阿维A与干扰素对人皮肤T细胞淋巴瘤细胞株Hut78细胞增殖及白细胞介素15表达的影响

目的：检测不同浓度阿维A、干扰素α（IFN?α）单独或联合应用对人皮肤T细胞淋巴瘤Hut78细胞株的增殖抑制作用及白细胞介素15（IL?15）表达的影响。方法将Hut78细胞分为空白对照组、阴性对照组、二甲基亚砜组（DMSO）和实验组,其中实验组分别用0.1~10μmol/L阿维A和5000~20000 IU/ml IFN?α单独或1.0μmol/L阿维A联合5000~20000 IU/ml IFN?α作用Hut78细胞,培养24、48、72 h后分别进行测定。CCK8法检测细胞增殖情况,ELISA检测药物处理后细胞的白细胞介素15（IL?15）表达情况。结果各浓度阿维A或联合作用组与DMSO组相比及IFN?α组与阴性对照组相比,Hut78细胞增殖和IL?15表达均受到明显抑制,抑制作用随浓度增加和时间延长而增强。重复测量方差分析显示,阿维A、IFN?α单独或联合作用不同时间,细胞增殖抑制率和IL?15表达差异均有统计学意义（P<0.05）,不同作用浓度之间差异亦有统计学意义（均P<0.05）,作用时间与药物浓度之间存在交互作用（均P<0.05）。1.0μmol/L阿维A＋10000或20000 IU/ml IFN?α组与相应浓度药物单独作用组比较,细胞抑制率差异在24、48、72 h时均有统计学意义（P<0.05）。1.0μmol/L阿维A+5000 IU/ml IFN?α组在24、48、72 h时与5000 IU/ml IFN?α组相比,IL?15表达差异均有统计学意义（P<0.05）;1.0μmol/L阿维A＋10000或20000 IU/ml IFN?α组与相应浓度药物单独作用组之间IL?15表达差异在24、48、72 h时均有统计学意义（P<0.05）。结论阿维A和IFN?α均对Hut78细胞的增殖有抑制作用,均可下调Hut78细胞IL?15的表达,这种作用随着浓度的增加和时间的延长而增强,且两者联合用药优于单独用药。     

不同底物间接免疫荧光对自身免疫性表皮下水疱病诊断的评价

【摘要】 目的 评价以正常人皮肤、猴食管及盐裂皮肤为底物的间接免疫荧光对自身免疫性表皮下水疱病的诊断价值。方法 选取2015年1月至2016年12月在中国医学科学院皮肤病研究所诊断的自身免疫性表皮下水疱病56例,其中大疱性类天疱疮（BP）47例,获得性大疱性表皮松解症（EBA）6例,线状IgA大疱性皮病2例,P200类天疱疮1例。对照组为70例天疱疮、15例慢性湿疹和15例健康成人。分别以正常人皮肤、猴食管及盐裂皮肤为底物行间接免疫荧光,观察荧光沉积情况,比较不同表皮下水疱病间接免疫荧光检测的敏感性和特异性。采用SPSS 13.0软件,计数资料比较采用χ2检验。结果 BP患者血清以正常人皮肤、猴食管为底物间接免疫荧光可见到荧光物质沿基底膜带线性沉积,盐裂皮肤间接免疫荧光可见BP患者荧光线性沉积于表皮侧,EBA和P200类天疱疮线性沉积于真皮侧。以正常人皮肤、猴食管及盐裂皮肤为底物间接免疫荧光对表皮下水疱病诊断的敏感性分别为73.2%、60.7%、94.6%,差异有统计学意义（χ2 = 18.2,P < 0.05）,特异性分别为98.0%、100%、97.1%,差异无统计学意义（P > 0.05）,以盐裂皮肤为底物时诊断的敏感性高于以正常人皮肤、猴食管为底物（χ2值分别为8.0、16.7,均P < 0.05）。结论 对于自身免疫性表皮下水疱病的诊断,盐裂皮肤为底物行间接免疫荧光检查优于以猴食管和正常人皮肤为底物。     

辣椒和乙醇对肉桂醛引起的兔耳刺激性接触性皮炎的影响

目的 探讨辣椒和乙醇对肉桂醛引起的皮肤刺激性接触性皮炎的影响.方法 日本大耳白兔32只,随机分成对照组、乙醇组、辣椒组、混合组,每组8只.对照组:生理氯化钠溶液日2次灌胃;乙醇组:50％乙醇、生理氯化钠溶液分别灌胃;辣椒组:10％辣椒液、生理氯化钠溶液分别灌胃;混合组:50％乙醇、10％辣椒液分别灌胃.剂量均为每次8 ml/kg,各组连续灌注7d.出现饮水量增加、大便干结、耳廓血管充血等辛辣刺激体征后,再将各组实验动物分成完整皮肤组和破损皮肤组,每组4只,按化妆品卫生规范进行皮肤刺激性实验,分别记录1、24、48和72 h刺激部位的情况,对皮肤刺激反应进行评分,组间差异比较采用方差分析.结果 辣椒与乙醇连续7d灌胃后,对照组症状体征评分为0.25±0.46,辣椒组为5.38±0.74,乙醇组为7.25±0.71,混合组为12.75±0.70,辣椒组、乙醇组及混合组的症状体征评分明显高于对照组,差异有统计学意义(P＜0.01).在家兔完整皮肤刺激状态评分中,混合组皮肤刺激反应强度在24、48 h显著高于灌注辣椒组(F=28.44、30.33,P＜0.05);在家兔破损皮肤刺激状态评分中,乙醇组、混合组皮肤刺激反应强度在24、48、72 h评分显著高于辣椒组(F=197.12、94.54、87.63,P＜ 0.01).结论 按照实验所用的剂量,辣椒、乙醇和混合物都能够在家兔诱导出辛辣刺激体征,辣椒和乙醇能明显增加皮肤对肉桂醛刺激的敏感性.     

长波紫外线照射对HaCaT细胞iNOS/NO表达的影响

目的 探讨不同剂量长波紫外线 （UVA） 照射HaCaT细胞后不同时间点诱导型一氧化氮合成酶（iNOS）的表达情况。方法 1 J/cm2、5 J/cm2和10 J/cm2 UVA照射HaCaT细胞后继续培养24 h、48 h和72 h,倒置相差显微镜下观察细胞形态的变化;分别用RT-PCR、Western印迹和Griess法检测HaCaT细胞iNOS mRNA、蛋白及NO的表达。结果 所有UVA剂量组HaCaT细胞iNOSmRNA在光照后24 h有表达,48 h达高峰,72 h后下降,各时间点间表达量差异有统计学意义（P < 0.05）;1 J/cm2 UVA照射后3个时间点均未见iNOS蛋白表达,而5 J/cm2和10 J/cm2 UVA照射后iNOS蛋白在24 h增加,48 h达高峰且显著高于24 h（P < 0.05）,照射后72 h无iNOS蛋白表达。所有UVA剂量组HaCaT细胞NO表达量在24 h升高,48 h显著升高,72 h平稳升高,3个时间点NO表达量均比正常对照组明显增加（P < 0.05）。对照组HaCaT细胞无iNOS mRNA和蛋白表达,NO表达量低。结论 HaCaT细胞iNOS和NO的表达变化与UVA照射存在时间和剂量关系。     

点阵Er:YAG激光照射对皮肤胶原增生的影响

目的 探讨点阵铒：钇铝石榴石（Er:YAG）激光定点多重脉冲照射对正常人皮肤的组织学影响。 方法 使用点阵Er:YAG激光连续多脉冲照射正常受试者的上臂皮肤,取照射后1 h、24 h、48 h、72 h、7 d、15 d及30 d的照射皮肤和未照射皮肤进行组织学观察,包括HE染色、抗热休克蛋白70、47免疫组化染色和Masson 染色。结果 点阵Er:YAG激光连续多脉冲照射正常受试者皮肤,刺激皮肤组织产生一系列点阵排列的局灶性锥状气化剥脱微损伤。照射后1 ~ 7 d,微损伤区皮肤产生炎症反应,表皮细胞上皮化;7 ~ 30 d局部真皮内胶原新生、重塑。热休克蛋白70在照射后72 h至第7天内呈峰值表达;热休克蛋白47在照射后15 ~ 30 d持续高表达。结论 点阵Er:YAG激光定点多重脉冲照射人体皮肤可以到达真皮深层,从而获得局灶性胶原增生的疗效。     

黄芩苷对中波紫外线诱导后BALB/c小鼠表皮光产物形成的干预作用

目的 观察中波紫外线（UVB）诱导BALB/c小鼠皮肤表皮细胞光产物的形成和清除情况,以及黄芩苷的干预作用。方法 将黄芩苷（1 mg/cm2）连续3 d外用于BALB/c小鼠表皮24 h后,采用免疫组织化学法及免疫印迹法检测180 mJ/cm2 UVB辐射后1 h、24 h和48 h小鼠表皮内环丁烷嘧啶二聚体（CPD）的产生和清除情况。结果 CPD仅在接受UVB辐射的小鼠皮肤内出现。相对于UVB辐射后1 h光产物的平均值,UVB辐射组1 h、24 h和48 h,CPD的信号强度分别为（100 ± 5.22）％、（75.34 ± 8.22）％和（42.11 ± 3.24）％;经过黄芩苷处理后的小鼠接受UVB辐射后1 h、24 h和48 h,CPD的信号强度分别为（81.45 ± 5.22）%、（32.14 ± 6.33）%和（5.21 ± 3.15）%;而丙酮处理后的小鼠接受UVB辐射后1 h、24 h和48 h,CPD的信号强度分别为（106 ± 8.21）%、（70.23 ± 4.13）%和（41.22 ± 4.21）%。结论 外用黄芩苷能抑制UVB辐射诱导光产物的形成,并在48 h内加速光产物的清除。黄芩苷是一种有效的紫外线防护剂。     

长波紫外线对皮肤成纤维细胞表达和分泌组织蛋白酶G的影响

目的 研究长波紫外线（UVA）照射对皮肤成纤维细胞组织蛋白酶G（CatG）表达和分泌的影响。 方法 原代培养的皮肤成纤维细胞来自儿童包皮,10代以内的细胞行后续实验。①以10 J/cm2 UVA照射皮肤成纤维细胞,24、48、72 h后提取照射组和对照组细胞蛋白和mRNA,并收集细胞上清液;②分别以10、20、30 J/cm2 UVA照射皮肤成纤维细胞,24 h后收集细胞和上清液。用RT-PCR 和Western印迹分别检测各组细胞CatG mRNA及蛋白的表达,ELISA检测细胞上清液CatG的含量。 结果 10 J/cm2 UVA照射后24、48、72 h,照射组细胞CatG mRNA表达分别为0.376 ± 0.014、0.308 ± 0.022和0.296 ± 0.032,对照组分别为0.183 ± 0.003、0.185 ± 0.005、0.182 ± 0.004;照射组细胞CatG蛋白灰度值分别为1.80 ± 0.12、1.41 ± 0.17和1.27 ± 0.09,对照组分别为0.96 ± 0.06、0.95 ± 0.22、1.00 ± 0.14;照射组细胞CatG mRNA、蛋白表达及细胞上清液CatG含量较相应的对照组升高（均P < 0.05）,且以照射后24 h表达最高。10、20、30 J/cm2 UVA照射后24 h,皮肤成纤维细胞CatG mRNA表达分别为对照组的1.90、2.51、3.04倍,细胞CatG蛋白表达分别为对照组的1.88、3.97、4.72倍,细胞上清液CatG含量分别为对照组的1.36、1.50、1.66倍,均随UVA剂量的升高而增加,其差异有统计学意义（均P < 0.01）。 结论 急性UVA照射促进皮肤成纤维细胞表达和分泌CatG。     

SIRT6对老年人皮肤成纤维细胞迁移能力和增殖能力的影响及机制研究

目的 探讨沉默信息调节因子2同源类似物6（SIRT6）对老年人皮肤成纤维细胞迁移能力和增殖能力的影响及机制。方法 在山西医科大学第二附属医院泌尿外科取不同年龄患者包皮环切术切取的包皮组织，其中老年组8例，青年组8例。用胶原酶消化法提取人皮肤成纤维细胞，Western印迹法检测不同年龄组人皮肤成纤维细胞中SIRT6和磷酸化p65蛋白（p-p65）的表达，划痕实验检测细胞迁移活性，CCK8法检测细胞增殖活性。将老年组皮肤成纤维细胞分成两组，一组用SIRT6慢病毒感染使其SIRT6表达增高作为SIRT6组，另一组以空病毒感染作为对照组，用上述方法分别检测SIRT6组和对照组细胞迁移活性、增殖活性和p-p65蛋白表达水平，实时PCR检测两组Ⅰ型和Ⅲ胶原蛋白、整合素亚基α3、α5和β1 mRNA的表达。采用GraphPad Prism 5软件进行统计学分析，t检验分析两组之间数据的差异。结果 老年组皮肤成纤维细胞SIRT6表达水平（0.434 ± 0.179）显著低于青年组（1.000 ± 0.067，t = 3.040，P = 0.012），p-p65表达水平（1.694 ± 0.148）显著高于青年组（1.000 ± 0.093， t = 2.949，P = 0.015），迁移率（43.81% ± 18.84%）显著低于青年组（94.63% ± 12.32%，t = 5.903，P = 0.003），24、48 h增殖活性也较青年组显著下降（P < 0.05）。老年组成纤维细胞过表达SIRT6后，与对照组相比，p-p65表达显著下降（P < 0.05），迁移能力和增殖能力显著提高（P < 0.05），同时Ⅲ胶原蛋白和整合素亚基α3、α5和β1的mRNA表达水平显著升高（P < 0.05）。结论 SIRT6可提高老年人成纤维细胞的迁移能力和增殖能力，其机制可能与抑制NF-κB通路有关。     

组织蛋白酶B在急性光损伤皮肤成纤维细胞中的表达及意义北大核心CSCD

目的 探讨组织蛋白酶B（CatB）在急性光损伤人皮肤成纤维细胞（HDF）中的表达变化及意义.方法 体外培养原代HDF,选取第4～8代细胞进行实验.实验设长波紫外线（UVA）照射组和不照射的对照组.CCK8法分别检测5、10、15、20和25 J/cm2 UVA照射后HDF的增殖率.用10 J/cm2 UVA单次照射致HDF急性光损伤,Western印迹及实时定量逆转录（RT）-PCR分别检测急性光损伤组HDF及对照组HDF照射后24、48、72 h CatB蛋白及mRNA表达;另外分别用10、15、20、25 J/cm2 UVA照射HDF,Western印迹及实时定量RT-PCR分别检测急性光损伤组及对照组HDF照射后48 h CatB蛋白及基因表达.数据采用SPSS 13.0统计软件行方差分析,两组间比较采用最小显著差异法（LSD）.结果 UVA照射导致HDF增殖率下降;当UVA剂量≤10 J/cm2时,细胞存活率均保持在85％以上,各照光组分别与对照组24、48、72 h时比较,均P＜0.05.Western印迹结果显示,急性光损伤组（10 J/cm2 UVA照射）CatB蛋白灰度值在照射后24 h（0.76±0.14）、48 h（1.34±0.38）、72 h（0.82±0.09）均较对照组（分别为0.35±0.01、0.45±0.12、0.61±0.06）升高（均P＜0.05）.实时定量RT-PCR显示,在急性光损伤组CatB的mRNA表达在照射后24 h（0.149±0.009）、48 h（0.173±0.009）、72 h（0.185±0.158）也较对照组（分别为0.089±0.015、0.091±0.010、0.111±0.017）上调（均P＜0.05）.10、15、20、25 J/cm2 UVA照射HDF后48 h,CatB的蛋白灰度值分别为0.99±0.07、1.49±0.14、1.89±0.08、2.07±0.06,均较对照组（0.60±0.05）升高（均P＜0.05）,CatB的mRNA表达也较对照组升高.结论 UVA致急性光损伤的皮肤成纤维细胞中CatB蛋白及mRNA表达均上调.     

热处理、UVB辐射及两者联合作用对人表皮黑素细胞合成热休克蛋白72的影响

目的 探讨热处理、中波紫外线（UVB）辐射及两者联合作用下,体外培养的人表皮黑素细胞热休克蛋白72（heat shock protein 72,HSP72）的变化。 方法 热处理（42 ℃,1 h/d,连续3 d）、UVB辐射（50 mJ/cm2,连续3 d）及两者联合（首先42 ℃,加热1 h,然后 50 mJ/cm2 UVB照射,连续3 d）处理正常人表皮（包皮）黑素细胞2 h、6 h后收集细胞,用实时荧光定量PCR技术和 Western 印迹技术分别检测不同组别间HSP72 mRNA及蛋白表达水平的差异。结果 热处理组（6.584 ± 0.871）及联合作用组（7.269 ± 0.454）与对照组（0.975 ± 0.089）相比,mRNA 表达水平明显增加（P < 0.001）;UVB辐射组（0.832 ± 0.084）与对照组相比,mRNA表达水平差异无统计学意义（P > 0.05）。Western 印迹结果显示,热处理组（2.022 ± 0.058）及联合作用组（2.080 ± 0.045）HSP72蛋白表达水平明显高于对照组（0.532 ± 0.033,P < 0.001）;UVB组（0.546 ± 0.021）与对照组相比,表达水平差异无统计学意义（P > 0.05）。析因设计方差分析结果显示,热处理与UVB辐射对HSP72 mRNA和蛋白表达均无交互作用（F = 2.106,1.399,P < 0.05）。结论 热处理引起HSP72表达显著增多,可能与热处理引起黑素细胞功能增强及UVB辐射后的损伤保护作用有关。     

盐裂皮肤间接免疫荧光技术在大疱性类天疱疮诊断中的价值评估

目的:评价盐裂皮肤间接免疫荧光（IIF-SSS）在大疱性类天疱疮（BP）诊断中的价值。方法:采用单中心临床回顾性研究。纳入2013年1月至2019年1月在中国医学科学院皮肤病医院就诊的初诊BP患者163例,对照组404例,包括天疱疮161例、湿疹67例、药疹26例、多形红斑23例、结节性痒疹18例等。于患者用药前采血,...     

盐裂皮肤间接免疫荧光及BP180 NC16a酶联免疫吸附测定在大疱性类天疱疮诊断中的意义

目的 探讨盐裂皮肤间接免疫荧光及大疱性类天疱疮（BP）180 NC16a-酶联免疫吸附测定（ELISA）检测在BP诊断中的意义。方法 收集2015年1月至2017年8月在中国医学科学院皮肤病医院用盐裂皮肤间接免疫荧光（IIF?SSS）和BP180 NC16a?ELISA检测BP患者174例和对照组129例血清。其中25例BP患者用直接免疫荧光（DIF）进行检测并与IIF?SSS和BP180 NC16a?ELISA敏感性进行比较。结果 IIF?SSS、BP180 NC16a?ELISA的敏感性分别为93.67%、96.55%;特异性分别为100%、96.12%。IIF?SSS与BP180 NC16a?ELISA相关系数0.147,为弱相关。其中25例BP患者血清学诊断方法（IIF?SSS,BP180 NC16a?ELISA）和DIF敏感性比较差异无统计学意义。结论 BP血清学诊断方法特异性强、敏感性高,值得临床推广应用。     

A comparative study of antibody titers of blister fluid and serum in patients with subepidermal immunobullous diseases

BACKGROUND: Subepidermal autoimmune bullous diseases (SABD) comprise several disorders, such as bullous pemphigoid (BP), cicatricial pemphigoid (CP), epidermolysis bullosa acquisita (EBA), herpes gestationis (HG), and linear immunoglobulin A (IgA) dermatosis (LAD), and are characterized by antibody production against the basement membrane structures of the skin and mucosa. Although indirect immunofluorescence (IIF) on serum is a routine test for the detection of basement membrane zone antibodies, there have only been a few studies related to IIF on blister fluid. Aim To perform IIF on blister fluid and to compare the results with those of serum. METHODS: IIF on salt-split skin was performed on the serum and blister fluid of 35 patients with SABD (25 bp, three EBA, three HG, three LAD, and one bullous systemic lupus erythematosus) with conjugated IgG, IgA, and C3. RESULTS: Twenty-eight of the 35 patients showed IIF-positive blister fluid with a titer similar or less than that of serum. In 25 patients with BP, the most common disease in this study, 23 cases (92%) had positive IIF on serum, 23 cases (92%) on blister fluid, and 24 cases (96%) on either serum or blister fluid. Immunoreactant titers in BP blister fluid and serum did not show significant differences (P > 0.05). Epidermal binding of immunoreactants was the most prevalent staining pattern of IIF on salt-split skin (92%) in BP. CONCLUSIONS: From the findings of this study, the blister fluid of patients with SABD can be used for IIF. Although IIF sensitivity on blister fluid is no more than that on serum, the performance of this test on blister fluid in addition to serum may reduce the number of false negative results of IIF found using either of these two substrates alone.     

盐裂皮肤直接和间接免疫荧光联合应用鉴别诊断表皮下大疱病的研究

本文探讨盐裂皮肤ＤＩＦ和盐裂皮肤ＩＩＦ联合应用鉴别诊断表皮下大疱病的价值。２０例两种盐裂皮肤免疫荧光均阳性的病例比较表明，盐裂皮肤ＩＩＦ示抗基底膜带抗体结合表皮侧或真皮侧与盐裂皮肤ＤＩＦ示免疫反应物（ＩｇＧ、ＩｇＡ、Ｃ＿３等）沉积于表皮例或真皮侧的部位基本一致。提示多数情况下盐裂皮肤ＤＩＦ或ＩＩＦ均可单独用于表皮下大疱病诊断，两者对比分析可以提高诊断率。     

Bullous pemphigoid in Liguria: A 2-year survey

BACKGROUND: The epidemiology of bullous pemphigoid (BP) is not clear because of the heterogeneity of the disease, and its possible association with internal malignancies has been under debate for many years. We report the findings of a 2-year study on incident BP cases in the Liguria region of Italy. SUBJECTS AND METHODS: Thirty-two patients with BP were collected over the 2-year period. Diagnosis was made based on clinical findings and confirmed by histology, direct immunofluorescence (DIF) and indirect immunofluorescence (IIF) with salt-split skin and monkey oesophagus, and immunoblotting (IB). All patients were thoroughly investigated for possible malignancies and all were followed up for 6 months to monitor the response to treatment. RESULTS: DIF showed linear deposits at the dermoepidermal junction in all but one patient. IIF gave positive findings for 15 sera tested with monkey oesophagus and 20 tested with salt-split skin. IB gave positive findings in 19 cases. There was a malignancy in six cases, but no clinical or immunological features that could be considered to predict this occurrence. CONCLUSIONS: The findings of this study are in accordance with most of the data found in the literature, including the fact that IgG serum levels did not predict the course of the disease. Contrary to previous indications, IgE levels were not indicative of disease course either. Mucosal lesions, erythema multiform-like lesions, negative IIF findings and antibodies to AgPB2 were not a prediction for the development of malignancy.     

Comparative sensitivity of indirect immunofluorescence to immunoblot assay for the detection of circulating antibodies to bullous pemphigoid antigens 1 and 2

Summary The sera of 263 patients with bullous pemphigoid (BP) were tested by indirect immunofluorescence (IIF) on salt-split skin (SSS) and immunoblot (IB) assay, in order to assess the diagnostic sensitivity of these techniques. Among the 263 sera tested. 198 sera (75%) contained antibasement membrane zone antibodies demonstrable by IIF reacting to the epidermal (98%) or both the dermal and epidermal sides (2%) of SSS. One hundred and eighty-two of the 263 sera (69%) reacted by IB with BP antigens (Ag), most commonly the BPAgl (93 cases, 51%), and a complex of BPAgl and the 180kDa minor BP antigen (BPAg2) (47 cases, 26%). BPAg2 alone was found in 42 cases (23%). A good correlation was found between the detection of autoantibodies by IIF and labelling of BPAg1 and/or BPAg2 by IB assay, in which 152 of 198 sera with an epidermal pattern in IIF identified a BP antigen. IB analysis of the 65 sera negative by IIF yielded positive results in 30 cases (46%). Thirty-one per cent (13 of 42) of sera recognizing by IB BPAg2, were negative by IIF, as compared with 12% (11 of 93) of those recognizing BPAg1 (P < 0.01). Comparing the sensitivity of the two tests, IIF (75%) was found to be more sensitive than IB (69%). Thirty-five of the 263 sera (13%) remained negative by both techniques. It can be concluded from this study that IIF on SSS appears to be a sensitive and reliable assay for screening BP;IB should be performed for the sera that are negative by IIF as it may reveal circulating antibodies, particularly to BPAg2.     

抗p200类天疱疮7例临床及免疫血清学特征分析

【摘要】 目的 总结抗p200类天疱疮患者的临床和免疫血清学特征。方法 收集2015年1月至2021年10月在中国医学科学院皮肤病医院确诊为抗p200类天疱疮的患者资料，回顾性分析其临床和免疫血清学特征。结果 纳入7例抗p200类天疱疮患者，盐裂皮肤-间接免疫荧光实验显示，7例患者血清IgG抗体均结合于盐裂皮肤的真皮侧，以真皮提取物为底物的免疫印迹显示，在相对分子质量200 000蛋白处有条带。4例呈经典型大疱性类天疱疮样损害，2例初起呈湿疹样损害，1例类似线状IgA大疱性皮病。6例循环IgG抗体可识别层粘连蛋白γ1-C端重组蛋白。4例患者接受不同剂量系统糖皮质激素治疗，其中1例对高剂量系统糖皮质激素（相当于泼尼松1.4 mg·kg-1·d-1）治疗抵抗；2例对米诺环素、氨苯砜治疗反应好；1例失访。4例患者在随访平均22.5个月时达到完全缓解停药；2例在随访平均8个月时达到最小剂量治疗保持完全缓解。结论 抗p200类天疱疮临床表现多样，重组Lnγ1-C端可作为可靠的抗原底物用于检测抗p200类天疱疮患者自身抗体；部分患者最终可达到完全缓解并停药。     

Defining target antigens in linear IgA disease using skin from subjects with inherited epidermolysis bullosa as a substrate for indirect immunofluorescence microscopy

Identification of target antigens in immunobullous disorders usually involves laborious techniques such as immunoblotting and immunoprecipitation, which do not always provide conclusive data. This is particularly true of linear IgA disease (LAD) in which the target antigen has often proved difficult to identify. As an alternative means of antigen identification in five adult patients with LAD, we performed indirect immunofluorescence (IIF) microscopy on a panel of skin samples taken from subjects with different forms of inherited epidermolysis bullosa (EB). Skin samples were selected that showed a complete absence of immunostaining for a specific basement membrane zone (BMZ) molecule (type VII collagen, laminin 5 or the 180-kDa bullous pemphigoid antigen BP180). In each case, the underlying genetic mutations had been defined and shown to consist of premature termination codons on both alleles of the particular gene, resulting in total ablation of the encoded protein. Two epidermal-binding LAD sera showed BMZ fluorescence on all substrates except BP180-deficient skin, suggesting that the target antigen was BP180, or a closely related molecule. In contrast, two dermal-binding LAD sera were positive on all substrates except the type VII collagen-deficient skin, suggesting that the target antigen was likely to be type VII collagen. One LAD serum sample, which showed combined dermal and epidermal fluorescence on normal salt-split skin, was also positive on all substrates tested, suggesting a target antigen other than type VII collagen, laminin 5 or BP180. The study confirms that LAD is a heterogeneous disorder and illustrates that IIF using a panel of skin samples which lack specific BMZ molecules, taken from subjects with inherited EB, is a relatively simple and useful tool to help identify target antigens in immunobullous disorders.     

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.     

Serological diagnosis of bullous pemphigoid (BP): comparison of the sensitivity of indirect immunofluorescence on salt-split to immunoblotting

Specialized immunological assays are required for the accurate diagnosis of bullous dermatoses such as bullous pemphigoid (BP), epidermolysis bullosa acquisita and bullous lupus erythematosus. The aim of this study was to analyse and compare the sensitivity of indirect immunofluorescence (IF) on salt-split skin and immunoblotting for the detection of circulating autoantibodies in BP. Of the BP patients selected for the study, 74/79 (94%) had circulating autoantibodies detected by at least one of the two methods. Both methods had comparable sensitivity and detected BP-specific autoantibodies in 82-85% of the patients. Because 20% of the patients were found to be positive by only one of the methods, both methods should be used in the diagnosis of BP. Indirect IF on salt-split skin is easier to perform and is preferable in routine analysis, but Western blotting may be used as a complementary assay with sera showing no reactivity on salt-split skin.