广泛存在着以非甲基化的胞嘧啶—鸟嘌呤核苷酸为核心的CpG基序。作为一种病原相关的分子模式（pathogen-associated molecular pattern, PAMP）,含有CpG基序的寡聚脱氧核苷酸（CpG oligodeoxynucleotides,CpG ODN）能激活包括B淋巴细胞和类浆细胞在内的多种免疫细胞,并诱导产生以Th1型细胞免疫反应为主的免疫应答。CpG ODN在哺乳动物细胞中的受体是Toll样受体（Toll-like receptors, TLRs）家族中的Toll样受体 9（TLR9）。TLR9所介导的免疫激活作用在某些传染病的预防、新型疫苗佐剂的开发及过敏性疾病和癌症的治疗中有着巨大的应用前景。     

TLR4在妊娠异常终止中的作用机制和研究进展

Toll样受体（toll-like receptors,TLRs）作为一种病原模式识别受体在免疫反应中起着重要的作用。TLR4是最早发现的TLRs成员,它是革兰氏阴性菌胞壁成分脂多糖（lipopolysaccharide, LPS）的识别受体。LPS与TLR4在胞外结合后,通过MyD88依赖和非依赖途径引起炎症反应。TLR4在生殖道炎症反应中具有重要作用,临床上可通过对利用TLR4信号转导通路的阻断来获得治疗效果。作者概述了TLR4的分子结构、组织表达和细胞内信号转导,以及在生殖系统病变状态下的表达,在此基础上提出了TLR4在生殖临床医学应用上可能的意义。     

Toll样受体及其对肠黏膜免疫的调节作用

Toll样受体(TLRs)是近年来备受关注的一种模式识别受体,在脊椎与非脊椎动物中具有病原体传感器的功能。TLRs对体内外特异性配体的识别是启动先天免疫的基础,并迅速增加对抗入侵病原体的保护性反应,最终激活适应性免疫。TLRs在肠道免疫对病原菌与益生菌的区分过程中发挥重要作用,同时TLRs可调控动物肠道上皮分泌抗菌肽杀灭病原菌,对肠道健康具有积极的作用。本文介绍了TLRs的种类、配体及相应的信号通路,探讨TLRs在肠道免疫调节中的关键作用。     

禽类Toll样受体研究进展

Toll样受体(TLRs)是模式识别受体家族的一员,在天然免疫反应中扮演重要角色。在禽类中至今已发现10种TLRs。研究表明在机体感染病原体时,将激活TLRs并引起一系列抗病毒相关因子的上调,参与宿主抗感染免疫反应。论文主要介绍禽类TLRs的基本特征及抗病毒信号转导通路,同时对TLRs的抗微生物作用进行分析,为TLRs的分子机制研究提供参考。     

Immunity and inflammation in the uterus

Microbes often infect the uterus and particularly the endometrium of animals. Infections are most commonly associated with natural service, pregnancy and the post-partum period, leading to inflammation with the elaboration of cytokines, chemokines and prostaglandins. Clinical diseases such as metritis, endometritis and abortion are important causes of infertility. The adaptive immune response to infection has been characterized previously, so the present review aims to highlight the emerging role for innate immunity in the endometrium. The detection of microbes and the innate immune response depends on the detection of pathogen-associated molecular patterns by pattern recognition receptors. The main families of pattern recognition receptors are Toll-like receptors (TLRs), nucleotide oligomerization domain-like receptors, retinoic acid-inducible gene I-like receptors and C-type lectin receptors. These receptors are most often expressed by hematopoietic cells, but the epithelial and stromal cells of the endometrium also possess functional receptors. For example, endometrial cells express TLR4 for recognition of the lipopolysaccharide endotoxin of Gram-negative bacteria, leading to secretion of IL-6, IL-8 and prostaglandin E(2) . It is likely that the epithelial and stromal cells provide a first line of defence in the endometrium to alert hematopoietic cells to the presence of microbes within the uterus.     

Identification of Card15/Nod2 mRNA in intestinal tissue of experimentally induced colitis in rats

Card15/Nod2 has been suggested to be an intracellular pathogen-associated molecular pattern (PAMPs) recognition molecule, which contains a leucine-rich repeat region similar to the Toll-like receptors (TLRs). Card15/Nod2 gene variants play an important role in the susceptibility to Crohn's disease. In this study, we examined the kinetics of Card15/Nod2 expression in intestinal tissue during inflammation in the 2, 4, 6-trinitrobenzenesulfonic acid (TNBS)-treated rat experimental colitis model. At 2 and 4 days after TNBS administration, the mononuclear cells remarkably infiltrated the mucosal layer and tunica muscularis, which was followed by a gradual decrease to resting levels at 14 days after TNBS administration. Card15/Nod2 mRNA expression increased and peaked at 4 days after the TNBS administration, followed by a gradual decrease in accordance with the amelioration of the inflammatory response. Expressions of Tlr2, Tlr4 and Myd88 were also upregulated in the inflamed colonic region, and in an in situ hybridization study, a positive signal for Card15/Nod2 was observed in the crypt of the epithelial cell layer and in the infiltrated cells of the submucosal and myenteric regions. These results suggest that in addition to the TLR recognition systems, Card15/Nod2 may contribute to the inflammatory process not only in the epithelial and submucosal layers but also in the tunica muscularis.     

Characterisation of antibodies to bovine Toll-like receptor (TLR)-2 and cross-reactivity with ovine TLR2

Host recognition of conserved pathogen-associated molecular patterns (PAMPs) and their interactions with pattern-recognition receptors, including the Toll-like receptors (TLR) is essential for innate immune response induction. The TLR1 family (TLR1, 2, 6 and 10) is involved in the recognition of gram-positive and gram-negative bacteria and heterodimers of TLR1 or TLR6 with TLR2 are crucial for the identification of several PAMPs. Studies on cell surface expression of TLR in ruminants are hampered by the lack of specific antibodies and no convincingly cross-reactive anti-human antibodies have been described so far. We describe herein four antibodies which recognise bovine TLR2. Differences in TLR2 expression were evident on bovine antigen presenting cells with high level expression on peripheral blood monocytes and monocyte-derived macrophages. Lower levels of expression were evident on dendritic cell populations derived in vitro and ex vivo, and on alveolar macrophages. One of the antibodies recognised TLR2 expression on ovine peripheral blood monocytes. The identification of antibodies specific for bovine and ovine TLR2 will facilitate studies of the role of this important PRR in the initiation of immune responses to important pathogens.     

Expression and function of TLR2, TLR4, and Nod2 in primary canine colonic epithelial cells

The gut maintains a delicate balance between the downregulation of inflammatory reactions to commensal bacteria and the capacity to respond to pathogens with vigorous cellular and humoral immune responses. Intestinal epithelial cells, including colonic epithelial cells (CECs) possess many properties of cells of the innate immune system, in particular the ability to recognize and respond to microbial antigens. Recognition of microorganisms by CECs is based upon their recognition of signature molecules, called microbe-associated molecular patterns (MAMP), by pattern recognition receptors (PRR) including membrane toll-like receptors (TLR) and cytosolic Nod2, an intracellular counterpart of TLRs. The purpose of this study was to determine whether primary CECs from normal dogs express a functional TLR2, TLR4, and Nod2 and whether they are regulated by inflammatory mediators. We show that canine primary CECs express TLR2, TLR4, and Nod2 that can be modulated in response to their respective MAMPs, lipopolysaccharides (LPS) or peptidoglycans (PGN). Furthermore, we demonstrate that these receptors are functional as evidenced by the induction of cytokine gene expression in response to LPS or PGN.     

Isolation,characterization, and functional analysis of ferret lymphatic endothelial cells

The lymphatic endothelium (LE) serves as a conduit for transport of immune cells and soluble antigens from peripheral tissues to draining lymph nodes (LNs), contributing to development of host immune responses and possibly dissemination of microbes. Lymphatic endothelial cells (LECs) are major constituents of the lymphatic endothelium. These specialized cells could play important roles in initiation of host innate immune responses through sensing of pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors (PRRs), including toll-like receptors (TLRs). LECs secrete pro-inflammatory cytokines and chemokines to create local inflammatory conditions for recruitment of naïve antigen presenting cells (APCs) such as dendritic cells (DCs) to sites of infection and/or vaccine administration. In this study, we examined the innate immune potential of primary LEC populations derived from multiple tissues of an animal model for human infectious diseases – the ferret. We generated a total of six primary LEC populations from lung, tracheal, and mesenteric LN tissues from three different ferrets. Standard RT-PCR characterization of these primary LECs showed that they varied in their expression of LEC markers. The ferret LECs were examined for their ability to respond to poly I:C (TLR3 and RIG-I ligand) and other known TLR ligands as measured by production of proinflammatory cytokine (IFNα, IL6, IL10, Mx1, and TNFα) and chemokine (CCL5, CCL20, and CXCL10) mRNAs using real time RT-PCR. Poly I:C exposure induced robust proinflammatory responses by all of the primary ferret LECs. Chemotaxis was performed to determine the functional activity of CCL20 produced by the primary lung LECs and showed that the LEC-derived CCL20 was abundant and functional. Taken together, our results continue to reveal the innate immune potential of primary LECs during pathogen-host interactions and expand our understanding of the roles LECs might play in health and disease in animal models.     

轮状病毒感染肠上皮细胞激活宿主先天免疫研究进展

轮状病毒(RV)是引起婴幼儿、幼畜禽急性肠胃炎的人畜共患病原,常与其他病原体混合感染,多以呕吐,严重水样腹泻,脱水为临床症状,感染后具有较高病死率,对人类公共卫生以及养殖业造成极大危害.RV病原相关分子模式(PAMP)可被肠上皮细胞(IECs)中一组可遗传的模式识别受体(PRR)识别,通过IECs、先天免疫细胞与RV互...