牙髓树突状细胞免疫组化标记

目的：利用免疫组化技术，观察人正常牙髓和早期龋坏牙牙髓中表达Ⅱ类分子的树突状细胞。方法：收集因正畸拔除的正常前磨牙和早期龋坏的阻生牙，取出完整冠髓，H－E染色和免疫组织化学SABC技术染色。分别选择抗HLA－DR和CD68的单克隆抗体与实验标本反应，显微镜下观察染色阳性细胞。结果：正常牙髓中可见少量的HLA－DR是性细胞和CD68阳性细胞。龋坏牙牙髓中HLA－DR阳性细胞和CD68阳性细胞明显增加。标记阳性的树突状细胞在形态上呈不同的外形，如圆形、椭圆形、不规则形、纺锤形以及树突状。结论：牙髓组织中存在表达Ⅱ类分子和具有典型特征的树突状细胞。     

免疫化疗对口腔鳞癌浸润T淋巴细胞及其亚群的影响

应用ABC免疫组化技术，检测口腔鳞癌患者接受免疫化疗（S－PVP）和单纯化疗（PVP）治疗前后肿瘤组织内CD3 、CD4 、CD3 和HLA－DR 细胞相对值，比较不同的诱导治疗方案对局部细胞免疫反应的影响．结果显示：S－PVP组治疗前后CD3 、CD4 、CD3 和HLA－DR 细胞分别为65．65、3660、35．93、54．04．和129．26、74．77、38．86、79．64。治疗后CD3 、CD4 细胞较治疗前明显增加（P＜0.05）．PVP组治疗前后CD3 、CD4 、CD3 和HLA－DR 细胞相对值无显著性差异．结果表明，S－PVP免疫化疗可使原发病杜内的T淋巴细胞数量增加，而单纯PVP化疗时局部细胞免疫反应的影响不甚明显。     

口腔扁平苔藓损害组织中树突状细胞亚群数量和比例

目的：研究口腔扁平苔藓（oral lichen planw,OLP）组织中树突状细胞（dendritic cell,DC）的亚群数量和比例,探索其在OLP疾病中的作用。方法：收集正常对照（HC组）,萎缩糜烂型OLP（E－OLP组）,斑纹型OLP（R－OLP组）的颊黏膜,通过流式细胞术染色和免疫组化染色,检测朗格汉斯细胞（CD1a＋CD11cint／lo CD207＋MHC II＋）、髓样DC （CD11c＋MHC II＋）、浆细胞样DC（CD11cint／lo CD123＋MHC II＋）和细胞毒性T细胞（CD3＋CD8＋）在组织中的浸润情况。结果：收集样本23例（HC组5例,E－OLP组7例,R－OLP组11例）,Bartlett检验各组数据方差不齐,采用Kruskal－Wallis非参数检验。朗格汉斯细胞的中位数比例分别为0．092％（HC）、0．564％（E－OLP）和0．541％（R－OLP）,3组间无统计学差异;髓样DC的中位数比例分别为0．311％（HC）、0．996％（E－OLP）和0．448％（R－OLP）,其中E－OLP组与HC组之间有统计学差异（P＜0．05）;浆细胞样DC的中位数比例分别为0．090％（HC）、3．490％（E－OLP）和2．010％（R－OLP）,2组OLP组与HC组之间均有统计学差异（P＜0．05）;CD8＋T细胞的中位数比例分别为0．126％（HC）、4．210％（E－OLP）和1．850％（R－OLP）,2组OLP组与HC组之间均有统计学差异（P＜0．05）。结论：DC在OLP中数量和比例增加,可能与疾病的自身免疫发病机制有关。     

年轻恒牙牙根发育过程中牙髓中Ⅲ型胶原的表达

目的：研究牙根发育不同阶段的人年轻恒牙牙髓中Ⅲ型胶原的表达变化：方法：根据牙根的发育情况，分为牙根刚开始发育、牙根发育中和根尖闭合共3组：采用SP免疫组化法。对各组牙髓标本的石蜡切片进行Ⅲ型胶原的免疫组化染色：结果：图像分析显示、3个组的冠髓中心染色强度之间有非常显著性差异，以牙根刚开始发育时染色最强，根尖闭合时染色最弱：在牙根刚开始发育组、牙髓中心区染色强于牙髓外层。结论：随着年轻恒牙牙根的逐渐发育，Ⅲ型胶原在牙髓中的表达逐渐减弱，因而可能参与牙根的发育。     

不同发育阶段牙髓组织中的基质金属蛋白酶-8的免疫组化研究

目的：探讨MMP-8在牙齿不同发育阶段的表达及作用。方法：将人不同发育阶段的牙髓(33例)按牙根形成情况分为3组，即：牙根开始发育组；牙根发育中组；根尖闭合组。对其石蜡连续切片进行MMP-8的免疫组化染色。结果：MMP-8均在牙髓中的成牙本质细胞胞质、成纤维细胞胞质、血管壁及细胞外基质中呈阳性表达。牙根刚开始发育时染色最强，随着牙根的发育染色强度减弱。结论：成牙本质细胞分泌MMP-8，参与前期牙本质矿化前胶原的降解；成纤维细胞在分泌胶原的同时也降解胶原，有维持牙髓中胶原稳定的作用；MMP-8参与牙齿成熟过程。     

正常及炎症牙髓组织中IL—6的表达

目的：通过检测ＩＬ－６在人牙髓组织中的表达，探讨ＩＬ－６与正常及炎症牙髓组织的关系，方法：采用免疫组化（ＡＢＣ）法，检测正常组８个牙及炎症牙髓组１０个牙中ＩＬ－６的表达。结果：正常人牙髓组织ＩＬ－６染色为阴性，炎症牙髓组织中ＩＬ－６染色 呈阳性表达细胞为单核－巨噬细胞，少量血管内皮细胞和成纤维细胞。结论：ＩＬ－６与牙髓组织的炎症反应密切相关，是重要的炎症介质之一。     

正常及炎症牙髓组织中IL-6的表达

目的：通过检测ＩＬ－６在人牙髓组织中的表达，探讨ＩＬ－６与正常及炎症牙髓组织的关系。方法：采用免疫组化（ＡＢＣ）法，检测正常组８个牙及炎症牙髓组１０个牙中ＩＬ－６的表达。结果：正常人牙髓组织ＩＬ－６染色为阴性，炎症牙髓组织中ＩＬ－６染色均呈阳性，主要表达细胞为单核－巨噬细胞，少量血管内皮细胞和成纤维细胞。结论：ＩＬ－６与牙髓组织的炎症反应密切相关，是重要的炎症介质之一。     

血管内皮生长因子在人年轻恒牙及成熟恒牙牙髓中的表达

目的：研究人年轻恒牙和成熟恒牙牙髓中血管内皮生长因子（vascular endothelial growth factor,VEGF）的表达及分布特征．探讨VEGF在恒牙牙根发育过程中的作用。方法：将因正畸或阻生拔除的健康牙分为2组,第1组年轻恒牙10例。第2组根尖闭合的成熟恒牙15例。对牙髓标本的石蜡切片进行VEGF的免疫组化染色,通过Imagepro-plus 5．1图像分析软件,对图像进行定量分析,采用SPSS13．0软件包进行χ^2检验、t检验、单因素方差分析及SNK-q检验。结果：VEGF在年轻恒牙牙髓成纤维细胞和成牙本质细胞胞质呈强阳性表达,显著强染于成熟恒牙（P〈0．05）;年轻恒牙根髓在喇叭状基顶区成纤维细胞VEGF染色强阳性,以此处向冠方及根方,VEGF表达逐渐减弱。结论：VEGF在人年轻恒牙和成熟恒牙牙髓组织中呈不同特征的表达．VEGF参与恒牙牙根的发育和成熟。     

血管内皮生长因子在人牙髓中的表达

目的：研究血管内皮生长因子（ vascular endothelial growth factor VEGF） 在正常、深龋或炎症牙髓中的表达及其意义。方法：采用免疫组化SABC法，对牙髓中的VEGF的表达进行组织学定位，并通过Image pro-plus 5.1图像分析软件对成牙本质细胞和牙髓成纤维细胞中VEGF染色进行平均光密度值（optical density OD）测定。利用SPSS13．0统计软件对各组数据进行单因素方差分析或秩和检验。结果：人牙髓中，VEGF主要表达在血管内皮细胞、成牙本质样细胞和牙髓成纤维细胞的胞浆中。正常组成牙本质细胞的VEGF表达较其它两组弱（P〈0.01）。与正常组相比，牙髓成纤维细胞中VEGF的表达在深龋组明显增强（P〈0．05），而在炎症组明显减弱（P〈0．05）。此外，VEGF在炎症组的某些炎细胞如中性粒细胞、浆细胞的胞浆中也有表达。结论：VEGF在龋病、牙髓炎中的变化可能与牙髓炎的发生、炎症发展有关，并且可能参与了成牙本质样细胞对牙髓损伤的修复过程。     

正常及炎性牙髓中T、B淋巴细胞的微波免疫组化研究

本研究旨在探讨T、B细胞与牙髓炎发生、发展及转归间的内在联系。采用微波免疫组化技术，对人正常及炎性牙髓中T、B细胞进行分类识别和定量分析、结果表明：正常牙髓中T细胞散在分布于基质中央区，CD细胞居多（CD／CD＝0．48），未检测出B细胞、炎性牙髓中T、B细胞增多，且主要集中在炎症中心。可复性牙髓炎时CD／CD＝0．58，出现B细胞（CD／CD＝11．38），不可复性牙髓炎时CD细胞占优势（CD／CD＝1．67），CD／CD　＝1．51，且有症状组中T、B细胞增加较无症状组多。提示T、B细胞介导的免疫反应在牙髓免疫病理改变中可能起重要作用。     

Immunocompetent cells in the pulp of human deciduous teeth

This immunohistological study sought to determine how the distribution and density of various immunocompetent cells change in the pulp of human deciduous teeth during the process of physiological root resorption. Forty-three extracted deciduous teeth at various stages of resorption were subjected to immunoperoxidase staining with the use of antibodies directed to HLA-DR, CD68, factor XIIIa and lymphocyte subsets. In intact deciduous teeth (group 0), all types of cells examined, except for CD20+ B lymphocytes, were detected. In teeth in which resorption was less than 1/3 of the root length (group 1), all types of cells showed a statistically significant increase compared with group 0 (P<0.05; Mann-Whitney's U-test). HLA-DR+, CD68+, and factor XIIIa+ cells with a dendritic profile kept their distribution in the periphery of the pulp, and oval and round, newly recruited macrophages accumulated in the central portion of the pulp and near the resorption sites. In teeth where resorption was 1/2 to 2/3 (group 2), all the cell types increased further. Aggregations of HLA-DR+, CD68+, and factor XIIIa+ cells were frequently seen in the central portion of the pulp, and T and B lymphocytes occasionally formed some clusters. Comparisons with group 1 revealed that the density of these cells, except for CD20+ cells, showed significant increases (P<0.05; Mann-Whitney's U-test). These results provided evidence showing that immunocompetent cells of deciduous tooth pulp increase with the progress of physiological root resorption, suggesting that immunocompetency of deciduous teeth is altered by this process.     

An immunocytochemical study of the inflammatory cell infiltrate and epithelial expression of HLA-DR in odontogenic cysts

The presence and distribution of lymphocytes, Langerhans cells, HLA-DR+ infiltrating cells and the expression of HLA-DR by lining epithelium was investigated in the walls of odontogenic cysts using an indirect immunoperoxidase method on acetone-fixed cryostat sections. The 23 cysts studied consisted of 14 dental (radicular) cysts, 5 keratocysts, 2 dentigerous cysts, 1 surgical ciliated cyst and 1 incisive canal cyst. The cell populations detected in the walls of all cysts were similar and consisted of HLA-DR+ macrophage-type cells and a mixture of T and B lymphocytes. Analysis of the T cell subsets revealed that in all cases the CD4+, Th/i subset predominated over the CD8+, Ts/c subset. 18/22 cyst linings contained cells expressing a Langerhans cell phenotype (CD1+). Cytoplasmic epithelial expression of HLA-DR was detected in 7/22 specimens. Neither the presence of HLA-DR+ epithelial cells nor LC were restricted to a given type of cyst. These findings indicate the occurrence of similar cellular processes irrespective of the proposed developmental or inflammatory aetiology of these lesions.     

舍格林综合征唇腺组织中CD45的表达及意义

目的：研究舍格林综合征(Sjogren’s Syndrome，简称SS)患者唇腺组织中过度浸润的淋巴细胞上的白细胞共同分化抗原(Leukocyte common antigen-45，CD45)的表达，以探讨淋巴细胞在其免疫反应中的作用。方法：通过HE染色观察淋巴细胞的浸润特点，采用免疫组织化学法检测CD45R、CD45RO在淋巴细胞上的表达。结果：唇腺组织过度浸润的淋巴细胞中CD45RO阳性表达的T细胞占65．74％，CD45R阳性表达的B细胞占27．38％．结论：在SS病变中是以细胞免疫为主、体液免疫为辅的免疫反应。     

Immunocompetent cells in the normal dental pulp

The existence and location of various immunocompetent cells in the human dental pulp were investigated. Pulp tissue for analysis was obtained both from clinically intact pre-molars and from third molars without restorations or caries. Frozen and acetone-fixed pulp tissue sections were subjected to indirect immunohistochemistry with monoclonal antibodies to the following cell types: all peripheral T cells, helper/inducer T cells, cytotoxic/suppressor T cells, macrophages, B cells, and Class II antigen-expressing cells. Dendritic cells expressing Class II antigens (HLA-DR, -DQ), indicating a capacity for presentation of antigen to T helper cells, were seen in the odontoblastic layer as well as in the central portions of the pulp tissue. T lymphocytes, divided into helper/inducer and cytotoxic/suppressor cells, were observed in all pulp specimens. B cells were not seen in any of the pulp samples examined. The data demonstrate that the human dental pulp is equipped with immunocompetent cells essential for the initiation of immunological responses.     

Relative proportions of mononuclear cell types in periodontal lesions analyzed by immunohistochemistry

In this study, we investigated the relative proportions of infiltrating mononuclear inflammatory cells in sections of granulation tissue from periodontitis lesions in both adult periodontitis (AP) and early onset periodontitis (EOP) patients. We utilised a set of cluster of differentiation (CD) antigen-specific monoclonal antibodies to detect different cell types within the tissues. These included anti-CD 20 (B cells), anti-CD 3 (pan T cells) and anti-CD 45RO (memory T cells), anti-CD 4 (helper T cells) anti-CD 8 (suppressor T cells) and anti-CD 68 (monocyte/macrophage). Biopsies of granulation tissue were obtained from 9 patients with adult periodontitis (AP), from 10 patients with early onset periodontitis (EOP) and for comparative purposes, biopsies of gingival tissue from 4 patients with AP. A significantly greater number of T cells (p < 0.05) were observed in EOP and gingival sections than in AP sections. In addition, a greater number of B cells were observed in the granulation tissues than in the gingiva (p < 0.05). The relative numbers of B cells (CD 20). T cells (CD 3) and macrophages (CD 68) were expressed as a percentage of their combined total for each of the patient groups and indicated that the proportion of B lymphocytes was greater in AP sections than in EOP or gingival sections (p < 0.02). The proportion of T cells was lower in the AP periodontitis sections than in the EOP periodontitis sections (p < 0.05). There were no significant differences in the proportion of macrophages between the 3 categories of tissue specimens. The relative ratios of B cells (CD 20) to T cells (CD 3) and B cells (CD 20) to memory T cells (CD 45RO) and macrophages (CD 68) to T cells (CD 3) and memory T cells (CD 45RO) were analyzed and indicated that there was a significant increase in the B to T cell ratio in AP sections compared to EOP and gingival sections (p < 0.02). There was also a significant increase in the macrophage to T cell ratio in AP sections as indicated by CD 68 to CD 3 ratios (p < 0.05). There were no differences regarding the relative proportions of memory T cells or in the ratios of CD 4+ to CD 8+ T cells in the different disease categories. In conclusion, these differences in the relative proportions of B cells, T cells and macrophages may reflect a difference in the immunopathology of AP and EOP.     

Immunohistochemical study of oral lichen planus associated with hepatitis C virus infection,oral lichenoid contact sensitivity reaction and idiopathic oral lichen planus

OBJECTIVES: Oral lichen planus (OLP) is a common mucocutaneous disorder and might be associated to a possible pathogenic relationship with hepatitis C virus (HCV) infection or hypersensitivity to dental alloy. We examined the clinical and immunohistochemical features of OLP associated with HCV infection (OLP-HCV), oral lichenoid contact sensitivity reaction (OLCSR), and idiopathic oral lichen planus (iOLP). The immunohistochemical expressions of CD4, CD8, B cells, Class II major histocompatibility complex antigen (HLA-DR), S-100, HSP60, Proliferating cell nuclear antigen (PCNA) and Ki-67 were compared to study the pathogenic differences of the three OLP groups. MATERIALS AND METHODS: Three groups of OLP patients, (I) OLP-HCV patients (n = 17), (2) OLCSR patients (n = 10) and (3) iOLP patients (n = 14) were retrieved from clinical records and tissues examined immunohistochemically by the avidin-biotin-complex technique. RESULTS: The patients with OLP-HCV showed widespread lesions. The proportion of CD8+ cells was found to be significantly higher in the lamina propria of the OLP-HCV patients and a significantly lower proportion of CD8+ cells of the OLCSR patients was noticed in the epithelium or the connective tissue papillae than in the iOLP patients. There were no significant differences in either the number of CD4+ cells or B cells between the three OLP groups. No significant differences in the number of HLA-DR+ cells were found between the three OLP groups and some OLP-HCV patients showed a significant increase of S-100+ cells in the epithelium compared with iOLP patients. There were no significant differences in either the number of PCNA+ or Ki-67+ cells between the groups. The patients showed similar weak expressions of HSP60 in the three OLP groups. CONCLUSION: The different distributions of the CD8+ cells that could have functionally different roles might be related to the distinct pathogenic mechanisms in the three OLP groups.     

小鼠牙髓干细胞同种异体移植后体内免疫反应初探

目的:初步探究小鼠牙髓干细胞同种异体移植后的全身免疫反应。方法:牙髓干细胞分离获得自C57BL/6 小鼠牙髓组织,培养至第3 代,与纳米羟基磷灰石(n-HA)复合培养形成复合物,扫描电镜观察其生长黏附情况。40只4周龄的C57BL/6 小鼠随机分成两组。实验组将复合物移植入上背部皮下,对照组将纳米羟基磷灰石植入上背部皮下,术后均使用外科方法严密缝合创口。两组动物术后均观察4周、6周处死,分离脾脏,制成脾淋巴细胞悬液,流式细胞仪测定CD4⁺、CD8⁺的值。结果:牙髓干细胞在体外与纳米羟基磷灰石结合良好;免疫组化染色实验组4周、6周均可见牙本质涎蛋白(dentin sialoprotein,DSP)阳性表达;实验组与对照组相比,术后4周、6周CD4⁺/CD8⁺值均无明显差异。结论:牙髓干细胞同种异体移植后,尚不能够引起机体的免疫排斥反应,其在体内仍具有低免疫原性,有望成为组织工程同种异体种子细胞来源。     

An immunocytochemical study of the inflammatory cell infiltrate and epithelial expression of HLA-DR in odontogenic cysts

The presence and distribution of lymphocytes, Langerhans cells, HLA-DR' infiltrating cells and the expression of HLA-DR by lining epithelium was investigated in the walls of odonlogenic cysts using an indirect immunoperoxidase method on acetone-fixed cryostat sections. The 23 cysts studied consisted of 14 dental (radicular) cysts, 5 keratocysts, 2 dentigerous cysts, 1 surgical ciliated cyst and 1 incisive canal cyst. The cell populations detected in the walls of all cysts were similar and consisted of HLA-DR+ macrophage-type cells and a mixture of T and B lymphocytes. Analysis of the T cell subsets revealed that in all cases the CD4⁺, T_h/i subset predominated over the CD8⁺, T_s/c subset. 18/22 cyst linings contained cells expressing a Langerhans cell phenotype (CD1⁺). Cytoplasmic epithelial expression of HLA-DR was detected in 7/22 specimens. Neither the presence of HLA-DR⁺ epithelial cells nor LC were restricted to a given type of cyst. These findings indicate the occurrence of similar cellular processes irrespective of the proposed developmental or inflammatory aetiology of these lesions.     

An immunohistochemical study of the distribution of immunocompetent cells, especially macrophages and Ia antigen-expressing cells of heterogeneous populations, in normal rat molar pulp.

The precise distribution of various immunocompetent cells in rat molar pulp was immunohistochemically examined by use of seven anti-rat monoclonal antibodies. It was demonstrated that rat molar pulp contained many OX6 (anti-Ia antigen)-positive cells and a large number of ED1 (anti-monocytes, macrophages, and dendritic cells)-positive, ED2 (anti-tissue macrophages)-positive, and/or OX35 (anti-macrophages and CD4+ lymphocytes)-positive cells. Macrophage-like cells predominated in the central portion of the pulp, while cells of dendritic appearance usually existed in the periphery of the pulp. Double-immunoperoxidase staining revealed that these cells showed some heterogeneity, but the majority could be classified as ED1+/OX6-/ED2+ cells, which may be Ia-histiocytes. Findings also suggested that true dendritic cells may be included in the ED1+/OX6+/ED2- category of cells. A small number of T lymphocytes and plasma cells were also detected. These results suggest that the normal dental pulp contains a variety of immunocompetent cells, with macrophages as the most dominating. Following the exogenous invasion of pathogenic stimuli in the pulp, these cells may participate in the defense reaction by acting as phagocytes or antigen-presenting cells, which are essential for the initiation of immune responses.     

恒牙牙根发育过程中牙髓中碱性成纤维细胞生长因子的表达

目的　研究人的年轻恒牙和成熟恒牙牙髓中碱性成纤维细胞生长因子(bFGF)的表达及分布特征,探讨 bFGF在恒牙牙根发育过程中的作用。方法　对牙髓标本的石蜡切片进行bFGF的免疫组化染色,并做图像定量分析。结果　bFGF在牙根刚开始发育、发育过程中及发育完成后的牙髓中的阳性表达呈逐渐减弱。结论　bFGF在人正常发育的牙髓组织中,随着牙根的逐渐发育而呈现不同特征的表达,因而参与牙髓的发育和成熟。