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。     

桥粒芯蛋白1酶联免疫吸附试验检测落叶型天疱疮血清抗体的评价

目的 探讨桥粒芯蛋白1（Dsg1）酶联免疫吸附试验（ELISA）检测落叶型天疱疮（PF）血清学抗体的意义。方法 将80例PF患者和132例对照人群的血清标本进行编盲,用ELISA法检测所有标本中抗Dsg1自身抗体,同时应用间接免疫荧光（IIF）法检测所有标本的抗体滴度,操作过程为随机检测,最后将两方法的结果进行比较。结果 75例PF患者和5例对照人群（包括1例不确定值,大疱性类天疱疮、SLE、皮肌炎、湿疹及健康者各1例）Dsg1 ELISA为阳性,71例PF患者和0例对照人群IIF为阳性。Dsg1 ELISA的敏感性为93.8%（95%可信区间0.85 ~ 0.98）,特异性为96.2%（95%可信区间0.91 ~ 0.99）。IIF的敏感性为88.8%（95%可信区间0.82 ~ 0.96）,特异性为100%（95%可信区间0.96 ~ 1.00）。两者相比,敏感性（P = 0.289）和特异性（P = 1.000）差异均无统计学意义。结论 Dsg1 ELISA是一种简便、敏感、特异的血清学检测方法,可作为诊断PF的一种辅助手段。     

噬菌体展示技术构建寻常型天疱疮患者桥粒芯糖蛋白3抗体文库

目的：用噬菌体展示技术构建寻常型天疱疮患者桥粒芯糖蛋白3抗体文库。方法：收集寻常型天疱疮患者外周血,分离淋巴细胞抽提RNA,采和RT－PCR反应以人的抗体特异性HuVHBACK、HuJHFOR和HuVkBACK、HuJkFOR引物扩增抗体重链可变区（VH）和轻链可变区（VL）基因片段,并将其与接头拼接成ScFv片段克隆于大肠杆菌。结果：抗体基因ScFv片段与噬菌体基因III的产物以融合蛋白的形式在噬菌体表面表达,构建了天疱疮患者的重组噬菌体抗体库,ELISA检测证明ScFv基因产物具有抗体活性。结论：天疱疮噬菌体抗体库的构建为天疱疮发病机理、诊断和治疗研究打下了基础。     

非桥粒芯糖蛋白抗体在寻常型天疱疮中的作用机制

既往发现寻常型天疱疮(pemphigus vulgaris,PV)发病机制与桥粒芯糖蛋白1(desmoglein,Dsg1)和Dsg3抗体有关,目前在PV患者中发现大量非Dsg自身免疫性抗体,包括桥粒胶蛋白1(desmocollin,Dsc1)和Dsc3、斑菲素蛋白1(plakophilin,PKP1)和PKP3、斑珠...     

抗构象表位桥粒芯蛋白抗体和乙酰胆碱受体抗体滴度与寻常型天疱疮病情活动度的相关性研究

【摘要】 目的 研究寻常型天疱疮患者血清中相关抗体滴度与病情严重程度和病情活动度的相关性。方法 收集2012—2015年于中国医学科学院皮肤病医院首次就诊的24例活动期寻常型天疱疮（PV）患者,评估患者活动期和稳定期天疱疮疾病面积指数（PDAI）,并采集血清标本。采用酶联免疫吸附实验（ELISA）检测血清标本中具有致病作用的抗构象表位桥粒芯蛋白（Dsg）抗体滴度、总Dsg抗体滴度和乙酰胆碱受体（AChR）抗体滴度。计量资料比较采用t检验,计数资料比较采用Fisher 精确检验法,相关性比较采用Pearson分析。结果 活动期患者的Dsg1抗体滴度（611.4 ± 136.8）与抗构象表位Dsg1抗体滴度（585.5 ± 134.7）差异无统计学意义（t = 0.13,P = 0.89）,Dsg3抗体滴度（708.6 ± 130.7）高于抗构象表位Dsg3抗体滴度（297.2 ± 54.4,t = 2.90,P < 0.01）。活动期患者的Dsg1抗体滴度及抗构象表位Dsg1抗体滴度与PDAI评分均呈正相关（r = 0.54、0.54,均P < 0.01）;Dsg3抗体滴度与PDAI评分无相关性（r = 0.11,P = 0.62）,抗构象表位Dsg3抗体滴度与PDAI评分呈正相关（r = 0.53,P < 0.01）。20例稳定期患者血清中Dsg1抗体与抗构象表位Dsg1抗体滴度与首次就诊时比较均明显下降。Dsg3抗体滴度仅7例明显下降;13例仍存在较高滴度的Dsg3抗体,其中6例抗构象表位Dsg3抗体滴度明显下降,5例AChR抗体滴度由阳性转为阴性。结论 Dsg1抗体及抗构象表位Dsg1抗体滴度都可以反映病情活动度。部分患者病情活动度与Dsg3抗体滴度不一致,抗构象表位Dsg3抗体或者AChR抗体可能有助于反映病情活动度。     

寻常型天疱疮患者及一级亲属抗体IgG亚型检测

目的观察寻常型天疱疮(pemphigus vulgaris,PV)患者和一级亲属血清中PV自身抗体(PV-IgG)亚型及其与桥粒芯糖蛋白(desmoglein,Dsg)反应性,探讨PV-IgG检测的临床意义和PV的发病机制。方法用间接免疫荧光法(indirect immunofluorescence,IIF)检测27例PV患者、40例一级亲属和20例正常对照者血清PV-IgG,并用免疫印迹法(Westernblot,WB)检测PV-IgG亚型及其与Dsg1、Dsg3反应性。结果PV患者的PV-IgG阳性率和平均抗体滴度明显高于其一级亲属者(P0.001),活动期患者的PV-IgG阳性率和平均抗体滴度也明显高于缓解期(P0.01);PV患者中PV-IgG阳性率(92.9%)明显高于一级亲属(17.5%)和正常对照组(0%)(P0.0001,0.001),PV患者中Dsg3反应性IgG4水平明显高于一级亲属(P0.01),且其在活动期病例中水平也明显高于缓解期(P0.05)。结论本病的发生与PV-IgG水平有关;Dsg3是PV的主要自身抗原;IgG4是PV的主要致病抗体。     

寻常型天疱疮信号通路研究进展

 寻常型天疱疮（PV）是一种罕见的自身免疫性大疱性皮肤病,其病理生理特征是自身抗体与桥粒芯蛋白（Dsg）结合导致棘层松解。尽管关于天疱疮发病机制信号通路的研究越来越多,但目前信号通路在天疱疮发病机制的作用尚未明确。p38丝裂原活化蛋白激酶（p38MAPK）、蛋白激酶C（PKC）和表皮生长因子受体（EGFR）等信号通路的活性改变被认为与PV发病密切相关。本文就相关信号通路阐述PV发病机制,为其寻找有效靶向药物治疗提供新的思路。     

桥粒芯糖蛋白抗体与天疱疮临床表型和疾病活动度的关系及其变化规律

【摘要】 目的 评估酶联免疫吸附试验（ELISA）检测桥粒芯糖蛋白1（Dsg1）、Dsg3抗体与天疱疮患者临床表型、疾病活动度的关系及变化规律。方法 收集2015年1月至2018年1月在中国医学科学院皮肤病医院就诊的天疱疮患者111例,按临床分型,采用ELISA测定患者初发时、控制阶段、维持阶段及复发时血清Dsg1、Dsg3抗体水平变化,并分析其变化规律。采用SPSS22软件,多组间比较采用单因素方差分析,组间两两比较采用LSD?t检验。结果 天疱疮初发时、控制阶段、维持阶段及复发时分别有92例、53例、33例、9例患者完成检测。92例初发患者中,36例落叶型天疱疮患者Dsg1和Dsg3抗体水平阳性率分别为100%、2.77%,10例黏膜型寻常型天疱疮分别为20%、80%,46例黏膜皮肤型寻常型天疱疮分别为97.82%、95.65%。初发时、控制阶段、维持阶段和复发时落叶型天疱疮患者Dsg1抗体水平分别为（137.43 ± 77.74）、（13.94 ± 14.81）、（21.50 ± 58.33）、（121.13 ± 86.89） U/ml;黏膜型寻常型天疱疮患者Dsg3抗体水平分别为（125.61 ± 94.81）、（34.5 ± 16.26）、0.6、258 U/ml;黏膜皮肤型寻常型天疱疮患者Dsg1抗体水平分别为（115.39 ± 70.62）、（15.74 ± 25.10）、（3.62 ± 12.09）、（78.60 ± 92.25） U/ml;Dsg3抗体水平分别为（137.98 ± 81.25）、（58.14 ± 63.46）、（29.26 ± 64.70）、（136.9 ± 101.47） U/ml。落叶型天疱疮Dsg1抗体水平和黏膜型寻常型天疱疮、黏膜皮肤型寻常型天疱疮Dsg3抗体水平在控制阶段、维持阶段均低于初发时、复发时（P < 0.05）。治疗过程中2例患者出现表位扩展现象,4例患者病情稳定期时出现Dsg抗体高滴度现象。结论 Dsg抗体谱与天疱疮临床表型相关,其ELISA值可用于监测疾病活动,并可对治疗的有效性作出评价。     

桥粒芯糖蛋白3刺激下寻常型天疱疮患者外周血T淋巴细胞增殖检测分析

目的 观察桥粒芯糖蛋白3（Dsg3）对寻常型天疱疮（PV）患者外周血T淋巴细胞增殖的影响。方法 PV患者12例,正常人22例,提取外周血单一核细胞（PBMC）,采用流式细胞仪技术分别检测Dsg3刺激下PBMC中T淋巴细胞亚群变化及T淋巴细胞增殖情况,分析PV患者与正常人之间存在的差异。结果 PV患者PBMC在Dsg3刺激下Th2型T淋巴细胞所占比例为12.17% ± 5.32%,Th1型T淋巴细胞所占比例为4.08% ± 1.50%;未经Dsg3刺激的阴性对照组分别为9.84% ± 5.41%和3.91% ± 1.38%。PV患者PBMC在Dsg3刺激和无Dsg3刺激时的Th2型T淋巴细胞所占比例均显著高于正常人（P < 0.05）;Dsg3刺激下PV患者T淋巴细胞发生增殖,CD4+ T淋巴细胞增殖率为4.65% ± 3.28%,显著高于正常人（P < 0.05）。结论 Dsg3可诱导PV患者CD4+ T淋巴细胞发生特异性增殖,且主要为Th2优势型。     

天疱疮患者桥粒芯糖蛋白抗体水平和疾病活动度关系及变化规律分析

目的 研究56例天疱疮患者疾病严重程度和桥粒芯糖蛋白1（Dsg1）和桥粒芯糖蛋白3（Dsg3）酶联免疫吸附试验（ELISA）指数之间的关系,探讨Dsg ELISA指数在不同型别天疱疮中转归的规律。 方法 用ELISA测定36例寻常型天疱疮和20例落叶型天疱疮患者治疗前、病情缓解且糖皮质激素开始减量时、糖皮质激素减量至相当于初始量1/2时、维持治疗开始时以及随诊2年时体内Dsg1和Dsg3 ELISA指数。 结果 Dsg ELISA指数与天疱疮疾病活动度相关,在疾病缓解时,Dsg ELISA指数下降,与治疗前差异均有统计学意义（P < 0.01）。在患者病情稳定使用维持量糖皮质激素、疗程到2年时,落叶型天疱疮中10例（50%）、寻常型天疱疮中7例（19.4%）Dsg1 ELISA指数出现阴性,只有1例（2.7%）寻常型天疱疮患者Dsg3 ELISA指数阴性。 结论 Dsg ELISA指数和天疱疮患者疾病严重程度相关,可能是一种评估病情的有用指标,可对治疗的有效性作出评价。     

Evaluation of desmoglein enzyme-linked immunosorbent assay (ELISA) in Indian patients with pemphigus vulgaris

OBJECTIVE: To evaluate the role of the enzyme-linked immunosorbent assay (ELISA) test for the detection of antibodies to desmoglein 1 (dsg1) and desmoglein 3 (dsg3) in the diagnosis of pemphigus vulgaris (PV), and its correlation with disease severity and clinical presentation (mucosal PV, cutaneous PV, mucocutaneous PV). METHODS: Twenty-seven active PV patients and 26 controls with other dermatologic disorders were included in the study. The severity of oral and cutaneous involvement was assessed and recorded. ELISA test for the measurement of anti-dsg1 and anti-dsg3 antibodies was performed (Medical and Biological Laboratories Co. Ltd., Nagoya, Japan). The cut-off ELISA value for both anti-dsg1 and anti-dsg3 was taken as 20. RESULTS: Of the 27 patients, 26 were ELISA positive for anti-dsg1 antibodies and 23 for anti-dsg3 antibodies. Of the controls, two were positive for anti-dsg1 and none for anti-dsg3 antibodies. The sensitivity and specificity of ELISA for anti-dsg1 in the diagnosis of PV were 96.3% and 92.3%, respectively. For anti-dsg3, they were 85.2% and 100%, respectively. The different morphologic types of PV could not be differentiated on the basis of antibody profile; however, a direct correlation between anti-dsg3 titers and the severity of oral disease was noted, and also between anti-dsg1 titers and the severity of cutaneous disease. CONCLUSIONS: ELISA (dsg1 and dsg3) is an efficient tool for confirming the diagnosis of PV. Specific antibody titers correlate with disease severity; however, desmoglein testing cannot differentiate between the various morphologic subtypes of PV.     

Evaluating efficacy of plasmapheresis for patients with pemphigus using desmoglein enzyme-linked immunosorbent assay

BACKGROUND: Pemphigus is an autoimmune bullous disease caused by circulating IgG autoantibodies against cell-cell adhesion molecules between keratinocytes: desmoglein (Dsg) 3 and Dsg1. Plasmapheresis is often used to treat severe cases of pemphigus. Enzyme-linked immunosorbent assays (ELISAs) against recombinant Dsg3 and Dsg1 have recently become available, allowing us to quantify IgG autoantibodies against Dsg3 and Dsg1. OBJECTIVES: Using ELISA against recombinant Dsg3 and Dsg1, to evaluate the efficacy of plasmapheresis in pemphigus. METHODS: Sera obtained from 10 patients with pemphigus vulgaris and one with pemphigus foliaceus following a total of 16 cycles of centrifugal plasmapheresis and 12 effluents from the plasmapheresis were subjected to ELISA against Dsgs. The percentage of IgG autoantibodies removed was calculated using two different formulae: one used serum titres before and immediately after plasmapheresis and the other used the absolute amounts of IgG autoantibodies in the effluents. The percentage fall of anti-Dsg antibody level was also calculated using the serum titres 1 day after plasmapheresis. RESULTS: Using serum titres immediately after plasmapheresis, there was a mean fall per treatment in anti-Dsg 3 antibody level of 43.0% (n = 12) and in anti-Dsg1 antibody level of 48.4% (n = 7). By contrast, calculated from the effluents, on average one treatment removed only 14.6% of anti-Dsg3 antibodies (n = 12) and 16.4% of anti-Dsg1 antibodies (n = 7). This should reflect the correct percentage as it is based on the absolute amounts of IgG autoantibodies removed. Using serum titres 1 day after plasmapheresis, there was a mean fall per treatment in anti-Dsg 3 antibody level of 12.9% (n = 2) and in anti-Dsg1 antibody level of 8.4% (n = 4). The percentage of IgG autoantibodies removed 1 day after plasmapheresis was lower than that found to be removed immediately after plasmapheresis (n = 6). CONCLUSIONS: One centrifugal plasmapheresis procedure eliminates about 15% of the IgG autoantibodies from the whole body. The percentage fall of anti-Dsg IgG antibody level differed depending on when the serum samples were obtained after plasmapheresis. The change in the percentage fall of anti-Dsg antibody level within 1 day after plasmapheresis is thought to be attributable to the passive diffusion of the IgG autoantibodies from the extravascular space to the intravascular space. Therefore, removal of IgG autoantibodies calculated using serum titres only should be evaluated carefully considering the equilibration of the IgG autoantibodies between the different body spaces.     

Desmoglein 1 and 3 enzyme-linked immunosorbent assay in Iranian patients with pemphigus vulgaris: correlation with phenotype, severity, and disease activity

BACKGROUND: Pemphigus vulgaris (PV) is a chronic autoimmune blistering disorder of the skin and mucosa characterized by the presence of autoantibodies against desmoglein3 (Dsg3). Some patients also have antibodies against desmoglein1 (Dsg1). The aims of this study were to evaluate the diagnostic value of Dsg enzyme-linked immunosorbent assay (ELISA) in Iranian PV patients, to assess its correlation with the clinical phenotype and severity of disease and to investigate the changes of these antibodies after treatment. METHODS: Seventy-three patients with PV (29 men, 44 women) presenting to the Pemphigus Research Unit at Razi Hospital, Tehran, Iran were enrolled. ELISAs were used to detect IgG autoantibodies reactive with the ectodomains of Dsg1 and Dsg3, and the correlation of antibodies with the clinical phenotype as well as oral and skin disease severity was assessed. In addition, the tests were repeated in 18 patients after treatment and the resulting remission. RESULTS: Anti-Dsg1 and anti-Dsg3 were detected in 56 (76.7%) and 69 (94.5%) patients, respectively. Anti-Dsg1 and anti-Dsg3 antibodies were present in 48 (94.1%) and 50 (98%) patients with mucocutaneous type, in 2 (12.5%) and 15 (93.7%) patients with mucosal type, and in 6 (100%) and 4 (66.7%) patients with cutaneous PV, respectively. The mean anti-Dsg1 index values were significantly higher in cutaneous and mucocutaneous phenotypes than mucosal PV (P < 0.001). The mean anti-Dsg3 index values were significantly lower in cutaneous and mucosal phenotypes than mucocutaneous PV (P < 0.01). The severity of skin lesions (but not oral lesions) was correlated with anti-Dsg1 antibody level (P < 0.001); on the other hand, the severity of oral lesions (P < 0.01) as well as skin lesions (P < 0.001) was significantly correlated with anti-Dsg3 antibody levels. Both anti-Dsg1 and anti-Dsg3 levels were significantly reduced after treatment and clinical remission (P < 0.001). CONCLUSION: Dsg ELISA is not only a sensitive tool for the diagnosis of PV, it can also serve as a predictive means for assessing the severity as well as for monitoring the disease activity. Although, in general, the clinical phenotype is related to the antibody profile, there are occasional cases with discordant phenotype and antibody profile. These discrepancies might be explained by genetic variations or the presence of possible minor antigens involved in the pathogenesis of pemphigus.     

Monitoring disease activity in pemphigus with enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3

BACKGROUND: Pemphigus is an antidesmoglein (Dsg) autoimmune disease that is divided into two major subtypes: pemphigus foliaceus (PF) and pemphigus vulgaris (PV). We previously developed enzyme-linked immunosorbent assays (ELISAs) using recombinant Dsg1 and Dsg3 to detect IgG autoantibodies in patients with pemphigus. The protocol for the ELISAs was optimized for serological diagnosis, but under the conditions used, these assays were not particularly useful for monitoring disease activity in certain patients. That is, the sera from some patients with high-titre antibodies continued to show high index values in the ELISA after clinical improvement. OBJECTIVES: In the study reported here, we modified the ELISA protocol to obtain 'true' index values that exhibit a better correlation with disease activity. METHODS: We tested two cases of pemphigus foliaceus (PF) and four cases of pemphigus vulgaris (PV), each with ELISA index values greater than 150 for Dsg1 or Dsg3. We ran an ELISA with sera from these patients serially diluted from 1 : 100 to 1 : 12,800. We then performed ELISA with a series of PV No. 1 sera diluted to 1 : 800 and PV No. 2-4 and PF No. 1-2 sera diluted to 1 : 1600, after which we plotted the ELISA index values against the time course of disease activity. RESULTS: In each of these cases, there was no apparent decline, over the course of the disease activity, in the ELISA index values at a serum dilution of 1 : 100, probably because the antigen-antibody reaction was saturated at that dilution. After running an ELISA with sera serially diluted from 1 : 100 to 1 : 12,800 we found that a linear dose-dependency between the dilution value and the index value was only observed when sera were diluted to 1 : 800 or more in one case (PV No.1) and to 1 : 1600 or more in the other five cases (PV No. 2-4, PF No. 1-2). After performing ELISA with these series as outlined above we plotted the ELISA index values against the time course of disease activity and found that the index values obtained from these appropriately diluted sera fluctuated in parallel with disease activity, and declined with clinical improvement. CONCLUSIONS: These findings indicate that when appropriate dilutions are used in Dsg1 and Dsg3 ELISA, these assays can provide useful serological information for assessing disease activity in PF and PV.     

In vitro keratinocyte dissociation assay for evaluation of the pathogenicity of anti-desmoglein 3 IgG autoantibodies in pemphigus vulgaris

Patients with pemphigus vulgaris (PV) have circulating anti-desmoglein (Dsg) 3 immunoglobulin G (IgG) autoantibodies that induce blister formation. We developed an in vitro quantitative assay to evaluate the pathogenic strength of anti-Dsg3 IgG autoantibodies in blister formation. To obtain intercellular adhesion mediated dominantly by Dsg3, we used primary cultured normal human keratinocytes expressing low level of Dsg2 in the presence of exfoliative toxin A that specifically digests Dsg1. After incubation with various antibodies, monolayers released by dispase were subjected to mechanical stress by pipetting, and the number of cell fragments were counted. When anti-Dsg3 monoclonal antibodies (mAb) obtained from pemphigus model mice were tested, pathogenic AK23 mAb yielded significantly higher number of cell fragments than AK7 or AK20 non-pathogenic mAb. Dissociation scores, defined with AK23 mAb as the positive control, were significantly higher with active stage PV sera (n=10, 77.4+/-21.4) than controls (n=11, 16.0+/-9.6; p=0.003). When pair sera obtained from 6 PV patients in active stage and in remission were compared, the dissociation scores reflected well the disease activity as those in active stage were four to 17 times higher than those in remission. When sera from different patients showing similar ELISA scores but different clinical severity were tested (n=6), the dissociation scores with sera from severe disease activity were significantly higher than those with sera in remission. These findings indicate that this dissociation assay will provide a simple and objective biological method to measure the pathogenic strength of pemphigus autoantibodies.     

The clinical transition between pemphigus foliaceus and pemphigus vulgaris correlates well with the changes in autoantibody profile assessed by an enzyme-linked immunosorbent assay

BACKGROUND: There are a number of reports of pemphigus with clinical shifting between pemphigus foliaceus (PF) and pemphigus vulgaris (PV). On the other hand, a novel enzyme-linked immunosorbent assay (ELISA) against recombinant baculoproteins of desmoglein 1 (Dsg1) (PF antigen) and Dsg3 (PV antigen) has been established and found to be extremely sensitive and specific. OBJECTIVES: To characterize the change in the antibody profiles in a series of pemphigus cases with mixed features of PF and PV by various methods, including the novel ELISA. Patients/methods Sera were obtained from eight cases undergoing a shift between PF and PV and three cases of coexistent PF and PV. The autoantigens were analysed by ELISA, as well as by immunofluorescence using normal human skin sections and immunoblotting using normal human epidermal extracts. RESULTS: The results of the ELISA, immunofluorescence and immunoblotting studies showed that the transition between PF and PV correlates well with the changes of autoantibodies against either Dsg1 or Dsg3. CONCLUSIONS: The clinical phenotype at each stage is defined by the anti-Dsg antibody profile in the serum of these pemphigus patients showing mixed features of PF and PV. In addition, ELISA using recombinant baculoproteins was particularly useful in distinguishing PF and PV.     

Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus

Pemphigus is an autoimmune blistering disease with two major subtypes, pemphigus vulgaris (PV) and pemphigus foliaceus (PF). Patients with pemphigus have circulating antidesmoglein (Dsg)1 and/or anti-Dsg3 IgG autoantibodies. We have previously developed enzyme-linked immunosorbent assays (ELISAs) using recombinant Dsg1 and Dsg3 expressed by baculovirus as a diagnostic tool for pemphigus. The purpose of this study was to evaluate the practical application of these ELISAs for clinical use with a large number of serum samples. We used 81 PV sera, 48 PF sera, 114 bullous pemphigoid (BP) sera, 124 collagen disease sera, nine sera of other non-pemphigus bullous diseases and 179 normal control sera. A cut-off value was determined by receiver-operating-characteristic plots. Forty-seven of 48 PF sera (97.9%) were positive in the Dsg1 ELISA and 79 of 81 PV sera (97.5%) were positive in the Dsg3 ELISA, while only two (1. 1%) and four (2.2%) of 179 normal sera were positive in Dsg1 and Dsg3 ELISAs, respectively. However, some disease control sera of BP and collagen diseases exceeded the cut-off value. Introduction of a grey zone helped to decrease the number of these false-positive sera. Furthermore, in three patients studied, the respective Dsg1 and Dsg3 ELISA scores showed parallel fluctuation with the disease activity along the time course. We conclude that Dsg1 and Dsg3 ELISAs provide a simple, sensitive and highly specific assay for the diagnosis of patients with PV and PF and that these ELISAs may be a valuable tool to monitor the disease activity. We also propose diagnostic criteria for pemphigus based on ELISA reactivity: if a serum is positive against Dsg3 it indicates a diagnosis of PV, regardless of reactivity against Dsg1; if a serum is negative for Dsg3 and positive for Dsg1, it indicates a diagnosis of PF.     

Application of the enzyme-linked immunosorbent assay (ELISA) in the serodiagnosis of syphilis.

The enzyme-linked immunosorbent assay (ELISA) technique, using an ultrasonicate of Treponema pallidum as antigen, has been evaluated as a serological test for syphilis. It is concluded that the test is simple, reliable, and relatively quick and that its sensitivity in all stages of syphilis is equal to the FTAABS test. Because its specificity is probably also high, ELISA might be used in future as a first-line screening test for the serodiagnosis of syphilis.