大疱性类天疱疮免疫学研究进展

大疱性类天疱疮是一种好发于老年人的自身免疫性表皮下大疱病,血清中存在针对靶抗原BP230和BP180的自身抗体,自身抗体识别的靶抗原表位主要位于BP180分子胞外的非胶原编码区NC16A区段.此外,细胞因子与补体的活化亦参与皮肤损伤和水疱形成.BP可能存在遗传易感性,HLA-II类基因HLA-DR和DQ在BP的发生和发展过程中起一定作用.     

大疱性类天疱疮患者血清抗BP180自身抗体识别抗原表位的研究

目的分析大疱性类天疱疮(BP)患者血清抗BP180IgG和IgE自身抗体所识别的抗原表位。方法利用柱亲和层析的方法从10例BP患者血清中提取抗BP180-NC16A自身抗体,用免疫印迹技术对所获IgG和IgE自身抗体分别进行抗原表位鉴定。结果所有10例患者的自身抗体均识别aa507-aa532区域内(NC16A-2和NC16A-2.5)至少一个抗原表位,而且来自同一患者的IgG和IgE自身抗体在该区域所识别的表位完全一致。BP自身抗体与NC16A-1、NC16A-3和NC16A-4区域的结合率较低,与NC16A-5片段不结合。结论BP致病性自身抗体所识别的主要抗原表位可能位于BP180分子NC16A片段的aa507-aa532区域内。     

大疱性类天疱疮免疫发病机制的研究进展

大疱性类天疱疮作为常见的自身免疫性疱病,国内外学者一直进行其发病机制的研究.目前认为,IgG是大疱性类天疱疮患者血清中主要的致病性自身抗体类型,与BP180等自身抗原结合后可同定并激活补体,导致肥大细胞脱颗粒,激活浸润的炎症细胞并使其释放蛋白水解酶,引起真表皮连接部位的重要蛋白结构受损,最终导致表皮下水疱的发生.研究表明,体液免疫和细胞免疫均参与大疱性类天疱疮的发病过程,但大疱性类天疱疮自身抗体及可能存在的自身抗原的来源及嗜酸粒细胞在发病过程中的作用尚不完全清楚.

Abstract：

Bullous pemphigoid (BP) is a common autoimmune blistering skin disease. Studies on its immunologic pathogenesis have been always carried out at home and abroad. IgG autoantibodies, which are the predominant type of pathogenic autoantibodies, can bind to multiple BP180 epitopes, result in the fixation and activation of complement, degranulation of resident mast cells, activation of infiltrating inflammatory cells and release of proteinases followed by the loss of cell-matrix adhesion structure at the dermal-epidermal junction, and finally cause the formation of subepidermal blisters. Although studies have shown that both humoral and cellular immunity are involved, little is known about the resource of autoantibodies, probalble existent autoantigens, and the roles of eosinophils in the pathogenesis of BP.     

大疱性类天疱疮患者血清总IgE与血清自身抗体的关系

目的 分析大疱性类天疱疮患者血清总IgE与抗BP180IgG抗体、抗BP230IgG抗体、抗表皮基底膜IgG抗体滴度(即间接免疫荧光滴度)的关系.方法 收集沈阳市第七人民医院2014年1月-2020年1月大疱性类天疱疮病例,进行回顾性分析,根据抗BP180IgG抗体、抗BP230IgG抗体阳性情况将患者分组,比较组间血...     

人源性抗大疱性类天疱疮抗原BP180单链抗体的体外功能鉴定

目的 探讨人源性抗大疱性类天疱疮抗原BP180单链抗体的生物学功能。方法 亲和层析方法纯化大疱性类天疱疮（BP）患者血清自身抗体,通过竞争性ELISA、竞争性免疫荧光和补体活化的竞争性抑制实验来观察所制备的抗BP180单链抗体对BP-IgG自身抗体结合人BP180抗原的竞争性抑制作用。结果 竞争性ELISA结果表明,在0 ~ 60 μg/ml范围内单链抗体对BP-IgG自身抗体的竞争性抑制作用呈剂量依赖关系,最大抑制率可达69.50%（与对照组相比,均有统计学意义,P < 0.01）。竞争性免疫荧光实验中,单链抗体浓度增加至40 μg/ml时,BP-IgG在基底膜带的沉积及其介导的补体C3活化作用均被抑制,呈阴性。结论 人源性抗BP180单链抗体在体外对BP致病性自身抗体与抗原结合及后续的补体活化具有一定的抑制作用。     

大疱性皮肤病

990295 疱液在诊断表皮下自身免疫性大疱病中的意义/周曙霞(上海二医大附属瑞金医院皮肤科)…人中间皮肤性病学杂志.-1998,12(5).-259为探讨这类疾病的池液中抗粘膜基底膜带(BMZ)自身抗体情况及在诊断中的意义,应用非盐裂皮肤及盐裂皮肤间接免疫荧光技术(IIF)检测38例表皮下自身免疫性大疱性患者疱液和血清中IgG、IgA、IgM及IgG亚型IgG_1～IgG_4抗BMZ抗体及滴度,并检测疱液和血清中结合补体C_3的抗特异性抗BMZ抗体。结果显示疱液和血清中各型抗BMZ抗体的滴度均无显著性差异,说明表皮下自身免疫性大疱病患者的疱液含有抗BMZ抗体,其浓度与血清中浓度无显著性差异,疱液可用于IIF检测患者循环抗BMZ抗     

SLE皮肤基底膜带自身抗体靶抗原的研究

目的：了解SLE皮肤基底膜带自身抗体（BMZ－Ab）靶抗原，并与自身免疫性大疱病相关抗原进行比较。方法：以人皮肤提取物为抗原，免疫印迹（IB）检测47例SLE及对照的14例大疱性类天疱疮（BP）、2例获得性大疱性表皮松解症（EBA）患者血清中IgG型自身抗体。结果：29／47例SLE血清IgG型自身抗体与表皮和／或真皮抗原反应，其中14／47例单纯结合180kD、145kD、130kD、97kD、85kD、75kD、66kD表皮抗原，8／47例单纯结合115kD、290kD真皮抗原，7／47例同时结合上述不同分子量表、表皮抗原。11／14例BP血清结合230kD、180kD、165kD、130kD、115kD、97kD表皮抗原。2例EBA血清结合290kD真皮抗原。结论：SLE BMZ－Ab靶抗原存在明显异质性。SLE与多种自身免疫性大疱BMZ抗原交叉，提示它们之间的内在相关性。     

大疱性类天疱疮患者血清抗BP180分子不同表位自身抗体定量分析及亲和力分析

目的 探讨大疱性类天疱疮（BP）患者血清抗BP180分子不同表位的自身抗体量。方法 制备BP180分子NC16A片段的不同抗原表位区NC16A-1、NC16A-2和NC16A-3,利用柱亲和层析的方法从10例BP患者血清中分别纯化抗不同表位区的自身抗体,定量,并用硫氰酸盐洗脱法测定抗NC16A-1、抗NC16A-2和抗NC16A-3自身抗体的相对亲和力。结果 经亲和层析纯化获得针对BP180-NC16A不同表位区的自身抗体,从20 mg总IgG中纯化抗NC16A-1、NC16A-2和NC16A-3自身抗体的产量分别为（49.0 ± 20.7） μg、（117.7 ± 22.4） μg和（39.5 ± 18.9） μg。抗NC16A-2自身抗体的量和亲和力水平均明显高于其他两个表位区的自身抗体。结论 BP致病性自身抗体所识别的主要抗原表位可能位于BP180分子的NC16A-2表位区（aa507-aa520）。     

血浆疗法治疗大疱性类天疱疮的临床疗效及自身抗体水平变化

目的:通过研究大疱性类天疱疮(bullous pemphigoid, BP)患者的临床资料,提高对大疱性类天疱疮的认识,并初步评价血浆疗法治疗大疱性类天疱疮的临床疗效,并探索其治疗机制。方法:2009年12月—2015年12月我院皮肤科收治的大疱性类天疱疮患者66例,28例患者接受血浆疗法联合糖皮质激素治疗,作为血浆疗法组(PT组),38例患者单纯应用糖皮质激素治疗,作为单纯糖皮质激素治疗组(HT组),比较两组的临床疗效,每组各采集16名患者的血清,以20名健康者血清作为对照,采用酶联免疫吸附(ELISA)法检测治疗前后血清中抗BP180抗体水平的变化以及抗BP180抗体亚型IgG1、IgG4水平的变化。结果:PT组有效率为92.86%,HT组为73.68%,PT组的治疗效果更为显著。两组患者治疗后抗BP180抗体滴度明显下降(P0.01),PT组较HT组下降明显(P0.01),PT组患者治疗后抗体亚型IgG1、IgG4测得的吸光度A值明显下降(P0.05)。结论:血浆疗法治疗大疱性类天疱疮有效,其机制可能与降低抗BP180抗体及抗体亚型IgG1、IgG4的水平有关。     

炎性细胞在大疱性类天疱疮发病机制中的作用研究进展

大疱性类天疱疮(BP)是由抗半桥粒抗原BPl80和BP230抗体所导致的自身免疫性水疱性疾病.补体活化、肥大细胞脱颗粒、中性粒细胞聚集等都在BP的发病机制中起着重要的作用.该文就炎性细胞在大疱性类天疱疮发病机制中的作用作一综述.     

抗BP180NC16A IgG亚型检测方法的建立以及在大疱性类天疱疮中的意义

【摘要】 目的 建立抗BP180NC16A IgG亚型的检测方法,并探讨其在大疱性类天疱疮（BP）中的意义。方法 原核表达GST-NC16A融合蛋白,并采用亲和层析法纯化。优化ELISA关键环节,建立抗BP180NC16A IgG各亚型的ELISA检测方法,并对10例未经治疗的BP、5例妊娠疱疹、1例成人线状IgA大疱性皮病、2例天疱疮患者血清分别进行检测。结果 通过方阵测定法确定GST-NC16A融合蛋白的包被浓度为500 μg/L,包被条件为4 ℃ 12 h,血清稀释倍数为1 ∶ 100,酶标二抗为1 ∶ 2000,孵育条件为37 ℃ 1 h,底物反应条件37 ℃ 20 min。10例大疱性类天疱疮患者10例IgG1阳性,9例IgG2阳性,5例IgG3阳性,9例IgG4阳性。2例寻常型天疱疮、1例成人线状IgA大疱性皮病均阴性。5例妊娠疱疹所有亚型均阳性,以IgG1和IgG3亚型为主。结论 抗BP180NC16A ELISA检测法特异性强、重复性好,是检测BP和妊娠疱疹患者抗BP180NC16A抗体亚型的半定量方法。     

Serum levels of IgE anti-BP180 and anti-BP230 autoantibodies in patients with bullous pemphigoid

BACKGROUND: Bullous pemphigoid (BP) is a subepidermal blistering disease characterized by autoantibodies against the hemidesmosomal proteins, BP180 and BP230. NC16A, a non-collagenous stretch of the BP180 ectodomain is the primary target of pathogenic IgG antibodies. Whereas IgG anti-BP180 autoantibodies play a primary role in the pathogenesis, there is a growing number of data regarding the potential pathogenic roles of IgE class autoantibodies in BP. OBJECTIVES: To examine the levels of IgG and IgE autoantibodies against BP180 and BP230, and to investigate mutual association and clinical relevance. METHODS: Sera obtained from 67BP patients and 36 healthy donors were subjected to ELISA assays to measure serum IgG and IgE levels of anti-BP180 and anti-BP230 antibodies. RESULTS: IgG anti-BP180 antibodies were positive in 63 (94%) of 67BP patients. IgG anti-BP230, IgE anti-BP180, and IgE anti-BP230 antibodies were found in 48 (72%), 20 (30%) and 45 (67%), respectively. IgG anti-BP180 levels were correlated with the affected areas. IgG anti-BP230 antibodies tended to increase in proportion to elongation of disease duration. IgE anti-BP230 levels showed a strong association with local eosinophil accumulation, while the levels were reversely related with the affected areas in BP. CONCLUSIONS: IgE autoantibodies to BP180 and BP230 are detected at high frequencies in BP. IgE anti-BP230 antibodies may have a role in attracting eosinophils to the skin lesions.     

Experimental models for the autoimmune and inflammatory blistering disease,Bullous pemphigoid

Bullous pemphigoid (BP) is a subepidermal skin blistering disease characterized immunohistologically by dermal-epidermal junction (DEJ) separation, an inflammatory cell infiltrate in the upper dermis, and autoantibodies targeted toward the hemidesmosomal proteins BP230 and BP180. Development of an IgG passive transfer mouse model of BP that reproduces these key features of human BP has demonstrated that subepidermal blistering is initiated by anti-BP180 antibodies and mediated by complement activation, mast cell degranulation, neutrophil infiltration, and proteinase secretion. This model is not compatible with study of human pathogenic antibodies, as the human and murine antigenic epitopes are not cross-reactive. The development of two novel humanized mouse models for the first time has enabled study of disease mechanisms caused by BP autoantibodies, and presents an ideal in vivo system to test novel therapeutic strategies for disease management.     

Absence of anti-BP180 antibodies in mothers of infants with bullous pemphigoid

BACKGROUND: It is not clear whether bullous pemphigoid (BP) of infancy is linked to maternal transmission of pathogenic autoantibodies. Objectives To search for anti-BP180 antibodies in the sera of infants with BP and their mothers, using sensitive and specific methods. METHODS: Four infants (<6 months) with BP and their mothers were tested for anti-BP180 antibodies by indirect immunofluorescence, immunoblotting and enzyme-linked immunosorbent assay (ELISA). RESULTS: We found anti-BP180 antibodies in the sera of the four infants with all methods. These antibodies reacted with the extracellular domain NC16A. In the serum of their mothers we found 180 and 160 kDa proteins, each in one case, but indirect immunofluorescence and ELISA were negative, suggesting the absence of anti-BP180 autoantibodies reacting with the extracellular domain NC16A. CONCLUSIONS: BP of infants is not due to maternofetal transmission of pathogenic autoantibodies. Other hypotheses for the pathophysiology of BP are discussed.     

Autoantibodies to the extracellular and intracellular domain of bullous pemphigoid 180, the putative key autoantigen in bullous pemphigoid, belong predominantly to the IgG1 and IgG4 subclasses

BACKGROUND: Autoantibodies to the extracellular domain (ECD) of bullous pemphigoid (BP) antigen 180 (BP180) are thought to play a crucial part in the pathophysiology of BP. OBJECTIVES: As the various IgG subclasses have different biological properties, we have sought to assess the relative isotype distribution of IgG to BP180 and their reactivity against the ECD and intracellular domain (ICD) of BP180. METHODS: The reactivity of 27 sera from patients with BP was assayed by immunoblotting against recombinant proteins covering the ECD and ICD of BP180. RESULTS: Twenty-seven (100%) and 21 (77%) of 27 BP sera, respectively, contained IgG1 and IgG4 autoantibodies binding to the ECD of BP180. Fourteen (82%) and six (35%) of the 17 BP sera that were reactive with the ICD of BP180 had autoantibodies of the IgG1 and IgG4 subclass, respectively. The profile of the isotype restriction appeared to be similar when the response to the ECD vs. that to the ICD was compared. IgG2 and IgG3 reactivity with BP180 was found less frequently. Patients with BP of longer duration showed a tendency to have, in addition to IgG1, an IgG4 response. CONCLUSIONS: Consistent with prior evidence indicating that subepidermal blister formation in BP is dependent upon complement activation, the frequent finding of complement-fixing IgG1 autoantibodies to both the ECD and ICD of BP180 might have pathogenic relevance in BP. These findings provide new insights relevant for our understanding of the immune response to BP180, the putative key autoantigen in BP.     

Bullous pemphigoid of childhood: autoantibodies target the same epitopes within the NC16A domain of BP180 as autoantibodies in bullous pemphigoid of adulthood

BACKGROUND: Bullous pemphigoid (BP) is an autoimmune subepidermal blistering disease of the elderly that rarely occurs in children. Most adult BP serum samples react with epitopes within the NC16A domain of BP180, a glycoprotein of the cutaneous basement membrane zone. OBJECTIVES: To characterize the autoimmune response in childhood BP using recombinant forms of BP180 and to determine the subclass distribution of autoantibodies and their correlation with disease activity. OBSERVATIONS: Serum samples from 2 infants with BP, aged 4 and 5 months, reacted by immunoblot analysis with 4 epitopes clustered within the N-terminal 45 amino acids of the NC16A domain. The same 4 epitopes have previously been shown to be the target in adult BP. Childhood BP antibodies to BP180 NC16A belonged to IgG1, IgG2, IgG3, and IgG4 immunoglobulin subclasses. IgE reactivity was not detected. Serum levels of antibodies targeting BP180 NC16A paralleled disease activity as detected by enzyme-linked immunosorbent assay. CONCLUSIONS: The fine specificity of autoantibodies to BP180 is the same in BP of childhood and adulthood. Childhood BP is a true variant of BP.     

IgG4 and IgE are the major immunoglobulins targeting the NC16A domain of BP180 in Bullous pemphigoid: serum levels of these immunoglobulins reflect disease activity

BACKGROUND: Bullous pemphigoid (BP) is an autoimmune blistering disease associated with autoantibodies against the hemidesmosomal glycoprotein BP180. The noncollagenous (NC)16A domain of BP180 has recently been shown to harbor major antigenic sites recognized by BP sera. OBJECTIVE: The purpose of this study was to characterize the subclass distribution and fine specificities of autoantibodies to BP180 NC16A present in the circulation of patients with BP before, and during the course of, therapy for this disease. METHODS: Eighteen BP sera were analyzed by immunoblotting and enzyme-linked immunosorbent assay for the presence of IgG1, IgG2, IgG3, IgG4, and IgE reactive with various sites on the BP180 NC16A domain. The sera were collected before treatment was started and at 4- and 8-week time points after initiation of treatment. RESULTS: We identified IgG4 and IgE as the major immunoglobulins that preferentially react with two distinct epitopes (MCW-1 and MCW-2) within BP180 NC16A. Levels of these autoantibodies correlated with disease activity in BP. During the course of disease, no change was observed with regard to the immunoglobulin subclass predominantly reacting with BP180 NC16A or the specific epitopes within this domain. CONCLUSION: Our data demonstrate that remission of BP is paralleled by a decrease of serum levels of IgE and the different IgG subclasses reactive with BP180 NC16A.     

Clinical significance of enzyme-linked immunosorbent assay for the detection of circulating anti-BP180 autoantibodies in patients with bullous pemphigoid

The NC16A domain of the 180-kDa bullous pemphigoid antigen (BP180) is the most immunogenic and, probably, pathogenic region in bullous pemphigoid (BP). In the present study, in order to determine whether serum level of circulating anti-BP180 autoantibodies is a valuable serum marker in BP, the immunoreactivity of sera against the NC16A domain of BP180 was measured using enzyme-linked immunosorbent assay (ELISA) in ten patients with BP. Serum levels of anti-BP180 autoantibodies correlated with the clinical course in BP patients, who received various therapeutic agents. The result suggests that this NC16A-ELISA is a useful method for evaluating the clinical course and efficacy of the therapy in patients with BP.     

Role of BP230 autoantibodies in bullous pemphigoid

Bullous pemphigoid (BP) is an autoimmune disease associated with subepidermal blistering due to autoantibodies directed against BP180 and BP230. BP180 is currently considered as the major pathogenic autoantigen. However, previous clinical findings suggested that anti-BP230 autoantibodies alone can cause skin lesions in animal models and many BP patients. The characteristics of BP230 and the pathogenic roles of anti-BP230 antibodies have been proposed. First, at the molecular level, BP230 mediates the attachment of keratin intermediate filaments to the hemidesmosomal plaque and interacts with other constituents of hemidesmosomes. Second, the presence of BP230 autoantibodies may correlate with specific clinical features of BP. The immunoglobulin (Ig)G autoantibodies from BP patients react mainly against the C-terminus of BP230, while the IgE autoantibodies are still inconclusive. Third, in vivo, autoantibodies against BP230 involved in the disease may not only induce the inflammatory response but also impair the structural stability of hemidesmosomes. This article reviews recently published work about the role of BP230 and its antibodies, including IgG and IgE, aiming to find clues of its clinical association and lay the foundation for the research on the pathogenicity of antibodies against BP230.