TOLL-like receptors linking innate and adaptive immune response

Invading pathogens are controlled by the innate and adaptive arms of the immune system. Adaptive immunity, which is mediated by B and T lymphocytes, recognises pathogens by rearranged high affinity receptors. However, the establishment of adaptive immunity is often not rapid enough to eradicate microorganisms as it involves cell proliferation, gene activation and protein synthesis. More rapid defense mechanisms are provided by innate immunity, which recognises invading pathogens by germ-line-encoded pattern recognition receptors (PRR). Recent evidence shows that this recognition can mainly be attributed to the family of TOLL-like receptors (TLR). Binding of pathogen-associated molecular patterns (PAMP) to TLR induces the production of reactive oxygen and nitrogen intermediates (ROI and RNI), pro-inflammatory cytokines, and up-regulates expression of co-stimulatory molecules, subsequently initiating the adaptive immunity. In this review, we will summarize the discovery and the critical roles of the TLR family in host defense, briefly allude to signaling mechanisms mediating the response to TLR ligands, and will provide an update on current knowledge regarding the ligand specificity of these receptors and their role in immunity of domestic animals, particularly cattle.     

Interaction of antigen presenting cells with mycobacteria

The interaction of mycobacteria with antigen presenting cells is a key feature in the pathogenesis of tuberculosis and the outcome of this interaction is pivotal in determining whether immunity or disease ensues. Human and mouse macrophages and dendritic cells (DC) have been shown to become infected with mycobacteria and to produce a response to infection that reflects their suggested role in immunity. Thus, macrophages elicit anti-microbial mechanisms for elimination of mycobacteria and DC up-regulate expression of molecules that aid their stimulation of T lymphocytes. We have examined the effects of infection with the avirulent strain Mycobacterium bovis BCG and with virulent M. bovis on bovine antigen presenting cells. Differences in the intracellular survival of bacteria within DC and macrophages were observed with higher numbers of bacteria maintained within DC following infection compared to macrophages. BCG was killed more effectively than M. bovis. Alterations in the expression of cell surface molecules involved in antigen presentation and the stimulation of T cells, including MHC II and CD40, were observed following infection of bovine antigen presenting cells. In addition infected DC secreted IL-12, TNF and IL-10 whereas macrophages produced TNF, IL-10 and little IL-12. Generally responses were more marked when virulent M. bovis was used compared to BCG. These studies indicate that infection of bovine antigen presenting cells by mycobacterial species results in the induction of both innate and adaptive immune responses that are critical for the outcome of infection.     

抗菌肽在哺乳动物防御系统中的作用

抗菌肽是近年来发现的广泛存在于自然界的一类阳离子抗菌活性肽，它们在宿主先天性免疫和适应性免疫中有重要作用。多数抗菌肽具有分子小、带正电、两亲性、抗菌谱广等共同特点。防御素和calhelicidins是哺乳动物的两大主要抗菌肽家族，它们通过抵抗致病菌入侵为宿主提供了第一道防线而对宿主具有先天的抗菌防御功能，其中一些多肽对未分化的树突状细胞、淋巴细胞有趋化性，另外还有诱导细胞因子生成、肥大细胞脱粒等作用，从而表明这些多肽能动员并增强宿主的先天性免疫和适应性免疫。本文主要对哺乳动物抗菌肽的一般性质、基因及其表达、在宿主防御中的作用、作用机理及研究前景进行了概述。     

Immune response to fungal infections

The immune mechanisms of defence against fungal infections are numerous, and range from protective mechanisms that were present early in evolution (innate immunity) to sophisticated adaptive mechanisms that are induced specifically during infection and disease (adaptive immunity). The first-line innate mechanism is the presence of physical barriers in the form of skin and mucous membranes, which is complemented by cell membranes, cellular receptors and humoral factors. There has been a debate about the relative contribution of humoral and cellular immunity to host defence against fungal infections. For a long time it was considered that cell-mediated immunity (CMI) was important, but humoral immunity had little or no role. However, it is accepted now that CMI is the main mechanism of defence, but that certain types of antibody response are protective. In general, Th1-type CMI is required for clearance of a fungal infection, while Th2 immunity usually results in susceptibility to infection. Aspergillosis, which is a disease caused by the fungus Aspergillus, has been the subject of many studies, including details of the immune response. Attempts to relate aspergillosis to some form of immunosuppression in animals, as is the case with humans, have not been successful to date. The defence against Aspergillus is based on recognition of the pathogen, a rapidly deployed and highly effective innate effector phase, and a delayed but robust adaptive effector phase. Candida albicans, part of the normal microbial flora associated with mucous surfaces, can be present as congenital candidiasis or as acquired defects of cell-mediated immunity. Resistance to this yeast is associated with Th1 CMI, whereas Th2 immunity is associated with susceptibility to systemic infection. Dermatophytes produce skin alterations in humans and other animals, and the essential role of the CMI response is to destroy the fungi and produce an immunoprotective status against re-infection. The resolution of the disease is associated with a delayed hypersensitive response. There are many effective veterinary vaccines against dermatophytoses. Malassezia pachydermatis is an opportunistic yeast that needs predisposing factors to cause disease, often related to an atopic status in the animal. Two species can be differentiated within the genus Cryptococcus with immunologic consequences: C. neoformans infects predominantly immunocompromised hosts, and C. gattii infects non-immunocompromised hosts. Pneumocystis is a fungus that infects only immunosupressed individuals, inducing a host defence mechanism similar to that induced by other fungal pathogens, such as Aspergillus.     

Toll样受体研究进展

Toll样受体(Toll-like receptors, TLRs)家族在病原体的识别和激活天然免疫方面发挥着非常重要的作用.激活TLRs不仅可以诱导天然免疫应答,而且有利于特异性免疫反应的发生,因而TLRs是天然免疫与获得性免疫之间的桥梁.不同TLRs可以有相同的功能,例如诱导炎性因子的产生或者上调辅助刺激分子的表达,也可以有特异的作用,例如具有诱导IFN-I产生的能力.这些作用不但在抗菌免疫反应中非常关键,而且也表现在自身免疫反应中.因而了解TLRs结构、分布、分类及功能可促进天然免疫机制研究的进一步深入,有利于对变态反应和自身免疫性疾病治疗措施的改进,也有利于解决诸如 CpG佐剂、DNA疫苗、预防过敏反应等实际应用过程中存在的问题.     

How the bovine viral diarrhea virus outwits the immune system

The interaction of bovine viral diarrhea virus (BVD virus) with its host has several unique features, most notably the capacity to infect its host either transiently or persistently. The transient infection stimulates an antiviral immune reaction similar to that seen in other transient viral infections. In contrast, being associated with immunotolerance specific for the infecting BVD viral strain, the persistent infection differs fundamentally from other persistent infections like those caused by lentiviruses. Whereas the latter are characterized by complex viral evasion of the host's adaptive immune response by mechanisms such as antigenic drift and interference with presentation of T cell epitopes, BVD virus avoids the immune response altogether by inducing both humoral and cellular immune tolerance. This is made possible by invasion of the fetus at an early stage of development. In addition to adaptive immunity, BVD virus also manipulates key elements of the host's innate immune response. The non-cytopathic biotype of BVD virus, which is capable of persistently infecting its host, fails to induce type I interferon. In addition, persistently infected cells are resistant to the induction of apoptosis by double-stranded RNA and do not produce interferon when treated with this pathogen-associated molecular pattern (PAMP) that signals viral infection. Moreover, when treated with interferon, cells persistently infected with non-cytopathic BVD virus do not clear the virus. Surprisingly, however, despite this lack of effect on persistent infection, interferon readily induces an antiviral state in these cells, as shown by the protection against infection by unrelated viruses. Overall, BVD virus manipulates the host's interferon defense in a manner that optimises its chances of maintaining the persistent infection as well as decreasing the risks that heterologous viral infections may carry for the host. Thus, since not all potential host cells are infected in animals persistently infected with BVD virus, heterologous viruses replicating in cells uninfected with BVD virus will still trigger production of interferon. Interferon produced by such cells will curtail the replication of heterologous viruses only, be that in cells already infected with BVD virus, or in cells in which the heterologous virus may replicate alone. From an evolutionary viewpoint, this strategy clearly enhances the chances of transmission of BVD virus to new hosts, as it attenuates the negative effects that a global immunosuppression would have on the survival of persistently infected animals.     

Influence of Enterococcus faecium SF68 Probiotic on Giardiasis in Dogs

Background: Giardiasis is a common, potentially zoonotic disease, and dogs often harbor and shed cysts without showing clinical signs. Treatment with the probiotic Enterococcus faecium SF68 has been shown to stimulate mucosal and systemic immunity in a variety of animal models and in young dogs, and to reduce giardial cyst and antigen shedding in rodents.
Hypothesis: Adult dogs with chronic naturally acquired giardiasis will have decreased giardial fecal cyst and antigen shedding and increased innate and adaptive immunity after 6 weeks probiotic treatment with E. faecium SF68.
Animals: Twenty adult dogs.
Methods: After a 6-week dietary equilibration period, dogs were randomized to receive E. faecium SF68 or placebo for 6 weeks, and then crossed over to the alternate treatment. We measured cyst shedding, fecal giardial antigen, fecal immunoglobulin A (IgA) concentration, and circulating leukocyte phagocytic activity at multiple timepoints to determine the effect of E. faecium SF68 on giardiasis and immune responses in these dogs.
Results: No differences were observed between placebo or E. faecium SF68 treatment for giardial cyst shedding, fecal antigen shedding, fecal IgA concentration, or leukocyte phagocytic activity.
Conclusions: Short-term treatment with E. faecium SF68 of dogs with chronic naturally acquired subclinical giardiasis fails to affect giardial cyst shedding or antigen content and does not alter innate or adaptive immune responses.     

Immunity in the mammary gland.

The ruminant mammary gland is an extremely important economic organ in that it provides a major nutrition source for a significant portion of the world's human population. The ruminant mammary gland is also responsible for providing protective immunity to neonates and for defending itself from invading pathogens. A wide array of humoral and cellular immune mechanisms are present in the mammary gland and actively participate in providing immunity to newborns and the mammary gland per se. The acute inflammatory response is essential in determining the outcome of intramammary challenge, and factors affecting innate and adaptive immunity in the context of mammary health are reviewed in detail. The ruminant mammary gland is also unique in that lymphocyte trafficking, which is essential to adaptive immunity, is shared with the peripheral immune system rather than the common mucosal immune system.     

Bovine uterine defense mechanisms: a review

Bovine uterine defense mechanisms during physiological and pathological conditions have been reviewed in this article. The initial uterine defense against bacterial infection is phagocytosis by uterine leucocytes (mainly neutrophils). The reported literature showed that very little work has been done on immunoglobulins and their role in the bovine uterine defense mechanisms; however, some investigators have found a positive correlation between gamma-globulin and the development of uterine infection after calving. Many explanations exist for the difference in susceptibility of the uterus to infection during the different phases of estrous cycle; however, most of the reports agreed that the uterine defense mechanism is inadequate during diestrus. The abnormal puerperium effects uterine defense mechanisms adversely and prolongs the time to complete uterine involution. Future treatment may utilise natural antimicrobial substances such as proteins or peptides derived from PMN, chemoattractant substances such as E. coli lipopolysaccharide or a bacteria-free filtrate of streptococci. Specific hyperimmunserum could also be used as opsonin for refractory cases of uterine bacterial infections.     

Immunoglobulin structure and function: genetic control of antibody diversity

Recent studies of the molecular biological characteristics of lymphoid cells have markedly increased our understanding of how millions of different antibodies can be synthesized by an individual mammal. In particular, studies have shown how antibody genes are arranged and rearranged within B-lymphocyte clones to provide each cell clone with antibody of defined specificity for antigen. The process involves the assembly, from disparate genetic elements, of a complete antibody gene that will code for an antibody protein. The assembly process, in itself, also provides mechanisms for generating the diversity of antibody variable region structure (that part of an antibody molecule that actually binds antigen) that is essential to a full role for humoral immunity in host defense mechanisms. Specifically, the diversity of structure characteristic of mature antibodies derives from 3 distinct mechanisms: innate variability of germ-line genes; mismatching of individual gene segments during their somatic rearrangement leading to junctional diversity; and somatic mutation in variable region genetic material during or after the rearrangement. Thus, it is now clearly understood that several processes are involved in explaining the origin of the antigen-combining diversity of antibody proteins. Certain "lottery-like" aspects of these genetic processes add to the combinatorial possibilities that are characteristic of the humoral immune system.     

犬的免疫介导疾病的诊断和治疗

我们经常能看到一些犬表现出干燥和皮屑剥落的皮肤,变薄变稀的皮毛,泪流不止,足部过度舔,慢性腹泻,多动,不能长胖或情绪波动.这些症状最常见的诊断是跳蚤过敏性皮炎,环境过敏或饮食中缺乏脂肪酸.通常多作为治疗皮肤疾病的原因前往看兽医.然而,所有这些症状往往指向一种免疫介导缺陷的障碍—不活跃的免疫系统（免疫缺陷）,或过度活跃的免疫系统（自身免疫性疾病）.繁育者将带有隐蔽免疫介导缺陷的犬用于育种繁殖,将带来一些亚临床性质的问题.而一些兽医对此类问题的出现并不很了解,忽略了涉及免疫系统的缺陷和自身免疫性疾病的对症治疗.本文提供了先天的和适应性免疫的实际概述,并介绍这些宿主的防御机制是如何涉及健康和疾病的.     

单核细胞的生物学特性及其应用

物种在长期进化过程中已经形成了比较完善的、抵御环境中病原微生物侵袭与感染的免疫系统,并随着物种的进化而进化。具有多能干细胞特性的外周血单核细胞,在非特异性免疫和特异性免疫过程中发挥着非常重要的作用,既能消灭侵入的病原微生物,又能够对病原微生物进行加工处理,将抗原呈递给其他免疫细胞,架起了先天免疫与获得性免疫的桥梁。单核细胞已经在药物靶向传送、消灭肿瘤、疾病监控、细胞治疗、抗病育种、新型疫苗研制、环境检测等领域发挥了重要作用,并获得了良好的实验效果。     

Immunology of the ocular surface.

The ocular surface immunity is a remarkable combination of the innate immune and adaptive immune systems, designed to prevent microbial invasion while minimizing damage to delicate ocular tissue. The innate immune system uses a variety of methods to minimize microorganism invasion, including mechanical tissue barriers and production of antimicrobial peptides. Tolerance of normal ocular flora is achieved by the presence of a minimal number of professional antigen presenting cells, immunosuppressive substances in tears, and the strategic intra- and intercellular location of the Toll-like receptors. Autoimmune diseases are common on the ocular surface, and with contributions of environmental and genetic factors, autoantigens are presented to the adaptive immune response. Toll-like receptors are the link between the innate and adaptive immune response, and are likely key components of the response of ocular tissue to infectious organisms and in the initiation and perpetuation of autoimmune disease.     

Bovine T cell responses to recombinant thioredoxin of Fasciola hepatica

Fasciolosis is an economically significant disease of ruminants, caused by infection with the digenetic trematodes, Fasciola hepatica and F. gigantica. Some vaccination trials using irradiated metacercariae or isolated proteins have been shown to afford significant protection. However, the mechanisms of specific immunity against this pathogen have not been elucidated. We have identified thioredoxin, a tegument antigen of F. hepatica, among several proteins that are common to both the juvenile and adult fluke within the mammalian host and have undertaken studies to characterize bovine T cell responses to recombinant thioredoxin protein (FH 2020). Peripheral blood mononuclear cells from immune cattle proliferated specifically to crude F. hepatica antigenic extract but not to FH 2020. However, after repeated stimulation of lymphocytes by alternating crude extract and FH 2020, FH 2020-specific proliferation by T cell lines was observed. T cell clones were subsequently generated and found to respond specifically but weakly to both crude antigen and FH 2020. Thioredoxin appears to be only weakly antigenic for bovine T cells and is, therefore, an unpromising candidate for inducing resistance to F. hepatica.     

Antigen dose and humoral immune response correspond with protection for inactivated infectious pancreatic necrosis virus vaccines in Atlantic salmon (Salmo salar L)

An enduring challenge in the vaccinology of infectious pancreatic necrosis virus (IPNV) is the lack of correlation between neutralizing antibodies and protection against mortality. To better understand the immunological basis of vaccine protection, an efficacy trial including Atlantic salmon (Salmo salar L.) vaccinated with a high antigen (HiAg) or low antigen (LoAg) dose vaccine was carried out in a cohabitation challenge model using the highly virulent Norwegian Sp strain NVI015. To pinpoint the immunological basis of vaccine protection, pathogenic mechanisms of IPNV were unraveled in control fish while obtaining feedback on mechanisms of protection in the vaccinated fish. During the incubation period, infection rates were highest in control fish, followed by the LoAg group with the lowest infections being in the HiAg group. Although both the liver and pancreas are target organs prone to tissue damage, infection in the liver was delayed until acute infection in most fish. A correlate of pathology determined as the cutoff threshold of viral copy numbers linked to tissue damage in target organs was estimated at ≥ 10^7.0, which corresponded with an increase in mortality. The kinetics of IFNα and Mx expression suggests that these genes can be used as biomarkers of IPNV infection progression. Mechanisms of vaccine protection involved reducing infection rates, preventing infection of the liver and reducing virus replication in target organs to levels below the correlate of pathology. Overall, the study shows that antigen dose corresponds with vaccine efficacy and that antibody levels can be used as a signature of protective immunity against pathological disease and mortality.     

Chlamydial vaccines

With few exceptions, immunity from chlamydiosis availed by current vaccines is limited and can even be detrimental. Possible reasons for failure include immunotype or strain differences, ill-defined virulence variation, presentation of deleterious antigens, and incorrect presentation of critical antigens to the body. Antigens that stimulate neutralizing antibodies active at 2 steps of infection have been identified. A third step, prevention of phagolysosomal fusion, needs to be further studied, and causal antigens need to be identified. A fourth possible stage for antibody participation is in antibody-dependent cell-mediated cytotoxicity. If chlamydial antigens are expressed on the surface of infected cells, this mechanism of destruction of infected cells and the antigens that elicit it will need to be more fully examined. Cell-mediated immune responses participating in eliminating chlamydial infections need to be further clarified. Activated macrophages are the best characterized effector mechanism of cell-mediated immunity thus far, regardless of the stimulatory cytokines involved. It is important to determine how sensitized lymphocytes recognize antigen(s) that cause them to release macrophage-activating cytokines. It must be determined whether chlamydial antigens are expressed on the surface of infected cells and then recognized by potential cytokine-releasing lymphocytes in context with major histocompatibility antigens (surface expression) or whether they are recognized on antigen-presenting cells functioning in more of a scavenging capacity. Membrane expression of antigen is also important in that it also defines whether cytotoxic T cells and antibody-dependent cell-mediated cytotoxicity have roles in resistance to chlamydial infection. Also, it is important to realize the possible limitation of these mechanisms to systemic sites of the body. If membrane expression does occur, it must be determined how it functions at mucosal sites, whether it occurs at the luminal surface of mucosal epithelial cells only, or whether there is expression of antigens at abluminal membrane surfaces perhaps more accessible to such immune effector mechanisms. Delivery of critical antigens to the individual is the final component in establishing effective vaccines. Carrier systems capable of stimulating long-lasting mucosal and systemic immunity are available and need to be further studied as protective immunogens become available.     

Immunopathogenic pathways in canine inflammatory myopathies resemble human myositis

Progress in the treatment of inflammatory myopathies is impeded by the lack of suitable animal models. Inflammatory myopathies occur spontaneously in the dog, are a heterogeneous group of disorders, and are more common than in humans. Clinical signs of weakness and muscle atrophy are reliably present, and there are histological and immunohistological similarities to forms of human myositis. In this study, microarray technology followed by quantitative real-time PCR and immunohistochemistry on muscle biopsy sections was used to investigate gene expression in cases of canine inflammatory myopathies. Several genes involved with innate and adaptive immunity were highly upregulated including those that participate in macrophage and dendritic cell activation and migration, and antigen processing and presentation. Other genes including those that participate in B cell growth, development, migration and activation, immunoglobulin genes, genes in pro-inflammatory and anti-inflammatory pathways, and genes involved with tissue remodeling were upregulated. In previous reports utilizing microarray technology in human myositis, there was activation of similar pathways involved in the immune response. This study strengthens the argument that forms of canine myositis may be important animal models of human myositis and suggests useful biomarkers for therapeutic response using the dog in pre-clinical trials.     

Antigen presentation in vaccine development

A variety of microorganisms, nutrients or toxins are generally intrude our body through mucosal tissues or skin, where equipment for both preventing their invasions and catching their information to activate internal immune systems for adapting surroundings is arranged. Among the equipment, cells in charge of innate immunity, particularly dendritic cells (DCs), having an excellent capacity for prompt recognition of invaded pathogens via toll-like receptors (TLRs) to alert B and T cells for establishing aquired/adaptive immunity by presenting their processed antigenic fragments, have been paid great attention. These TLR-activated, antigen captured DCs are divided into two groups; one is pathogen-retaining unit and the other is pathogen-controlling unit. The latter DCs present processed antigenic molecules from the pathogens to competent β T cells together with special containers, such as class I, class II MHC and CD1 to generate specific cellular immunity. The former two MHC molecules can present processed peptide antigens, whereas the last CD1 molecule can present glycolipid/lipid antigens. In contrast, B lymphocytes that captured antigens via their specific immunoglobulin (Ig) receptors present digested peptide fragments with their class II MHC to stimulate suitable CD4⁺ helper T cells which in turn secrete various cytokines to efficiently expand and maintain antibody production from that partner B cells to establish humoral immunity. These β T cells and antibodies, recognize either processed antigenic peptide or glycolipid fragments, and thus, identification of these epitopes enables us to generate artificial pathogen-specific vaccines. Based on the recent findings about precise mechanisms of antigen processing and presentation orchestrated at the surface compartment, future development of vaccines against various pathogens are discussed.     

Antimicrobial peptides and surfactant proteins in ruminant respiratory tract disease

In ruminants, respiratory disease is multifactorial and a leading cause of morbidity and mortality. Pulmonary innate immunity is the first line of defense for the respiratory tract. Alteration of regulation, expression, and function of these factors may be important to disease development and resolution. Many antimicrobial peptides and surfactant proteins are constitutively expressed in the respiratory tract and expression levels are regulated. Beta-defensins are cationic peptides with broad antimicrobial activity against bacterial, viral and fungal pathogens. Beta-defensins are primarily expressed in mucosal epithelia (and in some species leukocytes); where they may also participate in chemotaxis, wound repair and adaptive immune responses. Surfactant proteins A and D are secreted pulmonary surfactant proteins that have antimicrobial and immune regulatory activity. Anionic peptide is a constitutively expressed, aspartate-rich peptide that has antimicrobial activity and is most prominent during reparative epithelial hyperplasia. Regulation of these immune defense components by stress, pathogens, and inflammatory cytokines may play a role in the susceptibility to, severity and resolution of respiratory infection. The expression patterns of these molecules can be specific for host-species, class of pathogen and stage of infection. Understanding the regulation of antimicrobial peptide/protein expression will further enhance the potential for novel prophylactic and therapeutic modalities to minimize the impact of respiratory disease.