首次命名淀粉样变型类天疱疮并附一例报道

本文首次提出淀粉样变型类天疱疮的概念，并报道一例。患者中年女性，躯干、四肢苔藓样皮疹伴瘙痒17年，无水疱。组织病理示：真皮乳头大量嗜酸性无定形物质沉积，结晶紫染色：阳性；DIF：真皮乳头及基底膜IgG、IgM颗粒状沉积，C3、IgA阴性；IIF：类天疱疮循环抗体IgG 1∶320阳性。人皮肤盐裂IIF：循环抗体IgG表皮侧阳性；抗BP180抗体0 U/mL，抗BP230抗体169.2 U/mL。     

获得性大疱性表皮松解症误诊一例并文献复习

报道一例反复误诊的获得性大疱表皮松解症并对相关文献进行复习。患者,女,25岁。皮疹泛发全身,主要表现为张力性水疱,疱壁紧张,尼氏征阴性,在外院误诊为天疱疮、线状IgA大疱病。组织病理检查示:表皮下水疱;盐裂IIF：IgG沉积在真皮侧; DIF:表皮基底膜IgG、C3、IgM、IgA带状沉积,ELISA：BP180,BP230均阴性,诊断为获得性大疱性表皮松解症,给予甲泼尼龙、氨苯砜、人免疫球蛋白、吗替麦考酚酯等治疗,病情好转。     

盐裂皮肤间接免疫荧光及BP180 NC16a酶联免疫吸附测定在大疱性类天疱疮诊断中的意义

目的 探讨盐裂皮肤间接免疫荧光及大疱性类天疱疮（BP）180 NC16a-酶联免疫吸附测定（ELISA）检测在BP诊断中的意义。方法 收集2015年1月至2017年8月在中国医学科学院皮肤病医院用盐裂皮肤间接免疫荧光（IIF?SSS）和BP180 NC16a?ELISA检测BP患者174例和对照组129例血清。其中25例BP患者用直接免疫荧光（DIF）进行检测并与IIF?SSS和BP180 NC16a?ELISA敏感性进行比较。结果 IIF?SSS、BP180 NC16a?ELISA的敏感性分别为93.67%、96.55%;特异性分别为100%、96.12%。IIF?SSS与BP180 NC16a?ELISA相关系数0.147,为弱相关。其中25例BP患者血清学诊断方法（IIF?SSS,BP180 NC16a?ELISA）和DIF敏感性比较差异无统计学意义。结论 BP血清学诊断方法特异性强、敏感性高,值得临床推广应用。     

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     

BP180NC16a-ELISA对大疱性类天疱疮血清学诊断的评价

目的 评价BP180NC16a-ELISA对大疱性类天疱疮血清学诊断的效能.方法 BP患者42例,对照组42例(其中正常人对照24例,天疱疮18例),在患者用药前采血,比较BP180NC16a-ELISA和盐裂试验免疫荧光(IIF)检测的结果.结果 用BP180NC16a-ELISA检测时,BP患者中有1例呈阴性反应,其敏感性为97.62%;正常人对照组中有1例呈阳性反应,其特异性为97.62%,且BP180NC16a-ELISA法的A值与IIF滴度之间无相关性.结论 BP180NC16a-ELISA在疾病初始阶段是检测血清中抗BP180抗体的有效方法.     

NaCl分离皮肤为底物的IIF法鉴别诊断表皮下大疱病

对1989~1991年间,在本所就诊的100例表皮下大疱病,通过临床、组织病理、直接免疫荧光(DIF)、间接免疫荧光(IIF)法以及1M NaCl分离皮肤为底物的IIF法进行了诊断和评价.结果发现DIF检查的100例中有76例皮肤BMZ有免疫反应物沉积,另24例为阴性,但有典型的自身免疫性大疱病的临床表现;根据临床、DIF和分离皮IIF法修正诊断出大疱性类天疱疮(BP)54例、获得性大疱性表皮松解症(EBA)8例、线状IgA大疱性皮病(LABD)7例(成人4、儿童3),瘢痕性类天疱疮(CP)7例、水疱性红斑狼疮(BSLE)3例、迟发性皮肤卟啉症(PCT)2例、BP和寻常型天疱疮(PV)并发1例,BP和疱疹样皮炎(DH)并发1例,有17例未定.研究结果发现分离皮IIF法对提高表皮下大疱病的诊断水平有较重要的应用价值.但由于CP的抗BMZ抗体可分别出现在表皮侧或真皮侧,所以分离皮IIF法不能单独鉴别CP和BP、CP和EBA.应该结合临床、免疫印迹和免疫电镜等方法进一步诊断,从而也表明分离皮IIF法有一定的局限性.     

儿童线状IgA/IgG大疱性皮病一例

患儿,男,7岁.因躯干、四肢散发红斑、水疱伴瘙痒2个月就诊.发病前有泳池"暴晒"史.背部水疱组织病理:表皮下水疱,真皮乳头中性粒细胞及少许嗜酸粒细胞小脓肿,浅层中性粒细胞、嗜酸粒细胞、淋巴细胞浸润.正常人皮肤盐裂间接免疫荧光:循环抗体IgA、IgG于表皮侧均有沉积,局部区域IgG表皮真皮双侧沉积.综上诊断为儿童线状Ig...     

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.     

盐裂皮损周围皮肤直接免疫荧光鉴别诊断表皮下水疱病

应用盐裂皮损周围皮肤直接免疫荧光 (DIF)检查 2 2例表皮下水疱病的结果 :IgG免疫复合物沉积在表皮侧 1 7例 ,表皮真皮两侧均有 2例 ,均为大疱性类天疱疮 (BP) ;真皮侧 3例 ,2例为获得性大疱性表皮松解症 (EBA) ,1例为大疱性系统性红斑狼疮 (BSLE)。对照组均阴性。此法简便易行 ,经济实用 ,准确可靠 ,值得推广使用     

发生于特瑞普利单抗长期治疗后的大疱性类天疱疮二例

例1女,63岁,全身皮疹2个月伴瘙痒,表现为红斑基础上水疱大疱。2年前因直肠肛管恶性黑素瘤行肠周淋巴结清扫术,静脉注射特瑞普利单抗预防性治疗1年,停药后2周出现全身皮疹。上肢红斑处皮损直接免疫荧光示,IgG沿基底膜带沉积;血清盐裂间接免疫荧光,IgG沿表皮侧线状沉积。血清酶联免疫吸附试验示,BP180 > 200 U/...     

Comparative sensitivity of indirect immunofluorescence to immunoblot assay for the detection of circulating antibodies to bullous pemphigoid antigens 1 and 2

Summary The sera of 263 patients with bullous pemphigoid (BP) were tested by indirect immunofluorescence (IIF) on salt-split skin (SSS) and immunoblot (IB) assay, in order to assess the diagnostic sensitivity of these techniques. Among the 263 sera tested. 198 sera (75%) contained antibasement membrane zone antibodies demonstrable by IIF reacting to the epidermal (98%) or both the dermal and epidermal sides (2%) of SSS. One hundred and eighty-two of the 263 sera (69%) reacted by IB with BP antigens (Ag), most commonly the BPAgl (93 cases, 51%), and a complex of BPAgl and the 180kDa minor BP antigen (BPAg2) (47 cases, 26%). BPAg2 alone was found in 42 cases (23%). A good correlation was found between the detection of autoantibodies by IIF and labelling of BPAg1 and/or BPAg2 by IB assay, in which 152 of 198 sera with an epidermal pattern in IIF identified a BP antigen. IB analysis of the 65 sera negative by IIF yielded positive results in 30 cases (46%). Thirty-one per cent (13 of 42) of sera recognizing by IB BPAg2, were negative by IIF, as compared with 12% (11 of 93) of those recognizing BPAg1 (P < 0.01). Comparing the sensitivity of the two tests, IIF (75%) was found to be more sensitive than IB (69%). Thirty-five of the 263 sera (13%) remained negative by both techniques. It can be concluded from this study that IIF on SSS appears to be a sensitive and reliable assay for screening BP;IB should be performed for the sera that are negative by IIF as it may reveal circulating antibodies, particularly to BPAg2.     

Diagnostic features of pemphigus vulgaris in patients with bullous pemphigoid. Molecular analysis of autoantibody profile

BACKGROUND: The simultaneous presence of features of pemphigus vulgaris (PV) in patients with bullous pemphigoid (BP) has previously been reported in the literature. OBJECTIVE: The purpose of this retrospective study is to present 13 patients with an initial diagnosis of BP, who subsequently demonstrated coexistent serological features of both BP and PV. METHODS: The following information on each patient was documented, at the time of initial diagnosis: clinical profile on presentation, histology, direct immunofluorescence, indirect immunofluorescence (IIF) using monkey esophagus as substrate, salt-split skin (SSS) and an immunoblot assay. Since all 13 patients failed to respond to conventional systemic therapy, intravenous immunoglobulin (IVIg) was used as an alternative treatment modality. Prior to initiating IVIg therapy, in all 13 patients, serological studies were performed. In addition to IIF using monkey esophagus, an immunoblot assay and SSS, an enzyme-linked immunosorbent assay (ELISA) was performed to detect antibodies to desmogleins. These different assays were done to identify pathological autoantibodies typical of BP and PV. A control group of 25 healthy normal individuals, 37 patients with BP, 17 patients with PV and 12 patients with pemphigus foliaceus were used for comparison of serological studies. RESULTS: At the time of initial presentation, histological and immunopathological studies confirmed the diagnosis of BP in all 13 patients. Prior to the initiation of IVIg therapy, results of IIF using monkey esophagus as substrate demonstrated high levels of anti-intercellular cement substance (anti-ICS) or antikeratinocyte cell surface antibody. Sera of all 13 patients on SSS bound to the epidermal side of the split. In an immunoblot, using bovine gingival lysate as substrate, sera of 6 patients bound to both a 230-kD (BP Ag1) and 180-kD protein (BP Ag2), while 7 sera bound to only a 230-kD protein. All 13 patients had high levels of antibodies to desmoglein 3 on ELISA. In a pilot experiment, the anti-ICS antibody in sera from 6 random patients was found to be predominantly of the IgG4 subclass. Use of IVIg resulted in an effective clinical response and the maintenance of a prolonged clinical remission. CONCLUSION: In patients with BP, who are nonresponsive to conventional therapy, the presence of two autoimmune diseases or a dual diagnosis should be considered.     

Use of sodium-chloride separated human skin in detection of circulating anti-basement membrane zone antibodies

The sensitivity of the indirect immunofluorescence (IIF) technique for detection of circulating basement membrane zone (BMZ) antibodies was evaluated, employing NaCl-separated human skin and intact skin as substrate. Consecutive serum samples from 12 patients with clinically, histologically and immunohistologically verified bullous pemphigoid (BP) were investigated in parallel on both substrates, in dilutions ranging from 1:10 to 1:1,280. All BP sera showed linear deposits of IgG at the BMZ on intact skin, with titres ranging from 10 to 160. On NaCl-separated skin, all BP sera produced a linear epidermal fluorescent band for IgG, with titres ranging from 80 to 1,280. None of the sera showed deposits of IgM anti-BMZ antibodies. Sera from 5 healthy donors (dilutions 1:10) produced no fluorescence, either on intact or on NaCl-separated skin. The serum-titres of circulating anti-BMZ IgG antibodies in 2 patients with corticosteroid-resistant BP were significantly reduced (from 160 to less than 10) during treatment with plasmapheresis, when using NaCl-separated skin as substrate for IIF, whereas the serum-titres showed insignificant reduction (from 20 to less than 10), when using intact skin as substrate. We conclude that the IIF method is more sensitive for detection of circulating anti-BMZ antibodies, when NaCl-separated skin as compared with intact human skin is employed as substrate.     

The subclass distribution of IgG autoantibodies in cicatricial pemphigoid and epidermolysis bullosa acquisita

To study the subclass distribution of autoantibodies and their complement-fixing capacity in cicatricial pemphigoid (CP) and epidermolysis bullosa acquisita (EBA) we studied the sera from 23 patients by both indirect immunofluorescence (IIF) on 4-microns cryostat sections of normal human skin and immunoblotting of epidermal or dermal extracts. Monoclonal antibodies of strict specificity for human IgG subclasses were used. Sera from 20 patients with BP served as controls. In addition, total IgG subclass levels were determined by indirect competitive ELISA in all sera. Complement binding capacity was studied by IIF using antibodies to C3 after incubation of skin section with autoantibodies and source of fresh complement. CP autoantibodies reacting with the 230-240 kD and/or the 180-kD epidermal bands showed an IgG4/IgG1 subclass restriction, with a predominance of IgG4 in 10 cases, of IgG1 in four. In BP sera, IgG4 and IgG1 autoantibodies were detected with a similar frequency (100% and 83%, respectively). In EBA sera, autoantibodies reacting with the 290 kD and 145 kD dermal bands also showed an IgG1/IgG4 restriction. Concordant results were obtained by IIF. However, the IIF method had a lower sensitivity for the detection of IgG4 CP antibodies and IgG1 EBA antibodies than immunoblotting. Finally, when CP antibodies were analyzed for their complement-binding activity, it was found that sera containing IgG4 autoantibodies alone never fixed complement whereas all complement-fixing CP sera had IgG1 autoantibodies, suggesting that only this subclass of antibodies is capable of fixing complement.     

Development of a novel ELISA system for detection of anti-BP180 IgG and characterization of autoantibody profile in bullous pemphigoid patients

BACKGROUND: The NC16A immunodominant region of the bullous pemphigoid (BP) antigen BP180 has been used to develop several enzyme-linked immunosorbent assays (ELISAs) as diagnostic tools for BP autoantibody detection. OBJECTIVES: Because BP180 autoantibody reactivity is not restricted to NC16A, we have investigated the possibility of developing an ELISA based on selected epitopes additional to this immunodominant region. METHODS: Initially 78 BP sera were tested using an NC16A ELISA and IgG reactivity was detected in 64 BP sera (82%). The 14 NC16A-negative BP sera were then analysed by immunological screening against seven BP180-specific epitopes. Recombinant phages displaying BP180 epitopes were grown as plaques, blotted onto a nitrocellulose filter and incubated with BP sera. RESULTS: Three and five NC16A-negative BP sera reacted with epitopes AA 1080-1107 and AA 1331-1404 of the BP180 ectodomain, respectively. Thus, a novel ELISA with GST-1080 and GST-1331 (GST-1080/1331) was developed: 32 of 78 BP sera (41%) proved positive by this assay. The combined use of ELISAs with GST-NC16A and GST-1080/1331 detected IgG reactivity in 72 of 78 BP sera, increasing the sensitivity from 82% to 92%. In addition, autoreactivity against the three extracellular epitopes appeared to be related to the presence of both skin and mucosal involvement as assessed by Fisher's exact probability test. CONCLUSIONS: Our findings further characterize the autoimmune response in BP by identifying a subgroup of NC16A-negative patients who react with different BP180 extracellular epitopes. The developed ELISA system appears more sensitive than the ELISA based on NC16A alone and also informative about the epitope profile of BP patients.     

Heterogeneous bullous pemphigoid antibodies: detection and characterization by immunoblotting when absent by indirect immunofluorescence

We studied sera from 59 patients with bullous pemphigoid (BP) and 25 control subjects (normal volunteers, patients with pemphigus, psoriasis, eczema, or other dermatoses) by western blotting analysis on protein bands from normal human heat-separated epidermis. BP sera reacted with four protein bands that were not detected by control sera: two major bands at 220-240 and 165 kD and two faint bands at 190 and 95 kD. Three of these bands were significantly associated with BP: 220-240 kd (51% of the BP patients; p less than 0.001), 165 kD (49%; p less than 0.001) and 190 kD (20%; p less than 0.05). These results are consistent with a molecular heterogeneity of BP antibodies, because each individual BP serum showed a distinctive pattern of reactivity. Thirty out of the 59 BP sera contained anti-basement membrane zone antibodies demonstrable by indirect immunofluorescence (IIF). All these IIF positive BP sera reacted by immunoblotting with at least one protein band: 23 (77%) with the 220-240-kD band and 21 (70%) with the 165-kD band. Furthermore, 45% of the 29 IIF negative BP sera showed a reactivity with the 220-240-kD band and/or the 165-kD band. These results indicate that western immunoblotting might be a more sensitive method for the detection of circulating BP antibodies than IIF techniques, including IIF on salt split skin.     

Bullous pemphigoid in Liguria: A 2-year survey

BACKGROUND: The epidemiology of bullous pemphigoid (BP) is not clear because of the heterogeneity of the disease, and its possible association with internal malignancies has been under debate for many years. We report the findings of a 2-year study on incident BP cases in the Liguria region of Italy. SUBJECTS AND METHODS: Thirty-two patients with BP were collected over the 2-year period. Diagnosis was made based on clinical findings and confirmed by histology, direct immunofluorescence (DIF) and indirect immunofluorescence (IIF) with salt-split skin and monkey oesophagus, and immunoblotting (IB). All patients were thoroughly investigated for possible malignancies and all were followed up for 6 months to monitor the response to treatment. RESULTS: DIF showed linear deposits at the dermoepidermal junction in all but one patient. IIF gave positive findings for 15 sera tested with monkey oesophagus and 20 tested with salt-split skin. IB gave positive findings in 19 cases. There was a malignancy in six cases, but no clinical or immunological features that could be considered to predict this occurrence. CONCLUSIONS: The findings of this study are in accordance with most of the data found in the literature, including the fact that IgG serum levels did not predict the course of the disease. Contrary to previous indications, IgE levels were not indicative of disease course either. Mucosal lesions, erythema multiform-like lesions, negative IIF findings and antibodies to AgPB2 were not a prediction for the development of malignancy.     

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.     

Role of BP180NC16a-enzyme-linked immunosorbent assay (ELISA) in the diagnosis of bullous pemphigoid in China

Background Autoantibodies of bullous pemphigoid (BP) patients react with two components of the hemidesmosome of stratified epithelia: the BP antigen 230 (BP230) and the BP antigen 180 (BP180). Recently, strong evidence has been provided that autoantibodies to BP180 play a key role in subepidermal blister formation in BP patients, and NC16A contains an important antigen determinant of BP. Objective To study the role of BP180NC16a enzyme‐linked immunosorbent assay (BP180NC16a‐ELISA) in the diagnosis of BP in China. Methods Sera from BP patients (n = 42) and control subjects (normal controls, n = 24; pemphigus patients, n = 18) were measured by BP180NC16a‐ELISA. All BP sera were obtained at presentation from patients who had not received previous systemic treatment. The values of immunoglobulin G (IgG) antibody levels measured by ELISA were compared with those measured by indirect immunofluorescence (IIF) (gold standard for the diagnosis of BP) on salt‐split skin. Results Using BP180NC16a‐ELISA, 41 of the 42 BP sera were positive, whereas only one of the serum samples from 24 normal controls was positive and all the pemphigus sera showed a negative result. Thus, the sensitivity and specificity of BP180NC16a‐ELISA were both 97.62%. There was no correlation between the mean ELISA values and IIF titers. The ELISA and IIF results were further compared and analyzed using a 2 × 2 contingency table, which showed that they were not significantly different. Conclusions It is suggested that BP180NC16a‐ELISA is a useful tool for the diagnosis of BP.     

Anatomical distribution and immunological characteristics of epidermolysis bullosa acquisita antigen and bullous pemphigoid antigen

A Japanese patient with epidermolysis bullosa acquisita (EBA) was autopsied, and direct immunofluorescence (DIF) testing was performed. Using this patient's serum (EBA serum) and three bullous pemphigoid (BP) sera, the anatomical distribution and immunological characteristics of EBA antigen and BP antigen were investigated by indirect immunofluorescence (IIF). EBA antigen showed the same anatomical distribution as BP antigen in DIF and IIF studies; both antigens were limited to the skin, tongue, oesophagus, trachea, cornea and bladder. EBA antigen was located on the dermal side of both NaCl and PBS-separated skin, whereas BP antigen was limited to the epidermal side. Ethanol fixation abrogated the antigenic stability of BP antigen, but not that of EBA antigen. No difference was found when acetone or formalin fixation was used. The separation methods and prefixation in ethanol could be useful techniques applicable to the classification of the bullous disorders which manifest circulating anti-BMZ antibodies.