用石蜡切片行C3d、C4d免疫组化染色辅助诊断大疱性类天疱疮的意义

目的 探讨C3d、C4d免疫组化染色在石蜡包埋组织切片中辅助诊断大疱性类天疱疮的价值.方法 通过免疫组织化学SP法检测20例大疱性类天疱疮患者石蜡包埋组织切片中C3d、C4d的表达,并与家族性良性天疱疮、大疱性表皮松解症患者及正常皮肤进行对照.结果 20例大疱性类天疱疮患者石蜡包埋切片C3d、C4d真表皮交界基底膜处沉积率为95％(19/20),9例大疱性表皮松解症患者中真表皮交界基底膜处C3d、C4d阳性率0％(0/9),4例家族性慢性良性天疱疮患者基底膜带均为阴性.结论 石蜡包埋组织切片中,C3d、C4d免疫组化染色可以作为辅助诊断大疱性类天疱疮的方法之一.     

大疱性类天疱疮患者皮损中CXCR5的表达及意义

目的 检测滤泡辅助性T细胞（Tfh）趋化因子受体5（CXCR5）在大疱性类天疱疮患者皮损中的表达,分析和探讨CXCR5在大疱性类天疱疮发病机制中的作用。 方法 应用免疫组化法检测16例大疱性类天疱疮患者皮损及10例健康人皮肤中CXCR5的表达。 结果 CXCR5在健康人皮肤表达于基底细胞层,主要表达于细胞质、细胞膜处;在大疱性类天疱疮皮损中表达于基底细胞层与棘细胞层。CXCR5在健康人皮肤中阳性细胞表达均数为11.16 ± 4.47,在大疱性类天疱疮患者皮损中为35.70 ± 12.20,两组差异有统计学意义（t = 6.07,P < 0.01）。 结论 CXCR5可能参与大疱性类天疱疮的发病。     

寻常性银屑病并发成人型线状IgA大疱性皮病

报告l例寻常性银屑病并发成人型线状IgA大疱性皮病.患者男,36岁.因全身红色斑疹伴白色鳞屑反复发生20年.躯干、双上肢出现环状排列的水疱10d伴瘙痒就诊.皮损组织病理检查:表皮下水疱,疱内、真皮浅层和真皮乳头见中性粒细胞、嗜酸性粒细胞浸润;皮损周围皮肤直接免疫荧光显示基膜带Iga、IgG呈带状沉积;取患者血清行BP180NC16A(大疱性类天疱疮18 000抗原的近膜片段)-ELISA检查显示阴性;以盐裂正常人皮肤为底物,取患者血清行间接免疫荧光检查显示IgA、IgG呈带状沉积在真皮侧.诊断为寻常性银屑病并发成人型线状TgA大疱性皮病.     

自身免疫性大疱性皮肤病患者血清中IgM抗体所识别的皮肤抗原的研究

目的:研究自身免疫性大疱性皮肤病患者血清中IgM抗体所识别的皮肤抗原成分.方法:该研究共纳入10例大疱性类天疱疮患者、15例天疱疮患者(包括7例红斑/落叶型和8例寻常型).首先通过直接免疫荧光法分析受试患者皮肤中所沉积的免疫球蛋白或补体,再通过免疫印迹方法分析患者血清中的IgM抗体所识别的皮肤抗原成分.结果:在10例大疱性类天疱疮患者的皮肤中,C3、IgG、IgM单独沉积的例数分别是4例、2例、1例,IgG和C3共同沉积的2例,IgG、C3和IgA三者共同沉积的1例;在15例天疱疮患者中,C3、IgG单独沉积的例数分别是4例和2例,IgG和C3共同沉积的6例,IgM和C3共同沉积的3例.免疫印迹研究发现9例(9/10)大疱性类天疱疮患者、11例(11/15)天疱疮患者血清中的IgM抗体可以识别皮肤中分子量约80 kD的蛋白质.结论:IgM在自身免疫性大疱性皮肤病患者的皮肤中沉积的几率很低,但大多数患者血清中的IgM抗体都能够识别分子量约80 kD的皮肤抗原.     

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。     

湿疹中的循环基底膜带抗体

至少70％的全身性大疱性类天疱疮患者存在抗皮肤基底膜带(BMZ)的循环抗体,此抗体在正常人或其他大疱性皮肤病中不存在,除了妊娠疱疹及瘢痕性类天疱疮外,故对本病的诊断有重要的实际应用价值。作者报导用常规的免疫荧光技术处理,2例成人女性湿疹患者的皮损及其周围皮肤活检标本,其冷冻切片作直接免疫荧光检查,例1的BMZ处未检测到IgG;但7次检查中2次检测到有C_3,呈弱的线状型沉积。例2的BMZ处,2次检查无IgG,第3次检测到有很弱阳性的IgG呈线状型沉积;3次检查中1次查到C_3呈弱阳性线状型沉积。以正常人的皮肤、豚鼠唇、猴食道和病人自己的皮肤作为底物,作间接免疫荧光检查,发现2例患者的血清中含高滴度的抗BMZ的循环抗体,属IgG类抗体,免疫荧光的类型是线状型,与大疱性类天疱疮患者中所见的相同,未发现IgA或IgM类抗体。应用上述所     

扁平苔藓类天疱疮1例

报告1例扁平苔藓类天疱疮.患者男,36岁.因四肢紫红色扁平丘疹2个月,起水疱半个月就诊.皮肤科检查:四肢广泛分布紫红色扁平丘疹,上覆少许鳞屑;在红斑基础上及正常皮肤上散在分布米粒至黄豆粒大水疱,疱壁紧张,疱液清,尼氏征(-).皮损组织病理检查:兼具扁平苔藓及类天疱疮组织学改变.直接免疫荧光示表皮基膜C3强阳性;IgM、IgA、IgG弱阳性呈线状沉积.结合临床和组织病理检查符合扁平苔藓类天疱疮诊断.该病应与大疱性扁平苔藓及大疱性类天疱疮等鉴别.     

儿童大疱性类天疱疮1例

报告1例儿童大疱性类天疱疮,患儿女,8岁,2月前躯干、四肢皮肤出现红斑、水疱、大疱,尼氏征阴性。皮损组织病理检查示:表皮下水疱,疱腔内有嗜酸性粒细胞、中性粒细胞浸润;直接免疫荧光示:IgG、C3线状沉积于基底膜带。诊断为儿童大疱性类天疱疮,静注甲强龙治疗后效果良好,随访至今未复发。     

天疱疮大疱性类天疱疮病损组织中TNFα TNFRp55TNFRp75和iNOS的免疫组化检测

目的探讨TNFα、TNFRp55、TNFRp75、诱生型一氧化氮合成酶(iNOS)在天疱疮、大疱性类夫疱疮病损组织中所起的作用.方法应用免疫组化技术,对14例天疱疮、11例大疱性类天疱疮的皮损进行了检测.结果显示TNFα、TNFRp55、TNFRp75、iNOD在天疱疮和大疱性类天疱疮皮损中均有不同程度的表达,尤其在水疱的顶、底部表达更明显.结论TNFα通过与其受体TNFRp55和TNFRp75结合,在天疱疮表皮角质形成细胞的凋亡和水疱形成、大疱性类天疱疮基底膜带的破坏中可能起一定作用;TNFα可诱导细胞高表达jNOS,产生过量的一氧化氮(NO),可能引起组织的损伤,也可能是加剧天疱疮和大疱性类天疱疮组织损伤的一种因素.     

CD30+皮肤T细胞淋巴瘤并发大疱性类天疱疮

报告1例CD30+皮肤T细胞淋巴瘤并发大疱性类天疱疮.患者男,77 岁.因躯干四肢皮疹伴偶痒3 个月入院.皮损和淋巴结组织病理及免疫组化检查提示CD30+皮肤T细胞淋巴瘤.患者在入院期间出现局限性水疱,组织病理及免疫荧光检查证实为大疱性类天疱疮.给予泼尼松30 mg/d 口服;重组人干扰素α-2b 300万U 肌内注射,隔日1 次,皮损较前减轻.     

表皮下大疱病的鉴别诊断和抗原表达区域性差别的研究

通过间接免疫荧光和盐裂皮损周围皮肤直接免疫荧光（简称盐裂ＤＩＦ），分别研究正常人皮肤、类天疱疮（ＢＰ）及获得性大疱性表皮松解症（ＥＢＡ）抗原表达的区域性差别和表皮下大疱病鉴别诊断。 窝、肘窝、上背、下背、股内侧和下腹部皮肤ＢＰ抗原表达率较高；膝、阳窝、足背、肘、肘窝和下腹部皮肤ＥＢＡ抗原表达率较高。皮肤ＤＩＦ显示２５例表皮下大疱病中１６例（６４％）基底膜带有Ｃ３或ＩｇＧ或伴Ｃ３和ＩｇＡ沉积；盐裂ＤＩＦ表明２５例（１００％）均有ＩｇＧ或伴Ｃ３和ＩｇＡ沉积在表皮侧或真皮侧。结果提示，ＢＰ抗原高表达率与皮损好发部位相一致；ＥＢＡ抗原高表达率一部分与皮损好发部位一致。盐裂ＤＩＦ不仅提高ＤＩＦ阳性率，而且根据免疫反应物沉积部位可以鉴别出ＢＰ与ＥＢＡ以及大疱性系统性红斑狼疮。     

Two Cases of Atypical Bullous Disease Showing Linear IgG and IgA Deposition in the Basement Membrane Zone

Patients showing coexistent linear IgG and IgA deposition along the basement membrane zone on direct immunofluorescence have been described as either bullous pemphigoid, epidermolysis bullosa acquisita, linear IgA bullous dermatosis, or cicatricial pemphigoid, depending on the clinical features and laboratory findings. In the present report, we describe two cases showing atypical clinical features distinct from those of other known bullous diseases. No circulating antibodies were detected by indirect immunofluorescence of normal human skin. Indirect immunofluorescence of 1 M NaCl split skin revealed IgG and/or IgA antibodies reactive with the dermal side of the split. Immunoblotting of normal human epidermal and dermal extracts showed no apparent reactivity with known autoantigens. The results suggest that there may be a unique and distinct bullous disease with linear IgG and IgA deposition at the basement membrane zone.     

IgA-epidermolysis bullosa acquisita in a child resulting in blindness

Epidermolysis bullosa acquisita (EBA) is an acquired subepidermal bullous disease characterized by IgG autoantibodies directed against type VII collagen, the major component of anchoring fibrils. The classical phenotype of EBA is a non-inflammatory, mechanobullous disease resembling the dystrophic forms of inherited epidermolysis bullosa. Mucous membrane involvement is frequent but usually mild. We report a 1-year-old girl suffering from IgA-EBA, who presented with an initial eruption of disseminated urticarial lesions and tense blisters of the skin but subsequently developed severe oral and ocular lesions reminiscent of cicatricial pemphigoid. Direct immunofluorescence of the skin and buccal mucosa revealed linear IgA and C3 at the basement membrane zone (BMZ). IgA anti-BMZ autoantibodies stained the dermal side of salt-split skin by indirect immunofluorescence and recognized a dermal protein of 290 kDa co-migrating with type VII collagen by immunoblotting. Direct and indirect immunoelectron microscopy revealed IgA deposits overlying the anchoring fibrils. The ocular involvement led to total blindness in spite of intense treatment. This case of childhood IgA-EBA is particularly striking because of the cicatricial pemphigoid phenotype with severe ocular involvement which resulted in blindness. It reinforces the necessity to use modern immunological methods to classify autoimmune bullous diseases in order to allow early and appropriate treatment.     

Autoantibodies to bullous pemphigoid and epidermolysis bullosa acquisita antigens in an infant

We describe a 1-year-old boy with multiple tense blisters on the skin, who showed circulating autoantibodies directed to both bullous pemphigoid and epidermolysis bullosa acquisita antigens. The patient's serum IgG antibodies bound to the 290-kDa epidermolysis bullosa acquisita antigen with immunoblot analysis of human dermal extracts. Immunoblot analysis also demonstrated that the patient's serum autoantibodies were reactive with recombinant NC16a domain of the 180-kDa bullous pemphigoid antigen. This study confirmed the presence of circulating autoantibodies directed to both bullous pemphigoid antigen and epidermolysis bullosa acquisita antigen.     

A subepidermal blistering disease with histopathological features of dermatitis herpetiformis and immunofluorescence characteristcs of bullous pemphigoid: a novel subepidermal blistering disease or a variant of bullous pemphigoid?

A 64-year-old man presented with a bullous eruption which clinically and histopathologically resembled dermatitis herpetiformis. However, direct immunofluorescence analysis showed IgG deposits at the basement membrane zone, indicating a relationship with bullous pemphigoid or epidermolysis bullosa acquisita. Indirect immunofluorescence studies on salt-split skin showed binding of IgG mainly on the dermal side of the blister. Immunoblot analysis revealed a novel 200 kDa dermal antigen that could be associated with a major pathogen in this blistering a disease. The histopathological similarity to dermatitis herpetiformis and the immunofluorescence findings indicating bullous pemphigoid or epidermolysis bullosa acquisita seem typical of a distinct subepidermal blistering disease characterized by this 200 kDa antigen. However, the pathogenetic role of autoantibodies against this antigen should be further elucidated before confirming whether this case represents a novel subepidermal blistering disease or a special variant of bullous pemphigoid.     

DIFFERENTIATION OF BULLOUS PEMPHIGOID FROM EPIDERMOLYSIS BULLOSA ACQUISITA ON FROZEN SKIN BIOPSIES

Patients with bullous pemphigoid and epidermolysis bullosa acquisita may have similar clinical, histologic, and routine immunohistologic features. These two diseases can be distinguished by routine diagnostic studies either on a patient's serum tested by indirect immunofluorescence on salt-split normal skin or by obtaining a fresh perilesional skin biopsy, inducing a split at the lamina lucida, and testing for the site of IgG deposition by direct immunofluorescence. Often the serum studies are negative, while direct immunofluorescent studies yield the characteristic linear IgG staining of the basement membrane zone. To eliminate the need for a repeat biopsy to make a laboratory differential diagnosis, we studied the efficacy of salt-splitting perilesional skin biopsies that had been previously submitted and frozen for routine direct immunofluorescent studies. The biopsies were thawed, salt-split, and processed for direct immunofluorescence. Three epidermolysis bullosa acquisita biopsies and seven bullous pemphigoid biopsies examined demonstrated IgG staining at sites consistent with their respective diagnoses. The IgG appeared in the dermal side of the split biopsies in epidermolysis bullosa acquisita and predominantly, or exclusively, in the epidermal side in bullous pemphigoid. Thus the direct immunofluorescent study of previously frozen and subsequently salt-split skin biopsies may be used for the differential diagnosis of bullous pemphigoid from epidermolysis bullosa acquisita. In most cases, it may eliminate the need for a repeat biopsy.     

Human orf complicated by epidermolysis bullosa acquisita

Orf is a DNA parapoxvirus transmitted to humans by contact with infected goats and sheep. Many complications have been reported after orf infection, including erythema multiforme. A few cases of autoimmune bullous dermatosis complicating orf disease have been reported to date. They are usually characterized by tense blister eruptions with or without mucosal involvement; linear deposition of C3, IgG and/or IgA along the basement membrane; and negativity of indirect immunofluorescence analysis and enzyme‐linked immunosorbent assay (ELISA) (performed in four of 11 reported cases). These analyses have targeted antigens of bullous pemphigoid, mucous membrane pemphigoid or epidermolysis bullosa acquisita, except one case of mucosal pemphigoid with antilaminin‐332 antibodies. We describe the case of a patient who presented with an ulceration on his finger 10 days after direct contact with a lamb during Eid al‐Adha. Four weeks later he developed a severe tense blistering eruption associated with mucous membrane erosions. Indirect immunofluorescence analysis using the patient's serum revealed circulating antibasement membrane IgG that bound the dermal side of salt‐split skin. ELISA was positive for recombinant immunodominant NC1 domain of type VII collagen. We finally diagnosed epidermolysis bullosa acquisita complicating probable human orf infection.     

Autoimmune vesicles on the face and neck. A variant of Brunsting-Perry type localized bullous pemphigoid?

We report a case of blistering disease presenting a unique distribution of vesiculobullous lesions on the face and neck which is similar to Brunsting-Perry type of localized bullous pemphigoid (BP). Histopathology of a lesional skin biopsy demonstrated a subepidermal blister. Direct immunofluorescence demonstrated a strong linear deposition of IgG and IgA to the basement membrane zone, and a faint staining for C3. However, circulating antibodies were not detected by indirect immunofluorescence and immunoblotting. And the patient did not develop atrophic scars and was a relatively young woman. This case might be a variant of Brunsting-Perry type of localized BP or localized epidermolysis bullosa acquisita, presenting the clinical heterogeneity of subepidermal blistering diseases.     

The use of Michel's transport medium for immunofluorescence and immunoelectron microscopy in autoimmune bullous diseases

We report the use of Michel's solution, already a well established transport medium, in the combined use of direct immunofluorescence (IMF) and pre-embedding immunoelectron microscopy in 3 subepidermal bullous diseases - bullous pemphigoid (BP), epidermolysis bullosa acquisita (EBA) and dermal binding linear IgA disease (LABD). Our studies demonstrated that electron microscopy of normal skin maintained in Michel's medium for up to 28 clays showed remarkable preservation of all components of the basement membrane zone, including the ultrastructure of the basal keratinocytes, dermoepidermal junction and papillary dermis. However, epidermal cell cytolysis occurred after just 48 hours.
Immunoelectron microscopy using a gold probe has enabled us to localise the immunoreactants in bullous pemphigoid, epidermolysis bullosa acquisita and dermal-binding linear IgA bullous dermatoses. Our findings are comparable to and as equally reliable as those on immunoelectron microscopy of fresh skin biopsies with no loss of antigen deposition, and demonstrate an effective new use of a well established transport medium.