色素障碍性皮肤病

２００１１１３２白癜风患者外周血Ｔ淋巴细胞亚群和ｓＩＬ－２Ｒ水平变化的研究／汪新义（山东省皮防所）…／／临床皮肤科杂志．－２０００，２９（５）．－２５８～２６０ 采用 ＡＰＡＡＰ法和双抗体夹心 ＥＬＩＳＡ法，对１５０例自癜风患者外周血Ｔ淋巴细胞亚群和可溶性白细胞介素－２受体（ｓＩＬ－２Ｒ）进行了检测。结果显示寻常型白癜风ＣＤ３＋、ＣＤ４＋细胞数和ＣＤ４＋／ＣＤ８＋比值均显著低于正常对照（Ｐ＜０．０１），ｓＩＬ－２Ｒ活性明显高于节段型白癜风和正常人（Ｐ＜０．００１，Ｐ＜０．０１）；节段型自癜风ＣＤ３…     

白癜风患者外周血T淋巴细胞亚群和sIL-2R水平变化的研究

采用ＡＰＡＡＰ法和双缺本夹心ＥＬＩＳＡ技术检测１５０例白癜风患者外周血Ｔ淋巴细胞亚群及可溶性白介素－２受体（ｓＩＬ－２Ｒ）水平。结果显示：（１）寻常型白癜风外周血ＣＤ３＾＋、ＣＤ４＾＋细胞数、ＣＤ４＾＋／ＣＤ８＾＋比值明显低于正常对照（Ｐ〈０．０１）,ｓＩＬ－２Ｒ活性显著性高于节段型白癜风和正常人（Ｐ〈０．０１）;节段型白癜风ＣＤ４＾＋、ＣＤ４＾＋／ＣＤ８＾＋与正常对照组比较也显著减少（Ｐ〈０．０     

银屑病皮损中免疫细胞组分的研究

采用免疫组化技术检测３例寻常性银屑病患者皮损中免疫细胞的组分。结果发现银屑病皮损真皮上层中单个核细胞、ＨＬＡ－ＤＲ＋（活化）细胞和ＣＤ－１５＋（巨噬）细胞数均比非皮损部位显著增加（Ｐ＜０．０３，Ｐ＜０．００３和Ｐ＜０．０４）。ＣＤ－４５ＲＯ＋／ＲＡ＋（Ｔ／Ｂ）细胞数变化不大。研究结果提示银屑病发病与皮肤组织中积聚的免疫活化细胞在局部产生一系列引起表皮细胞异常增殖与分化的细胞因子有关。     

红斑狼疮皮损中浸润细胞免疫表型和角质形成细胞HLA-DR抗原表达的研究

应用抗ＨＬＡＤＲ、ＣＤ３、ＣＤ４、ＣＤ８、ＣＤ２０的单克隆抗体和ｓｔｒｅｐｔｒａｖｉｄｉｎｐｅｒｏｘｉｄａｓｅｓｔａｉｎｉｎｇ（ＳＰ）技术对１０名正常人皮肤,１６例ＳＬＥ皮损和１９例ＤＬＥ皮损进行了免疫组化研究。观察到正常人皮肤角质形成细胞未见ＨＬＡＤＲ抗原表达,而ＳＬＥ（６／１６）,ＤＬＥ（８／１９）皮损处角质形成细胞可以表达ＨＬＡＤＲ抗原。在ＳＬＥ、ＤＬＥ真皮内浸润细胞主要为Ｔ淋巴细胞（ＣＤ３＋浸润细胞）,且以ＴＨ细胞（ＣＤ４＋浸润细胞）占优势。另外,还发现在两种ＬＥ表皮角质形成细胞表达ＨＬＡＤＲ抗原处,真皮内可见ＣＤ３＋浸润细胞和激活的Ｔ淋巴细胞（ＨＬＡＤＲ＋浸润细胞）。讨论了ＬＥ皮损角质形成细胞ＨＬＡＤＲ抗原表达及其与病损内浸润细胞免疫表型的关系。ＬＥ皮损处ＨＬＡＤＲ＋角质形成细胞可能具有抗原递呈作用,而角质形成细胞异常表达ＨＬＡＤＲ抗原则可能与真皮内浸润单个核细胞或淋巴细胞释放的ＩＦＮα,ＴＮＦγ等有关。     

皮肤真菌病患者郎格罕细胞和T细胞亚群的检测

应用一组单克隆抗体，ＡＢＣ法对３０例体（股）癣病人外用血及皮损的郎格率细胞和Ｔ细胞亚群进行检测，结果表明病人外周血细胞，皮损中ＨＬＡ－ＤＲ＋细胞均高于正常对照（Ｐ＜０．０５～０．００１），细胞数目无明显改变（Ｐ＞０．０５），细胞在皮肤真菌病中可能递呈真菌抗原给Ｔ细胞。比率无明显改变（Ｐ＞０．０５），说明局部免疫功能基本正常。上述改变说明细胞免疫在皮肤真菌病的发病机理中起着重要作用。     

带状疱疹患者外周血T淋巴细胞亚群及NK细胞分析

对５０例带状疱疹患者外周血Ｔ淋巴细胞亚群及自然杀伤细胞（ＮＫ）用流式细胞仪进行检测,并与３０例正常人对照,结果带状疱疹组ＣＤ３下降不明显（Ｐ〉０．０５）,ＣＤ４下降明显（Ｐ〈０．０１）,ＣＤ８明显升高（Ｐ〈０．０１）,ＣＤ４／ＣＤ８下降明显（Ｐ〈０．０１）,ＮＫ细胞升高（Ｐ〈０．０１）,提示患者辅助性Ｔ细胞减少,抑制性Ｔ细胞增另,存在免疫抑制现象。     

银屑病皮损中γ干扰素的免疫组化分析

采用γ干扰素（ＩＦＮ－ ）单克隆抗体、双层ＡＰＡＡＰ染色法及显微分光光度测定技术对３０例银屑病患者皮损、１５例正常对照皮肤进行ＩＦＮ－ 抗原的测定。结果显示：除表皮基底层及表皮突下部 １～３层基层外，大量的ＩＦＮ－　抗原阳性染色弥漫分布于皮损全层表皮的角肮细胞间隙；正常对照皮肤的表皮中无明确的ＩＦＮ－　阳性反应；银屑病皮损表皮中ＩＦＮ－　的含量①与银屑病的活动性有关，进行期为２４．３８０８．２５０，静止期为１５．９５５　５．３２７（Ｐ＜０．０１）；②与皮损表皮中Ｔ细胞、ＨＬＡ－ＤＲ细胞的数量呈直线正相关（ｒ１＝０，６９４，ｒ２＝０．４３６，Ｐ＜０．０５）。     

银屑病家系与HLA致病单倍型研究

报告２３个多发银屑病家系及５９例无亲缘关系银屑病患者ＨＬＡ抗原频率分布。其增高的抗原为ＨＬＡ－Ａ１９，Ｂ１３、ＤＲ７（Ｐｃ＜０．００１），降低的抗原为ＨＬＡ－Ｃ７，ＤＱ１，（Ｐｃ＜０．００１），提示具有（Ｃｗ（Ｃ７）者对银屑病的发生具有某种防护作用。２３个家系中，健康同胞５５例，其中２０例与患病同胞有一条或两条单倍型相同，经６～８年观察，４例已发病；另３５例无相同单倍型者，未发病。提示对银屑病家系检测ＨＬＡ单倍型有助于对具有致病单倍型的健康同胞的临床监测。     

系统性红斑狼疮患者外周血单一核细胞CD23的表达

为了揭示Ｂ细胞中ＣＤ２３的表达与ＳＬＥ发生发展的关系及在ＳＬＥ发病机理中可能的作用，我们应用ＡＢＣ免疫组化法和斑点核酸杂交技术对ＳＬＥ患者外周血单一核细胞（ＰＢＭＣ）ＣＤ２３蛋白和ｍＲＮＡ表达进行了检测。结果显示：３０例ＳＬＥ患者ＰＢＭＣＣＤ２３蛋白表达显著增高（Ｐ＜０．０１），且与疾病活动呈正相关关系（ｒｓ＝０．３８１４，Ｐ＜０．０５）；具有不同ＡＮＡ、抗ｄｓＤＮＡ抗体水平，有无伴肾损、脑损的ＳＬＥ患者，ＰＢＭＣＣＤ２３表达均无显著性差异（Ｐ均＞０．０５）；单纯使用皮质类固醇激素治疗或和其它免疫抑制剂联合治疗的ＳＬＥ患者，ＰＢＭＣＣＤ２３表达亦无显著性差异（Ｐ＞０．０５）。２０例ＳＬＥ患者ＰＢＭＣＣＤ２３ｍＲＮＡ表达较正常人显著增高（Ｐ＜０．０１）。经治疗病情稳定后，ＣＤ２３蛋白和ｍＲＮＡ表达均降至正常（Ｐ均＞０．０５）。提示在ＳＬＥ活动期Ｂ细胞高度激活、增殖并大量表达ＣＤ２３，且该种表达与ＡＮＡ、抗ｄｓＤＮＡ抗体产生水平无直接关系     

银屑病患者HLA-DR基因分型的研究

利用多聚酶链反应技术及序列特异性寡核苷酸探针对３０例银屑病患者进行了ＨＬＡ－ＤＲ基因分型，并与２５５例北方汉族正常人ＨＬＡ－ＤＲ基因频率进行了比较。发现ＨＬＡ－ＤＲ７基因频率在银屑病患者较正常对照组明显增高（２５．０：１４．５，Ｐｃ＜０．０５，ＲＲ＝２．４４６），而ＨＬＡ－ＤＲ９基因频率却较正常对照组明显降低（３．３３： １２．４，Ｐｃ＜０．０５，ＲＲ＝０．２１８）。提示银屑病致病基因与ＨＬＡ－ＤＲ７关联。     

二期梅毒患者外周血淋巴细胞凋亡及其Fas、Bcl-2表达的研究

目的 检测二期梅毒患者外周血淋巴细胞(PBLC)凋亡及其Fas、Bcl-2的表达,探讨二期梅毒患者免疫功能异常与淋巴细胞凋亡的关系。方法 采用流式细胞仪检测33例二期梅毒患者和30例正常人PBLC细胞凋亡及其Fas、Bcl-2的表达。结果 梅毒患者PBLC及CD4⁺细胞Fas表达明显高于对照组(P<0.01),而Bcl-2表达明显低于对照组(P<0.01)。CD8⁺、CD19⁺细胞Fas、Bcl-2表达两组比较差异无统计学意义(P>0.05)。梅毒患者PBLC及CD4⁺细胞凋亡率显著高于对照组(P<0.01),CD8⁺及CD19⁺细胞凋亡率两组比较差异无统计学意义(P>0.05)。梅毒患者PBLC及CD4⁺细胞凋亡率分别与其Fas表达呈显著正相关(r=0.68,P<0.01;r=0.71,P<0.01),但与Bcl-2表达呈显著负相关(r=-0.82,P<0.01;r=-0.74,P<0.01)。结论 二期梅毒患者细胞免疫功能异常可能与淋巴细胞凋亡过度有关,而淋巴细胞凋亡过度与Fas、Bcl-2表达异常有关。     

The relevance of peripheral blood T-helper 1 and 2 cytokine pattern in the evaluation of patients with mycosis fungoides and Sézary syndrome

BACKGROUND: There is evidence that a T-helper (Th) 2 cytokine pattern dominates in the peripheral blood as well as in tissue of patients with Sézary syndrome (SS), and that the malignant clone is of Th2 phenotype. However, there are conflicting studies on the cytokine pattern in the peripheral blood in different stages of cutaneous T-cell lymphoma (CTCL). OBJECTIVES: To examine, by means of flow cytometry (FC), the Th1/Th2 cytokine profile [cytoplasmic interferon (IFN)-gamma/interleukin (IL)-4] in peripheral blood T cells from patients with mycosis fungoides (MF) and SS, the most common forms of CTCL, and to correlate their expression with clinical stage, clonality and T-cell immunophenotype changes in order to evaluate their relevance in CTCL progression. METHODS: We investigated by FC the percentage of CD3+ T cells expressing cytoplasmic IFN-gamma and IL-4 after stimulation in blood specimens of 43 CTCL patients (32 stage I-II and 11 stage III-IV), eight of whom were erythrodermic. Next, we compared cytoplasmic IFN-gamma and IL-4 expression between patients of different stages and controls, and correlated our findings to T-cell receptor (TCR)-gamma gene rearrangement, used as a marker of clonality, and changes in T-cell immunophenotype (CD4+, CD8+, CD4+/CD7-, CD4+/CD25+) and natural killer cells. Polymerase chain reaction amplification of the TCR-gamma gene was performed in 41 blood and 26 skin specimens. We also examined the cytokine expression pattern in patients with erythrodermic MF and SS. RESULTS: A significantly higher frequency of CD3+/IL-4+ T cells was found in late (III-IV) compared with early (I-II) CTCL patients (P = 0.002) or controls (P < 0.001). There were significant positive correlations between the percentages of CD3+/IL-4+ and the percentages of CD3+/CD4+ T cells (r = 0.385, P = 0.05), CD4+/CD7- T cells (r = 0.335, P < 0.05) and CD4+/CD25+ T cells (r = 0.433, P = 0.01); there was a negative correlation between the percentages of CD3+/IL-4+ and CD3+/CD8+ T cells (r = -0.463, P = 0.005) and a positive correlation between the percentages of CD3+/IFN-gamma+ and CD3+/CD8+ T cells (r = 0.368, P = 0.02). Increased percentages of CD3+/IL-4+, CD3+/CD4+ and CD4+/CD7- T lymphocytes were associated with the presence of clonality (P = 0.025, P < 0.001 and P = 0.0031, respectively). All independent variables showed a statistically significant difference between SS and erythrodermic MF patients, or controls, apart from cytoplasmic IL-4, which was high both in erythrodermic MF and SS patients compared with controls (P = 0.003 and P = 0.008, respectively). In multiple regression logistic analysis, the probability of belonging to advanced CTCL stages was associated only with increased cytoplasmic IL-4 (P = 0.007, odds ratio 1.13, 95% confidence interval 1.033-1.229). CONCLUSIONS: Increased T-cell cytoplasmic IL-4 is more frequent in late CTCL stages, correlates with T-cell immunophenotype changes found in advanced disease and is associated with clonality. Increased cytoplasmic IL-4 is frequent both in erythrodermic MF and SS patients, in contrast to other variables found increased only in SS, suggesting that IL-4 may be an early indicator of disease progression. Moreover, our results show that increased cytoplasmic IL-4 is the sole predictor of advanced CTCL disease and confirm the relevance of FC determination of IL-4 in the routine evaluation of CTCL cases.     

miRNA-146a对寻常性银屑病患者外周血CD4+ T细胞的调控研究

目的 研究miRNA-146a对寻常性银屑病患者外周血CD4+ T淋巴细胞的调控,探讨miRNA-146a在寻常性银屑病发病中的作用。 方法 收集寻常性银屑病患者和健康对照各30例。采用实时定量PCR检测外周血CD4+ T淋巴细胞miRNA-146a表达水平,ELISA检测血浆干扰素（IFN）-γ、白细胞介素（IL）-4表达水平。免疫磁珠法分选外周血CD4+ T淋巴细胞,将分离的细胞分为对照组、miRNA-146a组和miRNA-146a抑制物组,分别转染阴性对照miRNA、miRNA-146a模拟物和miRNA-146a抑制物后进行细胞培养,流式细胞仪检测Th1和Th2细胞数量,蛋白免疫印迹法和实时定量PCR分别检测IFN-γ受体α（IFN-γRα）、T细胞表达T盒（T-bet）、GATA-3蛋白和mRNA的表达,ELISA检测细胞培养液上清液IFN-γ、IL-4水平。 结果 寻常性银屑病患者外周血CD4+ T淋巴细胞miRNA-146a［（243.81 ± 94.32）％］与血浆IFN-γ水平［（27.69 ± 7.64） ng/L］较健康对照组［分别为（105.74 ± 22.93）％和（9.75 ± 2.81） ng/L］升高,差异均有统计学意义（t值分别为6.653和4.237,均P < 0.01）,且miRNA-146a的表达与血清IFN-γ呈正相关（r = 0.837,P < 0.01）。体外CD4+ T淋巴细胞培养结果显示,与对照组比较,miRNA-146a组Th1数量、T-bet蛋白和mRNA表达、培养上清液IFN-γ水平均显著增加,IFN-γRα蛋白表达降低,差异均有统计学意义（均P < 0.01）;但是,与对照组比较,miRNA-146a组和miRNA-146a抑制物组Th2细胞数量、GATA-3蛋白、GATA-3 mRNA表达以及上清液中IL-4水平差异无统计学意义（均P > 0.05）。结论 miRNA-146a可能通过影响Th1细胞的分化和功能参与对寻常性银屑病患者外周血CD4+ T淋巴细胞的调控,在银屑病发病过程中发挥一定的作用。     

CD70在SLE患者外周血T淋巴细胞中表达的研究

目的 通过研究共刺激分子CD70在SLE患者外周血T淋巴细胞的表达水平,以及DNA甲基化抑制剂氮杂胞苷对正常人外周血T淋巴细胞表达CD70分子的影响,探讨DNA甲基化在SLE患者外周血T淋巴细胞表达CD70分子中的调控作用。方法 用流式细胞仪检测了10例活动期、10例非活动期SLE患者和10例正常人对照外周血T淋巴细胞以及甲基化抑制组（氮杂胞苷处理正常人T淋巴细胞）的CD70+CD4+ T淋巴细胞阳性率;用RT-PCR方法测定了各组外周血T淋巴细胞的CD70 mRNA转录水平。结果 正常人外周血CD4+CD70+ T淋巴细胞阳性率为14.55% ± 5.49%,活动期、非活动期SLE分别为85.25% ± 14.08%和77.65% ± 18.77%,甲基化抑制组为81.54% ± 8.71%,与正常人对照组相比,活动期、非活动期SLE患者和甲基化抑制组CD70+CD4+ T淋巴细胞阳性率均明显升高（P < 0.01）,其外周血T淋巴细胞CD70 mRNA表达水平亦明显增加（P < 0.05）;SLE患者外周血CD70+CD4+ T淋巴细胞阳性率与SLE疾病活动度呈显著正相关（r = 0.72,P < 0.05）。结论 CD70的过度表达在SLE的免疫紊乱中起着重要的作用,并且CD70过度表达可以作为SLE疾病活动的一个指标。     

淋巴细胞活化在系统性红斑狼疮发病中的意义

目的 :　探讨系统性红斑狼疮 (SLE)患者外周血淋巴细胞HLA -DR以及植物凝集素 (PHA)刺激下的CD3+ CD4 + 、CD3+ CD8+ 、CD19+ 淋巴细胞的早期活化标志CD6 9的表达。方法 :　应用三色荧光标记流式细胞术检测活动期和非活动期SLE患者外周血CD3+ 、CD3+ CD4 + 、CD3+ CD8+ 淋巴细胞晚期活化标志HLA -DR以及在PHA刺激 4h后CD3+ CD4 + 、CD3+ CD8+ 、CD19+ 淋巴细胞的早期活化标志CD6 9的表达。结果 :　①PHA刺激 4小时后 ,活动期SLECD3+ CD4 + 、CD3+ CD8+ 细胞CD6 9的表达明显低于正常对照组 (P <0 .0 5 ) ;而非活动期SLE与正常对照组之间CD6 9表达无显著性差异 (P >0 .0 5 )。②PHA刺激前SLE活动组CD19+ 细胞表达CD6 9较正常对照组和SLE非活动期明显升高 (P <0 .0 5 ) ,PHA刺激 4小时后 ,活动期SLECD19+ 细胞表达CD6 9低于正常对照组和非活动期SLE ,存在显著性差异 (P <0 .0 5 ) ,而非活动期SLE与正常对照组之间CD19+ CD6 9+ 表达无明显差异 (P >0 .0 5 )。③活动期SLECD3+ HLA -DR+细胞明显高于正常对照组 (P <0 .0 1)和非活动期SLE(P <0 .0 1) ,CD3+ CD4 + HLA -DR+ 细胞、CD3+ CD8+HLA -DR+ 细胞明显高于正常对照组 (P <0 .0 5 ) ,而非活动期SLE与正常对照组之间 ,HLA -DR表达无显著性差异 (P >0 .0 5     

CD28/B7在银屑病皮损及外周血淋巴细胞中的表达

目的 探讨CD28/B7共同刺激分子在银屑病发病中的作用。方法 采用免疫组化ABC法检测22例银屑病皮损、流式细胞仪检测17例银屑病外周血淋巴细胞CD28、CD80、CD86的表达水平。15例整形外科手术患者的皮肤和外周血作为正常人对照。结果 免疫组化结果显示银屑病组皮损中CD28、CD80、CD86的表达明显增多,与正常人对照组相比差异均有显著性(P<0.01);进行期皮损中的表达显著高于静止期,差异均有显著性(P<0.05)。流式细胞仪检测显示CD28、CD80、CD86在银屑病组外周血淋巴细胞中的表达明显高于对照组,差异有显著性(P<0.01);进行期组与静止期组相比,CD28差异无显著性(P>0.05),CD80与CD86差异有显著性(P₁<0.01,P₂<0.05)。结论 CD28、CD80、CD86在银屑病发展过程中起一定作用。     

Low CD7 expression in benign and malignant cutaneous lymphocytic infiltrates: experience with an antibody reactive with paraffin-embedded tissue.

Loss of CD7 expression by neoplastic lymphocytes is considered a distinguishing characteristic of mycosis fungoides (MF) and cutaneous T-cell lymphoma. Reports to date examining for the CD7 immunophenotype in MF have been performed on fresh-frozen tissue. In this study, we used a paraffin-reactive antibody directed against CD7 to determine its range of expression in MF and to compare these results with those in controls. Examining 22 cases of MF and 61 controls, we found minimal CD7 expression by lymphocytes in MF and in a few cases of benign inflammatory dermatosis (BID). The lowest mean CD7 counts (as a percentage of total lymphocytes) were found in MF (patch stage: 5% +/- 5%, range: 0-10; plaque and tumor stages: 15% +/- 5%, range: 5-25), and these counts were significantly lower than those for BID (35% +/- 20%, range: 5-80; p = 0.001). By logistic regression analysis, low CD7 expression (<10% lymphocytes labeling) had sensitivity and positive predictive values of 80% and 72%, respectively, and specificity and negative predictive values of 93% and 96%, respectively, for the diagnosis of patch stage MF. False-positive results were found for spongiotic dermatitis. Moreover, spongiotic dermatitides exhibited a progressive decrease in mean CD7 counts from acute to subacute to chronic stages (50% versus 35% versus 30%, respectively). In conclusion, minimal CD7 expression is a specific finding for MF. Benign inflammatory infiltrates can also show low CD7 expression, however, which rarely matches that of patch stage MF. Progressive loss of CD7 expression in BID is the likely consequence of expansion of antigen-selected CD3+CD4+CD7- T cells. These inflammatory CD4+CD7- T cells may represent the physiologic counterpart to the neoplastic lymphocyte of MF.     

蕈样肉芽肿患者瘙痒与趋化因子CCL17相关性研究

【摘要】 目的 寻找参与蕈样肉芽肿（MF）患者瘙痒发生的相关分子。方法 纳入2009年10月至2021年8月于北京大学第一医院就诊的522例MF患者作为研究对象，统计瘙痒发生率。根据是否有瘙痒症状对患者进行分组，分析其中49例患者皮损活检组织的RNA测序数据，寻找有瘙痒症状与无瘙痒症状患者的差异表达基因；使用酶联免疫吸附试验和免疫组织化学技术分别检测88例MF患者血清和81例MF患者组织CC趋化因子17（CCL17）蛋白表达量；使用流式细胞仪检测46例患者外周血T淋巴细胞活化和分化标记，寻找与瘙痒症状相关的外周血淋巴细胞亚群。使用χ2检验、Mann-Whitney U检验、Spearman相关性检验进行统计分析。结果 522例患者中，男305例，女217例；早期MF 347例，进展期MF 175例。MF患者瘙痒发生率为67.2%（351例），进展期患者瘙痒发生率（81.7%，143/175）较早期患者（59.9%，208/347）上升（χ2 = 25.03，P＜0.001）。RNA测序结果显示，瘙痒MF患者CCL17 mRNA表达量高于无瘙痒MF患者（差异表达倍数 = 10.09，P＜0.001）。瘙痒患者血清CCL17浓度［1 017.05（377.12，4 831.80） pg/ml］较无瘙痒患者［361.66（180.47，500.08） pg/ml］上升（Z = -4.57，P＜0.001），且与瘙痒评分呈正相关（r = 0.57，P = 0.010）。在早期和进展期MF患者中，瘙痒患者血清CCL17浓度均高于无瘙痒患者（Z = -3.68，P＜0.001；Z = -2.54，P = 0.011）。瘙痒患者与无瘙痒患者CCL17免疫组化相对定量评分差异无统计学意义（Z = -1.84，P = 0.066）。瘙痒患者CD3+CD4+CD26-CCR4+恶性T细胞百分比高于无瘙痒MF患者（Z = -2.03，P = 0.043），且与血清CCL17浓度呈正相关（r = 0.49，P＜0.001）。结论 MF瘙痒患者皮损组织CCL17mRNA和血清CCL17浓度上升，CCL17与MF患者瘙痒发生相关，可能通过招募CD3+CD4+CD26-CCR4+恶性T细胞参与瘙痒发生。     

Perforin expression in peripheral blood lymphocytes and skin-infiltrating cells in patients with lichen planus

BACKGROUND: Current evidence suggests that lichen planus is a T-cell-mediated autoimmune disease in which cytotoxic mechanisms have been poorly investigated. OBJECTIVES: We investigated the expression of perforin in subpopulations of peripheral blood lymphocytes (PBL) in exacerbation and remission phases of the disease as well as in skin lesions. METHODS: We performed a simultaneous detection of perforin (intracellular molecule) and cell surface antigens on PBL by flow cytometry, and skin lesions were investigated by immunohistochemistry. RESULTS: The most interesting finding was a significant increase of perforin expression in cytotoxic T lymphocytes (CD3+ perforin+ cells) in the exacerbation phase of disease (P < 0.05), which was mostly located in the CD8+ subpopulation (CD8+ perforin+) (P < 0.01). Using immunohistochemistry we confirmed the infiltration of T lymphocytes in skin lesions, especially of CD4+ and CD8+ phenotypes, compared with uninvolved (P < 0.05) and healthy skin (P < 0.01). The expression of perforin was also significantly higher in lesional skin compared with nonlesional and healthy skin (P < 0.05). CONCLUSIONS: Our results clearly show the upregulation of perforin expression in peripheral blood as well as in lesions of patients with lichen planus and therefore suggest an important role for perforin in this autoimmune disease.     

特应性皮炎患者外周血T淋巴细胞Rho激酶活性变化及其意义

目的 观察特应性皮炎（AD）患者外周血T淋巴细胞中Rho激酶活化情况,分析其临床意义。方法 收集60例AD患者和60例健康儿童外周肝素抗凝血8 ml,分离提取T细胞和血清。分别用AD血清和健康对照血清培养AD患者T细胞或健康对照T细胞,分为患者T细胞 + 自身血清组、患者T细胞 + 健康对照血清组、健康对照T细胞 + 自身血清组、健康对照T细胞 + AD血清组。此外,分别用Rho激酶特异性抑制剂Y27632（Y27632组）、CD3/CD28单抗（CD3/CD28单抗组）、Y27632 + CD3/CD28单抗（Y27632 + CD3/CD28单抗组）处理AD患者T细胞,自身血清培养AD患者T细胞为患者T细胞组。采用Western印迹法检测各组Rho激酶活性,四甲基偶氮唑蓝比色法（MTT）检测T细胞增殖活性,ELISA检测白细胞介素（IL）6、IL-10水平。 结果 新鲜分离的AD患者外周血T细胞Rho激酶活性（2.47% ± 0.89%）显著高于健康对照组（0.65% ± 0.35%,t = 2.729,P < 0.05）。AD患者T细胞在体外经10%胎牛血清培养24 h后Rho激酶活性（0.70% ± 0.38%）较培养前显著降低（t = 2.658,P < 0.05）,但与培养24 h后健康对照T细胞（0.63% ± 0.32%）相比,差异无统计学意义（t = 1.010,P > 0.05）。与健康对照血清培养T细胞相比,AD血清培养T细胞会增加Rho激酶活性（F = 8.22,P < 0.001）,患者T细胞 + 自身血清培养24 h后Rho激酶活性（2.41% ± 0.87%）明显高于患者T细胞 + 健康对照血清组（0.76% ± 0.41%）,健康对照T细胞 + AD血清组（2.17% ± 0.85%）显著高于健康对照T细胞 + 自身血清组（0.64% ± 0.33%）,差异均有统计学意义（P < 0.05）。Y27632可显著抑制AD患者T细胞的增殖和IL-6的分泌（F = 18.68、22.95,P < 0.001）,Y27632组T细胞增殖率、IL-6的表达均显著低于患者T细胞组（均P < 0.05）,且Y27632 + CD3/CD28单抗组也显著低于CD3/CD28单抗组（均P < 0.05）,而Y27632对AD患者T细胞IL-10分泌无显著影响。 结论 AD患者T淋巴细胞存在Rho激酶信号活化异常,提示Rho激酶信号异常在AD发病过程中可能发挥着一定的作用。