分泌性免疫球蛋白A与肠道黏膜免疫的关系及其分泌的营养调控

分泌性免疫球蛋白A(SIgA)是由J链连接成的二聚体免疫球蛋白A(IgA)与分泌片段结合后形成的复合物。肠道内的SIgA是肠道黏膜免疫中的重要环节之一。SIgA可维持肠道黏膜内环境稳态,调控肠道内源微生态,并在黏膜上皮通过干扰病原体与上皮细胞受体结合,阻止病原体附着宿主细胞,影响病原菌毒力及免疫排异,从而阻止病原体的传播和进一步的感染,因而在免疫防御中起着非常重要的作用。营养因子(如脂肪酸、氨基酸、维生素等)在机体受到病原菌入侵或应激的情况下,可显著促进肠道SIgA的分泌,提高机体对病原菌的抵抗能力,在一定程度上改善应激动物的生产性能。     

Evaluation of systemic and secretory IgA concentrations and immunohistochemical stains for IgA-containing B cells in mucosal tissues of an Irish setter with selective IgA deficiency

Immunoglobulin A is the predominant secretory antibody at mucosal surfaces. In the dog, immunoglobulin A deficiency (IgAD) is characterized by low to absent serum IgA and normal to elevated serum immunoglobulin G (IgG) and immunoglobulin M (IgM) concentrations. However, studies comparing serum and secretory IgA in dogs have often documented a poor correlation, suggesting that serum concentrations should not be used to estimate mucosal secretion of this antibody. This report demonstrates the concurrent use of serum IgA, IgG, and IgM; secretory IgA (from bronchoalveolar lavage fluid); and immunohistochemical stains on bronchial and duodenal mucosa for IgA-containing B cells in a young Irish setter with recurrent respiratory and gastrointestinal signs.     

表面表达猪水肿病毒素保护性抗原重组大肠杆菌激发仔猪黏膜免疫反应的研究

以菌体表面表达猪水肿病毒素保护性抗原(SLT-ⅡeB)的重组大肠杆菌(PZBSPAX),对1日龄、15日龄仔猪口服免疫,用ELISA法对口服免疫后10 d、15 d、30 d仔猪肠黏液中特异性SIgA和血清中特异性IgA、IgG进行检测,结果显示口服免疫猪能产生较高水平的肠黏液特异性SIgA及血清特异性IgA和IgG,证明该重组菌能有效激发肠道黏膜免疫反应,同时也能激发全身性体液免疫反应.     

Quantitative Changes Associated with Calving in the Levels of Bovine Immunoglobulins in Selected Body Fluids I. Changes in the Levels of IgA, IgG1 and Total Protein

Levels of bovine IgA, IgG1 and total protein (TP) were determined in serum, saliva, tears and individual quarter lacteal secretions of six Holstein-Friesian cows sampled from six weeks before to four weeks after parturition. Hierarchal analyses of variance indicated significant variations among weeks, cows and quarters of the udder. A precipitous but non proportional drop in the levels of IgA and IgA1 in lacteal secretions occurred at calving. There was a concomitant increase in IgG1, and decrease in IgA, in serum. Correlation studies supported the concept of selective transport of IgG1 from serum to lacteal secretions in regulated amounts independent of serum IgG1 levels. Changes in the IgG1/TP ratio of serum and lacteal secretions supported the idea of a decrease in the selective transport mechanism. Correlation studies and estimations of secretory IgA (SIgA) in serum suggest that serum IgA is derived from IgA synthesized in secretory tissues. Highly significant correlations between IgA and IgG1 levels in all secretions postpartum suggest that local IgA synthesis and either IgG1 transport or local IgG1 synthesis are initiated by the same stimuli. Although some of the variation in the level reported for IgA and IgG1 in secretions resulted from protein dilution, much of the variation represents physiological differences between individual animals and tissues in the same animal. An IgG2/IgG1 ratio approaching that of serum occurred in a mastitic quarter of one cow. IgA was the principal immunoglobulin in saliva and tears, comprised a greater proportion of the immunoglobulin in milk whey than in prepartum lacteal secretions and was a minor immunoglobulin in bovine serum.     

Mucosal immunology: overview and potential in the veterinary species

A large part of the immune system is dedicated to protection from infection at mucosal surfaces. The concept of the common mucosal immune system has been investigated in several veterinary species where traffic of mucosally activated lymphocytes from induction to effector sites has been demonstrated. The dominant isotype found in secretions of the upper respiratory tract and gut of normal healthy and diseased animals is IgA. B lymphocytes have a relatively short half-life and there is continuous production of IgA at these sites, which is achieved by constant secretion from T helper and epithelial cells of cytokines that are critical for B cell maturation and IgA secretion. Specific stimulation of mucosal immune responses using intranasal presentation of live and inactivated antigens (with adjuvants active at mucosal surfaces) has shown great promise for inducing protective immunity to respiratory pathogens.     

Regulation of IgA responses in cattle, humans and mice

Secretory IgA (SIgA) constitutes the largest component of the humoral immune system of the body with gram quantities of this isotype produced by mammals on a daily basis. Secretory IgA (SIgA) antibodies function by both blocking pathogen/commensal entry at mucosal surfaces and virus neutralization. Several pathways of induction of IgA responses have been described which depend on T cells (T cell dependent or TD) pathways or are independent of T cells (T-independent or TI) and are mediated by dendritic cells (DCs) and/or epithelial cells. Many elements of IgA regulation readily cross species barriers (adjuvants, soluble and cognate factors) and are highly conserved whereas other pathways may be more specific to any given species and must be evaluated. Regulation of IgA production in cattle is not completely understood and thus we have focused in part on highly conserved factors such as transforming growth factor beta, Type I and Type 2 interferons, neuropeptides which interdigitate mucosal tissues (vasoactive intestinal peptide or VIP), and a small peptide (IgA inducing peptide or IGIP) which can serve as targets for modulation and increasing SIgA virus-specific antibodies. We have evaluated the potential utility of modulating these factors in vitro in regulation of qualitative aspects of antibodies of the IgM, IgG and IgA isotypes at mucosal surfaces and in secretions of the upper and lower respiratory tract to a virus of economic and public health importance, foot and mouth disease virus (FMDV). IgA responses in cattle are essential for host defense in response to various infectious agents. In cattle, IgA is not released into the colostrum, as is the case for other mammals but only IgG1 is selectively transported. In previous studies in cattle, IgA has been shown to be regulated by several cytokines including IFN-gamma, Type I interferons such as IFN-alpha and IFN-tau, transforming growth factor beta, IgA inducing peptide and other potential factors such as APRIL and BlyS which have not yet been fully evaluated in cattle. Many of these factors, namely TGF-beta and Type I interferons block cell cycle progression which is an essential component of Ig class switching and thus these factors require additional regulatory factors such as IL-2 to drive cells through cell cycle resulting in class switch recombination. Among these factors, IgA inducing peptide was originally identified from a bovine gut associated lymphoid tissue expression library and is highly conserved in pigs and humans at >90% at the amino acid level. The factor is regulated differently in various species but is consistently produced by dendritic cells.     

The neonatal Fc receptor (FcRn) is expressed in the bovine lung

In neonatal calves, maternal immunoglobulin (Ig) is transferred into respiratory secretion which contributes to protection against pathogens. The early predominance of IgG1 in respiratory tract secretions is progressively reduced in favor of IgA by age but in the lower, bronchoalveolar system secreted IgG remains the dominant secreted Ig even in adulthood. The trans-epithelial transport of secretory IgA into mucosal secretions is carried out by the polymeric Ig receptor. However, the mechanism by which IgG crosses epithelial cells to provide defense on mucosal surfaces is still unknown. In order to investigate the possibility that the neonatal Fc receptor, FcRn is involved in this transport we have first analyzed the localization of this receptor in the upper and lower respiratory tracts. Consistent with the in situ hybridization data, immunohistochemistry showed undetectable expression in the tracheal epithelial cells, relatively weak expression in epithelial cells of the bronchi, apparent staining those lining the bronchioli and randomly scattered signal over the alveolar tissue. The bovine FcRn may thus play a role in IgG transport across mucosal epithelial barriers as a trafficking receptor and ensure IgG predominance in the lower respiratory tract.     

Helicobacter pylori-specific immunoglobulin synthesis in gnotobiotic piglets: evidence for the induction of mucosal immunity in the stomach

Short term tissue biopsy cultures and paired, sera, bile and gastric and intestinal contents from Helicobacter pylori-infected gnotobiotic piglets were tested for the synthesis of H. pylori-specific immunoglobulin (Ig) isotype production by antigen-specific ELISA from post-infection days (PIDs) 2-28. Serum antibody levels in all three Ig isotypes were elevated from baseline values by PID 14, serum IgM levels reached peak levels on PID 14 and by PID 28 bile was strongly positive for IgA and IgG.Intestinal, but not gastric contents from infected piglets, contained IgA-specific antibody from PID 14 onward. Gastric mucosal epithelia adjacent to areas of inflammation in infected but not uninfected control piglets produced readily detectable amounts of porcine secretory component (SC); IgA-positive plasma cells were identified in gastric submucosa and lamina propria in these areas. Culture fluid supernatants, collected from explanted gastric cardia and antra and intestinal ilea of H. pylori-infected piglets had trace amounts of IgA as early as PID 2 in some animals, and strong IgA reactivity in all by PID 28. Supernatants also contained H. pylori-specific IgG by PID 14. A strong gastric lymph node IgA response contrasted with moderate IgA production in mesenteric lymph nodes and spleen. Mucosal biopsy production of H. pylori-specific IgG was more evenly distributed throughout the lymphoid system. These data support the contention that the Ig response to H. pylori is initiated within the gastric compartment and matures over time to a generalized IgA-dominated mucosal and IgG-dominated nonmucosal humoral immune response.     

Preparation of porcine IgA and its detection using the fluorescent antibody technic]

The preparation of immunoglobulin A (IgA) from porcine colostrum, intestinal content and serum is described. The best results were achieved with colostrum, from which an antigen of satisfactory purity was prepared by purification on Sephadex G-200, on DEAE cellulose and subsequent filtration on Sephadex G-200. The serum to this antigen raised in rabbits was adsorbed to an immunoadsorbent from porcine serum (PS) or porcine IgG. The adsorbtion of the serum against secretory IgA (SIgA) to PS removed its undesirable heterologous and nonspecific reactivity. The anti-SIgA serum adsorbed in this way still reacted with IgA from porcine serum. In the direct and indirect immunofluorescent staining we detected the main antigenic determinants of the SIgA molecule, i. e. the heavy chains and the secretory component.     

Perspectives in mucosal immunity: a ruminant model

All mammalian species have developed specialized defence mechanisms at the interface between mucosal surfaces and the environment and this system operates independently of the systemic immune system. Attachment of organisms to the mucosal epithelium is a primary prerequisite for infection and is an important virulence determinant for pathogens of these sites. The mucosal immune defences are thus characterized by production of IgA antibodies, the principal mode of action of which is to inhibit the adherence of pathogens to the mucosal epithelium. This introduction to the symposium on mucosal immunity will attempt to underline the role of local immunity in the intestine in providing local defence at that site, in addition to providing cellular molecular effectors for other mucosal sites.     

Local immunity in the respiratory tract of the chicken. II. The secretory immune response to newcastle disease virus and the role of IgA

The nature, specificity and characteristics of the secretory immune response in the respiratory tract of the chicken were investigated in young specific-pathogen-free chickens after vaccination with live lentogenic and inactivated Newcastle disease virus (NDV). Virus-neutralizing (VN) activity considerably exceeding transudation levels from serum were detected in lachrymal fluid, saliva and tracheal washes following infection by both ocular and oral routes. Heat inactivated virus inoculated into the trachea evoked neither serum nor secretory VN activity, whereas a commercial inactivated virus vaccine in mineral oil adjuvant stimulated high titres of serum antibody and some VN activity in tracheal fluids.Antibody in secretions limited, but did not prevent, reinfection of the trachea when birds were challenged 2 weeks later. In contrast to an elevation of circulating antibody titre, challenge induced only a repeated primary response of secreted antibody.All secretions contained IgA which, at least in saliva, accounted for 85% of its activity, the remainder being due to IgG. Fluorescent localization of immunoglobulin producing cells (IPC) demonstrated large numbers containing IgA in association with the upper respiratory tract, particularly in the Harderian gland which contained dense aggregations of plasma cells, many of which were producing IgA.It is concluded that the respiratory tract of the chicken possesses an antibody mediated secretory immune system analogous to that of mammalian species.     

动物多聚免疫球蛋白受体研究概况

多聚免疫球蛋白受体（polymeric immunoglobulin receptor,pIgR）是主要组织相容性复合物（MHC）家族中的重要成员,是分泌性免疫系统（SIS）的重要组成成分,在IgA/IgM的转运和黏膜免疫中发挥着重要的作用。作者结合近年来的研究综述了pIgR的分子结构、生物学功能和研究概况。     

猪支原体肺炎活疫苗（168株）肺内免疫机制研究

为研究猪支原体肺炎活疫苗(168株)的免疫机制,通过肺内接种免疫5 ～ 10日龄仔猪,并于免疫后不同时间点检测血清中IgG抗体效价、全血中淋巴细胞转化效率、呼吸道局部的IFN-γ浓度和特异性SIgA滴度,于免疫后28 d剖杀采集呼吸道上皮组织,通过扫描电镜法与原位杂交检测法观察疫苗株在呼吸道的存留以及对纤毛的影响情况.结果发现,免疫后猪血液中淋巴细胞转化增强1.52～2.01倍,支气管表面IFN-γ浓度和特异性SIgA滴度持续增加,但血清抗体一直未检测到.扫描电镜与原位杂交检测结果发现疫苗株能有效地黏附在支气管纤毛上皮细胞上,但对纤毛的影响较小.由此表明,猪支原体肺炎活疫苗(168株)通过肺内免疫可有效激活全身细胞免疫及呼吸道局部的黏膜免疫与细胞免疫反应,而且还可以通过黏附支气管纤毛上皮细胞产生占位效应而对上皮组织不产生损伤.     

生长抑素亚单位苗对鸡粘膜免疫的影响

在肌肉注射生长抑素 (SS)亚单位苗的基础上应用鸡新城疫弱毒苗进行消化道粘膜免疫鸡 ,显示和检查小肠粘膜中免疫球蛋白A (SIgA)阳性细胞和上皮内淋巴细胞 (iIEL)数量的变化。结果表明 ,应用SS亚单位苗首免后第 3周和第 4周 ,十二指肠中iIEL数量比对照组均显著增加(P <0 0 5) ;首免后第 3周 ,十二指肠、空肠和盲肠扁桃体中SIgA阳性细胞比仅用新城疫弱毒苗免疫组极显著增加 (P <0 0 1 ) ;首免后第 4周 ,十二指肠和空肠中SIgA阳性细胞比仅用新城疫弱毒苗免疫组极显著增加 (P <0 0 1 )。提示SS可抑制小肠粘膜中SIgA阳性细胞和iIEL的分化和增殖。SS亚单位苗可中和体内SS ,作为粘膜免疫促进剂预防家禽传染病     

大肠杆菌不耐热肠毒素对狂犬病病毒减毒疫苗株粘膜免疫效应的增强作用

将狂犬病病毒 SRV9减毒疫苗与大肠杆菌不耐热肠毒素 (L T)混合 ,分别经口腔滴入、灌胃、灌肠等途径免疫小鼠。通过检测外周血淋巴细胞特异性转化率、CTL反应、Ig G、Ig A、SIg A等免疫指标 ,并结合攻毒保护 ,探讨了狂犬病病毒经消化道不同途径接种产生的免疫效果以及 L T在粘膜免疫中的作用。结果表明 :肠道接种组的狂犬病病毒特异性免疫应答水平高于口腔和胃接种组的免疫应答水平 ,L T可增强狂犬病病毒减毒疫苗诱导的免疫应答反应     

分泌型IgA对肠道黏膜免疫的研究进展

分泌型IgA(secretory IgA,SIgA)作为一种包被于肠道黏膜的抗体,能保护肠道免受病原微生物和毒素的攻击而不引起炎症反应,对激活黏膜免疫和维持肠道内环境稳态起到重要作用。在动物肠腔中,SIgA能通过调节肠道上皮细胞受体的识别能力,阻断病原微生物侵入黏膜相关淋巴组织,随后在肠道蠕动和黏液绒毛的协助运动下,最终将病原微生物清除;且最近也有报道揭示SIgA在肠上皮胞吞转运作用下的新机制。因此,作者主要阐述SIgA在肠道黏膜免疫及其内环境方面发挥关键的生物学特征和功能,以及探讨胞吞转运机制下SIgA的潜在作用。     

IgA and secretory component (SC) in the third eyelid of domestic animals: a comparative study

Objective The third eyelid of domestic animals is important for the production and distribution of tears, in removing ocular debris and in protection of the globe, and has significant immunologic functions. Although it is known that tears contain antibodies of the immunoglobulin A (IgA) isotype which are produced mainly by plasma cells of the lacrimal gland, very little is known about the antibody repertoires in the third eyelid of domestic animals. To assess whether IgA is derived from local synthesis, we analyzed the location of IgA‐producing cells and the cellular distribution of secretory component (SC) in the third eyelid of domestic animals in a comparative study. Animal studied A total of 83 third eyelids of dogs, cats, pigs, cows, sheep, goats and horses were investigated in the course of this study. Procedures Third eyelids were obtained immediately after death, cut length‐wise, fixed overnight and processed for immunohistochemical detection of IgA and SC by the ABC technique. Results The results show that IgA‐producing plasma cells are densely populated in subepithelial spaces of the surface epithelium as well as in the nictitating gland in a species‐specific manner. In contrast, the SC could be demonstrated exclusively in glandular acinar and ductal epithelial cells and in different cell types of the surface epithelium, preferentially located on the bulbar side of the nictitating membrane. Conclusion It is suggested that most of the SC is locally produced by resident plasma cells and subsequently transferred through the surface epithelium and glandular duct cells by transcytosis. This indicates that the third eyelid is an important member of the secretory immune system in domestic animals.     

The metabolism and transport of bovine serum SIgA

Experiments were undertaken in six Holstein-Friesian cattle to determine whether secretory IgA (SIgA) could be transported from serum into exocrine body fluids. Preliminary data indicated that when IgG₁, IgG₂, IgM and SIgA were administered i.v., only SIgA and IgG₁ appeared in bile 90 min later at concentrations equal to or exceeding those in serum at the same time. Two hours post-injection, 70% of the SIgA recovered in bile was intact however only 30% co-precipitable with anti-secretory component (SC) while >90% of the administered IgA was precipitated by this method. All recovered IgG₁ was of low molecular weight. More detailed studies indicated that the IgA recovered in bile 7 h post-injection or in milk 3 h post-injection, was predominately lower molecular weight than intact SIgA. Most of this low molecular weight radioactivity was TCA precipitable and ca 50% was dialyzable; these data indicate that TCA-precipitability is an inadequate criterion for determining whether intact SIgA is transported. The radioactivity recovered in parotid saliva was almost entirely non-TCA precipitable and dialyzable. Almost all SIgA recovered in bovine serum remained intact and had a of 15.7 h. When transport into milk and bile was calculated from total, recovered radioactivity (i.e. 29% and 2.7, respectively), data compared favorably with those conducted in sheep in which dimeric IgA (without SC) was administered i.v. When we calculated transport on the basis of recovered intact IgA, only 1.47 and 0.54% of the injected dose had been transported into milk and bile, respectively, 24 h later. Most IgA in ruminant bile may be of serum origin although the same appears to be unlikely for the IgA in milk.     

猪SIgA分泌片的分离纯化及其多克隆抗体的制备

为制备抗分泌片多克隆抗体,采用分子筛凝胶层析、离子交换层析和硫酸铵盐析法从猪初乳中分离纯化分泌片和SIgA。SDS—PAGE鉴定表明,纯化的SIgA呈现分泌片、重链和轻链三条带,大小约为70ku;凝胶薄层扫描分析SIgA和分泌片纯度分别为90％和89％;琼脂扩散鉴定表明,SIgA和分泌片与抗分泌片单克隆抗体发生了特异反应。以纯化分泌片为抗原,制备抗分泌片多克隆抗体,琼脂扩散检测多抗的效价为1：32。高纯度分泌片的提取及高效价抗分泌片多抗的制备为粘膜负．疫研究奠定了物质基础。