鸡消化道的重要天然免疫效应分子-抗菌肽

抗菌肽广泛分布于动植物及昆虫体内,是天然免疫（非特异性免疫）防御系统的重要组成部分。抗菌肽不仅有广谱抗细菌能力,还可以抗真菌、被膜病毒等多种微生物及抗寄生虫,而且对一些恶性肿瘤细胞和艾滋病毒等也有杀伤作用。动物内源性抗菌肽是基因编码的阳离子多肽,在机体组织中分布广泛,尤其是那些与微生物密切接触的上皮组织如消化道、呼吸道、生殖道粘膜上皮和皮肤,抗菌肽在机体局部抗感染与免疫方面发挥着重要作用。     

Viral deubiquitinases and innate antiviral immune response in livestock and poultry

Among many of the pathogens, virus is the main cause of diseases in livestock and poultry. A host infected with the virus triggers a series of innate and adaptive immunity. The realization of innate immune responses involves the participation of a series of protein molecules in host cells, including receptors, signal molecules and antiviral molecules. Post-translational modification of cellular proteins by ubiquitin regulates numerous cellular processes, including innate immune responses. Ubiquitin-mediated control over these processes can be reversed by cellular or viral deubiquitinases (DUBs). DUBs have now been identified in diverse viral lineages, and their characterization is providing valuable insights into virus biology and the role of the ubiquitin system in host antiviral mechanisms. In this review, we briefly introduce the mechanisms of ubiquitination and deubiquitination, present antiviral innate immune response and its regulation by ubiquitin, and summarize the prevalence of DUBs encoded by viruses (Arteriviridae, Asfarviridae, Nairoviridae, Coronaviridae, Herpesviridae, and Picornaviridae) infecting domestic animals and poultry. It is found that these DUBs suppress the innate immune responses mainly by affecting the production of type I interferon (IFN), which causes immune evasion of the viruses and promotes their replication. These findings have important reference significance for understanding the virulence and immune evasion mechanisms of the relevant viruses, and thus for the development of more effective prevention and treatment measures.     

血红蛋白β亚基:一种多效性抗病毒天然免疫调节因子

血红蛋白是动物体内最重要的呼吸蛋白之一。在脊椎动物的红细胞内含量尤其丰富,可与氧分子呈可逆性结合。随着对血红蛋白研究的不断深入,相继有研究报道血红蛋白还具有储存能量、维持渗透压、抗菌和发挥类酚氧化酶活性等多种功能,现已成为研究蛋白质多重生物学功能的理想模式分子之一。近年来的研究发现,血红蛋白在抵抗病原体入侵、调控机体天然免疫应答过程中也发挥了重要作用,但其相关机制仍有待进一步阐明。     

Fowl adenoviruse-4 infection induces strong innate immune responses in chicken

Fowl adenovirus (FAdV), as the causative agent of hepatitis-hydropericardium syndrome (HHS), poses a significant threat to the poultry industry in China in recent years. In this study, we investigated the immunopathogenesis of a FAdV-4 strain HN/151025 in 60-day-old chickens. The virus was highly virulent in chickens, with a broader tissue tropism in chickens, causing 60 % mortality. Postmortem findings of dead chickens showed mild HHS and liver degeneration and necrosis. Importantly, FAdV-4 infection induced significant upregulation of genes encoding most toll-like receptors, some cytokines (interleukin-1β, 2, 6, 8, and 18, and interferon-γ), most of avian β-defensins, myeloid differentiation primary response protein 88, p38 mitogen-activated protein kinases, and inducible nitric oxide synthase, in tissues of infected chicken, especially in spleen and bursa of Fabricius. There was also a significant positive correlation between FAdV-4 genome load and the mRNA expression levels of most of these factors in specific infected tissues. The results indicated the potential role of these proteins in host immune response against FAdV-4 infection. However, overexpression of these proteins might contribute to tissue damage of FAdV-4 infected chickens, and eventually lead to chicken death.     

The innate antiviral immune system of the cat: molecular tools for the measurement of its state of activation

The innate immune system plays a central role in host defence against viruses. While many studies portray mechanisms in early antiviral immune responses of humans and mice, much remains to be discovered about these mechanisms in the cat. With the objective of shedding light on early host-virus interactions in felids, we have developed 12 real-time TaqMan(?) qPCR systems for feline genes relevant to innate responses to viral infection, including those encoding for various IFNα and IFNω subtypes, IFNβ, intracellular antiviral factor Mx, NK cell stimulator IL-15 and effectors perforin and granzyme B, as well as Toll-like receptors (TLRs) 3 and 8. Using these newly developed assays and others previously described, we measured the relative expression of selected markers at early time points after viral infection in vitro and in vivo. Feline embryonic fibroblasts (FEA) inoculated with feline leukemia virus (FeLV) indicated peak levels of IFNα, IFNβ and Mx expression already 6h after infection. In contrast, Crandell-Rees feline kidney (CrFK) cells inoculated with feline herpes virus (FHV) responded to infection with high levels of IFNα and IFNβ only after 24h, and no induction of Mx could be detected. In feline PBMCs challenged in vitro with feline immunodeficiency virus (FIV), maximal expression levels of IFNα, β and ω subtype genes as well as IL-15 and TLRs 3, 7 and 8 were measured between 12 and 24h after infection, whereas expression levels of proinflammatory cytokine gene IL-6 were consistently downregulated until 48h post inoculation. A marginal upregulation of granzyme B was also observed within 3h after infection. In an in vivo experiment, cats challenged with FIV exhibited a 2.4-fold increase in IFNα expression in blood 1 week post infection. We furthermore demonstrate the possibility of stimulating feline immune cells in vitro with various immune response modifiers (IRMs) already known for their immunostimulatory properties in mice and humans, namely Poly IC, Resiquimod (R-848) and dSLIM?, a synthetic oligonucleotide containing several unmethylated CpG motifs. Stimulation of feline PBMCs with dSLIM? and R-848 effectively enhanced expression of IFNα within 12h by factors of 6 and 12, respectively, and Poly IC induced an increase in Mx mRNA expression of 28-fold. Altogether, we describe new molecular tools and their successful use for the characterization of innate immune responses against viruses in the cat and provide evidence that feline cells can be stimulated by synthetic molecules to enhance their antiviral defence mechanisms.     

铅镉及铅镉联合胁迫致雏鸡红细胞免疫功能的影响

自红细胞免疫系统的新概念提出后[1],已证实红细胞有免疫粘附、杀伤抗原、清除循环免疫复合物[2]等多种功能,是机体免疫系统中的重要组成部分.铅、镉是极其重要的工业和环境污染物,均可造成免疫系统的损害,且铅和镉对环境的污染往往联合在一起.目前关于铅、镉单独对红细胞免疫功能影响的研究较多[6-9],而关于铅镉联合尤其是致禽类红细胞免疫功能的研究未见报道.     

Muscovy duck reovirus infection rapidly activates host innate immune signaling and induces an effective antiviral immune response involving critical interferons

Muscovy duck reovirus (MDRV) is a highly pathogenic virus in waterfowl and causes significant economic loss in the poultry industry worldwide. Because the host innate immunity plays a key role in defending against virus invasion, more and more attentions have been paid to the immune response triggered by viral infection. Here we found that the genomic RNA of MDRV was able to rapidly induce the production of interferons (IFNs) in host. Mechanistically, MDRV infection induced robust expression of IFNs in host mainly through RIG-I, MDA5 and TLR3-dependent signaling pathways. In addition, we observed that silencing VISA expression in 293T cells could significantly inhibit the secretion of IFNs. Remarkably, the production of IFNs was reduced by inhibiting the activation of NF-κB or knocking down the expression of IRF-7. Furthermore, our study showed that treatment of 293T cells and Muscovy duck embryo fibroblasts with IFNs markedly impaired MDRV replication, suggesting that these IFNs play an important role in antiviral response during the MDRV infection. Importantly, we also detected the induced expression of RIG-I, MDA5, TLR3 and type I IFN in Muscovy ducks infected with MDRV at different time points post infection. The results from in vivo studies were consistent with those in 293T cells infected with MDRV. Taken together, our findings reveal that the host can resist MDRV invasion by activating innate immune response involving RIG-I, MDA5 and TLR3-dependent signaling pathways that govern IFN production.     

免疫鸡群为何还会发病

目前,我国养禽业已由分散、零星、粗放型的放养逐渐向规模化、集约化的饲养转变,但由于综合防制技术落后,免疫监测手段滞后,免疫程序不当,老病未消灭,新病又不断出现等原因,严重威胁着养禽业的健康发展。笔者针对目     

Impact of nutrition on the innate immune response to infection in poultry

Viral and bacterial diseases remain a threat to the poultry industry and countermeasures to prevent and control them are needed due to production losses. With the continued threat of exotic and emerging diseases and concern over the use of antibiotics in animal production, there is a serious and urgent need to find safe and practical alternatives to prevent or control pathogens. Identification of new tools for the design of new immunological interventions or therapeutic antimicrobials to reduce microbial pathogens in poultry is now required more than ever. Immunological interventions to reduce microbial pathogens in poultry would be of great value to the poultry industry and to the consumer. We have been advocating boosting immunity and encouraging the host to utilize its innate immune system to control and clear infections. Our research has addressed the use of innate immune mechanisms and components to develop new immune modulators (prophylactic and therapeutic) and the characterization and production of antimicrobial peptides as potential immune modulators in poultry. Dietary bioactive food components that interact with the immune response have considerable potential to reduce susceptibility to infectious diseases. With this premise, this paper asks and answers a series of pertinent questions on the utilization of avian immunity for increasing resistance to a variety of potential pathogens problematic in today's commercial poultry industry. Using experimental data to provide answers to these questions, we hope to stimulate a dialog between avian immunologists and nutritionists that results in coordinating and integrating their expertise into specific practical solutions that will benefit the industry and improve the well-being of commercial poultry.     

The innate immune response of the bovine mammary gland to bacterial infection

Intra-mammary (IM) bacterial infection in cattle can result in clinical outcomes that range from being acute and life-threatening to those that are chronic and sub-clinical. The typical bacteria involved in IM bacterial infections activate the mammary immune system in different ways which can influence the severity of the outcome. A clear understanding of the mechanisms that activate and regulate this response is central to the development of effective preventative and treatment regimes. This review focuses on the different immune responses of the bovine mammary gland to common mastitis-causing pathogens. There is special emphasis on comparing the responses to Escherichia coli and Staphylococcus aureus infections, as these are typically associated, respectively, with acute/severe and chronic/sub-clinical forms of the disease.     

Aspects of the innate and adaptive immune responses to acute infections with BVDV

The immune response can be divided into innate and adaptive components that synergise to effect the clearance of pathogens. Recently, it has been realised that these arms of the immune system do not act independently, the magnitude and quality of the adaptive response is dependent on signals derived from the innate response. Here, we review the innate immune responses to bovine viral diarrhoea virus infections of cattle and relate these changes to immunosuppression and the subsequent development of the adaptive immune response.     

Porcine colon explants in the study of innate immune response to Entamoeba histolytica

Human amebiasis is caused by the protozoan Entamoeba histolytica. This protozoan is responsible for muco-hemorrhagic diarrhoea and liver abscess in affected populations. E. histolytica can be asymptomatic commensally confined to the intestinal lumen or can result in invasion of the colonic mucosa leading to ulceration and/or liver abscesses. Recently, human colonic explants have been identified as valuable in the study of host-parasite interactions. Here we investigated the potential of porcine colonic explants as an alternative to human tissues which are far less available. Porcine colonic explants were cultured with two strains of E. histolytica, one virulent (HM1:IMSS) and one avirulent (Rahman). Results from histopathological and real-time PCR analysis showed that porcine explants cultured with virulent ameba trophozoites react similarly to their human counterparts with an invasion of the tissue by the trophozoites and the triggering of typical innate immune response against the parasite. On the contrary, explants cultured with avirulent ameba trophozoites were preserved. The study open the way to the use of porcine colonic explants in the study of the complex interactions between the parasite and the host.     

影响鸡免疫应答因素分析

鸡的免疫类型分非特异性免疫和特异性免疫.特异性免疫又分为细胞免疫和体液免疫.细胞免疫是指来自于骨髓中的淋巴细胞在胸腺依赖区中被诱导分化成熟为T细胞,在受到抗原或有丝分裂源的刺激后,分化增殖转化为致敏淋巴细胞所表现出来特异性免疫.这类免疫应答不能通过血清传递,只能通过致敏淋巴细胞所传递.体液免疫是指来自于骨髓的淋巴细胞在法氏囊依赖区诱导分化为成熟的B细胞.在抗原物质的刺激下产生抗体以及血液性抗体与相应抗原接触后引起一系列抗原抗体反应的免疫应答方式[1].分析和掌握影响鸡免疫应答因素,对于制定适合现地养禽场的免疫程序和选择合理疫苗,保证该养禽场生物安全生产具有重要意义.     

Evaluation of innate immune responses in bovine forestomachs

Previous studies had indicated an active role of bovine forestomachs in the response to alimentary disorders as well as to inflammatory and infectious processes in both the gastro-intestinal (GI) tract and elsewhere. We investigated the potential of bovine forestomachs to receive, elaborate and produce signals and mediators of the innate immune response. Indeed, we detected the expression of Toll IL-1R8/single Ig IL-1-related receptor (TIR8/SIGIRR) and other receptors and cytokines, such as Toll-like receptor (TLR)4, interleukin (IL)-1β, IL-10 and Caspase-1 in the forestomach walls of healthy cows. Their presence suggests an active role of forestomachs in inflammatory disorders of the GI tract and other body compartments. Moreover, interferon (IFN)-γ was revealed in ruminal content. We confirmed and further characterized the presence of leukocytes in the rumen fluids. In particular, T-, B-lymphocytes and myeloid lineage cells were detected in the ruminal content of both rumen-fistulated heifers and diseased cows. An acidogenic diet based on daily supplements of maize was shown to inhibit leukocyte accumulation, as opposed to a control, hay-based diet, with or without a soy flour (protein) supplement. On the whole, results indicate that bovine forestomachs can receive and elaborate signals for the immune cells infiltrating the rumen content or other organs. Forestomachs can thus participate in a cross-talk with the lymphoid tissues in the oral cavity and promote regulatory actions at both regional and systemic levels; these might include the control of dry matter intake as a function of fundamental metabolic requirements of ruminants.     

Deciphering the involvement of innate immune factors in the development of the host response to PRRSV vaccination

The natural response of pigs to porcine reproductive and respiratory syndrome virus (PRRSV) infections and vaccinations needs to be altered so that better protection is afforded against both homologous and heterologous challenges by this pathogen. To address this problem, real-time gene expression assays were coupled with cytokine Elispot and protein analyses to assess the nature of the anti-PRRSV response of pigs immunized with modified live virus (MLV) vaccine. Although T helper 1 (Th1) immunity was elicited in all vaccinated animals, as evidenced by the genesis of PRRSV-specific interferon-gamma secreting cells (IFNG SC), the overall extent of the memory response was variable and generally weak. Peripheral blood mononuclear cells (PBMC) isolated from these pigs responded to PRRSV exposure with a limited increase in their expression of the Th1 immune markers, IFNG, tumor necrosis factor-alpha and interleukin-15 (IL15), and a reduction in the quantity of mRNAs encoding the innate and inflammatory proteins, IL1B, IL8 and IFNA. Efforts to enhance Th1 immunity, by utilizing an expression plasmid encoding porcine IFNA (pINA) as an adjuvant, resulted in a temporary increase in the frequency of PRRSV-specific IFNG SC but only minor changes overall in the expression of Th1 associated cytokine or innate immune marker mRNA by virus-stimulated PBMC. Administration of pINA, however, did correlate with decreased IL1B secretion by cultured, unstimulated PBMC but had no effect on their ability to release IFNG. Thus, while exogenous addition of IFNA during PRRSV vaccination has an impact on the development of a Th1 immune response, other alterations will be required for substantial boosting of virus-specific protection.     

鸡干扰素刺激基因12(ISG12)的克隆表达及抗病毒活性分析

干扰素刺激基因12(interferon-stimulated gene 12,ISG12)在病毒感染过程中已经被监测到,但是针对其抗病毒活性的研究却很少。本研究的主要目的是体外表达鸡的ISG12基因,以禽流感病毒(avian influenza viru,AIV)1215株为病毒模型,利用细胞培养检测其抗病毒活性。利用RT-PCR方法扩增鸡胚成纤维细胞(CEF)的ISG12基因,经测序分析,该基因序列与GenBank中登录的禽属ISG12基因序列相似性为100%。将该基因亚克隆至pET-32a载体构建重组原核表达质粒p32a-C12;工程菌p32a-C12/BL21经IPTG诱导表达,重组蛋白经Ni 2+琼脂糖凝胶柱层析纯化,获得相对分子质量约30 000的目的蛋白。利用ISG12蛋白预处理CEF和鸡外周血淋巴细胞(peripheral blood lymphocyte,PBL)后感染AIV 1215株,或者将该病毒与ISG12蛋白一起接种CEF细胞和PBL细胞,AIV在CEF中的复制能力与对照组无明显差异,而ISG12蛋白预处理鸡PBL细胞和同时感染AIV 1215株时的病毒含量均比对照低。体外抗病毒活性试验表明,ISG12蛋白能够抑制AIV 1215株在PBL细胞中的复制,但在CEF细胞上的作用不明显。     

不同品种绵羊和山羊主要先天性免疫指标的比较

对4个山羊品种和4个绵羊品种的7项主要先天性免疫学指标进行了检测并比较。对动物进行颈静脉采血,制备血清,用ELISA试剂盒测定IL-1、IL-2、IL-6、IL-18、IFN-γ、NK细胞、血清溶菌酶。利用生物统计学的方法和原理,以品种和品种来源地为应变量对检测结果进行比较分析。结果表明,山羊当地品种的NK和LYS高于引进品种（P〈0.05）;绵羊引进品种的IL-1、IL-2、IL-6、IL-18、IFN-γ、NK、LYS均明显高于地方品种（P〈0.01）。在不同品种之间,莱芜黑山羊NK和LYS水平高于崂山奶山羊、波尔山羊和鲁波山羊,萨福克羊的IL-1、IL-6、IL-18、IFN-γ、LYS水平最高且差异极显著（P〈0.01）。小尾寒羊IL-18、IFN-γ水平最低且差异极显著（P〈0.01）。说明不同种群绵羊、山羊的先天性免疫水平存在显著差异,为以后抗病育种提供合理的试验数据。     

鸡球虫病两种免疫方式的比较

本文简述鸡球虫病发病原因及几种常见鸡球虫病.并取爱拔益加肉鸡36 000羽公雏和36 000羽母雏,分为4栋鸡舍养殖(两栋公雏和两栋母雏),使用两种免疫方法分别对四栋鸡舍雏鸡进行免疫,并比较免疫效果.