天疱疮自身抗体的靶抗原研究

目的　了解天疱疮自身抗体的靶抗原及其在表皮的超微定位。方法　采用直接和间接免疫荧光 (IF)、免疫印迹 (IB)及包埋后金标记间接免疫电镜 (gold IIEM )等方法检测 2 7例天疱疮 (PV 2 2例 ,PF 5例 )血清与表皮抗原的结合情况。结果　 2 7例天疱疮直接和间接免疫荧光的阳性率分别为 10 0 %和 81.5 % ,表现为表皮细胞间有亮绿色荧光标记物呈网状分布。 2 1/ 2 2例PV血清与表皮提取物 13 0kD分子结合 ,2 / 5例PF血清与表皮提取物 160kD分子结合。对11例天疱疮 (9例PV ,2例PF)血清的间接免疫电镜研究结果显示 ,金颗粒均沉积在桥粒部位。结论　PV和PF的靶抗原分别是表皮 13 0kD和 160kD抗原 ,其超微结构均定位于桥粒。免疫印迹检查可作为鉴别PV和PF的重要辅助手段。     

落叶型天疱疮的唯一特殊表皮抗体

在1例落叶型天疱疮患者中发现有与仅存在于表皮角层下细胞间质的抗原起特异性反应的特有的抗体,这些抗体和寻常型天疱疮中那些存在于表皮全层的细胞间抗原起反应的抗体不同。所以,该患者的2份血清中的抗体仅与2份豚鼠食道和7份人皮肤的不同标本的角层下细胞间抗原相结合。与之相比,4例寻常型天疱疮的血清中的抗体却与这些标本中的整个鳞状细胞层的细胞间抗原相结合,用4份正常人血清则无细胞间的结合。这些特有的抗体在4例落叶型天疱疮中见到2例,在178份寻常型天疱疮的血清中均未见到,在各种皮肤病患者的339份血清中见到1份。与落叶型天疱疮抗体相结合的抗原浓度在豚鼠食道中较高,在全部试验标本中均可见到,位于最上面的2～4层细胞间隙内,与     

寻常型天疱疮抗原EC1-2表位的致病性

目的：通过寻常型天疱疮新生鼠模型的建立研究寻常型天疱疮抗原（PVA）致病性表位及其相应抗体的致病作用。方法：构建重组PVA细胞外区（EC）1－2片段分子，经亲和层析纯化后，免疫家兔，得到兔抗PVA EC1－2融合蛋白的抗血清，将提纯IgG抗体成分，被动转移到BALB／c新生鼠，15－18h后对新生鼠皮肤，血清进行组织病理，电镜和免疫荧光检查。结果：在对实验组新生鼠的评价中，其组织病理示表皮内水疱形成，棘细胞间棘突消失，棘 层松解；电镜示棘细胞间距离增宽，桥粒分离，溶解，消失，DIF和IIF均示棘细胞间免疫荧光沉积，面对照组小鼠除了IIF示棘细胞间微弱的荧光沉积外，其余检查均正常。结论：通过寻常型天疱疮新生鼠模型的建立，证明PVA分子中EC1－2表位为致病性表位，其相应的抗体为致病性抗体，另外，PVA重组分子的构建，新生鼠模型的建立为天疱疮以及其它自身免疫性疾病的诊断和治疗提供了一个模型 。     

表皮剥脱毒素A对天疱疮抗原的影响

目的 探讨表皮剥脱毒素A(ETA)对体外培养正常人角质形成细胞表面天疱疮抗原的影响及其作用机制。方法 人角质形成细胞复层化表达落叶型天疱疮抗原后与ETA作用,分别用寻常型或落叶型天疱疮抗体进行免疫荧光染色,提取培养细胞总蛋白做免疫印迹;测定ETA作用后细胞上清液中IL-1α,IL-6的浓度及其分解酪蛋白的活性。结果 ETA作用前后细胞间落叶型天疱疮抗体着色由强的连续发光转变为弱的断续发光,24h后恢复,寻常型天疱疮抗体着色无明显变化;免疫印迹检测显示ETA作用后落叶型天疱疮抗原被降解,寻常型天疱疮抗原未受影响。人角质形成细胞与ETA作用后IL-1α分泌受抑制,IL-6的分泌量低于检测水平;ETA作用后细胞上清液分解酪蛋白的活性24h内逐渐增加,36h后活性下降。结论 ETA的作用位点在落叶型天疱疮抗原,对寻常型天疱疮抗原无影响;ETA的这种作用具有可逆性,可能通过蛋白酶解作用降解落叶型天疱疮抗原。     

天疱疮和水疱脓疱性皮病的IgG和IgA抗细胞间自身抗体的独特型

比较了两种不同的IgA型天疱疮样抗体的结合型和真的IgG型天疱疮抗体的结合型。来自寻常型天疱疮和落叶型天疱疮患者血清的IgG抗体与正常人表皮细胞间隙联结,而仅寻常型天疱疮的抗体与来自人包皮的单层角朊细胞反应。1例落叶型天疱疮患者的IgG和IgA抗体与表皮的细胞间隙联结,但仅IgA抗体与培养的角朊细胞反应。1例水疱脓疱性发疹患者的IgA抗体(患者血清仅含IgA细胞间隙抗体)与表皮上部分联结,但不与单层角朊细胞联结。这些发现提示培养的单层人角朊细胞仅表达寻常型天疱疮抗原,而不表达落叶型天疱疮抗原,有至少二种独特的IgA细胞间隙抗体。     

大疱及疱疹性皮肤病

20 0 0 2 56 2　大疱性皮肤病自身抗体靶抗原定位研究 /杨春俊 (安徽医大一附院皮肤科 )…∥安徽医科大学学报 .- 1999,34(6 ) .- 4 51为进一步确定存在分岐的靶抗原在细胞间或皮肤基底膜带中的位置。验证有关大疱性皮肤病自身抗体靶抗原定位的不同研究结果。采用包埋后金标记直接和间接免疫电镜技术 ,结合免疫印迹技术分别对 5例天疱疮皮损组织中 Ig G的沉积部位、8例天疱疮患者血清中自身抗体 Ig G结合正常人皮肤的部位进行超微水平的研究。同样对 5例大疱性类天疱疮皮损组织中 Ig G的沉积部位及 8例大疱性类天疱疮患者血清中自身抗体 …     

用不同抗原来源进行免疫印迹法分析巴西落叶型天疱疮抗原

本文作者用人表皮提取物以及以牛鼻表皮提纯的桥粒体制剂为抗原,用免疫印迹法检查了27份巴西落叶型天疱疮(BPf)、13份日本散发性落叶型天疱疮(Pf)和7份寻常型天疱疮(Pv)血清.取其结果进行比较.作者用各型天疱疮血清进行间接免疫荧光法检     

The expression level of acetylcholine receptor in the patients with pemphigus

目的:研究不同型别天疱疮患者体内乙酰胆碱受体的表达状况,初步探讨其与病情活动度和天疱疮分型的关系.方法:主要采用免疫组化和ELISA方法.结果:乙酰胆碱受体IgG (acetylcholine receptor IgG,AChR IgG) 在 30例天疱疮患者中有8例患者为阳性,7例为寻常型天疱疮,1例为落叶型天疱疮,与正常对照组比较,差异没有统计学意义(P ＞ 0.05);在AChR IgG阳性的患者中,病情分级较重,当病情得到有效控制后,外周血AChR IgG的吸光度(A)值下降.AChRM3亚型在寻常型天疱疮中褐色的颗粒仍然分布在基底层,但颜色加深,颗粒变大,且大小不一致.AChRM3亚型在落叶型天疱疮中的表达模式与正常对照组无统计学差异.AChRα亚基在寻常型天疱疮和落叶型天疱疮中褐色的颗粒仍然分布在全层,但颜色加深,颗粒变大,且大小不一致.结论:在落叶型天疱疮和寻常型天疱疮中,AChR抗体的表达状况不同.AChR抗体更易出现在桥粒芯糖蛋白(Dsg)3抗体阳性患者的外周血中,在Dsg3阳性患者皮损的组织切片中,除了AChRα蛋白之外,AChRM3蛋白的染色模式也发生了改变.这提示Dsg3抗体更易导致AChR途径的异常.     

天疱疮发病机理的新进展

天疱疮抗原是桥粒性钙粘素超家族的成员，对人导细胞的粘附功能起重要作用，天疱疮是一种抗钙粘素的自身免疫性疾病。天疱疮抗原的特异性、结构和功能及桥粘素、纤维粘连蛋白在天疱疮发病机理中的作用有待进一步阐明。天疱疮抗体是天疱疮发病的关键因素，众多研究表明，天疱疮抗体ＩｇＧ４亚类是天疱疮的病原性抗体。天疱疮抗体与相应抗原结合后，磷脂酶Ｃ、１，４，５－三磷酸肌醇介导了细胞内的信息传导，细胞产生蛋白溶解酶，破坏     

大疱及疱疹性皮肤病

20120217 ELISA技术检测寻常型天疱疮桥粒芯蛋白3抗体水平研究/闫言(中国医科院、北京协和医院皮肤科),徐浩翔,晋红中…//临床皮肤科杂志.-2011,40(9).-529 ～531采用ELISA和间接免疫荧光法对106例寻常型天疱疮患者和106例对照人群进行检测.结果:ELISA法的敏感度、特异性分别为77.5％和94.3％,间接免疫荧光法分别为79.2％和94.3％,两种方法间差异无统计学意义.认为ELISA检测桥粒芯蛋白3抗体对寻常型天疱疮诊断是一种较好的辅助方法.     

Drug-induced pemphigus: autoantibodies directed against the pemphigus antigen complexes are present in penicillamine and captopril-induced pemphigus

Pemphigus is an autoimmune blistering disease characterized by circulating autoantibodies directed against the keratinocyte cell surface. The two variants, pemphigus foliaceus and pemphigus vulgaris, can be distinguished at the molecular level by immunochemical studies. The large majority of patients with pemphigus develop the disease spontaneously; however, there is a small group of patients who develop pemphigus after treatment with certain medications, of which penicillamine and captopril are the best documented. Most patients with drug-induced pemphigus have circulating and/or tissue bound epidermal cell surface autoantibodies; however, the molecular specificity of these autoantibodies has not been studied. We performed immunoprecipitation studies utilizing extracts of 125I-labeled suction blister epidermis and the sera of three patients with drug-induced pemphigus foliaceus (two due to penicillamine and one due to captopril) and one patient with captopril-induced pemphigus vulgaris. We found that the three patients with drug-induced pemphigus foliaceus had circulating autoantibodies that are directed against the pemphigus foliaceus antigen complex and that the one patient with drug-induced pemphigus vulgaris had circulating autoantibodies that are directed against the pemphigus vulgaris antigen complex. This study demonstrates that autoantibodies from drug-induced pemphigus patients have the same antigenic specificity, on a molecular level, as do autoantibodies from other pemphigus patients.     

Autoantibodies against desmoglein 2 are not pathogenic in pemphigus

BackgroundAnti-desmoglein 1 and 3 autoantibodies justify acantholysis in pemphigus; however, the pathogenesis of anti-desmoglein 2 is hypothetical.ObjectiveTo compare the participation of desmogleins 1, 2 and 3 through the production of serum autoantibodies, and protein and gene expression in the skin/mucosa of patients with pemphigus foliaceus and pemphigus vulgaris.MethodsThe autoantibodies were titrated by ELISA in 202 samples of pemphigus foliaceus, 131 pemphigus vulgaris, 50 and 57 relatives of patients with pemphigus foliaceus and pemphigus vulgaris, respectively, and 114 controls. Protein and gene expressions were determined by immunohistochemistry and qPCR in the skin/mucosa of 3 patients with pemphigus foliaceus and 3 patients with pemphigus vulgaris.ResultsHigher titers of anti-desmoglein 2 (optical density) resulted in pemphigus foliaceus and pemphigus vulgaris, when compared to controls (0.166; 0.180; 0.102; respectively; p < 0.0001). There was a correlation between anti-desmoglein 2 and anti-desmoglein 1 titers in pemphigus foliaceus (r = 0.1680; p = 0.0206). There was no cross-reaction of anti-desmoglein 2 with desmoglein 1 and 3. Protein overexpression of desmoglein 2 was observed in intact and lesional skin of patients with pemphigus compared to the skin of controls. Internalization granules of desmoglein 1 and 3, but not of desmoglein 2, were observed in lesions of pemphigus foliaceus and pemphigus vulgaris, respectively. Gene overexpression of desmoglein 2 was observed in the mucosa.Study limitationsSmall sample size for the statistical analysis of protein and gene expression.ConclusionAutoantibodies against desmoglein 2 are not pathogenic in pemphigus; protein and gene overexpression of desmoglein 2 in the skin and mucosa may be involved in acantholysis repair.     

The anti-desmoglein 1 autoantibodies in pemphigus vulgaris sera are pathogenic

Pemphigus vulgaris and pemphigus foliaceus are two closely related, but clinically and histologically distinct, autoimmune skin diseases. The autoantigens for pemphigus vulgaris and pemphigus foliaceus are desmoglein 3 and desmoglein 1, respectively. The anti-desmoglein 1 antibodies in pemphigus foliaceus and anti-desmoglein 3 antibodies in pemphigus vulgaris are pathogenic as determined by immunoglobulin G passive transfer animal models. More than 50% of pemphigus vulgaris sera also contain anti-desmoglein 1 autoantibodies; however, the pathogenicity of the anti-desmoglein 1 autoantibodies in pemphigus vulgaris remains unknown. In this study, we used soluble recombinant extracellular domains of desmoglein 1 and desmoglein 3 to obtain affinity-purified anti-desmoglein 1 and anti-desmoglein 3 autoantibodies from pemphigus vulgaris sera and examined the pathogenicity of each fraction separately using the passive transfer mouse model. By immunoprecipitation, the purified anti-desmoglein 1 and anti-desmoglein 3 showed no cross-reactivity. The anti-desmoglein 1 autoantibodies in pemphigus vulgaris induced typical pemphigus foliaceus lesions in neonatal mice, whereas the anti-desmoglein 3 fraction induced pemphigus vulgaris-like lesions. In addition, the pathogenic anti-desmoglein 1 and anti-desmoglein 3 autoantibodies in pemphigus vulgaris had predominant IgG4 subclass specificity. These findings suggest that the anti-desmoglein 1 antibodies in pemphigus vulgaris are pathogenic.     

Pemphiguserkrankungen bei Kindern und Jugendlichen

The pemphigus diseases, which include some of the most severe bullous autoimmune skin reactions, are seen predominantly in middle-aged and elderly individuals. Only endemic pemphigus foliaceus in South America most frequently affects juveniles and children. All non-endemic pemphigus diseases, including paraneoplastic pemphigus, have been reported to occur in adolescents and even very rarely in children younger than 10 years. Pemphigus vulgaris in pregnancy represents a frequently overseen medical problem and may result in fetal growth retardation, intrauterine death, premature delivery and – in about 30% – in neonatal pemphigus vulgaris of the newborn. Contrary to pemphigus vulgaris, the transplacental crossing of autoantibodies against desmoglein1 in pregnant women with pemphigus foliaceus hardly ever leads to neonatal skin lesions in the offspring. This phenomenon can be explained by differences in the distribution and cross-compensation of the pemphigus antigens desmoglein3 and 1 in neonatal and adult skin or mucosa, respectively.     

Common human leukocyte antigen alleles in pemphigus vulgaris and pemphigus foliaceus Italian patients.

Pemphigus refers to a group of autoimmune blistering skin diseases, mainly identified as pemphigus vulgaris and pemphigus foliaceus, both characterized by the presence of autoantibodies against keratinocyte adhesion molecules, leading to loss of cell-cell adhesion with consequent blister formation. Pemphigus vulgaris is reported to be associated with human leukocyte antigen DR4 and/or DR6 whereas no data are available on pemphigus foliaceus, except for the endemic Brazilian form (fogo selvagem), which is reported to be associated with DR1 and DR4. We here report human leukocyte antigen molecular typing on a total of 87 patients, 61 with pemphigus vulgaris and 26 with pemphigus foliaceus, versus 128 healthy matched controls. Generic typing showed an increase of DRB1*04 and DRB1*14 and a decrease of DRB1*07 in both pemphigus vulgaris and pemphigus foliaceus patients. Molecular subtyping of DR4+ and DR14+ subjects showed a highly significant association between the DRB1*1401 and both pemphigus vulgaris (p < 0.0001) and pemphigus foliaceus patients (p < 0.0001) together with a significant increase of the linked DQB1*0503 (pemphigus vulgaris p < 0.0001; pemphigus foliaceus p < 0.0001). Moreover, whereas the association between DRB1*0402 and pemphigus vulgaris (p < 0.0001) has been confirmed, no significant association between a specific allele of the DR4 group and pemphigus foliaceus, has been found. Therefore, at least in Italian patients, pemphigus vulgaris and pemphigus foliaceus share DRB1*1401 and DQB1*0503, as susceptible human leukocyte antigen alleles, whereas DRB1*0402 is only found associated with pemphigus vulgaris. The observation that both diseases, pemphigus vulgaris and pemphigus foliaceus, carry the same susceptible human leukocyte antigen alleles has been interpreted as a common genetic background predisposing to pemphigus as, like in other autoimmune disorders, it is not sufficient to explain the onset of the disease on the basis of the sole aforementioned alleles. Other linked genes and/or environmental factors should play a facilitating role in the outbreak of pemphigus, either pemphigus vulgaris or pemphigus foliaceus.     

Pemphigus foliaceus associated with absence of intercellular antigens in lower layers of epidermis.

A patient with pemphigus foliaceus was found to lack normal intercellular (IC) antigens in the lower layers of the epidermis. This was evidenced by the inability of her own IC antibodies, or of those from other patients with pemphigus vulgaris, to bind to the IC substance in the lower layers of her epidermis; whereas these same antibodies reacted to IC antigens in all layers of normal allogeneic skin and monkey and guinea pig esophagus. Lack of IC antigens in the lower layers of the epidermis may account for the subcorneal location of bullae in some patients with pemphigus foliaceus.     

Pemphigus

Pemphigus diseases comprise a group of autoimmune disorders which are characterized by intraepidermal blisters and autoantibodies to components of desmosomes. Desmosomes mediate adhesion between neighbouring keratinocytes. A common feature of pemphigus diseases are intercellular deposits of IgG or, less frequently, of IgA within the epidermis. The group of pemphigus diseases includes pemphigus vulgaris, pemphigus foliaceus, pemphigus vegetans, pemphigus herpetiformis, pemphigus erythematosus, paraneoplastic pemphigus, drug-induced pemphigus, and IgA pemphigus. Using molecular tools, some of the autoantigens in these diseases have been characterized. In pemphigus vulgaris, autoantibodies are directed to desmoglein 3 and in pemphigus foliaceus to desmoglein 1. Target antigens in IgA pemphigus are desmocollin 1 and desmoglein 3. In paraneoplastic pemphigus, autoantibodies react with a complex of various proteins, including desmoplakin 1 and 2, BP230, envoplakin, periplakin, plectin, desmoglein 3, and a yet uncharacterized 170 kD protein. This review summarizes new insights into the immunopathogenesis and diagnosis of pemphigus diseases.     

Ultrastructural localization of Brazilian pemphigus foliaceus (fogo seivagem) antigens in cultured human squamous cell carcinoma cells

The ultrastructural localization of Brazilian pemphigus foliaceus (BPF) (fogo selvagem) antigen(s) in cultured human squamous cell carcinoma cells was studied using immunogold electron microscopy. Five of six BPF sera, which showed positive cell-surface reactivity on immunofluorescence, bound to the cell-cell contact area of cytoplasmic projections. This binding pattern was apparently different from that of non-endemic pemphigus foliaceus and pemphigus vulgaris sera, and mouse monoclonal anti-buman E-cadherin antibody. The results suggest tbat BPF autoantibodies recognize a molecule(s) which is different from non-endemic pemphigus antigens, or different epitope(s) of a molecule identical with non-endemic pemphigus antigens, and that the epitope(s) to which BPF autoantibodies bind is expressed on cell-cell contact areas at a relatively early stage of cell-cell adhesion formation.     

Childhood Pemphigus Foliaceus with Exclusive Immunoglobulin G Autoantibodies to Desmocollins

Pemphigus refers to a group of potentially fatal blistering skin diseases that are often due to the deleterious effects of autoantibodies directed against desmosomal antigens. Although desmogleins have been mainly implicated as autoantigens in pemphigus, a steadily growing body of evidence suggests that other desmosomal proteins may be causally involved as well. Antibodies directed against desmocollin‐3 have been shown to play a direct role in the pathogenesis of several types of pemphigus. Here we describe the case of a child with localized pemphigus foliaceus and immunoglobulin G (IgG) reactivity exclusively directed to desmocollins. The present report suggests that autoantibodies against nondesmoglein antigens may play a role in the pathogenesis of superficial pemphigus, in addition to pemphigus vulgaris, paraneoplastic pemphigus, and IgA pemphigus.     

Dual diagnosis of Pemphigus and pemphigoid. Retrospective review of thirty cases in the literature.

BACKGROUND: Pemphigus and pemphigoid are two distinct groups of autoimmune blistering diseases. There are many reports of the simultaneous presence of clinical and serological features of both diseases in the same patient. OBJECTIVE: This study is a retrospective review of the present literature on reports of patients with features of both pemphigus and pemphigoid. We recommend that these patients be considered as having a dual diagnosis. METHODS: A review of the English language, peer-reviewed literature was conducted on patients described with features of pemphigus and pemphigoid. Available data on clinical profile, histology, immunopathology, treatment, follow-up and outcome were studied in 30 patients. They were divided into three groups: (1) bullous pemphigoid and pemphigus vulgaris, (2) mucous membrane or cicatricial pemphigoid and pemphigus vulgaris and (3) bullous pemphigoid and pemphigus foliaceus. RESULTS: In all three groups, most patients had a clinical phenotype resembling both diseases. In 17 patients with bullous pemphigoid and pemphigus vulgaris, 83% had a skin biopsy consistent with bullous pemphigoid, 70% had direct immunofluorescence studies typical of bullous pemphigoid and sera of 83% had antibodies typical of pemphigus vulgaris on indirect immunofluorescence. In 10 patients with mucous membrane or cicatricial pemphigoid and pemphigus vulgaris, a histology of mucous membrane pemphigoid was reported in 60% of the patients, direct immunofluorescence studies typical of mucous membrane pemphigoid were reported in 70% of the patients and in 80%, autoantibodies characteristic of pemphigus vulgaris were observed. In 3 patients with bullous pemphigoid and pemphigus foliaceus, the histologies were consistent with bullous pemphigoid, direct immunofluorescence was typical of pemphigus foliaceus and their sera had both autoantibodies. The majority of the 30 patients required long-term high-dose corticosteroids and immunosuppressive agents to control their disease. Three patients with bullous pemphigoid and pemphigus vulgaris (18%) died due to effects of prolonged immunosuppression. CONCLUSION: We characterize a group of patients who have clinical, histological and immunopathological features of bullous or mucous membrane or cicatricial pemphigoid with serological features of pemphigus. These patients did not achieve a prolonged clinical remission by conventional therapy. It is possible that early identification of these patients may improve their prognosis.