ELISA技术检测寻常型天疱疮桥粒芯蛋白3抗体水平研究

目的:评价酶联免疫吸附试验(ELISA)检测抗桥粒芯蛋白(desmoglein,Dsg)3抗体在寻常型天疱疮(PV)诊断中的意义.方法:对来自不同中心的106例PV患者和106例对照人群血清标本编盲后,进行ELISA检测抗Dsg3自身抗体和间接免疫荧光(IIF)法检测血清天疱疮自身抗体.结果:ELISA法检测Dsg3抗体敏感度为77.4%,特异性为94.3%;IIF法检测抗体敏感度为79.2%,特异性为94.3%.两组间的差异无统计学意义.结论:ELISA方法检测Dsg3抗体对于寻常型天疱疮的诊断是一种较好的辅助方法.     

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。     

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。     

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。     

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。     

落叶毒素引起角质形成细胞桥粒蛋白损伤的研究

落叶毒素由金黄色葡萄球菌分泌，是金黄色葡萄球菌性烫伤样皮肤综合征的主要致病因素。金黄色葡萄球菌性烫伤样皮肤综合征与落叶型天疱疮在临床及组织病理上非常相似，电镜发现落叶毒素作用于桥粒，近期研究揭示落叶毒素的作用底物为桥粒芯蛋白1。关于落叶毒素致病机制的理论主要有：落叶毒素的微观结构显示其类似于谷氨酸特异的丝氨酸蛋白酶，通过酶解作用裂解桥粒芯蛋白1；落叶毒素还可能具有超抗原活性，通过免疫反应破坏桥粒。对落叶毒素作用位点、作用机制的研究将有助于了解金黄色葡萄球菌性烫伤样皮肤综合征和天疱疮等相关疾病的发病机制，提高诊断与治疗水平。     

落叶毒素引起角质形成细胞桥粒蛋白损伤的研究

落叶毒素由金黄色葡萄球菌分泌 ,是金黄色葡萄球菌性烫伤样皮肤综合征的主要致病因素。金黄色葡萄球菌性烫伤样皮肤综合征与落叶型天疱疮在临床及组织病理上非常相似 ,电镜发现落叶毒素作用于桥粒 ,近期研究揭示落叶毒素的作用底物为桥粒芯蛋白 1。关于落叶毒素致病机制的理论主要有 :落叶毒素的微观结构显示其类似于谷氨酸特异的丝氨酸蛋白酶 ,通过酶解作用裂解桥粒芯蛋白 1;落叶毒素还可能具有超抗原活性 ,通过免疫反应破坏桥粒。对落叶毒素作用位点、作用机制的研究将有助于了解金黄色葡萄球菌性烫伤样皮肤综合征和天疱疮等相关疾病的发病机制 ,提高诊断与治疗水平     

天疱疮患者表皮中桥粒芯蛋白1的分布模式

目的:探讨桥粒芯蛋白1在天疱疮患者表皮中的分布模式.方法:用ELISA方法测定患者血清中的桥粒芯蛋白抗体滴度,同时用免疫组化的方法测定天疱疮患者皮肤中桥粒芯蛋白1的分布情况,并与其他表皮内发生棘刺松解的疾病以及正常人对照.结果:在20例特发性天疱疮中12例患者血清同时存在桥粒芯蛋白1和桥粒芯蛋白3的抗体,桥粒芯蛋白1在这12例患者皮肤中均呈现异常块状模式.8例患者的血清中只有桥粒芯蛋白1的抗体,其中4例患者表皮中桥粒芯蛋白1的分布为异常块状模式,4例为正常网状模式.6例药物诱发天疱疮中有1例患者血清中同时存在桥粒芯蛋白1和桥粒芯蛋白3的抗体,5例患者的血清中只有桥粒芯蛋白1的抗体,桥粒芯蛋白1在药物诱发天疱疮表皮中的分布模式为4例表现为正常网状模式,2例为异常的块状模式.结论:桥粒芯蛋白1在天疱疮患者表皮中的分布模式可出现正常的网状模式,但在血清中同时存在桥粒芯蛋白1和3抗体的患者中均呈现异常的块状模式.     

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...     

天疱疮抗体及PDAI与病情活动度关系的研究现状与展望

天疱疮疾病面积评分(Pemphigus disease area index,PDAI)是评估天疱疮病情活动度众多评分体系中的可靠指标;而抗桥粒芯糖蛋白(Desmoglein,Dsg)抗体,大多学者认为与病情活动度呈正相关,但目前对此尚未达成共识。因此或许可以通过验证PDAI值与抗Dsg抗体滴度的相关性来证实抗体滴度与病情活动度的关系,并进一步界定出病情轻中重度所对应的抗Dsg抗体滴度区间,指导治疗。国内尚无类似文献报道,还需要进一步的临床研究来证实。     

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.     

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.     

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.     

Neonatal pemphigus vulgaris with extensive mucocutaneous lesions from a mother with oral pemphigus vulgaris

The clinical phenotype of pemphigus is well explained by the combination of desmoglein (Dsg) 1 and Dsg3 distribution pattern and antiDsg autoantibody profile (Dsg compensation theory). It has been reported that neonatal skin has a similar Dsg distribution pattern to adult mucosal epithelia. We describe a newborn girl with mucocutaneous pemphigus vulgaris (PV) from a mother with mucosal dominant PV. The mother had had painful oral erosions for at least 7 months. Histopathological examination and direct and indirect immunofluorescence studies confirmed the diagnosis of PV and neonatal PV in the mother and daughter, respectively. The mother had a high titre of anti-Dsg3 IgG and a low titre of antiDsg1 IgG, while the neonate had only a high titre of anti-Dsg3 IgG, but no detectable antiDsg1 IgG. AntiDsg3 IgG, which caused the oral dominant phenotype in the mother, induced extensive oral as well as cutaneous lesions in the neonate. Our case provides clinical evidence for the Dsg compensation theory in neonatal 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.     

The detection of IgG and IgA autoantibodies to desmocollins 1-3 by enzyme-linked immunosorbent assays using baculovirus-expressed proteins, in atypical pemphigus but not in typical pemphigus

BACKGROUND: We have shown previously that human desmocollin (Dsc) 1 is recognized by IgA autoantibodies of subcorneal pustular dermatosis (SPD) type IgA pemphigus. However, the presence of IgG anti-Dsc autoantibodies is still controversial, and antibodies to Dsc2 and Dsc3 have not been clearly identified. OBJECTIVES: To investigate this by producing recombinant proteins consisting of the entire extracellular domains of human Dsc1, 2 and 3 in baculovirus, and to use them to establish an enzyme-linked immunosorbent assay (ELISA). METHODS: By this ELISA, we examined in total 165 cases of various types of autoimmune bullous diseases, as well as 23 normal controls. RESULTS: None of 45 sera of classical pemphigus showed either IgG or IgA antibodies to any Dsc. In contrast, one atypical pemphigus serum showed both IgG and IgA antibodies to Dsc1, which were adsorbed by incubation with Dsc1 baculoprotein. Furthermore, this ELISA detected both IgA and IgG anti-Dsc3 antibodies in one atypical case, and IgA antibodies to both Dsc2 and Dsc3 in another. This reactivity was confirmed by positive IgA immunofluorescence with Dsc2 and Dsc3 expressed on COS-7 cells. These results show that both IgG and IgA autoantibodies against all of Dsc1-3 are present in the sera of particular cases of nonclassical pemphigus, except for IgG antibodies to Dsc2, but that they are not detected in classical pemphigus. Unexpectedly, although IgA antibodies of all of eight SPD type IgA pemphigus sera reacted with Dsc1 expressed on COS-7 cells, only one serum was positive in Dsc1 ELISA for IgA. CONCLUSIONS: This result indicates either that Dscs expressed by baculovirus may not adopt the correct conformation or that Dscs may need association with other molecules to express all the epitopes for autoantibodies.     

Successful removal of pathogenic autoantibodies in pemphigus by immunoadsorption with a tryptophan-linked polyvinylalcohol adsorber

BACKGROUND: Autoantibodies against the glycoproteins desmogleins 1 and 3 which are components of the desmosomal adhesion complex have been shown to be responsible for the loss of epidermal adhesion characteristic of pemphigus. Elimination of these antibodies should clinically improve the pathology of this group of severe autoimmune blistering skin disorders. OBJECTIVES: To gather information about the efficacy of immunoadsorption in the reduction of pathogenic serum autoantibodies against desmogleins 1 and 3 and to evaluate the clinical benefit of immunoadsorption in the treatment of pemphigus. PATIENTS AND METHODS: Nine patients with pemphigus and detectable circulating desmoglein antibodies were included in this open trial. Two immunoadsorption treatments separated by a 48-h interval were performed per patient. Anti-desmoglein 1 and 3 antibodies in the patients' sera were monitored by enzyme-linked immunosorbent assay and indirect immunofluorescence before and following each immunoadsorption. In addition, the efficacy of the tryptophan-linked polyvinylalcohol adsorber in removing antidesmoglein antibodies was directly evaluated. RESULTS: IgG antibodies against desmogleins 1 and 3 were effectively eliminated from the patients' plasma upon passage through the adsorber and levels of serum autoantibodies were significantly reduced by immunoadsorption. A single immunoadsorption treatment led to a reduction of antidesmoglein autoantibodies of about 30%. Clinically, mucosal and cutaneous lesions improved allowing for a reduction of the systemic immunosuppressive treatment with glucocorticoids. CONCLUSIONS: Immunoadsorption with tryptophan-linked polyvinylalcohol adsorbers holds promise as a highly effective and safe adjuvant therapeutic regimen in pemphigus.     

Anti-desmoglein IgG autoantibodies in patients with pemphigus in remission

Background   Desmoglein (Dsg) enzyme-linked immunosorbent assay (ELISA) is a highly sensitive and specific method to detect anti-Dsg3 and anti-Dsg1 IgG autoantibodies in pemphigus vulgaris (PV) and pemphigus foliaceus (PF), respectively. Whereas ELISA index values fluctuate in parallel with disease activity, ELISA positivity during clinical remission has been observed.
Objective   To determine the prevalence of positive Dsg ELISA index values during clinical remission. To ascertain how positive Dsg ELISA scores during remission compare with those during active disease.
Methods   Dsg ELISA was performed on serum samples of PV and PF patients taken during remission (lesion-free ≥ 3 months on ≤ 15 mg or ≤ 5 mg/day prednisolone) and active disease. We used a modified ELISA protocol with optimal serum dilutions in sera with very high initial index values, as we previously described.
Results   When remission was defined as no eruption ≥ 3 months with ≤ 15 mg/day prednisolone, 20 of 43 PV patients (46.5%) and 4 of 12 PF patients (33.3%) showed Dsg3 and Dsg1 ELISA positivity, respectively. With ≤ 5 mg/day, 6 of 17 PV (35.3%) and 1 of 6 PF patients (16.7%) showed Dsg3 and Dsg1 ELISA positivity, respectively. The index value of each ELISA-positive remission serum was consistently lower than that of its corresponding active disease serum. We observed consistent correlation between ELISA index values and indirect immunofluorescence titres.
Conclusions   Circulating anti-Dsg IgG autoantibodies are found in a considerable percentage of pemphigus patients in remission, who have high levels of antibody production during active stages.     

A study of desmoglein 1 autoantibodies in pemphigus vulgaris: racial differences in frequency and the association with a more severe phenotype

BACKGROUND: Pemphigus vulgaris (PV) is characterized by pathogenic autoantibodies to desmoglein (Dsg) 3, but additional antibodies to Dsg1, the pemphigus foliaceus antigen, are detectable in some cases. OBJECTIVES: To investigate the clinical significance of the presence of both Dsg 1 and 3 antibodies. METHODS: In 79 subjects with PV, enzyme-linked immunosorbent assays were used to detect IgG autoantibodies reactive with the ectodomain of Dsg1 and Dsg3. RESULTS: There was a clear association between the clinical phenotype and the Dsg antibody profile. All subjects had Dsg3 autoantibodies and 61% had coexisting Dsg1 antibodies (Dsg3+/Dsg1+). PV limited entirely to the mucosal surfaces was seen only in Dsg3+/Dsg1- patients, while additional Dsg1 antibodies (Dsg3+/Dsg1+) predicted cutaneous in addition to mucosal involvement. Although minor cutaneous involvement was observed in most Dsg3+/Dsg1- patients, severe cutaneous involvement was seen only in Dsg3+/Dsg1+ patients. Dsg1 antibodies were detectable early in the course of disease and their appearance did not relate to the use of systemic therapy. The proportion of Dsg1+ patients was higher in those of Indian origin compared with white northern Europeans (P < 0.05). CONCLUSIONS: These data suggest that the presence of Dsg1 antibodies is predictive of a potentially more severe disease and that genetic factors may determine the Dsg antibody profile.     

Serum and salivary desmoglein 1 and 3 enzyme‐linked immunosorbent assay in pemphigus vulgaris: correlation with phenotype and severity

Background To the best of our knowledge there is only one report about salivary desmoglein (Dsg) 1 and 3 enzyme‐linked immunosorbent assay (ELISA) in pemphigus vulgaris (PV), whereas several studies have been performed on serum. Aims To find the sensitivity of serum and salivary anti‐Dsg1 and 3 antibodies in the diagnosis of PV, and to determine the relationship between disease severity and phenotype with antibody levels. Methods Fifty new patients with PV were included in this study. The diagnosis of PV was confirmed by histopathology and direct immunofluorescence. Demographical data, disease severity and phenotypes were recorded on questionnaire sheets. Dsg1 and Dsg3 ELISA were performed on serum and salivary samples of patients and controls. Results Thirty‐seven patients had mucocutaneous phenotype; whereas mucosal dominant and cutaneous dominant phenotypes were seen in 11 and 2 patients respectively. The sensitivities of serum anti‐Dsg3 and anti‐Dsg1 were 94% and 72% respectively. The sensitivities of salivary anti‐Dsg3 and anti‐Dsg1 antibodies were accordingly 94% and 70%. Compared with mucosal phenotype, serum and salivary anti‐Dsg1 antibodies were significantly higher in the patients with mucocutaneous phenotype. Serum Dsg1 antibodies were related with cutaneous and serum Dsg3 antibodies with mucosal severity scores. Salivary Dsg1 antibodies were significantly correlated with mucosal severity (P = 0.00); however there was no correlation between this antibody and cutaneous severity (P = 0.07). Salivary Dsg3 antibodies were not correlated with mucosal severity (P = 0.16). Conclusion Saliva Dsg ELISA could be used for diagnosis of PV. Salivary Dsg1 antibodies had a significant correlation with mucosal severity.