斑秃皮损内朗格汉斯细胞及CD8+ T细胞的数量及分布分析

【摘要】 目的 探讨斑秃患者脱发皮损中朗格汉斯细胞在斑秃病理进程中的分布以及与T细胞的关系。 方法 对29例斑秃患者（活动期16例,非活动期13例）头皮脱发皮损进行CD1a免疫组化染色,对其中17例斑秃患者行CD4、CD8免疫组化染色。荧光半定量PCR测定局部皮损浅层和深层CD1a和粒细胞巨噬细胞刺激因子（GM-CSF）的mRNA表达水平。 结果 斑秃患者表皮和真皮各处包括真皮浅层血管周围、毛囊周围,真皮深层血管周围、毛囊周围CD1a阳性LC数量均较健康对照显著增加（Z = 4.354,2.884,4.640,3.217, 3.496,均P ＜ 0.01）,活动期皮损表皮层、深层血管、深层毛囊CD1a阳性LC数量较非活动期皮损高（Z = 2.457, 2.130,1.954,P ≤ 0.05）。斑秃患者CD1a 、GM-CSF mRNA相对表达量在皮损真皮浅层虽与健康对照无差异,但在深层均高于健康对照（Z = 2.702,2.941,均P ＜ 0.01）。斑秃患者浅层血管周围LC与深层毛囊周围CD8+ T细胞数量呈正相关（r = 0.618,P ＜ 0.05）,活动组浅层血管周围LC与深层毛囊周围CD8+ T细胞数量分布呈正相关关系（r = 0.795,P = 0.01）,非活动组浅层血管周LC与深层毛囊周围CD8+ T细胞数量分布则无相关关系。 结论 斑秃患者皮损LC数量增加,且在活动期皮损升高更明显。活动期斑秃皮损中浅层血管周围LC与深层毛囊周围CD8+ T细胞数量呈正相关,推测LC在斑秃疾病进展中发挥作用。     

毛发疾病

20 0 4 0 387　斑秃患者皮损中 CD4 和 CD8 T细胞的检测 /肖风丽 (河北省医院皮肤科 )…∥临床皮肤科杂志 .- 2 0 0 3,32 (7) .- 396～ 397采用免疫组化的方法检测。结果发现斑秃皮损中 CD4 和 CD8 细胞明显增多 ,且以 CD4 T细胞为主 ,主要浸润于血管、毛囊周围以及毛囊外根鞘 ,表皮有散在分布。同时发现斑秃活动期CD4 T细胞数和 CD4 /CD8 值明显高于稳定期和正常头皮 ,提示斑秃活动期 CD4 T细胞可能起较重要作用。CD8 T细胞数在斑秃活动期和稳定期无明显差异 ,但均明显高于正常对照组 ,提示CD8 T细胞可能在斑秃…     

早期斑秃皮损处炎症细胞因子及凋亡因子的异常表达北大核心CSCD

目的 探讨早期斑秃皮损浅层、深层及斑秃生长期毛囊凋亡因子及炎症细胞因子的表达.方法 收集25例早期斑秃皮损及15例正常头皮病理取材组织标本,通过荧光定量PCR检测标本中凋亡基因p53、caspase3、Fas和生存素、bcl-2以及炎症细胞因子白细胞介素（IL）-4、IL-10、干扰素γ（IFN-γ）、IL-12 mRNA表达水平,免疫组化检测皮损中生长期毛囊P53的蛋白质表达.结果 斑秃生长期毛囊促凋亡因子caspase3、P53、Fas mRNA表达水平较对照组升高,变化倍数分别为6.78、8.01、9.74,差异有统计学意义（均P＜0.05）.抑制凋亡因子bcl-2、生存素mRNA较正常降低,变化倍数分别为0.08、0.03（均P＜0.01）,炎症细胞因子无改变.斑秃浅层Th1因子IFN-γ、IL-12 mRNA较正常水平升高,变化倍数分别为2.75、85.67（P值分别＜0.05、0.01）.Th2因子IL-10表达水平降低,变化倍数为0.002（P＜ 0.01）.而且,斑秃皮损浅层IL-10及IL-12 mRNA改变幅度高于深层（P值分别＜ 0.01、0.05）.免疫组化显示斑秃皮损生长期毛囊每100个细胞中p53阳性细胞数较对照组升高（t=23.79,P＜ 0.01）.结论 斑秃生长期毛囊凋亡因子升高,抑制凋亡因子表达下降提示凋亡因子在斑秃发病中起一定作用.     

斑秃共聚焦激光扫描显微镜影像学特征分析

目的 通过在体共聚焦激光扫描显微镜（CLSM）采集斑秃的微解剖图像,研究斑秃的CLSM影像学特征。方法 2010年1月至2011年5月,临床诊断斑秃患者46例。选定患处皮损及31例皮损附近正常皮肤行CLSM检查,并与斑秃的组织病理象作比较。结果 CLSM显示,单位面积内毛囊数目斑秃进展期［（134.856 ± 18.301）/cm2］及静止期［（147.159 ± 17.536）/ cm2］均显著低于正常对照组［（301.613 ± 35.317）/cm2,P值均 < 0.05］,毛囊内毛干缺失,毛囊周围、毛囊及真皮浅层毛细血管周围可见炎细胞浸润,静止期炎细胞浸润程度较进展期减轻;恢复期［（227.778 ± 16.861）/cm2］毛囊数目较进展期增加,但仍少于正常对照（P < 0.05）,可见毳毛及终毛生长,炎细胞浸润进一步减少。结论 斑秃CLSM图像与常规组织病理表现相符合,CLSM描述斑秃细胞水平的成像特点,可用于斑秃的临床观察及动态随访。     

斑秃毛囊退行性变及局部炎症细胞浸润的研究进展

斑秃是一种非瘢痕性的炎症性脱发性疾病,病情多能自限,但容易复发。斑秃发病机制不明,组织病理上表现为生长期毛囊周围炎症浸润及毛囊退行性变两个部分。目前研究认为,内外源因素作用于遗传易感人群引起生长期毛囊深层周围炎症细胞浸润,浸润的炎症细胞及细胞因子、神经肽等形成恶性循环,循环结局为毛囊上皮细胞凋亡,大批毛囊同时陕速进入退行期,导致斑秃发生。     

某些细胞因子在天疱疮皮损中的表达

目的:检测Th2类细胞因子在天疱疮皮损中的表达.方法: 选用单克隆抗体CD4,CD8,CD25,CD54和CD124等,对40例天疱疮患者的皮损及皮损周围皮肤标本用S-P法进行免疫组化研究,同时以20例正常人皮肤组织为对照.结果: CD4+和CD8+的T细胞见于全部标本真皮浸润的单一核细胞内,且以CD4+的T细胞为主(P<0.01),在表皮CD8+的T细胞在寻常型天疱疮和红斑型天疱疮比较有显著性差异(P<0.01),27例病人显示CD25染色阳性,与皮损周围相比CD54在皮损处表达增多(P<0.05).某些CD124阳性细胞见于表皮及真皮.对照组中CD4+和CD8+T细胞稀少,CD4/CD8比值为1.4,在皮损及皮损周围的皮肤中CD25的表达无显著性差异(P>0.05),而角质形成细胞不表达CD25.CD54的表达见于血管内皮细胞和真皮血管周围的单一核细胞,与皮损周围相比,皮损处CD54+增加(P<0.05).结论:细胞免疫在天疱疮的发病中有重要作用.     

斑秃皮损中CD4、CD8、FasL和穿孔素表达的研究

目的:通过研究T淋巴细胞杀伤靶细胞的途径和机制,探讨T淋巴细胞在斑秃发病中的作用.方法:采用免疫组化ABC法,检测斑秃和正常组织皮损中CD4、CD8、FasL 和穿孔素的表达.结果:在正常组织中CD4、CD8阳性表达,FasL 和穿孔素阴性表达.在斑秃皮损中CD4、CD8、FasL均阳性表达,而穿孔素为阴性表达.结论:T淋巴细胞在斑秃发病中起着重要作用,T 淋巴细胞可能通过Fas-FasL结合途径引起细胞凋亡,细胞凋亡参与了斑秃的发病.     

亲毛囊性蕈样肉芽肿四例分析

报道4例亲毛囊性蕈样肉芽肿。男3例,女1例,年龄32 ~ 52岁。3例以头面部及颈部受累为主,密集多发性暗红色不规则浸润性斑块、结节、肿块、毛囊性丘疹及痤疮样皮损,躯干、四肢表现为斑片、轻度浸润的斑块、毛囊性丘疹及痤疮样皮损。1例头面部未受累,全身密集毛囊性丘疹。组织病理显示真皮内、血管周围及毛囊周围大量淋巴样细胞为主的团块状浸润,部分细胞异形,浸润细胞移入毛囊上皮是他们共同而显著的特征,而亲表皮现象不明显。其中2例毛囊内充满黏蛋白,阿新蓝染色阳性。免疫组化显示浸润细胞主要为CD4阳性T细胞。对常规用于经典蕈样肉芽肿的治疗反应差,病情难以完全缓解,即使部分缓解后也极易复发。     

痤疮皮损的免疫细胞化学研究

本文应用碱性磷酸酶—抗碱性磷酸酶技术,对15例痤疮皮损中的浸润细胞进行免疫细胞化学表现型分型。结果表明:细胞浸润主要位于真皮浅层、毛囊和皮脂腺周围,以单个核细胞为主,T细胞以T_H细胞明显增高,T_H:T_S比值为2.57:1。作者认为细胞免疫参与了寻常痤疮的致病过程。     

局部应用FK506:有效的斑秃免疫疗法?利用Dundee实验秃毛大鼠模型进行研究

斑秃(AA)是一种自身免疫性疾病。Dundee实验秃毛大鼠(DEBR)是理想的AA实验动物模型,其皮损特征与人AA十分相似,于毛发生长初期的毛囊周围有单个核细胞(MNC)的浸润,CD4~+细胞与CD8~+细胞的比为     

Characterization of infiltrating T cells in human scalp explants from alopecia areata to SCID nude mice: possible role of the disappearance of CD8+ T lymphocytes in the process of hair regrowth

T cells may play a role in the pathogenesis of alopecia areata (AA). We attempted to elucidate the linkage between infiltrating T cells and hair regrowth processes by grafting scalp skin from the affected region of patients with AA onto severe combined immune deficiency (SCID) nude mice. When the AA scalp was grafted into the mice, the grafts were accepted, and normal hair regrowth was observed. Before grafting, CD4+ and CD8+ T cells had infiltrated into the peribulb area. After grafting, the telogen hair shifted to anagen hair, and the CD4+ and CD8+ T cell infiltrates in the bulb area decreased in all cases. CD8+ T cells had almost disappeared from all portions of the follicles. It has been suggested that CD8+ T cells play a crucial role in the pathogenesis of AA. The absence of CD8+ T lymphocytes that responded to follicular autoantigens may induce hair regrowth in the grafted skin. In addition, the CD4+ human T cells that had infiltrated or still remained in the upper-middle portions including the bulge area accompanied the HLA-DR expression after grafting. Infiltrating or surviving T cell phenotypes and locations changed during the hair cycle in the grafts. These results indicate that the location of infiltrated T cells and their phenotypes may participate not only in hair loss but also in regrowth of hair in AA.     

毛囊干细胞在盘状红斑狼疮性脱发和斑秃中的变化及意义

目的了解毛囊干细胞在常见瘢痕性脱发和非瘢痕性脱发中的变化及意义。方法采用SP免疫组化法检测毛囊干细胞常用标志分子CK15、CD200在常见瘢痕性脱发和非瘢痕性脱发即盘状红斑狼疮(DLE)和斑秃皮损中的表达。每组8例患者,以同等数量正常人头皮做对照。结果毛囊干细胞标志CK15、CD200在正常对照的阳性率均为100%,而在DLE患者头皮中的阳性率分别为37.5%和12.5%,同正常对照相比均明显减少(P<0.05);斑秃患者毛囊中CK15阳性表达率为100%,CD200阳性表达率为50%,CD200阳性表达率与正常对照相比明显减少(P<0.05)。结论毛囊干细胞在DLE患者中明显受损,可能与其永久性脱发有关;而在斑秃中毛囊干细胞无减少。CD200在毛囊表达减少可能与斑秃及DLE的发病相关。     

Induction of cellular immunity against hair follicle melanocyte causes alopecia

Alopecia areata (AA) is generally regarded as an organ-specific autoimmune disease. Although it has been hypothesized that the autoimmunity is mediated by T cells and that hair follicle melanocyte is one of the targets, definitive evidence is lacking. We here demonstrate that AA-like lesions can be induced in mice by inducing CD8⁺ T-cell-mediated immunity to hair follicle melanocytes. We found that hair loss was induced in mice-bearing interleukin-12-producing B16 melanoma cells by the depletion of CD4⁺ T cells, accompanied by vitiligo-like coat color change. The alopecic lesions varied in size from pachy to extensive. In many instances, hair loss developed and was followed by the regrowth of white hairs. Histological analysis revealed that mononuclear cells infiltrated in and around the bulb region of hair follicles. Furthermore, immunohistochemical examination clearly showed the intra-follicular infiltration of CD8⁺ T cells. Neither the vitiligo-like coat color nor AA-like lesions were induced when CD8⁺ T cells were codepleted. These observations indicate that the induction of CD8⁺ T-cell-mediated immunity against hair follicle melanocytes causes alopecia. It is thought that there are many types of AA with different mechanisms, targets etc. Although hair follicle melanocytes have long been thought to be one of the targets of AA, evidence to support the hypothesis is sparse. Therefore, we believe that our observation is significant to support the hypothesis.     

Strong expression of CD40, CD54 and HLA-DR antigen and lack of evidence for direct cellular cytotoxicity are unique immunohistopathological features in alopecia areata

The pathological role played by T cells infiltrating hair follicles in lesions of alopecia areata (AA) is unknown. We examined the expression in cryostat sections of scalp skin obtained from a total of 28 patients with AA and from five normal control subjects of (1) molecules related to the induction of cell death including Fas, Fas ligand (FasL), perforin, granzyme B (GB), and TIA-1, (2) molecules related to antigen presentation including CD1a, CD40, CD54, CD80, and CD86, and (3) molecules induced by interferon gamma (IFN-gamma) including CD40, CD54, Fas, and HLA-DR. CD3(+) T cells infiltrated perifollicularly, perivascularly and in the hair structure and there was a predominance of CD4(+) over CD8(+) cells. Antigen-presenting cells expressing CD1a, CD40, CD54, or HLA-DR were also seen. Expression of CD40, CD54, HLA-DR and CD95 was also seen in the hair structure including the dermal papilla. Consistent with these observations, IFN-gamma-producing cells were also detected in the perifollicular infiltrate. In contrast, few Fas-L(+), perforin(+), GB(+) or TIA-1(+) cells were found adjacent to the follicles. Apoptotic cells were recognized only in the outer root sheath of catagen hairs. These findings suggest that infiltrating T cells interact with perifollicular or follicular antigen-presenting cells to produce IFN-gamma, which deprives dermal papilla cells of their ability to maintain anagen hair growth.     

IL-17-positive mast cell infiltration in the lesional skin of lichen planopilaris: Possible role of mast cells in inducing inflammation and dermal fibrosis in cicatricial alopecia

Lichen planopilaris (LPP) is a primary cicatricial alopecia characterized by the infiltration of lymphocytes in the upper portion of hair follicles. Inflammation around the bulge region of hair follicles induces destruction of hair follicle stem cells and tissue fibrosis, resulting in permanent hair loss. Treatment is still challenging, and the precise pathophysiology of this disorder is unknown. To clarify the pathogenesis of LPP, we performed histological and immunohistochemical analysis on specimens obtained from LPP patients. Formalin-fixed and paraffin-embedded samples were evaluated by staining with haematoxylin and eosin (HE), toluidine blue stain, immunohistochemistry and immunofluorescence. The immunohistochemical analysis demonstrated that CD4-positive T cells preferentially infiltrated into the follicular infundibulum in the LPP lesions. Toluidine blue stain detected a large number of mast cells in the inflammatory lesions of LPP. Interestingly, immunohistochemical analysis demonstrated that the mast cells harboured IL-17A- and IL-23-producing activity and expressed the IL-23 receptor. The number of IL-17A-positive mast cells was significantly higher in the LPP lesions than in normal scalp. Moreover, the IL-17 receptor was expressed exclusively in the follicular epithelial cells in the LPP lesions. These results suggested that mast cells infiltrating hair follicles might play a role in the pathogenesis of LPP via the IL-23/IL-17 axis.     

Type 1 interferon signature in the scalp lesions of alopecia areata

Background Autoimmune attack of the bulbar region of anagen phase hair follicles by CD8+ T cells and Th1 cytokines has been proposed to result in hair loss in alopecia areata (AA). The initiating stimuli are unknown. As interferon‐α therapy may trigger AA, we propose that type 1 interferons are involved in the induction of disease. Objectives To compare lesional scalp from patients with AA with scalp lesions of cutaneous diseases associated with local type 1 interferon‐related protein expression. Methods Lesional scalp of patients with AA, discoid lupus erythematosus, lichen planopilaris and androgenetic alopecia was examined by immunohistochemistry for expression of the type 1 interferon‐inducible myxovirus protein A (MxA), the chemokine receptor CXCR3, and the cytotoxic proteins granzyme B (GrB) and T‐cell intracytoplasmic antigen 1 (TiA‐1). Results MxA was expressed in the intradermal and subcutaneous compartments of the hair follicle including sebaceous glands in inflammatory AA similar to lesions of cicatricial alopecia (discoid lupus erythematosus, lichen planopilaris) but not in the epidermal compartment of AA, and not at all in noninflammatory AA or androgenetic alopecia. The location of CXCR3‐expressing cells correlated with MxA expression. The inflammatory cells around the hair follicle in AA included a lower number of GrB+ and TiA‐1+ cells compared with cicatricial alopecia and demonstrated predominant TiA‐1+ expression. Conclusions We demonstrate the expression of type 1 interferon‐related proteins in the inflammatory lesions of AA. The distribution pattern of the interferon signature and cytotoxicity‐associated proteins in AA differs from cicatricial alopecia.     

CXCL10 produced from hair follicles induces Th1 and Tc1 cell infiltration in the acute phase of alopecia areata followed by sustained Tc1 accumulation in the chronic phase

BackgroundAlopecia areata (AA) is an organ-specific and cell-mediated autoimmune disease. T lymphocytes densely surround lesional hair bulbs, which is histologically referred to as “swarm of bees”. However, pathomechanisms of “swarm of bees” are still uncertain.ObjectiveWe investigated the pathological mechanisms of “swarm of bees”, focusing on T-cell chemotaxis so that inhibition of chemotaxis may be strong candidate of novel treatments for AA.MethodsWe investigate the expression of chemokine receptors on T cells obtained from peripheral blood mononuclear cells (PBMCs) and skin infiltrating cells in AA patients. In addition, real-time chemotaxis assay was also demonstrated.ResultsIn PBMCs, the frequency of CXCR3⁺CD4⁺ T cells (Th1) was significantly higher in acute-phase AA than in chronic-phase AA or healthy control, while CXCR3⁺CD8⁺ T cells (Tc1) were significantly increased in chronic-phase AA. In the skin lesions of acute-phase AA, CXCR3⁺CD4⁺ and CXCR3⁺CD8⁺ T cells infiltrated in the juxta-follicular area. In chronic-phase AA, CXCR3⁺CD8⁺ T cells dominated the infiltrate around hair bulbs, possibly contributing to the prolonged state of hair loss. Lymphocytes obtained from a lesional skin of acute-phase AA contained CXCR3⁺CD4⁺ and CXCR3⁺CD8⁺ T cells at higher percentages than those of PBMCs, suggesting preferential emigration from the blood. Immunohistochemical and real-time RT-PCR studies demonstrated that hair follicles of acute-phase AA expressed a high level of Th1-associated chemokine CXCL10. By chemotaxis assay, freshly isolated PBMCs from acute-phase AA patients had a strong velocity of chemotaxis toward CXCL10 with increased expression of F-actin.ConclusionsThese results suggest that the increased production of CXCL10 from hair follicles induces preferential infiltrates of highly chemoattracted Th1 and Tc1 cells in the acute phase of AA, and Tc1 infiltration remains prolonged in the chronic phase.     

Immunohistological study of the development of the cellular infiltrate in the pelage follicles of the DEBR model for alopecia areata

The Dundee experimental bald rat (DEBR) undergoes hair loss associated with perifollicular infiltrates of mononuclear cells (MNC), a pathological characteristic of human alopecia areata (AA). To investigate further the pathogenesis of the disease in this animal model, we have studied the development, composition and extent of the perifollicular MNC infiltration in young (6-week-old), prelesional (3-month-old), active lesional, and established lesional DEBR rats, using 6-week- and 6-month-old Wistar rats as normal controls. The proportions of hair follicles showing infiltration by MNC and their main subsets were determined using immunohistochemical staining of serial cryostat sections of flank skin biopsies. There was a good correlation between the degree of leucocyte (OX-1⁺) infiltration of anagen hair follicles and the development of hair loss. In 6-week-old DEBR skin, there were few perifollicular cells expressing MHC class II, with positively stained dendritic cells in the dermis above the sebaceous gland. There was a sparse perifollicular distribution of CD4⁺ cells (W3/25) and macrophages (ED-1⁺). No CD8⁺ cells (OX-8⁺) were seen associated with DEBR hair follicles, and only small numbers were present in Wistar rats. In prelesional DEBR rats there was an increased perifollicular presence of MHC class II⁺ cells, macrophages, and particularly of CD8⁺ cells, with little change in CD4⁺ cells. Active and established lesional rats, i.e. animals with overt loss of hair, showed a significant increase in the degree of MNC infiltration and the proportion of infiltrated follicles, the majority of which were in dystrophic anagen. In the perifollicular infiltrate the CD4⁺:CD8⁺ ratio was approximately 2:1. An intrafollicular infiltrate was prominent, and was composed of CD8⁺ cells and macrophages, with bulbar and suprabulbar keratinocytes expressing MHC class II antigens. CD4⁺ cells were not detected in follicular epithelium. ICAM-1 expression correlated with MNC infiltration. These results show marked similarities to lesional human AA. They also focus on a possible active role for CD8⁺ cells in the pathogenesis of hair loss in the DEBR rat.