Th1 cytokines in oral lichen planus

BACKGROUND: Cell-mediated immune responses in oral lichen planus (OLP) may be regulated by cytokines and their receptors. METHODS: In situ cytokine expression and in vitro cytokine secretion in OLP were determined by immunohistochemistry and ELISA. RESULTS: The majority of subepithelial and intraepithelial mononuclear cells in OLP were CD8+. In some cases, intraepithelial CD8+ cells were adjacent to degenerating keratinocytes. CD4+ cells were observed mainly in the deep lamina propria with occasional CD4+ cells close to basal keratinocytes. Mononuclear cells expressed IFN-gamma in the superficial lamina propria and TNF-alpha adjacent to basal keratinocytes. Basal keratinocytes expressed TNF-alpha as a continuous band. TNF R1 was expressed by mononuclear cells and basal and suprabasal keratinocytes. There was variable expression of TGF-beta1 in the subepithelial infiltrate while all intraepithelial mononuclear cells were TGF-beta1-. Keratinocytes in OLP stained weakly for TGF-beta1. Unstimulated OLP lesional T cells secreted IFN-gamma in vitro. TNF-alpha stimulation down-regulated IFN-gamma secretion and up-regulated TNF-alpha secretion. IL-4, IL-10 and TGF-beta1 secretion were not detected. CONCLUSIONS: These data suggest the development of a T helper 1 immune response that may promote CD8+ cytotoxic T-cell activity in OLP.     

慢性牙周炎病人基础治疗前后Th17型细胞因子及RORC的表达变化

目的:研究慢性牙周炎(CP)病人牙周基础治疗前后龈沟液(GCF)中的Th17型细胞因子及其外周血特异性转录因子RORC的表达变化规律。方法:选择CP病人30例,使用ELISA检测牙周基础治疗前后GCF中Th17型细胞因子(IL-17、IL-21)的水平变化,并利用荧光定量PCR检测牙周基础治疗前后外周血CD4+T细胞中特异性转录因子RORC的表达水平。结果:牙周基础治疗后龈沟液中的Th17型细胞因子IL-17、IL-21水平以及外周血CD4+T细胞中特异性转录因子RORC表达水平均较治疗前显著下降,差异均有统计学意义(P<0.05)。结论:Th17型细胞因子在慢性牙周炎发生发展中发挥重要的致炎作用。     

Oral lichen planus: immunohistology of mucosal lesions.

BACKGROUND: Current evidence suggests that immunological mechanisms are involved in oral lichen planus (OLP) pathogenesis. The events implicate activated epithelia that comprise antigen-presenting Langerhans cells, immunocompetent keratinocytes and subepithelial inflammatory infiltrate. Also, the presence of a high density of leucocyte cells may occur for the expression of a variety of adhesion molecules. The aim of this study was to analyse the immunoexpression of some adhesion molecules as well as lymphocytic markers in order to determine the disease pathogenesis in a Venezuelan population. METHODS: The 18 OLP and 10 normal oral mucosa biopsies were immunostained for CD4, CD8, CD1a, LFA-1, VCAM-1 and ICAM-1. RESULTS: The results showed an increased number of CD4+, CD8+, CD1a+ cells in OLP. Serial sections showed CD4+ and CD8+ cells also expressed LFA-1. The expression of ICAM-1 and VCAM-1 were significantly higher in OLP. CONCLUSIONS: The immunological reaction begins with Langerhans cells activation, which presents an antigen to CD4+ lymphocytes. Those cells through ICAM-1 and LFA-1 promote epithelial destruction. Afterwards, cytokine production, ICAM-1 and VCAM-1 expression can activate CD8+ lymphocytes leading to the chronic form of the disease.     

慢性牙周炎与口腔扁平苔藓相关性研究进展

慢性牙周炎是发生在牙周支持组织的慢性感染性疾病,口腔扁平苔藓是口腔黏膜最常见的慢性炎症性疾病之一。糜烂型口腔扁平苔藓更常伴随慢性牙周炎的发生,牙周局部刺激因素也可加重其症状。流行病学上二者均高发于中老年人群,且在发病机制上慢性牙周炎与口腔扁平苔藓都与免疫、遗传和微生物感染等因素密切相关。本文将对慢性牙周炎与口腔扁平苔藓的相关性作一综述。     

IL-17在OLP病损中的表达及其作用机制的研究

目的：进一步探索IL-17在OLP局部病变中的表达及其作用机制.方法：16位OLP患者和5位健康志愿者被募集参与本试验.采用免疫荧光染色法检测IL-17 （interleukin-17）在OLP病损组织中的表达.进一步采用免疫组织化学染色法检测IL-17受体（interleukin-17 receptor,IL-17R）在OLP病损组织中的表达情况.结果：IL-17＋细胞散在分布于OLP病损组织黏膜固有层中.OLP局部组织中均存在IL-17R的表达,IL-17R主要表达于上皮基底角质形成细胞;同时在上皮下淋巴细胞浸润层中也可见相当数量的炎症细胞表达IL-17R.结论：在OLP病损局部IL-17可与表达于上皮基底角质形成细胞和上皮下淋巴细胞浸润层中部分炎症细胞表面的特异性受体IL-17R结合,进而参与OLP的局部炎症反应.     

Oral lichen planus and chronic junctional stomatitis: differences in lymphocyte subpopulations

Objective. Oral lichen planus (OLP) is an oral counterpart or oral manifestation of the common skin disease lichen planus. Chronic junctional stomatitis (CJS) is a relatively unknown condition characterized by a stromal lymphocyte infiltrate, which is also a diagnostic feature of OLP. The differential diagnosis of OLP and CJS is unclear and they have been suggested to represent variants of the same disease. Material and methods. To investigate possible differences in lymphocyte (sub)populations between these two conditions, we immunostained 10 OLP and 10 CJS specimens for CD1-a, and the lymphocyte markers, CD3, CD4, CD5, CD8, and CD20. We scored the staining results by a four-step grading system and used the Fisher exact test to analyze them statistically. Results. The proportional amount of (CD20 positive) B lymphocytes was higher in CJS than in OLP and the predominance of CD4 positive T lymphocytes over CD8 positive T lymphocytes was stronger in OLP than in CJS. The differences were statistically significant. Conclusion. The results reflect differences in the lymphatic infiltrate between OLP and CJS. Their significance needs further investigation.     

Th17相关细胞因子在大鼠实验性根尖周炎表达的研究

目的辅助性T细胞17（Th17）是-种CD4阳性T细胞新亚群,本研究探讨Th17相关细胞因子与根尖周炎的炎症反应及牙槽骨破坏的关系。方法选取5周龄雄性Wistar大鼠,实验组用球钻将下颌第1磨牙开髓,根管锉清理牙髓,髓腔暴露于口腔中,14d后断颈处死。以未开髓的上颌第1磨牙作为对照组。取上下颌骨,采用Micro—CT观察根尖病损的形成情况;实验组和对照组标本均为6例,标本固定、脱钙,冰冻切片、组织学方法观察根尖病损的特点,采用抗CD68和抗TCRαβ的单克隆抗体进行免疫组化;用酶组织化学染色（TRAP染色）进行破骨细胞染色;实验组和对照组根尖组织提取总RNA,RT-PCR及荧光实时定量PCR方法定量检测根尖组织中Th17相关细胞因子IL-17、IL-23、IL-β、IL-6、TGF—β,以及破骨细胞分化因子RANKL的mRNA表达情况。结果Micro．CT显示,14d实验组有明显的根尖病损形成;组织学结果表明,实验组均存在根尖脓肿,有大量中性粒细胞、CD68阳性的巨噬细胞和散在的TCRctl3阳性T细胞浸润;病损骨面有较多的TRAP阳性破骨细胞,表明有活跃的骨吸收;RT—PCR及实时定量PCR结果显示,实验组根尖组织中Th17相关细胞因子IL-17、IL-23、IL1β、IL-6,及破骨细胞分化因子RANKL的mRNA均显著高于对照组,而TGF-β无显著性变化。结论Th17相关细胞因子在实验性根尖周炎中均有高表达,提示它们可能参与炎症过程和骨破坏。     

Overexpression and varied clinical significance of Th9 versus Th17 cells in distinct subtypes of oral lichen planus

ObjectiveOral lichen planus (OLP) presents with large numbers of T lymphocytes accumulating beneath the epithelium of the oral mucosa; however, its aetiology remains obscure. A potential role for an emerging novel T cell subset, Th9, in OLP has recently been suggested but remains to be clarified. The current aim was to investigate the expression and potential clinical significance of Th9 cells in distinct subtypes of OLP.Materials and methodsPeripheral blood samples were collected from 41 OLP patients and 18 healthy controls (HCs). Flow cytometric analysis was used to detect the CD4+ T helper subset Th9 (IL-9⁺IL-17⁻CD4⁺ Th cells) and Th17 (IL-9⁻IL-17⁺CD4⁺ Th cells) expression levels.ResultsFlow cytometry results showed significantly elevated levels of Th9 cells in reticular and erosive OLP compared to HCs. Th9 expression in erosive OLP was less than in reticular OLP, indicating that Th9 but not Th17 cells may play a predominant role in reticular disease. However, in erosive OLP patients, we found much higher levels of Th17 cells compared to reticular OLP patients and HCs, indicating that Th17 dominates in erosive OLP. Statistical analysis showed positive correlations of Th9 cells and Th17 cells in patients with reticular or erosive OLP but none in HCs.ConclusionsTh9 and Th17 cells may take the predominant roles in reticular and erosive OLP respectively, and their numbers were positively correlated in reticular and erosive OLP patients. Elevated circulating Th9 cells may help maintain immune balance in OLP immunopathogenesis, which requires further investigation.     

IL-35在牙周炎及口腔扁平苔藓中的表达及相关性分析

目的:探讨IL-35在牙周炎及口腔扁平苔藓免疫机制中的作用及相关影响,以及牙周炎与口腔扁平苔藓之间的相互关系.方法:按纳入标准选择单纯口腔扁平苔藓患者20例,单纯牙周炎患者20例,牙周炎伴口腔扁平苔藓患者20例和健康者12例;记录患者的一般信息及牙周探诊深度、临床附着丧失、牙龈指数、龈沟出血指数,并收集静脉血清及龈沟液样本,采用酶联免疫吸附测定血清及龈沟液中IL-35的表达水平.采用SPSS19.0软件包对数据进行统计学分析.结果:单纯口腔扁平苔藓组、单纯牙周炎组、牙周炎伴口腔扁平苔藓组龈沟液及血清中IL-35浓度显著高于健康对照组(P＜0.01),牙周炎伴口腔扁平苔藓组龈沟液及血清中IL-35浓度显著高于单纯口腔扁平苔藓组及牙周炎组(P＜0.05);各组牙周临床指标与龈沟液及血清中IL-35表达水平呈正相关关系.结论:牙周炎及口腔扁平苔藓中IL-35表达水平显著升高,其升高程度与两者呈正相关.     

口腔扁平苔藓患者Th1/Th2的表达及临床意义的研究

目的 观察Th1型和Th2型细胞因子在口腔扁平苔藓(OLP)外周血中的表达。方法 密度梯度离心法分离20例充血糜烂型,16例光滑型OLP病例和19例正常对照组人群的外周血单个核细胞,逆转录-聚合酶链反应法检测各组外周血单个核细胞中IFN-gamma mRNA和IL-4 mRNA的表达;ELISA法检测各组血清中IFN-gamma和IL-4的表达。结果 充血糜烂型和光滑型OLP患者中IL-4 mRNA和蛋白的表达均高于正常对照组,差异有统计学意义(P<0.01);而充血糜烂型和光滑型OLP患者中IFN-gamma mRNA和蛋白的表达均低于正常对照组,差异有统计学意义(P<0.01)。但表达结果在充血糜烂型及光滑型OLP患者组间的差异无统计学意义(P>0.05)。结论 Th1/Th2失衡表达与OLP发病机制密切相关,为临床治疗OLP提供新的思路。     

Changes in the expression of stem cell markers in oral lichen planus and hyperkeratotic lesions

Despite the pivotal role of stem cells in homeostasis of oral epithelia the location of this cell population within the tissue is uncertain. How disease influences these cells in vivo also remains to be elucidated. In this study we have used six molecular markers to identify stem cells in normal and diseased buccal mucosa. Samples of normal oral mucosa (NOM), hyperkeratosis (OHK) and oral lichen planus (OLP) were immunostained for alpha6 and beta1 integrins, keratin 15 (K15), melanoma-associated chondroitin sulphate proteoglycan (MCSP), NG2 the rat homologue of human MCSP and notch 1. K15, NG2 and beta1 staining was continuous in the basal layer of NOM whilst alpha6 and MCSP were limited to basal cells at the tips of connective tissue papillae. K15 was downregulated in OLP whereas alpha6, beta1 and MCSP were upregulated in both OLP and OHK. NG2 remained unchanged and notch 1 was absent in all samples. Therefore, the stem cell phenotype in OLP and OHK maybe altered in response to pathological signaling. Classification of these changes is essential to understand the role of adult stem cells in the pathogenesis of oral diseases characterised by abnormal keratinocyte proliferation and differentiation.     

口腔扁平苔藓与牙周炎关系的研究进展

牙周炎为发生在牙周组织的慢性感染性疾病,同时口腔扁平苔藓也是口腔粘膜的常见疾病。临床上大多数口腔扁平苔藓患者牙周状态差,并且牙周基础治疗对其有一定疗效;在发病机制上二者都与免疫、遗传及心理精神因素等有关。牙周炎与口腔扁平苔藓这两种慢性炎症疾病可同时存在或伴随患者终身,维护好牙周健康已成为口腔医学必须面对的现实。本文对近年来牙周炎与口腔扁平苔藓的关系进行回顾总结,为牙周病和口腔扁平苔藓的进一步研究奠定基础。     

口腔扁平苔藓患者Th1／Th2失衡表达的研究

目的：观察口腔扁平苔藓（oral liclaen planus,OLP）患者外周血中Th1、Th2型细胞转录因子T-het和GATA-3以及Thl、Th2型细胞因子的表达,进一步探索OLP的发病机制。方法：通过密度梯度离心法分离20例充血糜烂型,16例光滑型OLP病例和19例正常对照组人群的外周血单个核细胞,逆转录-聚合酶链反应（RT—PCR）法检测各组外周血单个核细胞中T—betmRNA和GATA-3 mRNA的表达;ELISA法检测各组血清中干扰素-γ（interferon gamma,IFN-1）和白细胞介素-4（interleukin-4,IL-4）的表达。结果：充血糜烂型和光滑型OLP患者中T—betmRNA、IFN-γ的表达均低于正常对照组,差异有统计学意义（P〈0．01）;而充血糜烂型和光滑型OEP患者中GATA-3 mRNA、IL-4的表达均高于正常对照组,差异有统计学意义（P〈0．01）。T—bet mRNA和GATA-3 mRNA的表达在充血糜烂型及光滑型OLP患者组间的差异有统计学意义（P〈0．01）;而IFN-γ和IL-4的表达在充血糜烂型及光滑型OLP患者组间的差异无统计学意义（P〉0．05）。结论：Th1／Th2失衡表达与OLP发病机制密切相关,为临床治疗OLP提供参考。     

Perforin and granzyme B involvement in oral lesions of lichen planus and chronic GVHD

J Oral Pathol Med (2010) 39 : 741–746 Background: Oral lesions of lichen planus and chronic graft‐vs.‐host disease (cGVHD) have similar clinical and histological features, but distinct etiology. Apoptosis induced by cytotoxic T lymphocyte has been proposed as a mechanism of keratinocytes death. Cytotoxicity can be mediated by granules containing granzyme B and perforin. Since common features can reflect similarities in immunological mechanisms, we studied the role of those molecules in both diseases. Methods: We analyzed 29 cases of oral lichen planus and 27 of oral cGVHD. The sections were studied on H&E, perforin and granzyme B staining. Results: The total means (epithelium plus connective tissue number) of the granzyme B‐ and perforin‐positive cells were significantly higher in cGVHD than in oral lichen planus lesions (P < 0.05). Also, it was found that the higher the number of perforin+ cells, the higher the number of granzyme‐B+ cells in the epithelium and in the connective tissue for both groups (P < 0.05). In oral lichen planus, the number of single apoptotic bodies had a positive correlation with connective tissue granzyme immunostaining and a negative correlation with perforin (P < 0.01). On the contrary, in oral cGVHD, the number of apoptotic body clusters presented a positive correlation with connective tissue perforin (P < 0.01). Conclusions: Our findings indicate that apoptosis in oral lichen planus seems to be correlated with granzyme B release, while in oral cGVHD, perforin seems to be more important. Although these diseases present clinical and histological similarities, subtle differences seem to exist in their pathogenetic mechanisms.