Peripheral T-cell subsets in patients with reticular and atrophic-erosive oral lichen planus

OBJECTIVE: Oral lichen planus (OLP) presents with various clinical forms that can be classified into 2 major types: reticular OLP and atrophic-erosive OLP. Our objective was to investigate immunologic differences between these 2 types. STUDY DESIGN: We investigated possible immunologic differences between 26 patients with reticular OLP and 26 patients with atrophic-erosive OLP. RESULTS: No differences were detected in serum Ig levels or complement levels. However, the mean proportions of CD4+CD45RO+ and DR+ lymphocytes were significantly higher in patients with atrophic-erosive OLP than in patients with reticular OLP, whereas the mean proportion of CD8+CD45RA+ lymphocytes was significantly lower in patients with atrophic-erosive OLP. CONCLUSION: These findings suggest that the 2 clinical types of OLP might have different immunopathogenic mechanisms.     

口腔扁平苔藓细胞凋亡及凋亡蛋白Bcl-2、Bax表达的变化

目的观察口腔扁平苔藓（oral lichen planus, OLP）病损区口腔黏膜超微结构的变化,以及Bcl-2 、Bax和凋亡细胞的表达水平,探讨细胞凋亡在OLP发生、发展中的作用。方法选取35例OLP患者和10例健康志愿者（对照组）。采用免疫组织化学法检测0LP患者和正常对照组口腔黏膜组织中Bcl-2 、Bax的表达,应用TUNEL法及流式细胞术检测上皮细胞的凋亡情况,应用透射电镜技术观察口腔黏膜超微结构的变化。采用SPSS17.0软件包对数据进行统计学处理。结果免疫组织化学检测显示,Bcl-2在OLP中的阳性表达率与正常口腔黏膜相比无显著差异（P>0.05）,但在淋巴细胞浸润带处表达增强。Bax在OLP中的阳性表达率和表达强度与正常口腔黏膜相比显著增高（P<0.01）。OLP病变组织的上皮细胞凋亡指数显著高于对照组（P<0.01）,与流式细胞术测定结果一致（OLP组上皮细胞凋亡百分率显著高于对照组）。透射电镜观察发现,OLP组口腔黏膜凋亡细胞增多,染色质边聚,并见凋亡小体。结论凋亡细胞表达上调以及病损区超微结构改变,病损区淋巴细胞浸润带处Bcl-2 与上皮基底层角质形成细胞中Bax的表达增强,证实细胞凋亡异常与OLP 的发生、发展有密切关系。     

转录因子RORγτ和FOXP3在口腔扁平苔藓病损组织中的表达及意义

目的:口腔扁平苔藓(oral lichen planus,OLP)是一种发生于口腔黏膜的T淋巴细胞介导的自身免疫性疾病。辅助性T细胞(helper T lymphocytes, Th)在OLP的发病过程中具有重要作用,本研究旨在进一步探索OLP患者局部病损组织中辅助性T细胞亚群Th17细胞和Treg细胞的作用。方法:纳入43例OLP患者和13例健康志愿者。采用实时定量PCR法检测局部病损组织中Th17和Treg细胞的特征性转录因子RORγτ和FOXP3的表达,采用GraphPad Prism 5 软件对其表达差异进行统计学分析。结果:OLP局部病损组织中转录因子RORγτ和FOXP3的表达显著高于正常黏膜组织,而且均与OLP的临床分型密切相关。萎缩糜烂型OLP组病损组织中RORγτ/FOXP3比值显著高于网状型OLP组和健康对照组,而网状型OLP组的RORγτ/FOXP3比值虽然高于对照组,但差异无显著性。结论:Th17细胞和Treg细胞均参与OLP的局部免疫反应;同时,Th17/Treg失衡也参与了重症型OLP的致病过程,且表现为Th17细胞优势。     

The Role of Factors Associated With Apoptosis in Assessing Periodontal Disease Status

Background: Little is known about the release of apoptotic proteins during periodontal breakdown. This pilot study investigates the presence of factors associated with apoptosis in serum, saliva, and gingival crevicular fluid (GCF) and their association with periodontal disease severity and activity. Methods: GCF, whole saliva, and serum were obtained from 47 adult patients with chronic periodontitis (CP) and 10 healthy controls. Clinical measurements, including probing depth (PD), clinical attachment level (CAL), and radiographs, were used to classify patients into healthy, mild, and moderate/severe CP groups. Enzyme‐linked immunosorbent assays were used to measure apoptosis or DNA fragmentation in GCF and active caspase‐3, soluble Fas (sFas), and sFas ligand (sFasL) in saliva and serum. Western immunoblotting was used to detect Fas, FasL, sFasL, and caspase‐3 expression in GCF. Results: DNA fragmentation was positively correlated with PD and CAL regardless of patient disease status (P <0.001). sFas and sFasL were present in saliva and serum, but there were no differences between groups. In GCF, the greater odds of detecting Fas, sFasL, and caspase‐3 increased with increasing PD and CAL (P <0.05). In addition, sites with inflammation and PD ≥5 mm had significantly greater odds of exhibiting Fas, sFasL, and caspase‐3 expression compared with sites without inflammation and PD <5 mm (P <0.05). Caspase‐3 was not detected in saliva or serum. At the patient level, only FasL and disease status were significantly correlated (P <0.05). Conclusion: Factors associated with apoptosis were detected in GCF in patients with CP.     

Serum cytokine levels in patients with oral mucous membrane disorders

Serum cytokine levels were examined in 18 oral squamous cell carcinoma (SCC), 26 lichen planus (OLP), 20 recurrent aphthous stomatitis (RAS), 8 herpetic gingivostomatitis (HGS), 16 pseudomembrane candidiasis (PMC) and 19 acute bacterial infection (ABI) cases. All SCC and most PMC patients possessed clear serum IL-3. No clear increase of IL-4 was observed in most cases though over 20 pg/ml were found in a few OLP, RAS and ABI. ABI exhibited the highest IL-6, and the cytokine level was lower in RAS, PMC, HGS and OLP in this order. Suppressed IL-6 activity was elevated with improvement of HGS lesion. TNF-alpha increased in 9 OLP, but the levels were below 100 pg/ml in all cases. Most SCC possessed higher GM-CSF activity than the controls. Increase of the cytokine corresponding with improvement of the oral lesion was seen in HGS, but not in OLP. From these results, each serum cytokine seems to reflect a characteristic pathophysiology of individual oral disorder.     

口腔扁平苔藓患者血清IL-2、IL-4、IL-6水平与临床症状的相关性

目的 探讨不同类型口腔扁平苔藓(OLP)患者血清IL-2、IL-4、IL-6在OLP发病及治疗中的意义。方法 采用ELISA方法检测60例各型OLP患者血清IL-2、IL-4、IL-6的水平，记录患者的疼痛程度、充血和糜烂面积。结果 OLP患者血清IL-2、IL-4水平较对照组无显著差异，而各型OLP患者IL-6显著升高。糜烂型OLP患者IL-4水平显著高于对照。血清IL-2、IL-4水平与疼痛程度、充血面积和糜烂面积之间无相关性，而IL-6与三者呈正相关。结论 本研究支持自身免疫机制是OLP的发病机制之一的观点；在OLP的不同阶段患者的免疫状态是变化的；IL-6在OLP的发病中发挥了一定作用。血清IL-6水平也许可以作为动态监测OLP疾病进展的指标和疗效观察的指标。     

口腔扁平苔藓固有层淋巴细胞中Fas及Fas配体的表达变化

目的 检测Fas及Fas配体(Fas ligand,FasL)在口腔扁平苔藓(oral lichen planus,OLP)固有层淋巴细胞中的蛋白表达,探讨Fas、FasL和活化诱导的细胞死亡(activation-induced cell death,AICD)与OLP发病的关系.方法 采用脱氧核糖核苷酸末端转移酶介导的原位缺口末端标记法检测31例OLP和10例正常口腔黏膜(normal oral mucosa,NOM)中淋巴细胞凋亡情况;分别使用免疫组化法检测组织总淋巴细胞及CD8~+、CD4~+T细胞Fas、FasL的表达.结果 OLP与NOM固有层淋巴细胞凋亡率[分别为(1.9±1.8)%、(11.5±9.0)%]差异有统计学意义(P=0.013).OLP淋巴细胞Fas、FasL表达与NOM组相比明显增强[阳性表达率分别为52%(16/31)、71%(22/31),P值分别为0.005、0.000].OLP中CD8~+与Fas~+细胞双阳性表达率为10%,与NOM组相比无明显升高(P=0.313),而CD8~+与FasL~+细胞双阳性表达率[58%(3/31)]显著升高(P=0.002).CD4~+与Fas~+细胞双阳性表达率为35%(11/31),较NOM组显著升高(P=0.031),其中网纹型的表达明显升高(阳性表达为8/19,P=0.019),但糜烂、萎缩型的表达无显著升高(阳性表达为3/12,P=0.097).CD4~+与FasL~+表达率为16%(5/31),与NOM组相比无明显增强(P=0.182).结论 OLP淋巴细胞凋亡低下;OLP中T细胞亚群Fas、FasL表达不均衡,CD8~+细胞和萎缩、糜烂型OLP中CD4~+细胞可能逃逸AICD,与炎症的持续和进展有关;网纹型OLP中部分CD4~+T细胞可能经历AICD.     

Osteopontin expression in oral lichen planus

To explore circulation levels of osteopontin (OPN), tumour necrosis factor (TNF)-α and transforming growth factor (TGF)-β1 from patients with oral lichen planus (OLP) for clinical application. A group of 26 subjects with OLP were compared with 26 sex- and age-matched control (NC) subjects. Local lesion tissue was examined for OPN by immunohistochemical analysis. And, serum OPN, proinflammatory TNF-α and TGF-β1 levels were measured by enzyme-linked immunoabsorbent assay. The serum concentrations of OPN and TNF-α were significantly higher in OLP patients than the NC group ( P  < 0.05). Although serum concentrations of TGF-β1 increased slightly, they were not statistically significant. Erosive-form OLP exhibited significantly elevated TGF-β1 serum levels, compared with reticular-form OLP. The above results suggest that the production of OPN is associated with the inflammatory process of OLP development, and may serve as a potential disease marker of OLP.     

Ruolo delle epatopatie nella fase acuta del lichen planus orale

ObjectivesOral lichen planus (OLP) is considered an extrahepatic manifestation of chronic liver disease (CLD). To survey the incidence of OLP clinical exacerbations (OLPCE) in atrophic-erosive lesions, we assessed functional status of their liver in OLP patients by serological assays.Material and methodsWe selected 96 patients with CLD-related OLP-CLD (mean age 62.28 years ± 7.42, range 48-78 years; female:male ratio 2.3:1) from a cohort of 476 OLP patients. As intervention group, forty-eight out of 96 patients OLP patients had chronic HCV infection. As controls, forty-eight of 96 OLP-CLD patients had a chronic drug-induced hepatitis. The other 48 had a chronic drug-induced hepatitis (control group). Clinical signs were scored according to the criteria reported by Thongprasom et al. Serum samples from each patient were analyzed to determine the following: serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), lactate dehydrogenase (LDH), gamma-glutamyl transpeptidase (γ-GT), pseudo-cholinesterase (PCHE), albumin/globulin ratio (AGR), alkaline phosphatase (AP), and blood glucose (BG) levels. Statistical analysis was based on the unpaired Student t-test. Differences were considered significant when p was < 0.05 (two-tailed).ResultsSGOT, SGPT, γ-GT, PCHE and BG were resulted significantly altered in 33/48 and 9/48 patients in the intervention group as well as control group, respectively. In both groups, the clinical score of OLP lesions was higher in patients with alterations of blood tests, mainly liver enzymes. The incidence of OLPCE was also significantly higher among OLP-CLD patients with alterations of the laboratory data. The atrophic-erosive form was the one most frequently observed.ConclusionsThe status of HCV infections and of toxic hepatitis alone does not seem to be the cause of OLPCE. Altered liver function parameters may trigger OLPCE with both types of CLD. Interdiscilinary cooperation between oral pathologists and hepatologists is vital whenever elevated transaminases levels are observed in OLP patients, to improve the clinical course of disease.     

OLP患者唾液和血清中骨桥蛋白等细胞因子的表达

目的：研究口腔扁平苔藓（OLP）患者唾液和血清中骨桥蛋白（OPN）等细胞因子的含量及其相互关系。方法：经病理学检查确诊的OLP患者26例,其中充血糜烂型12型,网纹/斑块型14例。酶联免疫吸附试验（ELISA）检测患者唾液和血清中OPN,肿瘤坏死因子-α（TNF-α）,转化生长因子-β1（TGF-β1）的含量。以26名健康志愿者作为正常对照。结果：OLP患者唾液和血清OPN,TNF-α含量与正常对照组相比均有升高（P〈0.05）;充血糜烂型OLP的TGF-β1唾液和血清水平显著高于网纹/斑块型OLP（P〈0.05）。三种抗体在唾液和血清中表现出明显的相关关系。结论：OPN在OLP患者唾液和血清中含量升高,唾液和血清中OPN含量具有相关性。唾液作为无创伤性检测标本有望有限地代替血清标本应用于OLP与口腔白斑的临床检测。     

Serum immunoglobulins and autoantibodies in patients with oral lichen planus

A qualitative and quantitative determination of serum immunoglobulins was made in 34 patients (27 female, 7 male) with histologically verified oral lichen planus (OLP). The subjects ranged in age from 33 to 80 years and 28 of the cases had OLP changes of atrophic-erosive type. The possible presence of rheumatoid factors (RF), antinuclear antibodies (ANA) or antibodies to mitochondrias, renal glomeruli and smooth muscles was studied in 30 of the patients. The investigation also comprised a control group of 23 patients (17 female, 6 male) without mucosal changes. Immunoelectrophoresis showed an abnormal pattern in 10 OLP cases (29%), the most common change being a polyclonal increase of IgG in 8 persons. Elevated levels of serum IgG (greater than 15 g/l) were seen in a total of 9 OLP patients (26%), but only in 1 of the control cases (4%). The mean IgG value was significantly higher in the OLP than in the control group, 13.4 and 10.3 g/l, respectively. Regarding levels of IgA and IgM, no difference was noted between the groups. Autoantibodies were registered in 27% of the OLP patients and in 9% of the controls. The immune changes could be correlated in some respect to positive fungal cultures, allergies and decreased unstimulated saliva, but not to age, endocrine disorders or joint diseases.     

Periodontal status in patients with oral lichen planus: a study of 90 cases

OBJECTIVE: This study was carried out to relate periodontal status to the oral lesions in patients with oral lichen planus (OLP). MATERIALS AND METHODS: Periodontal status was evaluated in 90 patients with OLP and in 52 controls, in terms of the plaque index (PII), simplified calculus index (CIS) and periodontal disease index (PDI). RESULTS: No significant differences were observed between the two groups as regards the different periodontal indices. The plaque and calculus indices were higher in the more extensive forms of OLP (P = 0.02 and P = 0.012, respectively), and in the presence of gingival involvement (P = 0.004 and P = 0.04). A significant association was also observed between the presence of atrophic-erosive lesions and increased periodontal deterioration (P = 0.037). CONCLUSIONS: Increased plaque and calculus deposits are associated to a significantly higher incidence of atrophic-erosive gingival lesions in patients with OLP.     

High proliferative activity and chromosomal instability in oral lichen planus

The study aimed to assess the proliferative activity and karyotype in Oral Lichen Planus (OLP) lesions. G-banding chromosomal analysis of short-term primary cultures, and immunohistochemical expression of Ki67 and p53 were applied in 30 consecutive OLP patients divided into two groups according to clinical presentation of the lesions, and in nine subjects as negative controls. Mean values of Ki67 and p53 expression differed significantly (P<.01) between controls and patients groups with reticular or atrophic-erosive forms of OLP, whereas there was no significant difference between the two groups of patients with reticular or atrophic-erosive lesions. Six OLP patients showed clonal chromosome alterations, four of them associated with p53 overexpression. In conclusion, OLP is characterized by a high cellular turnover in most patients irrespective of clinical disease presentation. The genetic instability found in some patients should be interpreted as a consequence of the enhanced epithelial turnover, although we cannot rule out the possibility that some of the cytogenetic non-random anomalies observed represent early steps in cancer development.     

PD-L1mRNA与PD-L2mRNA在口腔扁平苔藓中的表达特点及可能作用

目的:研究PD-L1mRNA、PD-L2mRNA在口腔扁平苔藓(oral lichen planus,OLP)患者外周血与损害组织,以及在不同临床类型与病理分型中的表达特点。方法:采用实时定量PCR方法检测60例OLP患者(斑纹型35例,充血糜烂型25例;不伴异常增生38例,伴轻度异常增生22例)PD-L1mRNA、PD-L2mRNA表达水平,并以10名正常人外周血与黏膜组织作为对照,比较不同临床类型和病理类型OLP患者间PD-L1mRNA、PD-L2mRNA表达水平差异,并分析PD-L1mRNA、PD-L2mRNA在OLP患者外周血与损害组织中表达的相关性。利用SAS6.12软件包中的非配对两样本均数比较的Wilcoxon秩和检验,比较不同组别间的差异。结果:PD-L2mRNA在OLP患者外周血中表达降低,损害组织中表达升高,且在不同临床类型OLP患者之间存在显著差异(P<0.05)。PD-L1mRNA在OLP组与正常对照组之间的表达差异无统计学意义(P>0.05)。结论:PD-L2分子可能在OLP患者全身与损害局部对OLP的发生、发展产生不同的影响。     

P53 and bcl-2 immunoexpression in patients with oral lichen planus and oral squamous cell carcinoma

Objective: The aim of this study was to determine by immunohistochemistry the presence and significance of p53 and bcl-2 proteins in oral lichen planus (OLP) and oral squamous cell carcinoma (OSCC). Study Design: We used 21 cases diagnosed as OLP 16 diagnosed as OSCC and four normal gingival biopsies taken from healthy patients were used as controls. Slides were processed for immunohistochemistry using anti-p53 and anti-bcl-2 monoclonal antibodies. Results: We found p53 immunoexpression in 71.4% OLP cases and 68.7% OSCC cases, with no immunoexpression in control cases. Bcl-2 was negative for all OLP and OSCC cases, and mild positivity was observed in normal tissue. We found significant correlation among p53 expression and OSCC malignancy. Conclusions: Our results suggest that TP53 system mainly promotes a hyperproliferative state by cell cycle arrest of the OLP epithelial cells for repairing damaged DNA nor apoptosis and that anti-apoptotic action of bcl-2 is not important in this disease. Key words:Oral lichen planus, oral squamous cell carcinoma, p53, Bcl-2, carcinogenesis, malignant transformation.     

Cell cycle regulating mechanisms in oral lichen planus: molecular bases in epithelium predisposed to malignant transformation

OBJECTIVE: Expression of p53, p21, ki-67, Bcl-2 and caspase-3 proteins in oral lichen planus (OLP) was studied to investigate cell cycle regulation mechanisms in this disease. MATERIAL AND METHODS: Oral biopsies were obtained from 51 patients with OLP and 26 controls for immunohistochemical analysis (peroxidase antiperoxidase) to quantify expression of the proteins under study (-: 0%, +: <10%, ++: 10-25%, +++: 26-50%, ++++: >50% positive cells). RESULTS: Basal expression of caspase-3 was negative in 22 cases (46.8%) and positive in <10% of basal cells in 22 cases (46.8%); caspase-3 expression in inflammatory infiltrate was negative in 22 cases (46.8%) and positive in <10% of lymphocytes in 20 cases (42.5%). Basal expression of Bcl-2 was negative in 35 cases (74.5%); Bcl-2 was expressed in inflammatory infiltrate in 34 cases (72.3%) and was positive in <25% of lymphocytes in 14 of these (29.7%). Basal expression of p53 and p21 was positive in 32 (67.9%) and 23 (48.8%) cases, respectively. Basal expression of ki-67 was positive in 45 cases (95.7%), of which 20 (42.5%) showed positivity in >25% of cells; ki-67 was expressed in inflammatory infiltrate in 23 cases (48.9%). Significant associations were found between basal expressions of p53 and ki-67 (p<0.001) and between Bcl-2 expression in infiltrate and basal expression of ki-67 (p<0.001). No association was observed between basal expressions of p53 and caspase-3 (p=0.08). Bcl-2 expression in infiltrate and basal expression of ki-67 were independently associated with presence of OLP. CONCLUSIONS: Epithelial cells in OLP do not preferentially develop apoptosis but rather cycle arrest or an increased proliferation rate, which may create a suitable substrate for malignant transformation.     

凋亡蛋白Bcl-2、Bax在白斑、口腔扁平苔藓中的表达

目的:观察凋亡蛋白Bcl-2、Bax在白斑、口腔扁平苔藓上皮细胞中的表达,探讨其在口腔白斑、口腔扁平苔藓癌变过程中的作用机制。方法:采用免疫组化法检测10例正常口腔黏膜上皮、18例口腔扁平苔藓、23例白斑、22例口腔鳞癌上皮组织中凋亡相关蛋白Bcl-2、Bax的表达水平。结果:Bcl-2在白斑、口腔扁平苔藓上皮细胞层无异常表达,但在口腔扁平苔藓淋巴细胞浸润带过度表达。Bcl-2在鳞癌组织中呈高表达,与正常黏膜相比有显著性差异(P<0.05)。Bax在上皮单纯增生、轻度、中度不典型增生和低分化鳞癌及糜烂型口腔扁平苔藓组织中呈过度表达,与正常黏膜相比有显著性差异(P<0.05)。结论:Bax参与了口腔白斑癌变的早期事件,而Bcl-2在不典型增生转化为鳞癌的阶段并未发生作用。口腔扁平苔藓的发病机制可能与Bcl-2抑制淋巴细胞凋亡,使细胞免疫亢进,从而刺激上皮细胞Bax过度表达,诱导角朊细胞凋亡有关。     

P53 and bcl-2 immunoexpression in patients with oral lichen planus and oral squamous cell carcinoma

Objective: The aim of this study was to determine by immunohistochemistry the presence and significance of p53 and bcl-2 proteins in oral lichen planus (OLP) and oral squamous cell carcinoma (OSCC). Study Design: We used 21 cases diagnosed as OLP 16 diagnosed as OSCC and four normal gingival biopsies taken from healthy patients were used as controls. Slides were processed for immunohistochemistry using anti-p53 and anti-bcl-2 monoclonal antibodies. Results: We found p53 immunoexpression in 71.4% OLP cases and 68.7% OSCC cases, with no immunoexpression in control cases. Bcl-2 was negative for all OLP and OSCC cases, and mild positivity was observed in normal tissue. We found significant correlation among p53 expression and OSCC malignancy. Conclusions: Our results suggest that TP53 system mainly promotes a hyperproliferative state by cell cycle arrest of the OLP epithelial cells for repairing damaged DNA nor apoptosis and that anti-apoptotic action of bcl-2 is not important in this disease.     

Systemic and topical corticosteroid treatment of oral lichen planus: a comparative study with long-term follow-up

BACKGROUND: Topical corticosteroids are the mainstay treatment for oral lichen planus (OLP), but some authors suggest that systemic corticosteroid therapy is the only way to control acute presentation of OLP. METHODS: Forty-nine patients with histologically proven atrophic-erosive OLP were divided into two groups matched for age and sex. The test group (26 patients) was treated systemically with prednisone (50 mg/day), and afterwards with clobetasol ointment in an adhesive medium plus antimicotics, whereas the control group (23 patients) was only treated topically with clobetasol plus antimycotics. RESULTS: Complete remission of signs was obtained in 68.2% of the test group and 69.6% of the control group, respectively (P = 0.94). Similar results were obtained for symptoms. Follow-up showed no significant differences between the two groups. One-third of the patients of the test group versus none in the control group experienced systemic side-effects (P = 0.003). CONCLUSIONS: The most suitable corticosteroid therapy in the management of OLP is the topical therapy, which is easier and more cost-effective than the systemic therapy followed by topical therapy.     

细胞凋亡相关蛋白Bcl-2对OLP病损区淋巴细胞聚集的影响

目的:探讨凋亡相关蛋白Bcl-2在口腔扁平苔藓(OLP)病变中的生物学行为及对OLP皮损中浸润淋巴细胞聚集、分布的影响,进一步了解OLP的发病机制。方法:采用免疫组织化学检测法,与正常口腔黏膜对照,观察分析25例OLP皮损中Bcl-2蛋白的表达水平及淋巴细胞聚集、分布。结果:OLP病损区T细胞中Bcl-2过度表达,固有层淋巴细胞异常克隆、聚集,浸润增加。结论:OLP病损区中T细胞高密度聚集可能是Bcl-2诱导的局部淋巴细胞过度增殖后关联反应。