程序性死亡分子1及其配体在口腔鳞状细胞癌患者外周血中的表达及临床意义

目的 检测口腔鳞状细胞癌患者外周血中程序性死亡分子1（PD-1）及其配体（PD-L1）的表达水平,探讨其生物学及临床意义。方法 选取82例口腔鳞状细胞癌患者（口腔鳞癌组）和25例健康对照者（对照组）为研究对象,收集研究对象的外周血,采用流式细胞术检测外周血T淋巴细胞表面PD-1、PD-L1的表达及T淋巴细胞亚群计数,采用酶联免疫吸附方法检测血清中可溶性PD-1（sPD-1）和可溶性PD-L1（sPD-L1）的表达水平。采用SPSS 17.0软件对数据进行检验和相关性分析。结果 口腔鳞癌组外周血CD8+ T淋巴细胞百分数明显高于对照组（P<0.05）,而CD3+、CD4+T淋巴细胞百分数及CD4+/CD8+T亚群百分数比值均低于对照组（P<0.05）。口腔鳞癌组外周血CD4+、CD8+ T淋巴细胞表面PD-1、PD-L1的阳性表达率均高于对照组（P<0.01）。口腔鳞癌组血清sPD-L1水平高于对照组（P<0.05）,而sPD-1水平二者差异无统计学意义（P>0.05）。sPD-L1的表达与临床分期、肿瘤分化程度及淋巴结转移状态相关（P<0.05）,与性别、年龄、肿瘤部位及大小无关。结论 口腔鳞状细胞癌患者外周血T淋巴细胞亚群存在不同程度的免疫功能抑制,CD4+和CD8+ T淋巴细胞表面PD-1及PD-L1表达显著升高。异常升高的sPD-L1可能与口腔鳞状细胞癌的发生发展有关。     

复发性阿弗它溃疡患者外周血中CD4+/CD45RA+细胞亚群的表达及意义

目的探索复发性阿弗它溃疡(recurrent aphthous ulcer,RAU)患者T淋巴细胞亚群CD4+/CD45RA+细胞的数量变化.方法应用流式细胞术检测RAU患者及正常对照人群CD4+/CD45RA+亚群细胞的数量.结果 RAU患者的CD4+/CD45RA+细胞数约占CD4+细胞总数的14.24%,正常对照约占总数的21.56%,两者有显著性差异(P＜0.01).结论 RAU患者外周血T淋巴细胞亚群中CD4+/CD45RA+细胞量减少,提示CD4+/CD45RA+细胞可能与RAU的发病有关.     

小鼠正畸加力过程中脾脏T淋巴细胞亚群变化

目的:观测小鼠磨牙加力不同时间点脾脏CD3+、CD8+、CD4+T淋巴细胞及Th细胞亚群的比例变化,分析正畸加力对免疫系统中T淋巴细胞变化的影响.方法:4周龄雄性C57BL/6小鼠随机分为1 d、3 d、5 d和7 d加力组以及相应时间的对照组.加力组向磨牙施加25 g力;对照组安置相同加力装置,力值为0.使用流式细胞...     

沙利度胺对复发性阿弗他溃疡患者外周血淋巴细胞亚群的影响

目的：观察沙利度胺对复发性阿弗他溃疡（ RAU）患者外周血淋巴细胞亚群的影响及治疗疗效。方法：选择轻型RAU患者（轻型RAU组）85例,健康对照组80例,随访观察服用沙利度胺25 mg／d治疗轻型RAU的疗效;同时比较健康对照组,轻型RAU组治疗前后外周血淋巴细胞亚群检测结果。结果：和健康对照组相比,轻型RAU患者CD4＋T淋巴细胞数目减少,CD8＋T淋巴细胞增多,淋巴细胞亚群比例失调;经沙利度胺治疗后,轻型RAU患者淋巴细胞亚群比例异常改善,同时患者溃疡间歇期明显延长,溃疡数目明显减少,差异均具有统计学意义（P＜0．05）。结论：轻型RAU患者存在外周血T淋巴细胞亚群比例异常,沙利度胺可有效调节CD4＋、CD8＋T淋巴细胞亚群比例,延长溃疡间歇期,减少溃疡发作,临床疗效满意。     

牙龈卟啉单胞菌表面相关物质对T淋巴细胞凋亡的影响

目的 研究牙龈卟啉单胞菌表面相关物质(SAM)对人外周血中T淋巴细胞活化及凋亡的作用。方法 选取10名全身及牙周组织健康受试者,分离外周血中T淋巴细胞,在有/无SAM情况下培养0～96h,用荧光探针(AnnexinV-FITC、PI、CD69)进行标记,并进行流式细胞仪检测。结果 T淋巴细胞+SAM组AnnexinV+/PI-细胞百分数在48h、72h、96h分别为17.36±2.53,26.48±2.26,46.79±4.53。T淋巴细胞组AnnexinV+/PI-细胞百分数在48h、72h、96h分别为7.15±2.13,6.78±2.15,11.56±2.48,T淋巴细胞+SAM组与T淋巴细胞组之间存在明显差异(P<0.01)。CD69+T淋巴细胞+SAM组和CD69+T淋巴细胞组AnnexinV+/PI-细胞百分数除24h外的4个时间点上都无明显差异(P>0.05)。CD69+淋巴细胞+SAM组AnnexinV+/PI-细胞百分数在各个时间点上都明显高于T淋巴细胞+SAM组(P<0.01)。结论 SAM能够诱导人外周血中T淋巴细胞活化,并且能够通过活化促进其凋亡。     

髓样来源的抑制细胞在舌癌小鼠中的表达及意义

目的 评价舌癌小鼠外周血和病变部位髓样来源的抑制细胞（MDSC）的变化及意义。方法 建立4NQO舌癌小鼠模型,观察MDSC细胞在外周血的表达,同时观察T细胞亚群的表达,评价MDSC细胞与T细胞亚群及CD4⁺/ CD8⁺比例变化的相关性。观察MDSC细胞在舌黏膜病变部位的表达,并检测舌组织精氨酸酶1（ARG-1）mRNA的表达。结果 4NQO小鼠外周血中MDSC比例随着肿瘤进展显著升高（P<0.01）,外周血中MDSC比例与相应CD3⁺CD8⁺T细胞的比例呈正相关,与CD4⁺/CD8⁺比呈负相关关系。4NQO小鼠异常增生区域的MDSC细胞较正常对照组有所增加,在舌癌组织内及与正常组织的交界处,均浸润了大量的MDSC细胞,而且ARG-1 mRNA水平显著升高（P<0.01）。结论 MDSC细胞在舌癌组织和外周血中的扩增可能是肿瘤进展的重要原因。MDSC细胞可能成为口腔癌免疫治疗的潜在靶点。     

口腔扁平苔藓患者外周血T淋巴细胞亚群的观察

T 淋巴细胞亚群是一个具有多功能的细胞群,各细胞群之间相互协调,维持其平衡是人体免疫功能正常运行的必要条件。本实验应用 OKT 系单克隆抗体改良微量细胞毒实验对36名口腔扁平苔藓患者及30名健康对照者外周血 T 淋巴细胞亚群进行了观察,结果显示:CD_3~+%CD_4~+%患者与健康对照无明显差异,CD_8~+%患病组高于健康对照(P<0.05),CD_4~+/CD_8~+比值低于对照组(P<0.05)。说明口腔扁平苔藓患者存在着 T 细胞亚群平衡紊乱。CD_8~+的细胞毒性 T 淋巴细胞增多对口腔扁平苔藓病变形成具有重要意义,支持 DLP 为异常的细胞介导的免疫反应的发病机理。     

复发性口疮患者T细胞亚群在中医辩证分型和Lehner分类中的表现差异与临床意义

目的　选择 166例复发性口疮患者按中医实证、虚证分型及 Lehner分型讨论它们之间外周血 T细胞亚群的变化差异。方法　 T淋巴细胞亚群采用单克隆抗体 APAAP法检测。结果　实证组、虚证组病人 CD3、CD4、CD4/CD8减少 ,而 CD8的升高只在实证组有显著差异 ( P<0 .0 1)。在 Lehner分类中四型的 CD3、CD4、CD4/CD8均减少 ,而 CD8的升高只在腺周型和白塞氏型才有显著统计学意义 ( P<0 .0 1)。结论　复发性口疮患者无论以中医分型或以 Lehner分型它们之间的 T细胞紊乱均存在差异 ,有临床指导意义。     

曲安奈德和帕夫林联合治疗老年糜烂型OLP疗效观察

目的：探讨局部注射曲安奈德配合口服帕夫林治疗老年人糜烂型OLP的疗效。方法：对34例老年糜烂型OLP患者用药治疗前后检测T淋巴细胞亚群的变化及观察临床疗效指标。结果：患者经治疗后临床有效率91.18%,T淋巴细胞亚群检测CD3＋、CD4＋T细胞升高（P〈0.01）,CD8＋T细胞下降（P〈0.01）,CD4＋/CD8＋治疗后上升（P〈0.01）。结论：口腔糜烂型OLP经治疗后,临床疗效肯定,血中T淋巴细胞亚群状态有明显改善。     

根尖牙乳头干细胞外泌体对舍格伦综合征小鼠治疗效果的实验研究

目的    探索根尖牙乳头干细胞来源的外泌体（exosomes derived from stem cells from apical papilla,SCAP-Exo）对舍格伦综合征（Sjögren syndrome,SS）小鼠的治疗效果。方法    酶消化法提取根尖牙乳头干细胞,超速离心法提取SCAP-Exo,并应用透射电镜、纳米颗粒跟踪技术及Western Blot进行鉴定。选择10周龄雌性健康ICR小鼠6只作为对照组;选择10周龄雌性NOD小鼠12只随机分为NOD组和SCAP-Exo组,每组6只。饲养2、4周后,SCAP-Exo组小鼠尾静脉注射SCAP-Exo,其他组小鼠尾静脉注射PBS。于饲养6周后检测各组小鼠唾液流率。随后收集小鼠外周血,流式细胞术检测外周血中辅助性T细胞17（T helper 17,Th17）/CD4+ T细胞比例,ELISA检测血清中白细胞介素-17A（interleukin-17A,IL-17A）表达水平。最终处死各组小鼠,通过HE染色观察下颌下腺中淋巴细胞浸润水平。结果    透射电镜下可见SCAP-Exo呈杯状的囊泡结构;纳米颗粒跟踪技术分析SCAP-Exo直径为30 ~ 150 nm;外泌体特异性标记蛋白CD9、Alix呈阳性表达,不表达细胞内质网特异性蛋白Calnexin。3组小鼠唾液流率、外周血Th17/CD4+ T细胞比例及IL-17A表达水平总的比较,差异均有统计学意义（F值分别为59.169、293.217、189.583,均P < 0.05）。其中,与对照组相比,NOD组小鼠唾液流率明显降低、外周血Th17/CD4+ T细胞比例及IL-17A表达水平明显升高;而相较于NOD组,SCAP-Exo组小鼠唾液流率明显增加、外周血Th17/CD4+ T细胞比例及IL-17A表达水平明显降低;差异均有统计学意义（均P < 0.05）。相较于NOD组,SCAP-Exo组小鼠下颌下腺淋巴细胞浸润水平显著降低,仅存在轻度散在的淋巴细胞浸润或出现极个别的淋巴细胞浸润灶。结论    SCAP-Exo能有效治疗SS小鼠,可能与其调节Th17细胞转化有关。     

Lymphocyte changes in peripheral blood,spleen, and liver in DMBA-induced squamous cell carcinoma of mouse cheek skin

The peripheral blood, spleen, and liver lymphocyte subsets of mice with experimental cheek skin carcinoma were determined. The carcinoma was induced by the topical application of 2% (w/v) 9,10-dimethyl-1,2-benzanthracene (DMBA) to cheek skin twice a week for 12 weeks, and it was examined macroscopically and histopathologically. The composition of lymphocyte subsets (T cells, B cells, CD4⁺ single-positive [SP] T cells, and CD8⁺SP T cells) in peripheral blood, spleen, and liver was determined by flow cytometry at 3-week intervals for up to 24 weeks. Spleens and livers were assessed by determining their content of natural killer (NK)T cells. The results showed histopathological progression of the skin lesions from papilloma to squamous cell carcinoma at week 12. Body weight was significantly reduced from weeks 15 to 24, and spleen weight was significantly increased at weeks 21 and 24, but liver weight was not significantly different from the control. The lymphocyte subset composition of peripheral blood showed significant elevation of T cells at weeks 6 and 9, followed by reduced levels at weeks 21 and 24, with significant reduction of B cells at weeks 6 and 9, followed by elevation at weeks 21 and 24. CD4⁺SP T-cell content was elevated at weeks 6, 9, and 12, and reduced at weeks 21 and 24. CD8⁺SP T-cell content was significantly reduced at weeks 6, 9, and 12, and elevated at weeks 21 and 24. The composition of the lymphocyte subsets in the spleen was similar to their composition in peripheral blood. The composition of both T and B cells in the liver was significantly different from that in the corresponding control group, but no significant differences were found in either CD4⁺SP or CD8⁺SP T cells. These findings revealed that the DMBA-induced cheek skin carcinoma in mice affected not only the lymphocyte subsets in peripheral blood, but the cells in the spleen and liver as well.     

Differential regulation of IL-2 and IL-4 in patients with tobacco-related oral squamous cell carcinoma

AIM: The aim of the study was to investigate the systemic immunity in terms of major lymphocyte subsets and the expression of IL-2 and IL-4 in T-cell subsets from peripheral blood of patients with tobacco-related intraoral squamous cell carcinoma. METHODS: CD3+, CD4+ and CD8+ T-cell subsets and CD16+ CD56+ natural killer cells, and intracellular cytokines in T-cell subsets were determined by two-colour flow cytometry and confocal microscopy. RESULTS: Oral cancer patients showed a significantly reduced (P < 0.001) CD3+ and CD4+ T-cell subsets with a lower CD4/CD8 ratio when compared with the normal controls. The frequency of CD3+ IL-4+ and CD8+ IL-4+ T cells were significantly higher (P < 0.001) while CD4+ IL-2+ were significantly lower (P < 0.02) in patients when compared with the normal controls. Late stage of the tumour was associated with reduced expression of IL-2 in both CD4+ (P < 0.05) and CD8+ (P < 0.03) subsets. CONCLUSIONS: The tobacco-related intraoral squamous cell carcinoma seems to be associated with multiple systemic immune defects particularly, an impaired CD3+ and CD4+ T cells in the peripheral blood as well as a differential regulation of IL-2 and IL-4 in CD4+ and CD8+ T-cell subsets. The cytokine response in these patients seems to be skewed from protective Th1 to immunosuppressive Th2 type. Thus these patients could be ideal candidate for immunomodulation therapy.     

The effect of dental alloys on mouse lymphocyte subpopulations

The aim of the present study was to determine the effect of nickel‐containing alloys on lymphocyte subsets in an experimental setting. Plates of alloys containing nickel (Ceramalloy, Talladium, Cerillium, Rexillium) or gold (Orion) were implanted subcutaneously into mice. The levels of CD4⁺ and CD8⁺ T‐lymphocyte subpopulations and of Smig⁺ B lymphocytes were determined at various intervals following implantation, using monoclonal antibodies and flow cytometry. No changes were detected in the proportion of the lymphocyte subsets tested. One month after implantation, the mean fluorescence intensity of CD4, CD8 or Smig, in the peripheral blood lymphocytes (PBL) of the nickel alloy‐implanted animals, was significantly higher than that prior to this procedure. Only a mild increase in CD4 and CD8 was noted after implantation of the gold alloy. The observed effects are most likely attributable to the surgical trauma, and do not indicate that nickel‐containing dental alloys influence T cell subsets in this murine model.     

The proportion of suppressor-inducer T-lymphocytes is reduced in recurrent aphthous stomatitis

A flow cytometric analysis of peripheral blood lymphocytes was undertaken in recurrent aphthous stomatitis patients. The project aimed at detecting differences within lymphocyte subsets using type-specific monoclonal antibodies. Peripheral blood samples were taken from RAS patients in both active and remission phases of the disease and from a group of healthy control subjects. There were no statistical differences between the active and remission phases within any of the lymphocyte subsets examined. There was, however, a significant difference between the RAS group and the control group. RAS patients have depressed CD4+ cell numbers and elevated CD8+ cell numbers. The CD4:CD8 ratio is also depressed. A dissection of the CD4+ subset shows raised numbers of CD4+, 4B4+ lymphocytes and depressed numbers of CD4+, 2H4+ lymphocytes. Previous studies have shown disruption of peripheral blood lymphocyte numbers in Beh?et's syndrome. A similar pattern has now been shown in uncomplicated cases of minor RAS.     

The proportion of suppressor-inducer T-lymphocytes is reduced in recurrent aphthous stomatitis

A flow cytometric analysis of peripheral blood lymphocytes was undertaken in recurrent aphthous stomatitis patients. The project aimed at detecting differences within lymphocyte subsets using type-specific monoclonal antibodies. Peripheral blood samples were taken from RAS patients in both active and remission phases of the disease and from a group of healthy control subjects. There were no statistical differences between the active and remission phases within any of the lymphocyte subsets examined. There was, however, a significant difference between the RAS group and the control group. RAS patients have depressed CD4+ cell numbers and elevated CD8+ cell numbers. The CD4: CDS ratio is also depressed. A dissection of the CD4+ subset shows raised numbers of CD4+, 4B4+ lymphocytes and depressed numbers of CD4+, 2H4+ lymphocytes. Previous studies have shown disruption of peripheral blood lymphocyte numbers in Behcets syndrome. A similar pattern has now been shown in uncomplicated cases of minor RAS.     

Lymphocyte numbers and function in relation to periodontitis and smoking

BACKGROUND: T and B lymphocytes play important roles in periodontitis. Smoking is considered a risk factor for periodontitis and may exert its negative effects through leukocytes. Taking smoking into consideration, the aim of this study was to analyze numbers of circulating T (CD3+) cells and their CD4+ and CD8+ subpopulations, B (CD19+) cells, and T-cell proliferative capacity in periodontitis. METHODS: Lymphocyte immunophenotyping for T cells, their CD4+ and CD8+ subsets, and B cells was performed on peripheral blood from 76 periodontitis patients and 36 controls. Proliferative capacity of T cells was determined in whole-blood lymphocyte culture assays after mitogenic stimulation. RESULTS: Total T cells, CD4+ and CD8+ subpopulations, and responsiveness to specific T-cell stimuli did not differ between patients and controls; in addition, B cells were not significantly elevated in periodontitis patients. However, more periodontal breakdown in smoking patients was associated with higher numbers of CD3+ T cells, as well as with CD4+ and CD8+ T-cell subsets, and increased T-cell proliferation. Numbers of B cells were not affected by smoking. CONCLUSIONS: The increased numbers of T-cells and elevated T-cell responsiveness in patients who smoke may be one of several explanations why smoking is a risk factor for periodontitis. The mechanism of how T-cell function contributes to increase the severity of periodontal breakdown in smoking periodontitis patients needs to be investigated further.     

Myeloid-derived suppressor cells contribute to oral cancer progression in 4NQO-treated mice

Oral Diseases (2011) 18 , 67–73 Objective: Abnormal myelopoiesis especially the expansion of myeloid‐derived suppressor cells (MDSCs) is increasingly recognized as an important reason for the escape of tumor from immune surveillance. This study aims to investigate the role of this specific population of cells in oral cancer progression. Materials and Methods: 4‐Nitroquinoline 1‐oxide (4NQO) was used to induce oral cancer in C57BL/6 mice. The tongue mucosa was examined by hematoxylin and eosin staining. The distribution of MDSCs in the spleen and peripheral blood and T cell subsets in the spleen was analyzed by flow cytometry. The expression of MDSCs in the tongue tissues was investigated by immunohistochemical staining, and the expression of arginase‐1 (ARG‐1) and NOS‐2 in the tongue tissues was detected by real‐time PCR. Results: We found that during tumor progression, significantly increased frequency of MDSCs was observed in the spleens and peripheral blood of 4NQO‐treated mice, and the frequency of MDSCs in the spleens was positively correlated with systemic CD3⁺CD8⁺ T cells. Moreover, 4NQO‐treated mice showed significantly higher MDSCs infiltration and ARG‐1 mRNA level in the tumor site. Conclusions: Myeloid‐derived suppressor cells contribute to oral tumor progression and represent a potential target for immunotherapy of oral cancer.     

A comparison of T4:T8 lymphocyte ratio in the periodontal lesion of healthy and HIV-positive patients.

Previous reports describe a characteristic, rapidly progressive, periodontitis that is unique to patients who are seropositive for HIV antibody (Western blot +). The purpose of this study was to compare the T4 and T8 lymphocyte subpopulations in the peripheral blood and periodontal lesions of these HIV patients with those of healthy controls. T-cell subsets in peripheral blood were quantified by flow cytometry. The values from this analysis were used to calculate the peripheral T4:T8 lymphocyte ratio for each patient. Gingival tissue (papilla) was obtained from 8 HIV+ patients and from 6 healthy HIV- control patients during routine gingival surgery. The T-cell subpopulations in the gingival tissue were determined using serial cryostat sections that were labeled with monoclonal antibodies for T4 and T8 cells and developed using an avidin-biotin-peroxidase system. Six sections were taken from each of the 14 tissue specimens (one per patient). The sections were examined at 450 x and the mean number of T4 and T8 cells calculated for each section. These mean values were then used to determine the T4:T8 lymphocyte ratio for each tissue specimen. The peripheral blood analysis revealed a mean serum T4:T8 ratio of (2.07 +/- 0.455) for the controls and (0.58 +/- 0.26) for the HIV patients. The significantly lower T4:T8 ratio in HIV patients is consistent with their diagnosis. Although the results indicated that the mean T4:T8 lymphocyte ratio in the gingiva of controls was highly variable (2.70 +/- 1.344), the gingiva of HIV patients consistently exhibited a complete absence of T-cells.(ABSTRACT TRUNCATED AT 250 WORDS)