组织常驻记忆性T细胞与银屑病复发的研究进展

银屑病是慢性复发性炎症性皮肤病,经常在原皮损消退的部位复发。组织常驻记忆T细胞(tissue resident memory T cells, TRM)能够长期驻留在皮肤中。近年来许多研究表明TRM是银屑病复发的重要原因,有效地抑制TRM可能是控制银屑病复发的关键。但TRM细胞具有抗损伤和抗凋亡特征,给控制复发带来困难。本文就皮肤TRM的产生、驻留以及TRM与银屑病复发的研究进展进行综述。     

银屑病患者皮损处γδT细胞受体γ链V区取用模式的初步研究

目的探讨银屑病患者皮损处γδT细胞的取用模式及其在银屑病发病中的作用。方法通过实时荧光定量PCR的方法检测18例寻常性银屑病患者和18例正常人皮肤及外周血PBMC中γδT细胞受体γ链V区基因(TRGV,T cell receptor gamma variable,亦称Vγ,TCRGV)的表达情况。结果寻常性银屑病组皮损中Vγ9的表达率为对照组皮肤的4.3倍,而对照组外周血PBMC中Vγ9的表达率为寻常性银屑病组的2.7倍,差异均有统计学意义(P=0.00003,0.0002)。对照组皮肤Vγ11的表达率为寻常性银屑病组皮损的3.5倍,差异有统计学意义(P=0.0003)。银屑病组和对照组比较外周血PBMC中Vγ11的表达率、皮肤及外周血PBMC中Vγ1-8和Vγ10的表达差异均无统计学意义(P均0.05)。结论寻常性银屑病患者皮损中γδT细胞Vγ9较正常人高表达,Vγ9+γδT细胞可能是银屑病患者真皮中有致病作用的γδT细胞的主要亚型;银屑病患者皮损中γδT细胞Vγ11较正常人低表达。对这一现象有必要进行深入研究。     

学术动态-2014（5）380.1

许多临床试验采用针对白介素（IL）-17的抗体治疗银屑病取得成功,然而,关于皮肤炎症反应中IL-17的来源和动态变化趋势目前尚有争议。研究者通过筛选健康志愿者建立两组人体模型模拟银屑病样的皮肤炎症反应,观察表达IL-17的免疫细胞的动态变化趋势。结果两组模型都明确揭示中性粒细胞和T细胞的流入情况,并显示IL-17主要由中性粒细胞和肥大细胞表达,同时中性粒细胞还表达IL-17相关转录因子RORγt并且能形成细胞外陷阱。在皮肤炎症反应过程中,肥大细胞保持相对稳定,而中性粒细胞有一个明显的动态变化趋势,因此说明IL-17+/RORγt+中性粒细胞影响机体皮肤炎症反应的发生,并可能参与炎症性皮肤病（银屑病）的发病。但是,T细胞中只有少数表达IL-17,这使皮肤炎症反应中IL-17主要与T细胞相关的传统观点遭遇挑战。［J Invest Dermatol, 2014, 134（5）: 1276-1284.］     

银屑病皮损CD8αα+ T细胞表型鉴定

目的 探究银屑病皮损局部浸润的CD8+ T细胞表型及其在银屑病发病中的作用。方法 收集2017年1 - 12月第四军医大学西京皮肤医院明确诊断的8例进行期斑块状银屑病患者皮损,男女各4例,年龄24 ~ 50岁。8例健康对照皮肤来源于整形外科手术剩余皮肤,男女各4例,年龄23 ~ 46岁。采用免疫荧光技术检测皮损CD8+ T细胞的分布及亚群比例,鉴定其免疫学表型及炎症因子白细胞介素（IL）17A的表达。结果 8例银屑病皮损真、表皮均可见CD8+ T细胞浸润,其中CD8αα+ T细胞表型占88.48% ± 7.39%,8例健康人皮肤局部仅有个别CD8+ T细胞浸润,其中CD8αα+ T细胞表型占14.43% ± 13.14%,两组比较,t = 11.5,P < 0.01。银屑病皮损表皮CD8αα+ T细胞表达组织局部定植标志CD103,真皮CD8αα+ T细胞不表达CD103。银屑病皮损CD8αα+ T细胞表达CD45RA-CCR7-效应记忆性T细胞表型,不表达CD8+ 调节性T细胞标志Foxp3、CD25和CD122。银屑病皮损CD8αα+ T细胞中分泌IL?17A的细胞比例为24.85% ± 4.25%,对照组CD8αα+ T细胞不分泌IL?17A,两组差异有统计学意义（t = 5.853,P < 0.01）。结论 银屑病皮损浸润的CD8αα+ T细胞是效应记忆性T细胞,可能通过分泌IL?17A参与银屑病的发生发展。     

Th22及其细胞因子在斑秃患者外周血的表达和意义

目的：探讨Th22及其细胞因子在斑秃患者外周血中的表达及其临床意义,分析T淋巴细胞在斑秃发病机制中的作用。方法选取上海市闵行区中心医院2015年1月至2016年5月收治的38例斑秃患者作为研究对象,设为斑秃组;同时取同期健康体检38例作为健康对照组,用流式细胞仪检测Th22细胞水平,用酶联免疫吸附检测血清中IL?22、IL?17的水平。结果斑秃组外周血Th22、Th17、IL?17+IL?22+CD4+、IFN?γ+IL?22+CD4+T细胞表达水平均高于健康对照组（P<0.01）。斑秃组血清IL?22、IL?17水平均高于健康对照组（P<0.05）;与轻型斑秃患者和稳定期患者相比,重型斑秃和活动期患者Th22、Th17、IL?17+IL?22+CD4+、IFN?γ+IL?22+CD4+T细胞及血清IL?22、IL?17的表达进一步升高（P<0.05）。结论 Th22及其细胞因子可能参与斑秃的发生、发展、预后。     

咪喹莫特诱导小鼠银屑病样皮损机制的研究进展

咪喹莫特作为治疗人乳头瘤病毒感染的药物,在临床应用中发现可引起银屑病样皮损并能加重银屑病患者病情,而被广泛用于银屑病发病机制及治疗靶点的研究.由咪喹莫特诱发的小鼠银屑病样模型具有操作简单、易于成模的优点.应用咪喹莫特后,小鼠体内有包括Toll样受体信号通路等多条通路被激活,其中部分为协同作用.激活的信号通路进一步诱导角质形成细胞、树突细胞、Th17、γδT细胞、中性粒细胞等产生一系列炎性因子(如白细胞介素1、23、17,干扰素α等),这些炎性因子对银屑病样皮损的形成起到重要的作用.     

不同证候银屑病患者外周血Th1/Th17细胞表达差异的研究

目的 探讨不同证候银屑病的发病与外周血Th1/Th17细胞轴漂移的相关性。方法 用流式细胞仪检测CD4+T细胞内细胞因子干扰素?酌（IFN-?酌）、白介素17A（IL-17A）的表达情况,双抗体夹心法（ELISA）检测不同证候银屑病患者血清中Th１型细胞因子IFN-?酌和Th17型细胞因子IL-17A的水平。 结果 血热组银屑病患者外周血单一核细胞（PBMC）中CD4+IFN-?酌+水平显著高于血瘀组及健康对照组（P < 0.05）,而血瘀组与健康对照组外周血PBMC中CD4+IFN-?酌+水平差异无统计学意义。血热组银屑病患者PBMC中CD4+IFN-?酌+水平与PASI评分呈正相关（P < 0.05）,血瘀组不具相关性（P > 0.05）。血热组、血瘀组银屑病患者及健康对照组PBMC中CD4+IL-17A+三组间差异无统计学意义。血热组银屑病患者PBMC中CD4+IL-17A+水平与PASI评分不具相关性（P > 0.05）,而血瘀组呈正相关（P < 0.05）。ELISA结果示,血热组银屑病患者外周血清中IFN-?酌水平显著高于血瘀组及健康对照组（P < 0.05）,而血瘀组与健康对照组差异无统计学意义（P > 0.05）;血热组银屑病患者血浆中IFN-?酌水平与PASI评分呈正相关（P < 0.05）,血瘀组银屑病患者呈负相关（P < 0.05）。血热组、血瘀组银屑病患者及健康对照血清中IL-17A水平三组间差异无统计学意义（P > 0.05）,血热、血瘀组银屑病患者血清中IL-17A水平与PASI评分不具相关性（P > 0.05）。 结论 Th1细胞在血热型银屑病的发病中占主导地位,当血热型银屑病向血瘀型转化或正常转归时,Th1细胞的表达下降,而Th17细胞在不同证候银屑病患者中差异无统计学意义。     

Notch1信号对银屑病患者外周血Th17细胞分化和功能的影响

目的 探讨Notch1信号对银屑病患者外周血Th17细胞分化和功能的影响。方法 取35例寻常性银屑病患者与32例健康对照外周血,流式细胞仪检测单个核细胞（PBMC）中Th17细胞占CD4+ T淋巴细胞的比例,实时荧光定量反转录PCR检测PBMC中维A酸相关孤儿核受体γt（RORγt）、白介素17（IL-17）、Notch1及发状分裂相关增强子1（Hes-1）mRNA表达水平,酶联免疫吸附试验检测血清及培养上清液中IL-17含量。分析Notch1 mRNA的表达与银屑病皮损面积和疾病严重程度评分（PASI）、Th17细胞比例、RORγt mRNA、IL-17 mRNA及蛋白表达水平的相关性。将银屑病患者PBMC分为0（对照组）、2.5、5.0、10.0、20.0 μmol/L γ分泌酶抑制剂（DAPT）处理组,检测DAPT阻断Notch1信号对PBMC中Th17细胞比例、RORγt及IL-17表达水平的影响。结果 银屑病患者PBMC中Th17细胞占CD4+ T细胞比例为2.863% ± 0.969%,RORγt、Notch1、Hes-1、IL-17 mRNA表达水平分别为5.255 ± 0.998、6.743 ± 1.756、6.384 ± 1.665、6.944 ± 1.626,IL-17血清含量为（36.444 ± 5.936） ng/L,均高于健康对照组［分别为0.604% ± 0.124%、1.530 ± 0.485、1.731 ± 0.456、1.627 ± 0.485、1.698 ± 0.329、（11.762 ± 2.260） ng/L,P < 0.01］。银屑病患者Notch1 mRNA表达水平与PASI、Th17细胞比例、RORγt mRNA表达水平、IL-17 mRNA表达水平及血清含量均呈正相关（r值分别为0.584、0.544、0.518、0.549、0.511,均P < 0.05）。5种不同浓度DAPT处理组银屑病患者PBMC中Th17细胞比例、RORγt mRNA表达水平、IL-17 mRNA表达水平及培养上清液含量差异有统计学意义（F值分别为79.527、82.239、78.086、80.558,均P < 0.01）,2.5、5.0、10.0、20.0 μmol/L DAPT组上述指标均低于对照组（均P < 0.05）,且随着DAPT作用浓度的增加,各指标呈降低趋势。结论 Notch1信号可促进银屑病患者外周血Th17细胞的分化及RORγt 、IL-17、Hes-1的表达。     

寻常性银屑病患者外周血CD4+CD25+调节性T细胞的功能研究

目的 测定银屑病患者外周血中的CD4+CD25+调节性T细胞功能的改变,探讨调节性T细胞在其发病中的作用。方法 寻常性银屑病患者12例,银屑病面积和严重度指数（PASI）评分15 ～ 34.5分,均为慢性斑块状。健康对照组6例,为健康献血者。通过免疫磁珠体外分离外周血中的CD4+CD25+T细胞和CD4+CD25- T细胞,纯度达90％以上。采用［3H］-脱氧胸腺嘧啶苷方法检测CD4+CD25+ T细胞的增殖和抑制功能,ELISA法测定细胞因子和RT-PCR检测Foxp3 mRNA的表达。结果 多克隆刺激后CD4+CD25+ T细胞无明显增殖,IL-2可逆转此无反应状态。健康对照组CD4+CD25+ T细胞对CD4＋CD25－ T细胞增殖的抑制率可达60％,而银屑病组抑制率仅为19.5％（P < 0.01）。银屑病患者CD4＋CD25－ T细胞单独培养及与CD4＋CD25+ T细胞共培养后,分泌IFN-γ水平分别为（179.66 ± 48.97） ng/L和（109.55 ± 48.04） ng/L,健康对照组分别为（87.36 ± 33.36） ng/L和（32.55 ± 15.69） ng/L,两组比较,P值均 < 0.05;对IFN-γ分泌的抑制率银屑病患者组为40％,健康对照组为63％（P < 0.05）;健康对照组CD4＋CD25+ T细胞和CD4＋CD25－ T细胞单独培养,TGF-β分泌分别为（3025 ± 769） ng/L和（2450 ± 431） ng/L（P < 0.05）。两组之间,无论单独培养或两种细胞共培养,IL-10、TGF-β的分泌差异均无统计学意义（P > 0.05）。CD4+CD25+ T细胞表达高水平的Foxp3, 银屑病患者和健康人对照调节性T细胞和效应T细胞的Foxp3表达相似。结论 银屑病患者外周血中调节性T细胞抑制功能的减弱导致效应T细胞增殖失控,可能参与了银屑病的发病。     

银屑病患者外周血CD4+T细胞内干扰素γ、白介素4表达与发病的关系

目的：探讨干扰素γ（IFN－γ）、白介素4（IL－4）表达在银屑病发病中的作用。方法：银屑病患者和正常人外周血经碘诺霉素和佛波酯刺激（同时加莫能呈抑制细胞因子向细胞外转移）后用三色荧光抗体染色；用流式细胞仪检测CD4^ T细胞内细胞因子IFN－γ、IL－4的表达情况。结果：未加刺激物时，银屑病患者与正常人外周血CD4^ ／IF－γ^ 细胞数量分别占0．79％、0．88％，CD4^ ／IL－4^ 细胞数量分别占1．92％、1．88％，两组间IFN－γ、IL－4的表达差异无显著性（P＞0．05）；经碘诺霉素和佛波酯刺激后银屑病患者CD4^ ／IFN－γ^ 细胞数量占21．69％，明显高于正常人对照组10．26％（P＜0．001），CD4^ ／IL－4^ 细胞数量分别占4．24％、4．40％，两组间差异无显著性（P＞0．05）。结论：未活化的CD4^ T细胞分泌的细胞因子极少。经抗原刺激后银屑病患者外周血Th1、Th2细胞分化失衡，导致IFN－γ异常表达，其在银屑病发病机制中起重要作用。     

Cytokines and their effects on maturation,differentiation and migration of dendritic cells

In this review the role of cytokines in the maturation and migration of phenotypically and functionally diverse dendritic cell (DC) subpopulations is discussed and their role in the progress of differentiation from bone marrow progenitors to lymphoid DC is described. GM-CSF is the most important cytokine for the development of functional DC and acts in concert with a varying mixture of other cytokines such as IL-4, IL-1 and TNF-α to direct the development of individual DC subpopulations. Received: 18 July 1996     

Dendritic cells and their role in skin-induced immune responses

This artide gives a brief review on dendritic cells (DC) with regard to their origin, life cyde and functions. The regulation of immune responses by DC functioning as antigen-presenting cells is discussed. Special attention is given to epidermal DC, e.g. Langerhans cells. The perspectives of DC-based therapy are also mentioned.     

Limited reliability of E-cadherin as a specific marker for in vitro-generated Langerhans cells

Langerhans cells (LC) are a unique population of dendritic cells (DC) found in the epidermis where they can be identified by the expression of CD1a, E-cadherin and cytoplasmic Birbeck granules (BG) as their hallmark. Over the past years many techniques have been described to generate LC in vitro from either monocytes or CD34⁺ hematopoietic cell progenitors. Antibodies against Lag and Langerin (two epitopes associated with BG) and E-cadherin (a Ca²⁺-dependent homophilic adhesion molecule) have been used to detect in vitro-generated LC. In this study we investigated whether the expression of E-cadherin on in vitro-generated CD1a⁺ from either CD34⁺ cells or monocytes is able to discriminate LC from other DC. Our results demonstrate that E-cadherin alone is not a reliable marker to specifically identify in vitro-generated LC.     

二期梅毒患者外周血Th1/Th2及Tc1/Tc2的研究

目的探讨CD4+T细胞(Th细胞)和CD8+T细胞(Tc细胞)亚群在梅毒发病机理中的作用及其相互关系。方法采用流式细胞术检测经三色荧光抗体染色的二期梅毒患者外周血CD4+/IFN-γ+(Th1),CD4+1/IL-4+(Th2),CD4+/IFN-γ+/IL-4+(Tho),CD8+/IFN-γ+(Tc1),CD8+/IL-4+(Tc2),CD8+/IFN-γ+/IL-4+(Tc0)细胞含量。结果梅毒组Th1细胞含量、Th1/Th2,Tc1/Tc2比值均明显低于正常对照组(P<0.01),而Tc0和Tc2细胞含量显著高于正常对照组,其他细胞两组间无显著性差异(P>0.05)。结论二期梅毒患者Th1/Th2及Tc1/Tc2比值失衡,可能是机体不能完全清除梅毒螺旋体造成其潜伏体内形成长期感染的重要因素之一。     

Indeterminate dendritic cell neoplasm of the skin: A 2‐case report and review of the literature

Indeterminate dendritic cell neoplasm (IDCN) is an exceedingly rare and mostly cutaneous histiocytosis, frequently associated with other hematopoietic malignancies. We report 2 cases of multilesional cutaneous IDCN. A 55‐year‐old male with no associated malignancy and complete response to ultraviolet phototherapy; and a 72‐year‐old male with chronic myelomonocytic leukemia (CMML). Both cases showed histiocytoid cytology, positivity for CD1a and no expression of langerin or BRAF^V600E. With our patients, the literature describes 79 cases of IDCNs, including 65 (82%) with only skin involvement, 7 cases (9%) with involvement of skin and a second site, 5 cases (6%) involving lymph nodes only, 1 splenic lesion and 1 systemic disease. Seventeen cases (22%) were associated with other hematopoietic malignancies, most commonly CMML (6 cases), follicular lymphoma (4 cases) and acute myeloid leukemia (3 cases). All IDCNs associated with myeloid malignancies were limited to the skin, while most cases associated with lymphoma were limited to lymph nodes. Reported responses of cutaneous lesions to ultraviolet phototherapy are encouraging, while systemic chemotherapy is appropriate for clinically aggressive cases and treatment of associated malignancies. Recognition of the clinico‐morphologic spectrum of IDCNs should prevent misdiagnoses and prompt investigation of possible associated neoplasms.     

皮肤Merkel细胞癌伴鳞状细胞癌1例

患者女,76岁。左面颊部肿物1年,明显增大1个月。皮损特点为紫红色球形肿物。皮损组织病理示:表皮见团块状鳞状上皮中-重度异型增生,突破基底膜;真皮全层见形态一致的肿瘤细胞浸润,巢状或片状分布,圆形或卵圆形,胞质少、嗜碱性,核大、散点状分布染色质,核仁明显,可见多数核分裂相。免疫组化:CK20(部分+),P63及CKH(鳞状分化的细胞+),Syn(+),CgA(+),CD56(+),LCA(-),CK7(-),S100(-)。诊断:皮肤Merkel细胞癌伴鳞状细胞癌。     

内皮细胞生长因子促进细胞增殖的研究

用高效液相色谱分析技术获得纯化内皮细胞生长因子 ( ECGF) ,体外培养发现 ECGF能提高人脐静脉内皮细胞及 L92 9成纤维细胞的分裂增殖能力     

Granulomas en dermatopatología: principales entidades. Parte I

This series of 2 articles on dermatopathologic diagnoses reviews conditions in which granulomas form. Part 1 clarifies concepts, discusses the presentation of different types of granulomas and giant cells, and considers a large variety of noninfectious diseases. Some granulomatous diseases have a metabolic origin, as in necrobiosis lipoidica. Others, such as granulomatous mycosis fungoides, are related to lymphomas. Still others, such as rosacea, are so common that dermatologists see them nearly daily in clinical practice.     

Histogenesis of pure and combined Merkel cell carcinomas: An immunohistochemical study of 14 cases

The histogenesis of Merkel cell carcinoma (MCC) has remained unresolved. Moreover, one of the questions is whether pure MCC and combined MCC represent the same histogenesis and entity. The existence of combined MCC suggests that MCC likely arise from pluripotent stem cells. Merkel cells (MC) localize within the bulge area, which is populated by hair follicle stem cells. We used hair follicle stem cell markers to investigate whether MCC share certain characteristics of these stem cells. Fourteen MCC specimens were examined histologically and immunohistochemically. There were six pure MCC and eight combined MCC. In six combined MCC, both MCC components and squamous components at least focally shared the expression of one or more of cytokeratin (CK)15, CK19 and CD200, which are hair follicle stem cell markers. On the other hand, four cases of pure MCC showed partially distinct CK19 expression, but did not show CK15 and/or CD200 expression. There was a distinct difference between pure MCC and combined MCC on the expression of hair follicle stem cell markers. The normal skin expressed CK15, CK19 and CD200 in the bulge area, whereas CK15 and CD200 were absent in the MC‐rich glabrous skin and touch domes. The results led us to hypothesize that combined MCC originate from the hair follicle stem cells. We postulate that combined MCC undergo multidirectional differentiation into squamous, glandular, mesenchymal and Merkel cells. Further investigation is warranted to confirm the histogenesis of pure MCC and combined MCC.