天疱疮患者血清中Dsg1及Dsg3抗体水平检测及意义

目的 研究18例天疱疮患者血清中抗桥粒芯糖蛋白抗体1(Dsg1)及抗桥粒芯糖蛋白抗体3(Dsg3)的表达水平和治疗前后的变化以及抗体指数与病情严重程度的相关关系,以探讨Dsg1及Dsg3在天疱疮患者的诊断、监测病情变化及治疗中的应用.方法 对18例患者通过病理及直接免疫荧光进行诊断,并用ABSIS评分标准对患者的病情进行评估,应用酶联免疫吸附试验(ELISA)法检测患者及20例正常人血清中Dsg1及Dsg3抗体的水平,最后通过统计分析判断各指标间的关系及意义.结果 18例患者中Dsg1阳性13例,Dsg3阳性10例;治疗后的Dsg抗体指数较治疗前下降,治疗前后的Dsg1差值为3.13±13.52,Dsg3差值为3.81±10.05,差异有统计学意义(P<0.05);20例正常人血清中,Dsg1阳性2例,阴性18例;Dsg3阳性1例,阴性19例.Dsg1及Dsg3抗体检测的灵敏性分别为72.22％及55.56％,特异性分别为90％及95％.治疗前后的Dsg抗体指数与病情评分分别具有相关性(均为P<0.05).结论 天疱疮患者血清中Dsg1及Dsg3中抗体检测具有较高的灵敏性和特异性,其抗体水平与疾病的严重程度具有一定的相关性,可作为天疱疮的常规辅助检测,并用于监测病情变化及指导治疗.     

天疱疮患者血清抗桥粒芯糖蛋白抗体与疾病临床特征的相关性研究

目的：探讨天疱疮患者血清抗桥粒芯糖蛋白（Dsg）抗体与疾病临床特征的相关性。方法：选择49例天疱疮患者,比较其抗Dsg类别、亚型、滴度与疾病特征的相关性。结果：寻常型天疱疮（PV）患者活动期抗Dsgl、Dsg3抗体阳性率明显高于稳定期（P〈0．05）,落叶型天疱疮（PF）患者活动期抗Dsgl抗体阳性率高于抗Dsg3抗体（P〈0．05）;活动期患者两种抗体滴度随病情加重而明显升高（P〈0．05）;活动期患者血清两种抗体IgG4亚型滴度明显高于IgG1,稳定期患者两种抗体IgG4亚型滴度明显下降且远低于IgG1（P〈0．05）。结论：天疱疮患者血清抗Dsg抗体与疾病表型、病情、活动性存在明显相关性,测定其存在和滴度对临床诊断与治疗有重要指导意义。     

天疱疮患者抗桥粒芯糖蛋白与疾病相关性分析

目的：探讨酶联免疫吸附测定（ELISA ）检测血清桥粒芯糖蛋白（Dsg ）1和Dsg3在天疱疮诊断及治疗中的应用价值。方法选择2014年1～12月在该科确诊为天疱疮的患者47例（观察组）,同期排除天疱疮的患者52例为对照组。间接免疫荧光法（IIF）检测抗表皮棘细胞桥粒抗体,ELISA检测Dsgl、Dsg3;分析其与天疱疮病情的相关性。结果 ELISA法的敏感性为95．74％,特异性为92．31％。IIF法敏感性为93．62％,特异性为86．54％。二者比较差异无统计学意义（P＞0．05）。30例寻常型天疱疮患者16例Dsg1、Dsg3均阳性,8例红斑型和5例落叶型天疱疮患者均只有Dsg1阳性,2例增殖型天疱疮Dsg1、Dsg3均为阳性。寻常型和增殖型天疱疮Dsg1、Dsg3滴度分别为130．85±86、112．30±85．05,疾病活动性评分（5．10±1．86）分,相关系数分别为 r＝0．476（P＝0．008）、r＝0．816（P＝0．001）。落叶型和红斑型天疱疮Dsg1滴度142．59±78．52,疾病活动评分平均为（2．77±0．92）分,r＝0．800,P＝0．001。结论 ELISA检测Dsg1、Dsg3敏感性、特异性高,有助于天疱疮的诊断及病情严重程度的评价。     

经桥粒芯糖蛋白ELISA检测确诊由寻常型天疱疮向落叶型天疱疮转变的的3例报道

作者报道3例经组织学及直接、间接免疫荧光检测确诊的寻常型天疱疮(PV)患者,在发病2～4年后向落叶型天疱疮(PF)转变。在疾病晚期对这3例患者的血清进行桥粒芯糖蛋白(Dsg)ELISA检测显示抗Dsg1存在,抗Dsg3抗体缺如。在这些患者进行血清检测时已开始强的松龙及免疫抑制治疗,由寻常型天疱疮向落叶型天疱疮的转变是恒定不变或是由低于特定阈值的抗Dsg3抗体的选择性抑制所致尚不明确。作者对以往报道的由PV向PF以及由PF向PV转变的病例也作了概述。经桥粒芯糖蛋白ELISA检测确诊由寻常型天疱疮向落叶型天疱疮转变的的3例报道@Ng P.P.L$…     

免疫吸附快速去除重度天疱疮患者的抗桥粒芯糖蛋白自身抗体后临床缓解期延长

背景:天疱疮的表皮松解是由循环自身抗体分别与寻常型天疱疮(PV)的桥粒钙黏素桥粒芯糖蛋白3(Dsg3)和落叶型天疱疮(PF)的桥粒芯糖蛋白1(Dsg1)结合引起。研究表明免疫吸附(IA)治疗性去除抗Dsg自身抗体可使临床病情缓解。目的:该项干预性研究旨在评估肽基Globaffin吸附系统治疗重度天疱疮患者的有效性和安全性。患者和方法:作者对4例PV和2例PF患者应用IA治疗,这些患者均伴有严重慢性疾病且对传统免疫抑制治疗抵抗。IA连续治疗4d为1疗程,继而为4周的治疗间歇期。每日IA治疗前后分别采取血清标本检测血清IgG和抗Dsg1/Dsg3IgG自身抗体…     

寻常型天疱疮血清学诊断中抗桥粒芯蛋白抗体与间接免疫荧光检测的比较

背景:间接免疫荧光法(IIF)是检测天疱疮自身抗体的标准方法。最近可用酶联免疫吸附试验(ELISA s)检测血清抗桥粒芯蛋白1(D sg1)抗体及抗桥粒芯蛋白3抗体(D sg3)。结果显示以黏膜表现为主的寻常型天疱疮(PV)患者仅检出抗D sg3抗体,伴黏膜皮肤损害的PV患者有抗D sg1及抗D sg3抗体,落叶型天疱疮(PF)患者仅有抗D sg1抗体。目的:比较IIF与ELISAs诊断天疱疮及其亚型的敏感性及特异性。方法:研究了33例寻常型天疱疮及5例落叶型天疱疮患者,并与50例健康个体或患有不相关皮肤病的患者进行比较。猴食管用作IIF的底物。结果:32例PV患者中…     

8例缓解期高滴度血清抗体的天疱疮患者临床分析

目的 评估缓解期血清抗桥粒芯蛋白(Dsg)抗体呈现高滴度的天疱疮患者的临床表现、诊治过程及实验室检查结果,为此类患者的治疗提供依据。方法 回顾性分析2013年1月至2020年9月皮肤科诊治的天疱疮患者临床资料,收集确诊为天疱疮并且缓解期血清抗Dsg抗体水平(酶联免疫吸附测定法测定)≥150 U/ml或较初始治疗时下降≤20%(以最大检测上限150 U/ml计算)的患者资料。结果 在此期间收治并获得随访的134例天疱疮患者中,共有8例患者符合纳入标准,随访时长21~85个月,持续缓解时间18~70个月,均以每日口服≤10 mg泼尼松相当量糖皮质激素维持治疗,血清抗Dsg抗体均呈高滴度。8例患者首诊时Dsg-1抗体阳性和Dsg-3抗体阳性者各为7例,其中2例患者分别在病情缓解后19和21个月Dsg-1抗体仍为高滴度,5例患者在病情缓解的18~70个月Dsg-3抗体持续保持高滴度,1例患者两种抗体均呈高滴度状态,以Dsg-1抗体更为明显。Dsg-1和Dsg-3抗体均持续高滴度的患者在19个月的缓解期后出现复发,余患者仍处于临床缓解状态。结论 天疱疮患者治疗后,部分患者抗体滴度呈现高水平状态,以Dsg-3抗体持续不转阴更常见,且与复发无明显相关性,而Dsg-1抗体不下降较少见,患者复发可能性较Dsg-3不下降者大。对于这类临床长期缓解但抗体呈高滴度的天疱疮患者,应注意根据抗体类型并结合患者实际情况调整糖皮质激素用量。对于Dsg-1抗体持续高水平的患者,激素减量应适度放缓。     

1例大疱性类天疱疮患者的抗BP180和抗桥粒核心糖蛋白3自身抗体滴度的平行波动

作者报道1例73岁日本女性,出现抗BP180和抗桥粒核心糖蛋白3(D sg3)自身抗体。患者在四肢末端出现张力较大的大疱、无明显的黏膜受累并且皮肤活检显示表皮下水疱伴嗜酸细胞性海绵水肿。临床和组织学特征提示大疱性类天疱疮的诊断,而抗Dsg3IgG可能并不显示明显的致病作用。有趣的是,在整个病程中抗Dsg3IgG滴度的波动与抗BP180IgG滴度平行。尽管这些自身抗体产生的具体机制尚不清楚,但是由于针对这两种独立皮肤抗原的IgG自身抗体产生有着密切联系,从而提示有一种针对皮肤自身抗原的共享的免疫调节机制。1例大疱性类天疱疮患者的抗BP180…     

天疱疮患者临床特征和Dsg1及Dsg3抗体结果分析

  目的  研究天疱疮患者的临床特征,分析桥粒芯糖蛋白1抗体(desmoglein 1,Dsg1)和桥粒芯糖蛋白3抗体(desmoglein 3,Dsg3)ELISA检测结果与天疱疮分型及严重程度之间的关联。  方法  选取2021年1—12月安徽医科大学第一附属医院收治的50例天疱疮患者,根据临床表现、实验室指标对天疱疮进行分型和分级,对诱发因素、伴发疾病、皮肤感染情况、Dsg1及Dsg3抗体指数等进行回顾性分析。  结果  50例天疱疮患者中寻常型天疱疮39例、红斑型天疱疮8例、落叶型天疱疮2例和副肿瘤性天疱疮1例;轻度28例,中度11例,重度11例;常见诱发因素和糖皮质激素不规范使用有关,占比20%(10/50);最常见皮肤感染菌为金黄色葡萄球菌,检出率为18%(9/50)。不同类型天疱疮患者Dsg3抗体指数差异有统计学意义(Z=-3.481,P＜0.001);不同严重程度天疱疮患者Dsg1及Dsg3抗体差异有统计学意义(P值分别为0.002、0.024)。  结论  天疱疮患者治疗中要注重糖皮质激素的正确使用,皮肤感染在天疱疮患者中较为常见,Dsg1及Dsg3抗体检测在天疱疮患者中具有较高的阳性率,Dsg1及Dsg3抗体指数可作为天疱疮病情评估的指标。      

儿童天疱疮抗体IgG亚型及与天疱疮抗原的反应性

目的观察寻常型天疱疮（Pemphigus vulgaris,PV）患儿和一级亲属血清中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 immunofluorescenceI,IF）检测5例PV患儿、12例一级亲属和15例正常对照者血清PV-IgG,并用免疫印迹法（Western blot,WB）检测PV-IgG亚型及其与Dsg1、Dsg3反应性。结果 PV患儿的PV-IgG阳性率和平均抗体滴度明显高于其一级亲属者（P〈0.01）,活动期患儿的PV-IgG阳性率和平均抗体滴度也明显高于缓解期（P〈0.01）;PV患儿中PV-IgG阳性率（80.0%）明显高于一级亲属（25.0%）和正常对照组（0.0%）（P〈0.01）,PV患儿中Dsg3反应性IgG4水平明显高于一级亲属（P〈0.01）,且其在活动期病例中水平也明显高于缓解期（P〈0.05）。结论寻常型天疱疮的发生与PV-IgG水平有关;Dsg3是PV的主要自身抗原I;gG4是PV的主要致病抗体。     

Effects of chimeric molecule of recombinant Dsg3EC1-2 with toxin PE40 on T and B lymphocytes in Pemphigus Vulgaris

Objective:To observe the effects of the recombinant chimeric toxin Dsg3EC1-2PE40 on T and B lymphocytes isolated from Pemphigus Vulgaris (PV) patients to further study its biological therapeutic function for PV. Methods :Recombinant chimeric toxin Dsg3EC1-2PE40 was first identified, expressed and purified, and then its effects on T and B lymphocytes of PV patients in vitro were detected and quantified by ELISPOT assay and MTT assay. Results :The purity of the expressed protein Dsg3EC1-2PE40 was up to 80%. In ELISPOT assay, with Dsg3EC1-2PE40, the overall number of B cells that produce anti-Dsg3 antibodies among PV patients was only about 60% of the comparable number with Dsg3EC1-2. The proliferation of T cells of PV patients was inhibited markedly by Dsg3EC1-2PE40. There was significant difference between the different groups with Dsg3EC1-2PE40 and Dsg3EC1-2. Conclusion:The recombinant chimeric toxin Dsg3EC1-2PE40 decrease the number of B cells that produce anti-Dsg3 antibodies in PV patients and can inhibit or kill T cells of PV patients in vitro.     

T细胞介导的细胞免疫在寻常型天疱疮中的作用

寻常型天疱疮(PV)是一类危及生命的自身免疫性大疱性皮肤黏膜病,以往认为PV的发生与体液免疫有关,目前认为,除了B细胞的功能异常产生致病性抗体外,T细胞所介导的细胞免疫异常介导了PV患者出现病理性免疫应答。文章介绍了与PV发病相关的HLA-II类抗原,Dsg3特异性的自身反应性T细胞,以及调节性T细胞的最近研究发现。     

人皮肤组织桥粒芯糖蛋白4片段的克隆和在寻常型天疱疮中免疫识别的初步研究

目的 扩增人皮肤组织桥粒芯糖蛋白4(desmoglein 4,Dsg4)胞外区域EC1、EC2、EC3和EC4的核酸序列,为寻常型天疱疮(pemphigus vulgaris, PV)发病机制的研究提供依据.方法 根据GenBank中的Dsg4序列(登录号为AY177664)分析,通过逆转录聚合酶链反应(RT-PCR)法从人正常皮肤组织中抽提RNA,逆转录合成cDNA,聚合酶链反应(PCR)扩增皮肤组织桥粒芯糖蛋白4胞外区域EC1、EC2、EC3和EC4目的基因;应用基因重组技术将目的基因分别与质粒pET32a在T4 DNA连接酶作用下相连接,转化E.coli DH5α感受态细菌,用含氨苄青霉素的LB培养基平板筛选转化菌,阳性重组子DNA序列测定鉴定;重组融合蛋白经ELISA法与PV患者、正常对照组血清进行反应.结果 RT-PCR扩增产物经凝胶电泳得到均约为350 bp的4条条带;质粒载体pET32a连接的目的基因经序列测定后与在GenBank登录的Dsg4胞外区域EC1、EC2、EC3和EC4核酸序列完全一致,4个Dsg4胞外区域cDNA读码框架正确;Dsg4 EC1、EC2、EC3和EC4与患者血清反应,而不与正常对照组血清反应.结论 Dsg4胞外区域多肽片断在PV发病中可能有作用.     

Prospects for autoimmune disease: Research advances in pemphigus

Pemphigus is an autoimmune disorder, known to be caused by autoantibodies directed against critical adhesion molecules of squamous epithelial cells, the desmogleins. These autoantibodies induce blistering of skin and mucosal surfaces and lead to severe morbidity and, potentially, death. Key factors include associated major histocompatibility complex class II genes, the structure of the desmoglein antigens, and the role of autoantibody in impairing cellular adhesion. This article discusses the precise structure of the major histocompatibility complex class II gene-peptide-T-cell receptor complex involved and of the environmental and genetic factors that induce autoimmunity against desmoglein 1. Discovery of antigen-specific immunotherapy and insight into environmental factors that initiate autoimmunity in genetically susceptible individuals are needed.     

Effects of mycophenolate mofetil on cutaneous lupus erythematosus in (NZB × NZW) F1 mice

BackgroundFew studies have evaluated the effects and precise molecular mechanism of mycophenolate mofetil (MMF) in the treatment of human cutaneous lupus erythematosus (CLE). Our findings shed light on the therapeutic effects of MMF in a UVB-induced NZB × NZW (NZBW) F1 CLE mouse model.MethodsContinuous MMF treatment (60 mg/kg/day) was administered up to Day 50 from the beginning of UVB induction (Day 0; 20 weeks old), as the pathologic features of CLE are present after 50 days. The therapeutic effects of MMF treatment in NZBW lupus mice were examined by comparing histopathological changes, lupus band test (deposition of immune complexes at the dermal–epidermal junction) and colocalization of autoantibodies with a dermal autoantigen Dsg3, and by evaluating the associations of local matrix metalloprotease activities.ResultsMMF improved survival in the NZBW lupus mice from 35.7% to 81.8%. The proteinuria, blood urea nitrogen, and interleukin 6 levels were significantly reduced after MMF treatment. The dermal lymphocytic infiltration, deposition of immune complexes at the dermal–epidermal junction, colocalized autoantibodies with Dsg3, and epidermal matrix metalloprotease activity were also attenuated in MMF-treated NZBW F1 mice.ConclusionThe results confirmed that MMF could substantially attenuate skin damage due to CLE in the NZBW F1 mouse model.     

寻常型天疱疮与大泡性类天疱疮皮损角质形成细胞Fas和Fas-L表达的研究

目的 :探讨 Fas和 Fas- L在寻常型天疱疮 (PV)和大泡性类天疱疮 (BP)皮损发生发展中的作用。方法 :采用免疫组化方法研究两病患者皮损 Fas和 Fas- L的表达情况。结果 :1角质形成细胞 Fas/Fas- L表达的部位 :PV,疱内分离的及疱周围的棘层细胞 ;BP,水泡上方及附近的棘层细胞 ;2寻常性天疱疮、大泡性类天疱疮表皮细胞 Fas/Fas- L表达都明显高于正常对照组(均 P <0 .0 1)。结论 :角质形成细胞 Fas/Fas- L表达异常与 PV和 BP皮损的发生可能有密切联系     

Role of autoantibodies against the linker subdomains of envoplakin and periplakin in the pathogenesis of paraneoplastic pemphigus

Background The presence of autoantibodies against multiple epidermal proteins is an important feature in paraneoplastic pemphigus （PNP）. Circulating anti-desmoglein 3 autoantibody, the major pathogenic autoantibody in pemphigus vulgaris （PV）, has been proved pathogenic in PNP. Because of many clinical differences between PNP and PV, we speculate about the involvement of other autoantibodies in the pathogenesis of PNP. Envoplakin （EPL） and periplakin （PPL） are recognized by most PNP sera. Their linker subdomains are highly homologous and necessary for the association of intermediate filaments. Methods We characterized the autoantibodies against the linker subdomains of EPL and PPL in PNP patients＇ sera and their associated tumors by enzyme-linked immunosorbent assay （ELISA） and immunofluorence. We also applied the purified autoantibodies against EPL and PPL from PNP sera to cultured human epidermal keratinocytes （HEK）, to evaluate the changes of cell-cell adhesion. Results Autoantibodies against EPL and PPL were detected in most PNP patients by ELISA, and the decrease of these autoantibodies after removal of the tumors was roughly comparable to the improvement of clinical symptoms. Cultured tumor cells from PNP patients secreted these autoantibodies. Specific immunoglobulin receptors for EPL and PPL were found on B lymphocytes in tumors from PNP. Furthermore, purified anti-EPL and anti-PPL autoantibodies from PNP sera were capable of dissociating cultured human epidermal keratinocytes. Conclusion Autoantibodies against EPL and PPL may also be pathogenic in PNP.