营养不良型大疱性表皮松解症1家系COL7A1基因诊断和产前诊断

目的分析常染色体显性遗传的营养不良型大疱性表皮松解症(DDEB)1家系的致病基因COL7A1基因突变位点,并在此基础上探讨产前诊断的可行性。方法采用聚合酶链扩增反应及双向直接测序的方法对DDEB家系中2例不同表型的患者进行COL7A1基因突变检测,以家系中的2例健康者及50例无亲缘关系的健康个体作对照。确定致病突变后,对家系中的2例高危胎儿分别进行了产前遗传学分析和胎儿绒毛组织DNA产前诊断。结果该家系2名不同表型患者COL7A1基因的87号外显子第6859位碱基鸟嘌呤G被腺嘌呤A替代(c.6859GA,p.Gly2287Arg),家系中健康对照及无亲缘关系的健康个体均未发现该突变。先证者姐姐怀孕4周,被证明未携带致病突变,继续妊娠并诞生一个健康男婴。先证者配偶怀孕11周,产前诊断胎儿携带与患者相同的突变。先证者夫妇选择治疗性引产术后,取胎儿皮肤标本行基因诊断,结果与产前诊断相同。结论甘氨酸替代突变c.6859GA,(p.Gly2287Arg)是该DDEB家系的致病原因,同一家系患者呈现不同表型。COL7A1基因分析可以快速且准确地进行DDEB的基因诊断和产前诊断,有利于指导生育健康后代。     

反常性痤疮一家系nicastrin基因突变研究

目的 研究一家系2例汉族反常性痤疮患者γ分泌酶基因的突变.方法 提取家系中5名成员(2例患者、先证者父亲、2例目前未发病者)的外周血DNA,扩增nicastrin蛋白(NCSTN)、早老素(PSEN)1、早老素增强子(PSENEN)、前咽缺陷蛋白(APH1)基因所有外显子和侧翼序列进行测序,并以100例无关系健康人作为对照.同时比较先证者皮损与4个健康对照NCSTN基因mRNA表达差异.结果 检测到2例患者血样DNA存在NCSTN基因中第477位碱基发生C→A的杂合突变,即c.477C＞A,其余3名家系成员及健康对照未发现相应突变;查询美国国家生物技术信息中心网站单核苷酸多态性数据库也未发现此突变.另外,先证者皮损NCSTN mRNA水平较健康对照明显减少.结论 NCSTN基因新的无义突变c.477C＞A是该反常性痤疮家系的致病突变,并可能通过无义介导的mRNA降解途径导致该基因功能失活.     

显性营养不良型大疱性表皮松解症一家系调查

报告1例显性营养不良犁大疱性表皮松解症患者的家系调查结果.先证者男,43岁.双小腿水疱、丘疹伴瘙痒33年.皮损组织病理学表现为表皮下裂隙形成,真皮中上部有角质囊肿形成.该家系5代35名成员中有该病患者10例(男4例,女6例),属常染色体显性遗传.     

五个汗孔角化病家系的临床和遗传特点

目的 调查5个3种类型汗孔角化病家系的临床和遗传资料,进而分析3种类型汗孔角化病的临床表现和遗传特点。方法 当5个家系先证者的临床诊断明确后(主要诊断依据是特征性皮损和组织病理),现场调查家系资料,对每个家系的家庭成员进行相关临床检查和遗传调查。这5个家系包括3个播散性浅表光线性汗孔角化病(DSAP,共266人,其中患者100例)家系,1个散发性跖汗孔角化病及播散性汗孔角化病(PPPD,共90人,其中患者26例)家系和1个经典斑块型(PM,共34人,其中患者17例)家系。结果 5个家系都为常染色体显性遗传,都具有以边缘呈堤状隆起、中央萎缩的典型皮损,有角化不全细胞柱的病理变化。可分为3种亚型,每一类型各有自身特色。DSAP型是汗孔角化病的一种常见类型,疾病初发年龄较其他类型早(一般8-20岁),初发皮损在颜面部。PPPD型初发年龄为14-20岁,初发皮损为掌跖部或面部,初发皮损为掌跖部者病情较重,初发皮损为面部者病情较轻。PM型的初发年龄较迟,为20-30岁,初发皮损为前臂、手背、耳际和前额等非特定部位。PPPD型家系和PM型家系都有汗孔角化病不同类型的共存现象。结论 汗孔角化病是一种具有不同表现度的常染色体显性遗传病,具有多种临床类型和不同的遗传基础,不同临床类型间既存在一致性也存在差异性,而且不同临床类     

大疱及疱疹性皮肤病

20110597痒疹样显性遗传真皮型大疱性表皮松解症一家系调查/李丽娜(河南省医院皮肤性病科),朱钵,李建国…∥中华皮肤科杂志.-2010,43(9).-659先证者男,21岁。双下肢起丘疹、结节伴瘙痒15年。该家系4代共26名成员,发现14例患者,男6例,女8例,皮损形态基本一致,但临床表现轻重不一,所有患者皮损主要分布于胫前,前胸后背均有散在丘疹;家系中患者最     

SASH1基因新杂合错义突变与遗传性泛发性色素异常症一家系相关

目的 探讨一家系遗传性泛发性色素异常症的临床特征和分子遗传学发病基础.方法 对该家系进行详细的遗传学调查、体检,并对先证者进行反射式共聚焦显微镜检查和组织病理学检查.采集8例家系成员(其中患者5例,正常个体3例)和200例家系外无亲缘关系的健康对照的血样,提取基因组DNA,PCR扩增SASH1和ABCB6基因的全部外显子及其侧翼序列,并进行DNA直接测序.结果 该家系符合常染色体显性遗传模式.家系中9例患者均在出生后1年内发病,皮损最初累及面部,后逐渐泛发于全身.健在的8例患者中,7例皮损表现为不规则淡褐色至深褐色色素沉着斑,其余皮肤呈均匀的色素减退.仅先证者大姐皮损表现为泛发性网状色素沉着斑和色素减退斑.患者的口腔黏膜、指(趾)甲、牙齿和一般健康状况均正常.DNA测序显示,5例患者SASH1基因第15号外显子上均出现1个杂合错义突变(c.1761C＞G,p.Ser587Arg),而家系中3例正常个体及家系外200例健康对照均未检测到此突变.此外,均未发现ABCB6基因突变.结论 遗传性泛发性色素异常症的临床特征和分子遗传学发病基础存在异质性.SASH1基因的p.S587R突变可能是该家系患者发病的原因.     

痒疹样营养不良型大疱性表皮松解症一家系调查

报告痒疹样营养不良型大疱性表皮松解症一家系调查结果。该家系共5代34名成员,其中患病者15例(男8例,女7例),属常染色体显性遗传。先证者,女性,18岁。于1岁左右双踝部出现数个水疱,双胫前皮肤在外伤、搔抓后形成圆形或卵圆形丘疹、结节,似米粒至花生米大,暗红色,质地较硬,部分皮损表面有痂壳。双足多个趾甲增厚或脱失。皮损组织病理学表现为多处表皮下裂隙形成,真皮内散在较多的表皮样囊肿,并有少量淋巴细胞浸润。该家系中其他患者的皮损与先证者类似。     

角化性皮肤病

20042907　一播散性浅表性汗孔角化病家系BNC、CSK基因突变/胡正茂(中南大学医学遗传学国家重点实验室)…//中华皮肤科杂志.-2004,37(2).-111该家系是一个遗传了3代共37人的湖南家系,有完整的家系图,可推断为常染色体显性遗传。对成员II1,进行了皮损活检,对30个成员采集血样,选取先证者进行突变检测。经过筛选UCSC数据库,在所定位区间内选了2个基因作为DSAP2的候选基因。结果仅在9号外显子发现1个C→T变异,并非致病基因,未能检测到BNC,CSK两基因的突变,提示这两个基因不是导致DSAP2的原因。将进一步收集家系,精细定位,有助于确…     

色素异常性皮肤淀粉样变病一家系GPNMB基因突变检测

目的 对1个色素异常性皮肤淀粉样变病家系的3例有临床表型患者及5例无症状家系成员的GPNMB基因进行变异分析，明确其致病原因与遗传模式。方法 收集先证者血液、口腔拭子样本及部分家系成员口腔拭子样本并提取DNA,对先证者行高通量全外显子测序寻找致病变异，并采用Sanger测序对家系内部分成员进行致病变异的检测。结果 该家系存在GPNMB基因的第5外显子c.565C>T(p.Arg189X)突变，其中先证者GPNMB基因存在c.565C>T(p.Arg189X)纯合突变，其父亲、母亲、姑妈、两位姐姐GPNMB基因的第5外显子均存在c.565C>T(p.Arg189X)杂合无义变异。结论 GPNMB基因第5外显子c.565C>T(p.Arg189X)纯合变异为该色素异常性皮肤淀粉样变病家系患者的致病原因，遗传模式为不完全显性，且存在不同表现度，该突变位点的新遗传模式在国内尚未查到有相同的报道。     

以严重掌跖角化过度为主要表现的表皮松解性角化过度症一家系报告及基因突变分析

目的:报告一个以严重掌跖角化过度为主要表现的表皮松解性角化过度症(epidermolytic hyperkeratosis,EHK)家系,并检测其基因突变情况.方法:收集1个EHK家系的临床资料,取先证者皮损行组织病理检查.提取先证者及其亲属外周血DNA,应用PCR扩增角蛋白1(KRTl)、角蛋白10(KRT10)和角蛋白9(KRT9)基因编码区的全部外显子及其侧翼序列并行双向DNA测序,以100名健康志愿者外周血DNA作正常对照.结果:先证者皮损组织病理符合表皮松解性角化过度,所有患者的KRT1基因第1436位碱基发生T→C错义突变,导致其第479位氨基酸从异亮氨酸(Ⅰ)变为苏氨酸(T) (p.I479T).家系中未受累者和100名正常对照者未检测到该突变.在该家系所有成员中未检测到KRT10和KRT9基因突变.结论:KRT1基因的错义突变(p.I479T)可能是导致该家系患者临床表型的病因.     

The predominance of IgG4 in prodromal bullous pemphigoid

Background Prodromal bullous pemphigoid (PBP) and bullous pemphigoid (BP) demonstrate immunoglobulin G (IgG) and/or C3 deposition at the basement membrane zone (BMZ) on direct immunofluorescence. BP‐180‐specific IgG1, IgG4, and IgE antibodies have been detected in BP. However, the distribution of IgG subclasses is unknown in PBP. Objectives We will describe the role of anti‐BMZ IgG subclasses in PBP and we will correlate these findings to better understand the pathogenesis of PBP. Methods Skin biopsies and serum samples were obtained from 45 patients who had PBP. The skin tissue was processed for direct immunofluorescence studies. Sera were analyzed by indirect immunofluorescence for the presence of circulating anti‐BMZ IgG antibodies (by standard IIF) and IgG subclasses antibodies (by sandwich double antibody immunofluorescence [SDAI]). Sera were also analyzed for antibodies against BP‐180 and BP‐230 antigens by enzyme‐linked immunosorbent assay (ELISA). Results Thirty‐two patients (71%) had IgG and C3 staining at the BMZ, while 13 patients (29%) had isolated C3 staining at the BMZ on direct immunofluorescence. All patients demonstrated staining on the epidermal side of the salt‐split skin. Of the seven skin specimens that were available for C5‐9 SDAI testing, all were found to be positive along BMZ area. Standard IIF studies demonstrated the presence of circulating BMZ antibodies in 11 of the 30 patients (36.6%). When SDAI for IgG subclass differentiation was utilized, 17 of 30 (56.6%) patients were found to have circulating anti‐BMZ antibodies. All of these 17 patients had IgG4 subclass antibodies. Thirteen patients did not have detectable IgG subclass anti‐BMZ antibody on SDAI. Sixteen of 30 patients had detectable anti‐BP‐180 or anti‐BP‐230 antibodies, while 12 (40%) did not have detectable antibody against BP antigens on ELISA. Conclusions IgG4 is the initial and predominant anti‐BMZ antibody subclass detected in PBP. Demonstration of linear C5‐9 at the BMZ enhances the early diagnosis of PBP. Predominance of IgG4 and the initial presence of IgG4 on skin lesions as well as the presence of only IgG4 subclass anti‐BMZ antibody suggest that IgG4 subclass antibody could be the initial immunologic event encountered in patients with PBP.     

Chondroitin 6-sulfate proteoglycan but not heparan sulfate proteoglycan is abnormally expressed in skin basement membrane from patients with dominant and recessive dystrophic epidermolysis bullosa

Two distinct groups of proteoglycans, chondroitin 6-sulfate (C6-S) proteoglycan and heparan sulfate proteoglycan (HSPG), have been recently shown to reside within the lamina densa of normal human skin basement membrane (BM). To determine whether either or both antigens are normally expressed in one or more forms of epidermolysis bullosa (EB), a disease known to have specific alterations in skin BM, we have examined by indirect immunofluorescence 31 specimens of clinically normal skin from 28 EB patients (simplex, 5; junctional, 8; dominant dystrophic [DDEB], 9; recessive dystrophic [RDEB], 9) with monoclonal antibodies to C6-S and HSPG. HSPG was normally expressed in all EB and control skin specimens, whereas C6-S was absent along the dermoepidermal junction of 9 of 9 RDEB and 7 of 9 DDEB, and reduced in 2 of 9 DDEB cases. In contrast, C6-S was normally expressed in 5 of 5 EB simplex, 5 of 6 junctional EB, and all control skin specimens. We have subsequently extracted a greater than 400 kD C6-S proteoglycan from normal skin BM and have found that the core protein may also contain heparan sulfate side chains. Our findings suggest that 3B3 monoclonal antibody recognizes a hybrid proteoglycan in human skin, and that its absent or reduced binding in dystrophic EB skin BM may reflect either absence of associated core protein or posttranslational alterations in the proteoglycan side chains.     

Identification of an epidermal basement membrane defect in recessive forms of dystrophic epidermolysis bullosa by LH 7:2 monoclonal antibody: use in diagnosis

LH 7:2 is a monoclonal antibody that was raised against an extract of human epidermal cells and identifies an epitope within the lamina densa of the basement membrane of stratified squamous epithelia. Using indirect immunofluorescence we found intense labelling with LH 7:2 at the epidermal basement membrane (EBM) of normal skin, and in skin samples from patients with simplex, junctional, dominantly inherited dystrophic and acquired forms of epidermolysis bullosa (EB), as well as bullous pemphigoid. Staining was absent or only very faint in generalized recessive dystrophic EB (RDEB), and patchily reduced in the localized form of RDEB. We conclude that LH 7:2 recognizes an EBM antigen which may be important in the pathogenesis of RDEB. Moreover, the antibody provides a useful probe for the rapid diagnosis of RDEB and is of special value in helping to discriminate between localized RDEB and typical dominant dystrophic EB--conditions which closely resemble each other clinically and which cannot be distinguished by means of transmission electron microscopy.     

A unique epithelial basement membrane antigen defined by a monoclonal antibody (KF-1)

The basement membrane zone (BMZ) of human skin is a complex structure which contains several well-defined components including bullous pemphigoid antigen, laminin, type IV collagen, and proteoglycan. Characterization of additional basement membrane (BM) constituents has been limited by their relative inaccessibility, insolubility, and low tissue concentration. We have produced a murine monoclonal antibody that has enabled us to define a unique constituent of the BMZ of human stratified squamous epithelia. The monoclonal antibody (KF-1) was raised by standard techniques using suction blister-derived trypsinized human epidermal cells as the antigen. Indirect immunofluorescence and immunoperoxidase staining of human and rhesus monkey tissues with KF-1 produced linear BMZ staining of stratified squamous epithelia. Glandular and vascular BMs were not stained. Immunoelectron microscopic studies of normal human skin and esophagus showed specific binding of KF-1 to the lamina densa of the BMZ, a localization identical to that of type IV collagen. However, unlike type IV collagen, which is not species specific and is found in all BMs, the antigen defined by KF-1 is collagenase-resistant and is specific for primate stratified squamous epithelia. These findings confirm the existence of regional variation in BM composition, and demonstrate for the first time that the lamina densa of stratified squamous epithelial BMs contains a constituent other than type IV collagen.     

IgE and its related phenomena in bullous pemphigoid

This study was designed to analyse IgE and its related phenomena in bullous pemphigoid (BP). We analysed 17 BP sera by indirect immunofluorescenee (IIF) and immunoblotting (IB) using a monoclonal antibody to IgE. In addition, inflammatory cells in lesional skin from 11 patients with BP were analysed by the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique using monoclonal antibodies to IgE and Fc?RII/CD23. IgE class anti-basement membrane zone (BMZ) autoantibody was detected in nine of 17 sera (52.9%) by IIF. IgG class anti-BMZ antibody could block the BMZ-binding reactivity of IgE class antibody. Titres of IgE class autoantibody in the sera ranged from 1:40 to 1:320, and statistically correlated with serum IgF levels. Two of 11 sera contained an IgE class autoantibody which recognized a 230-kDa BP antigen by IB. By radio-allergosorbent test (RAST), IgE-specific antibodies to an extended series of common inhalant and food allergens were detectable in six sera with higb concentrations of total IgF(over 3300 IU/ml). IgE-bearing and Fc?RII-expressing cells were demonstrated in the upper dermis and along the BMZ in seven of 11 biopsy specimens by tbe APAAP technique. The distribution and number of IgE-bearing cells in the lesions were similar to those of the FcEERII-expressing cells. Tbese results suggest that botb IgE-mediated immune responses and autoimmunity characterize BP as distinctive features.     

Detection of a novel basement membrane antigen by GDA-J/F3 anti-human sperm fibrous sheath monoclonal antibody

Basement membrane zones (BMZ) of human epithelia were stained with GDA-J/F3 monoclonal antibody, which was originally raised against sperm cells. Using indirect immunofluorescence and immunoperoxidase techniques, the antibody reacted with the BMZ of stratified squamous epithelia (skin and its appendages, tongue, lip, oesophagus and cervix). It also stained the BMZ of trachea, nasal ciliated mucosa, some mammary ducts of lactating and resting breast, amnion and ureter but failed to react with that of stomach, ileum, colon, rectum, kidney, liver, fallopian tube, lung or their blood vessels. In testes, the antibody did not react with the BMZ of the seminiferous tubules although the sperm tails were stained. Split-skin immunofluorescence and immunoelectron microscopy localized GDA-J/F3 antigen to the inferior border of the lamina densa of the BMZ. In human foetuses, the epidermally associated antigen was detected at an estimated gestational age of 9 weeks, and in the amnion at 15 weeks. The antibody reacted with tissues from monkey but not from mouse, rat, cow or pig suggesting the late appearance of the antigen during evolution. Although the GDA-J/F3 was difficult to characterize biochemically, its tissue distribution, ontogeny and ultrastructural localization suggests that this antigen may be a type VII collagen-associated protein, whose expression is altered in recessive dystrophic epidermolysis bullosa. This disease could represent abnormalities in type VII collagen structure, assembly, transport or interaction with associated proteins.     

The localization of target antigens and autoantibodies in linear IgA disease is variable: correlation of immunogold electron microscopy and immunoblotting

Linear IgA disease (LAD) of adults and children is a dapsone-responsive, autoimmune subepidermal blistering disease characterized by linear IgA deposits at the basement membrane zone (BMZ) of the skin and mucosa. Circulating IgA antibodies to BMZ components are often present. In this study we investigated the ultrastructural localization of the antigens and autoantibodies in six patients with LAD (five adults and one child). Using a direct postembedding immunogold electron microscopy (EM) technique, three different patterns of IgA antibody deposition were seen in the skin of four patients with LAD. The IgA deposits localized within the uppermost part of the lamina lucida and to the basal surface of the hemidesmosome in two patients, to the lamina lucida in one, and to the lamina densa in the fourth patient. Using an indirect immunogold EM technique and serum or purified blister fluid from two additional LAD patients, we showed that the serum autoantibodies of one patient bound to the hemidesmosome of the BMZ, while the autoantibodies in the blister fluid of the other patient bound to the lamina densa and sublamina densa including the anchoring fibrils in a labelling pattern similar to that of the monoclonal antibody (LH7.2) to collagen VII. All the autoantibodies binding to the hemidesmosome or lamina lucida recognized a protein in epidermal extracts of molecular weight 180 kDa or its breakdown product of 97 kDa, 200 kDa or 230 kDa. The antibodies binding to the lamina densa recognized proteins of 180 and 285 kDa. The antibodies that bound to the lamina densa and anchoring fibrils recognized collagen VII. In this immunogold EM study we have shown four patterns of IgA labelling in six patients with LAD, associated with five different antigens as recognized by immunoblotting. These results, together with our previous immunofluorescence and immunoblotting findings add support to the contention that LAD is a heterogeneous disease as regards both the target antigens and epitopes.     

Vesiculobullous systemic lupus erythematosus: Report of two cases and a review of the literature

Vesicles and bullae complicating systemic lupus erythematosus (SLE) are relatively uncommon. Two young women with SLE presented with vesiculobullous eruptions on sun-exposed areas that resembled dermatitis herpetiformis (DH) histologically. There were active visceral manifestations of SLE in both patients, including mesangioproliferative glomerulonephritis. Granular deposits of IgG and/or IgM, along with IgA, were demonstrated along the basement membrane of skin by direct immunofluorescence microscopy. Review of fifteen additional cases of vesiculobullous SLE reported in the literature suggests that this cutaneous manifestation of SLE is associated with a high incidence of IgA deposits in skin and glomerulonephritis. The following criteria for the diagnosis of a distinct subset of vesiculobullous skin lesions occurring in patients with SLE are proposed: (1) a diagnosis of SLE based upon American Rheumatism Association (ARA) criteria; (2) vesicles and bullae arising upon but not limited to sun-exposed skin; (3) histopathology compatible with DH; (4) negative indirect immunofluorescence for circulating basement membrane zone (BMZ) antibodies; (5) direct immunofluorescence reveals IgG and/or IgM and often IgA at the BMZ.     

Childhood vulval lichen sclerosus: autoimmunity to the basement membrane zone protein BP180 and its relationship to autoimmunity

Lichen sclerosus (LS) is associated with autoimmune disease in female children and adults. In adult women, there are antibody and T‐cell responses to proteins in the basement membrane zone (BMZ). The aim of this study was to investigate reactivity to the BMZ in girls with LS. Nine girls with vulval LS were studied clinically and serologically. The presence of circulating BMZ autoantibodies was investigated. Autoimmunity was assessed by personal and family history of autoimmune diseases and autoantibodies. We detected circulating BMZ antibodies in four of the nine children, all with IgG responses. Three patients were positive by indirect immunofluorescence, one had a positive ELISA reaction to bullous pemphigoid antigen (BP)180, and three had a positive reaction on BP180 immunoblots. There was no association with autoimmune disease or clinical features. To our knowledge, this is the first study to find BMZ autoantibodies in children with vulval LS. The autoantibodies were directed at BP180 and were exclusively of the IgG class.     

Subclass distribution of IgG autoantibodies in bullous pemphigoid

The distribution of IgG subclasses in the antibasement membrane zone autoantibody of pemphigoid in skin and serum was analyzed by use of monoclonal antibodies to human IgG subclasses. The predominant subclass was IgG4 which was present in 23 of 24 skin biopsies, IgG1 was next and IgG3 was found only occasionally. In 3 of 24 biopsies IgG4 was the only IgG subclass detected, C3 was absent in 2 of these, the third contained IgM and C3. Serum autoantibodies were similarly analyzed by indirect immunofluorescence (IIF) when again IgG4 autoantibody was the dominant subclass. No IgE autoantibody was detected by IIF.