免疫荧光试验在天疱疮临床缓解期病人中的应用

寻常性天疱疮(PV)是一种自身免疫病。直接免疫荧光(DIF)检查可见表皮中有自身抗体, 间接免疫荧光(IIF)检查血清中有循环抗体。DIF是诊断PV的可靠检查方法,IIF检查的抗体滴度常与疾病的严重程度有关。本研究试图探讨在PV临床缓解期免疫荧光检查对判断疗效及预后的指导意义。共研究24例处于临床缓解期的PV病人,其中男性13例(年龄59～78岁),女性11例(年龄     

类天疱疮发病机理的新进展

类天疱疮又称大疱性类天疱疮(Bullous pemphigoid,BP),是一组侵犯人的复层鳞状上皮组织的、自身免疫性的表皮下大疱性疾病。其特征为:1.临床上呈现张力性的厚壁水疱;2.组织学上为表皮下水疱和以嗜酸细胞为主的白细胞浸润;3.直接免疫荧光(DIF)检查皮损附近基底膜带(BMZ)有线状IgG或IgM及C_3沉着;4.间接免疫荧光(IIF)检查血清中有循环IgG抗BMZ抗体;5.超微结构定位:自身抗体和补体位于BMZ电子透明板。和红斑狼疮一样,本病也具有光谱样特征,BP和疤痕性类天疱疮     

用免疫荧光技术观察表皮与血清中天疱疮自身抗体

对11名落叶型(PF),8名寻常型天疱疮(PV)进行了免疫荧光法检查.全部PF与PV表皮ICS在DIF检查中均出现IgG与C₃沉积,其中84.6%患者血清中天疱疮抗体(PAb)阳性,多数滴度为1:40-1:1280,45.4%PF患者在DIF染色中其IgG主要或仅只沉积在表皮浅层ICS部位,而PV则否.著者在讨论中提出:(1)在DIF检查中表皮ICS有IgG沉积即可确诊为天疱疮;(2)若此沉积在表皮浅层之强度较深层伪强,超过一个“+”时可确定为PF;(3)在大疱性皮肤病患者血清中,若PAb滴度较高时(>1:40),可以确定为天疱疮的诊断.     

获得性大疱性表皮松解症抗原和大疱性类天疱疮抗原的解剖学分布和免疫学特性

作者对一例获得性大疱性表皮松解症(EBA)的尸检标本作直接免疫荧光法(DIF)检查,以EBA及大疱性类天疱疮(BP)病人血清作间接免疫荧光(IIF)检查,测定此两病抗原的免疫学特征和分布。方法和结果如下:(1)各个器官组织标本取自     

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.     

天疱疮实验室诊断的研究进展

天疱疮是由血液循环中的天疱疮抗体引起的以皮肤粘膜松弛性大疱为特征的自身免疫性皮肤病 ,其诊断主要依靠临床表现、组织病理和免疫学检查。由于天疱疮抗体滴度水平与病情相关 ,所以临床上以各种免疫学技术检测血清中的天疱疮抗体作为诊断、病情监测及疗效判断的依据。近年来 ,随着分子生物学技术的快速发展 ,天疱疮的实验室诊断有不少新进展 ,使诊断的特异性和敏感性大为提高。1　直接免疫荧光和间接免疫荧光随着免疫病理诊断方法的建立 ,直接免疫荧光 (ＤＩＦ)和间接免疫荧光 (ＩＩＦ)逐渐成为天疱疮诊断必不可少的检查。ＤＩＦ发现Ｉｇ…     

寻常型天疱疮食道受累:临床、组织学及免疫病理学研究

作者对11例新诊断的和1例复发性寻常型天疱疮患者进行了上胃肠道的内窥镜检查。其中女9例,男3例,年龄25～85岁。除1例外,均不接受全身性治疗。活组织检查标本取自食道下、上段(分别为35和20cm处)。每份标本一分为二,一份供组织病理学检查,另一份冰冻切片进行直接免疫荧光检查(DIF)。皮肤活检标本应用同样技术作DIF。病人血清标本作间接免疫荧光检查(IIF),查IgG类天疱疮抗体。结果:内窥镜检查显示5例患者食道受累,3例食道上端有水疱或糜烂,2例整个食道有纵行的线状红斑。这5例中3例有食道     

结节性类天疱疮:2例报告

角化过度性结节是类天疱疮的一种罕见表现,有时不易诊断。作者报告2例确诊的和1例可疑的结节性类天疱疮病例。 例1,49岁,印度籍妇女。瘙痒性皮疹9个月。体检:躯干、四肢、头皮有色素性丘疹、结节,并有脱屑;无大疱;口腔、眼无损害;大腿皮疹活检,未见水疱或裂隙。直接免疫荧光(DIF)检查IgG、C_3在真、表皮结合处(DEJ)呈线状沉积。以分离皮肤为底物,间接免疫荧光(IIF)检查,循环抗基底膜IgG抗体在表皮侧结合。强的松龙每日30mg口服,瘙痒很快减退。一年后病情复     

天疱疮抗体致棘刺松解量化研究

观察1例寻常型天疱疮不同浓度天疱疮抗体（Pab)致表皮棘刺松解的作用，及对组织尿激酶型纤溶酶原（uPA)表达的影响。结果提示间接免疫荧光（IIF）检出Pab量接近Pab致棘刺松解量，IIF阴性可作为病情缓解的可靠指标，而直接免疫荧光（DIF）更适于作为活动性天疱疮的诊断指标。     

异质性大疱性类天疱疮抗体:当间接免疫荧光阴性时,用免疫印迹技术检测及其特性

作者以热分离的正常人表皮的提取物作为抗原来源,用Western免疫印迹技术分析蛋白区带,对59例大疱性类天疱疮(BP)患者的血清进行了研究。59例BP患者中,30例血清用间接免疫荧光(IIF)检查循环抗基底膜带(BMZ)抗体阳性,     

间接ELISA和ABC-ELISA法对天疱疮抗体检测的研究

目的 建立一种新的血清中的天疱疮抗体(PAb)检测方法.方法 利用PI₁天疱疮抗独特型单抗,用亲和层析法纯化患者血清中的PAb;通过间接ELISA法和ABC-ELISA法建立一种PAb的ELISA检测体系.结果 纯化的PAb为IgG4亚类,间接ELISA和ABC-ELISA法检测患者血清中PAb的敏感性和特异性较高,与间接免疫荧光法比较无显着性差异,并初步得到了检测纯化PAb浓度的标准曲线.结论 本研究建立的ELISA法是一种较好的PAb定性分析法,对天疱疮的诊断具有较好的临床应用前景,并为进一步对患者血清中PAb进行定量分析打下了基础;本检测体系针对于PAb的IgG4亚类,为进一步研究IgG4在天疱疮发病机理中的意义提供了一种较好的方法.     

Calcium enhances the sensitivity of immunofluorescence for pemphigus antibodies

Indirect immunofluorescence (IF) to detect pemphigus and pemphigoid autoantibodies is commonly performed with monkey esophagus (ME) as substrate and phosphate-buffered saline (PBS) as a diluent. The purpose of this study was to evaluate comparative IF titers using human skin (HS) as substrate with variations in the buffers employed. Substrates (ME or HS) were incubated in PBS, Tris-acetate-buffered saline (TAS), TAS with 5 mM CaCl+2 (TAS-Ca+2), and PBS or TAS with 1 mM EDTA, prior to incubation with pemphigus or pemphigoid sera for indirect IF. We examined sera from 11 patients with pemphigus vulgaris (PV), 10 patients with Brazilian pemphigus foliaceus (BPF), and 4 patients with bullous pemphigoid. In 20 of 21 pemphigus sera, endpoint indirect IF titers were highest on normal skin with TAS-Ca+2. Six sera (2 PV and 4 BPF) had endpoints that were 5 double dilutions higher than the endpoints obtained with ME and PBS. Six sera (3 PV and 3 BPF) were 4 double dilutions higher, 7 sera (3 PV and 4 BPF) were 2-3 double dilutions higher, and 2 PV sera were equivalent with both substrate/buffers. Preincubation of either tissue with EDTA prior to indirect IF abolished PV and BPF antibody binding completely. Exposure to EDTA after the tissue was incubated with PV or BPF sera did not affect indirect IF titers. In the presence of Ca+2, the antigen was resistant to trypsin in concentrations of 0.001%; however, in the absence of added Ca+2 it was destroyed by 0.0001% trypsin. These differences were not observed with bullous pemphigoid sera; all 4 sera had similar endpoint indirect IF titers. This study shows a significant increase in the sensitivity of indirect IF assays for pemphigus autoantibodies by the use of Ca+2-supplemented buffers on human skin. This finding may also have implications for procedures designed to purify and/or detect pemphigus antigens.     

Immunoblot and immunoelectronmicroscopic analysis of endemic Tunisian pemphigus

Tunisian pemphigus is a newly described form of endemic pemphigus whose clinical, histological and epidemiological characteristics have recently been detailed. The objective of this study was to analyse the binding properties of autoantibodies present in sera from patients with endemic Tunisian pemphigus using immunoblotting and indirect immunoelectron microscopy (IEM). Thirty patients with pemphigus foliaceus (PF) and six with pemphigus vulgaris (PV) seen in the dermatology department of Tunis Hospital between 1992 and 1994 were selected for this study. Seven of 30 (23%) and six of 12 (50%) PF sera tested bound to the 160 kDa band of desmoglein 1 when tested on bovine tongue and human epidermal extracts, respectively. Two of six and two of three PV sera tested bound to the 130 kDa desmoglein 3 in these two extracts. Immunoblot and indirect IEM showed that 24 of 30 (80%) PF sera contained IgG1, IgG3 or IgG4 antibodies that bound to a 185-kDa polypeptide localized on the desmosomal plaque. This immunological analysis showed that most endemic Tunisian pemphigus sera correspond to PF sera and are characterized by a high frequency of autoantibodies directed against a recently identified 185-kDa antigen of the desmosomal plaque.     

天疱疮抗体结合靶抗原的定位研究

目的 研究天疱疮抗体铺皮细胞间抗原在超微结构水平的部位。方法 采用ＬＲＷｈｉｔｅ树脂为包埋剂,用金标记包一直接和间接免疫电镜技术,观察天疱疮患者皮损中ＩｇＧ的沉积部位和患者血清中ＩｇＧ型自身抗体结构正常人皮肤的部位。结果 寻常型天疱疮和落叶型天疱疮的直接和间接免疫电镜均在表皮细胞间的桥粒部位觅金颗粒沉积,在非桥数部位的角质形成细胞间未金颗粒沉积。结论寻常型天疱疮和落叶型天疱疮的靶抗原均是桥粒成分,     

天疱疮患者病情与抗桥粒芯蛋白抗体水平的相关性研究

目的:研究天疱疮患者血清中抗桥粒芯蛋白(desmoglein,Dsg)1和抗Dsg3抗体水平与其皮肤、口腔黏膜损害严重程度的相关性,同时对间接免疫荧光(IIF)检测的天疱疮抗体滴度与治疗中使用皮质类固醇控制剂量的相关性进行分析。方法:采用酶联免疫吸附试验(ELISA)试剂盒测定55例天疱疮患者血清中抗Dsg1和抗Dsg3抗体水平。结果:抗Dsg1抗体水平与患者皮肤损害严重程度有显著相关性(P<0.01),抗Dsg3抗体水平与口腔黏膜损害严重程度有显著相关性(P<0.01)。天疱疮患者血清IIF滴度与抗Dsg1抗体水平相关(P<0.01),寻常型天疱疮患者IIF滴度与抗Dsg1和抗Dsg3抗体水平均有相关性(P分别<0.01和<0.05)。寻常型天疱疮患者皮质类固醇控制剂量与抗Dsg1抗体水平和IIF滴度显著相关(P<0.05)。结论:ELISA方法检测天疱疮患者抗Dsg1和抗Dsg3抗体对天疱疮的临床诊断、分型、衡量口腔黏膜和皮肤损害严重程度具有一定意义。     

Comparison of desmoglein ELISA and indirect immunofluorescence using two substrates (monkey oesophagus and normal human skin) in the diagnosis of pemphigus

A prospective study was performed to assess the usefulness of desmoglein enzyme-linked immunosorbent assay testing compared with indirect immunofluorescence in the diagnosis of new cases of pemphigus, as well as to compare the relative sensitivities of monkey oesophagus and normal human skin as substrates for indirect immunofluorescence. These tests were performed on the sera of 29 consecutive new cases of pemphigus diagnosed over a 2-year period based on clinical, histological and direct immunofluorescence findings. Desmoglein enzyme-linked immunosorbent assay was positive in all patients whereas indirect immunofluorescence was positive in only 25 of 29 patients. All four patients with negative indirect immunofluorescence had positive antinuclear antibodies or cytoplasmic fluorescence that could have masked the anti-intercellular antibodies. Desmoglein enzyme-linked immunosorbent assay appeared to reflect the disease activity better than indirect immunofluorescence in a few patients who had active disease of recent onset. Monkey oesophagus was found to be superior or equal to human skin as a substrate for indirect immunofluorescence in both pemphigus vulgaris and foliaceus.     

Sensitivity of indirect immunofluorescence test in the diagnosis of pemphigus

Indirect immunofluorescence testing of sera from patients with pemphigus produces a positive intercellular staining on a variety of epithelial substrates with different sensitivity. We aimed to determine the sensitivity of indirect immunofluorescence (IIF) test in detecting pemphigus antibodies, using two different substrates: guinea pig lip and human skin. IIF detected antibodies in 66 out of 109 patients with different types of pemphigus. Sensitivity of IIF performed with guinea pig lip was 40%, while with human skin it increased to 69%. However, we found that neither human skin nor guinea pig lip was sensitive enough to make an IIF test reliable for the diagnosis of pemphigus.     

天疱疮患者273例组织病理和免疫病理分析

目的 分析组织病理、DIF、IIF和ELISA四种实验室检测方法在天疱疮患者诊断中的临床意义.方法 回顾性分析273例天疱疮住院患者的组织病理、DIF、IIF、抗Dsgl和抗Dsg3抗体的检查结果.结果 组织病理检查显示,186例(82.30％,186/226)表现为表皮内水疱形成或细胞松解;DIF检查显示,170例(...     

Clinical, histologic and immunologic features of pemphigus in Bangladesh

We studied the clinical, histologic and immunologic features of 32 pemphigus patients evaluated in Dhaka. The diagnosis was pemphigus vulgaris (PV) in 72% of cases, and pemphigus foliaceous (PF) in 28%, with male to female ratios of 1.3 : 1 and 1.25 : 1, respectively. Only 25% of PV patients had vesicles or bullae involving mucous membranes as the initial presentation. Direct (DIF) and indirect (IIF) immunofluorescence tests were positive in all 32 patients. The DIF testing of tissue demonstrated intercellular substance (ICS) deposition of IgG (59%) or IgG plus C3 (41%). Serum ICS antibody titers ranged from 1 : 20–40 in PV and from 1 : 10–20 in patients with PF. To our knowledge, this is the first report concerning the characteristics of pemphigus in Bangladesh.