免疫球蛋白在自身免疫性大疱病的研究进展

免疫球蛋白作为人体内具有活性的免疫效应分子,在自身免疫性大疱病的发病、诊断及治疗中发挥着重要的作用。人免疫球蛋白可以分为IgG、IgA、IgM、IgD、IgE5大类,除IgD外其他都在自身免疫性大疱病的发病过程中存在并发挥不同的作用。IgG可通过激活补体、活化白细胞、释放蛋白水解酶等诱发水疱形成,其不同亚型也有所区别。IgA可引起粒细胞迁移从而导致水疱脓疱的发生。IgE与荨麻疹样红斑、嗜酸性粒细胞浸润相关。IgM多见于巴西落叶型天疱疮。这些都为自身免疫性大疱病的诊断和治疗提供了新的思路。     

线状IgA大疱病基底膜带自身抗体的靶抗原研究进展

线状IgA大疱病在临床表现、靶抗原的种类及超微定位上表现为异质性 ,其IgA型基底膜带自身抗体可结合多种基底膜蛋白 ,这些抗原成分分布在真表皮基底膜的不同部位 ,不同的抗原表位可被基底膜带自身抗体识别。部分线状IgA大疱病拥有与其它自身免疫性大疱病相同的靶抗原及表位。     

自身免疫性大疱病IgA型基底膜带抗体的分析

大疱性类天疱疮(BP)、线状IgA大疱病(LABD)、获得性大疱性表皮松解症(EBA)和大疱性系统性红斑狼疮(BSLE)等疾病是一组自身免疫性表皮下大疱病.目前普遍认为,IgA型抗体线状沉积于基底膜带(BMZ)是LABD的特征.近来研究发现BP、EBA和BSLE等亦存在IgA型基底膜带抗体(BMZ-Ah)^[1],因此我们应用免疫印迹法对这组疾病的IgA型和IgG型BMZ-An进行分析.     

自身免疫性表皮下大疱病诊疗共识（2022）

【摘要】 自身免疫性表皮下大疱病主要包括大疱性类天疱疮、黏膜类天疱疮、瘢痕性类天疱疮、妊娠类天疱疮、扁平苔藓类天疱疮、线状IgA大疱性皮病、获得性大疱性表皮松解症、抗p200/层黏连蛋白γ1类天疱疮及疱疹样皮炎,不同的疾病靶抗原和致病性自身抗体存在差异,临床表现有相似之处也有明显不同,诊断与鉴别诊断有赖临床、病理、免疫病理检查及血清抗体检测等。为规范此类疾病的临床诊疗,中华医学会皮肤性病学分会和中国医师协会皮肤科医师分会组织本领域专家,依据近年国内外临床研究数据和指南共识,制订本共识 。     

表皮下自身免疫性大疱病相关抗原研究进展

综述了表皮下自身免疫性大疱病表皮基底膜带相关抗原的研究进展,有ＢＰＡＧ１、ＢＰＡＧ２、Ⅶ型胶原、Ⅳ型胶原、板层素５和６、９７ｋＤ蛋白、４５０ｋＤ蛋白、１２０ｋＤ蛋白、４５ｋＤ蛋白、２００ｋＤ蛋白、１０５ｋＤ蛋白、１００ｋＤ和１４５ｋＤ蛋白等。讨论它们各自在发病中的作用,并就研究方向提出展望。     

生物疗法在自身免疫性大疱病中的应用

自身免疫性大疱病(AIBD)是指一组发生在皮肤和黏膜、以水疱和大疱为基本损害,重者可危及生命的自身免疫性皮肤病。目前,AIBD治疗方法仍首选糖皮质激素或联合免疫抑制剂,但长期应用不良反应多,且部分患者疗效欠佳或不能耐受。随着对AIBD分子病理学机制认识的不断深入,针对这类疾病不同病理过程的生物疗法不断出现,该文将对其在自身免疫性大疱病中的应用及进展作一综述。     

自身免疫性疱病与银屑病的相关性

自身免疫性疱病（AIBD）和银屑病的发病机制有部分相似性。近年来,多项研究报道银屑病与AIBD之间具有相关性,以大疱性类天疱疮多见,还包括寻常型天疱疮、红斑型天疱疮、线状IgA大疱性皮病等,多数AIBD在银屑病发病后发生,部分患者两病同时出现或AIBD先出现。本文综述AIBD与银屑病发病的相关性及可能存在的机制。     

自身免疫性表皮下大疱病基底膜带自身抗体的检测

目的 比较免疫印迹（ＩＢ）和盐裂皮肤间接免疫荧光（ＩＩＦ）检测自身免疫性表皮下大疱病（ＳＡＢＤ）基底膜带自身抗体（ＢＭＺ－Ａｂ）的敏感性和特异怀。方法 分另应用ＩＩＦ和ＩＢ技术对９７例ＳＡＢＤ患者血清中ＩｇＧ型或ＩｇＡ型ＢＭＺ－Ｚｂ进行检测。结果 ＩＩＦ法阳性率７５．３％,免疫印迹法阳性率７９．４％,两者在检测灵敏度上无显著性差异。结论 二者均可用于ＳＡＢＤ中ＢＭＺ－Ａｂ的检测,二者联用对于提高Ｂ     

表皮下自身免疫性大疱病相关抗原研究进展

综述了表皮下自身免疫性大疱病表皮基底膜带相关抗原的研究进展,有BPAG1、BPAG2、Ⅶ型胶原、Ⅳ型胶原、板层素5和6、97kD蛋白、450kD蛋白、120kD蛋白、45kD蛋白、200kD蛋白、105kD蛋白、100kD和145kD蛋白等。讨论它们各自在发病中的作用,并就研究方向提出展望。     

Blister fluid for the diagnosis of subepidermal immunobullous diseases: a comparative study of basement membrane zone autoantibodies detected in blister fluid and serum

The subepidermal immunobullous diseases bullous pemphigoid (BP), cicatricial pemphigoid (CP), pemphigoid gestationis (PG) and linear IgA disease (LAD) are characterized by circulating and in vivo deposition of antibodies to antigens in the cutaneous basement membrane zone (BMZ). Indirect immunofluorescence (IMF) of serum is a routine diagnostic test to detect circulating BMZ antibodies in these diseases. We have compared the titres of IgG and IgA and their subclasses, also of IgM and IgE BMZ antibodies in serum and aspirated blister fluid in 35 adult patients with subepidermal immunobullous diseases: BP ( n  = 30), PG ( n  = 2), CP ( n  = 1), and LAD ( n  = 2), by indirect IMF on intact and salt-split skin. The antibody titre in blister fluid was the same or one dilution less than serum in most cases and there was no significant difference between these results ( P  > 0.05). IgG1 and IgG4 were the predominant subclasses in both blister fluid and serum in BP. Indirect IMF of serum and blister fluid was also carried out on trypsinized epidermal cells in a subgroup of patients with BP ( n  = 19). Typical polar fluorescence was obtained in all 14 cases which had positive indirect IMF on intact and split skin. Our findings demonstrate that blister fluid can be used as an alternative to serum for indirect IMF in subepidermal immunobullous diseases. This avoids the need for venesection and has a practical application in children and those with poor venous access.     

Assessment of skin basement membrane zone antibodies in the urine of patients with acquired subepidermal immunobullous diseases

BACKGROUND: In bullous pemphigoid (BP), cicatricial pemphigoid (CP) and linear IgA disease (LAD), autoantibodies to the basement membrane zone (BMZ) are found in skin and mucosa, blood and blister fluid. OBJECTIVES: To assess whether BMZ antibodies might also be detected in urine. METHODS: Urine and serum samples from 62 patients (32 with BP, 17 with CP and 13 with LAD) were analysed for antibody isotypes and subclasses by indirect immunofluorescence, and urine and serum samples from 40 patients (25 with BP, eight with CP and seven with LAD) were screened for target antigens using immunoblotting. RESULTS: Fourteen of 32 patients with BP had detectable levels of IgG BMZ autoantibodies in their urine, and all 32 had positive sera. Of these 14 BP patients, 13 had epidermal-binding serum autoantibodies at a titre > 1 : 160, and one had dermal-binding serum antibodies at a titre of 1 : 40. BMZ autoantibodies were not detected in the urine of the CP or LAD patients, but the corresponding sera were of low titre or negative. IgG subclasses (IgG1-4) were less frequently detected in urine than in serum. IgG4 was the predominant subgroup found (10 urine samples and all 14 sera), followed by IgG1 (two urine samples and 12 sera); IgG2 was detected in a single urine sample and three sera, and IgG3 was not detected. Eight of 25 BP and one of eight CP urine samples were positive on immunoblotting, and bound BP230 and/or BP180 with IgA and/or IgG autoantibodies. IgA autoantibodies were not detected in the urine of the seven LAD patients. The corresponding sera were often more positive, with 21 of 25 BP, five of eight CP and six of seven LAD sera immunoblotting the major BP antigens. CONCLUSIONS: The detection of IgG autoantibodies from urine samples using indirect immunofluorescence correlated with a high titre of IgG autoantibodies in the serum. IgG and IgA autoantibodies in the urine were detected by immunoblotting, although less frequently than in serum. The finding of BMZ antibodies in the urine of many BP patients may have clinical relevance, and may have a restricted application in the diagnosis of immunobullous disease.     

Scanning electron microscopy of a blister roof in dystrophic epidermolysis bullosa

In dystrophic epidermolysis bullosa the genetic defect of anchoring fibrils leads to cleavage beneath the basement membrane, with its consequent loss. We performed scanning electron microscopy of an inverted blister roof of a case of dystrophic epidermolysis bullosa, confirmed by immunomapping and gene sequencing. With a magnification of 2000 times a net attached to the blister roof could be easily identified. This net was composed of intertwined flat fibers. With higher magnifications, different fiber sizes could be observed, some thin fibers measuring around 80 nm and thicker ones measuring between 200 and 300 nm.     

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.     

Immunofluorescence testing in the diagnosis of autoimmune blistering diseases: overview of 10-year experience

BACKGROUND

Immunofluorescence testing is an important tool for diagnosing blistering diseases.

OBJECTIVE

To characterize the immunofluorescence findings in patients diagnosed with autoimmune blistering skin diseases.

METHODS

We retrospectively analyzed immunofluorescence results encompassing a 10-year period.

RESULTS

421 patients were included and divided into 2 groups: group 1- intraepidermal blistering diseases (n=277) and 2- subepidermal blistering diseases (n=144). For group 1, positive DIF findings demonstrated: predominance of IgG intercellular staining (ICS) and C3 for pemphigus foliaceus-PF (94% and 73% respectively), pemphigus vulgaris-PV (91.5%-79.5%) and paraneoplastic pemphigus-PNP (66%-33%); ICS IgA in 100% of IgA pemphigus cases, and IgG deposits in the basement membrane zone (BMZ) along with ICS in one Hailey-Hailey patient. The IIF findings revealed mean titers of 1:2.560 for PV and 1:1.280 for PF. For paraneoplastic pemphigus, IIF was positive in 2 out of 3 cases with rat bladder substrate. In group 2, positive DIF findings included multiple deposits at basement membrane zone for epidermolysis bullosa acquisita-EBA (C3-89%,IgG-79%,IgA-47%,IgM-21%) mucous membrane pemphigoid-MMP (C3,IgG,IgA,IgM-80%) and bullous pemphigoid-BP (C3-91%,IgG-39%,IgA-11%,IgM-6%), and IgA at basement membrane zone for IgA linear disease (99%) and dermatitis herpetiformis-DH (dermal papillae in 84.6%). For lichen planus pemphigoides, there was C3 (100%) and IgG (50%) deposition at basement membrane zone. indirect immunofluorescence positive findings revealed basement membrane zone IgG deposits in 46% of BP patients, 50% for EBA, 15% for IgA linear dermatosis and 50% for LPP. Indirect immunofluorescence positive results were higher for BP and EBA with Salt-Split skin substrate.

CONCLUSION

Our results confirmed the importance of immunofluorescence assays in diagnosing autoimmune blistering diseases, and higher sensitivity for indirect immunofluorescence when Salt-split skin technique is performed.     

Bovine gingival lysate: a novel substrate for rapid diagnosis of autoimmune vesiculo-bullous diseases. A preliminary observation

Immunoblot assays have been developed to characterize the autoantigens and to detect autoantibodies in muco-cutaneous autoimmune vesiculo-bullous diseases using different substrates. However the results have been inconsistent, because availability and standardization of different substrates has been a major problem. The aim of this study was to develop an immunoblot assay using bovine gingival lysate as substrate because it is easily and readily available as well as inexpensive. Sera from patients with different vesiculo-bullous diseases were studied. These included 25 patients with pemphigus vulgaris (PV), 8 with paraneoplastic pemphigus (PNP), 12 with pemphigus foliaceus (PF), 25 with bullous pemphigoid (BP), and 22 with cicatricial pemphigoid (CP). Serum samples from 40 normal human volunteers were also studied. The autoantibody titers were determined based on the binding pattern of each disease and compared to those obtained by routine indirect immunofluorescence (IIF). Our observations suggest that the titers from immunoblot assays were significantly higher than titers obtained by IIF (P<0.0001). When the autoantibody titers were compared using bovine gingival lysate and human epidermal lysate as substrate, statistically significant differences were not observed. The use of bovine gingival lysate as a substrate will facilitate the rapid and early serological diagnosis of patients with vesiculobullous diseases. It may also be of benefit to laboratory investigators studying these autoantibodies.     

Effectiveness of Cytochrome c Oxidase Subunit 1 Gene Nested Polymerase Chain Reaction Compared to Dermoscopy or Microscopy Alone for the Detection and Diagnosis of Sarcoptes scabiei var. hominis Infection

BackgroundBackground: While microscopy (MS) evaluation of skin scrapings has a 100% positive predictive value and specificity by definition for scabies diagnosis, it has low sensitivity. Dermoscopy (DS) has not yet been widely accepted for diagnosis, and long-term clinician training is required.ObjectiveTo evaluate the diagnostic validity of cytochrome c oxidase subunit 1 (cox1) gene nested polymerase chain reaction (PCR) as an adjunctive method for diagnosing scabies.MethodsThis was a prospective, single institution study, conducted on a total of 302 skin lesions from 50 patients suspected of scabies at Kangdong Sacred Heart Hospital in Seoul, Korea. DS, MS, and cox1 nested PCR were performed on all patients.ResultsOf the 302 lesions, 145 (48.0%) were obtained at first visit and 157 (52.0%) were identified in the course of follow-up visits after treatment. For all lesions, DS and MS sensitivity levels were 55.9% (73/136) and 55.2% (75/136), respectively, with cox1 gene nested PCR considered as 100%. The results of DS and MS identification showed no difference between each other and showed significant difference from that of cox1 gene nested PCR.ConclusionNested PCR detecting cox1 may be prospectively used to comprehensively diagnose lesions of scabies in clinical practice.