口腔肿瘤患者外周血单核细胞CD_(69)的表达及意义  

The Expression of CD_(69) on PBMC from the Patients with Oral Tumours

作　　者：胡雁[1] 章小缓[1] 李梅[2] 曾翔舒[2] 许曼波[3] 艾伟健[4] 

机构地区：[1]中山大学光华口腔医学院牙周黏膜科,广东广州510600 [2]广东省口腔医院医务科 [3]广东省口腔医院实验室 [4]广东省口腔医院颌面外科

出　　处：《口腔医学研究》2005年第6期649-651,共3页Journal of Oral Science Research

摘　　要：目的:测定外周血单核细胞CD69分子的表达,探索口腔肿瘤患者的T细胞的活化功能。方法:实验对象10位健康人作为对照组;8位口腔良性肿瘤患者为实验组Ⅰ;10位口腔鳞状细胞癌的患者为实验组Ⅱ。每位实验对象在实验的当天,抽取10 mL的静脉血。用F icoll-Paque剃度离心法分离PBMC。将PBMC悬浮液分别与TCM,Con A和PHA培养4 h和20 h,分别收获细胞,用双色和单色单克隆荧光抗体CD4-FITC/CD8-PE和CD69-PE标记,CD69的表达用流式细胞仪和相应的分析软件SYSTEMTM II SOFTWARE分析。结果:在PBMC经Con A刺激4 h后,口腔鳞状细胞癌患者的CD69表达的增长率(44.5%)明显低于对照组(67.8%)和实验组Ⅰ(70.8%),P<0.05。同样,经PHA刺激后4 h,口腔鳞状细胞癌患者的CD69表达的增长率也低与其他两组(P<0.05),而且,PB-MC的CD69表达率也低与其他两组(P<0.05)。结论:本研究提示这种免疫反应力的下降可能是由于淋巴细胞的活化或增殖功能低下的原因。由PHA诱导的PBMC的CD69表达可能成为口腔癌患者免疫监视系统的指标之一。Objective： Previous studies showed that the decreased counts of CD19 and NK cells could play a role in the pathogenesis of oral cancers. T ceils are also proved to be a critical mediator in immunity to tumours in animal models. The purpose of this study is to demonstrate the activation of T cells from patients with oral cancers by investigating the expression of activation inducer molecule （ AIM ） CD69 driven by Con A （ concanavalin A ） and PHA （ phytohemagglutinin - M ） （Pharmacia Biotech, Sweden） on peripheral blood mononuclear cells （PBMC）. Methods： 10ml venous blood was collected from each object at the same time on each day of experimentation. PBMC were isolated by density gradient centrifugation with Ficoll - Paque Plus （ Pharmacia Biotech, Sweden）. After washing twice with RPMI - 1640, the cells were adjusted to 3 × 10^6 cells/ml. 100μl PBMC were then added into the wells of 96 - well plates with or without 100μl Con A or PHA at 20μg/ml. All plates were incubated at 37℃for4, and 20 hours in a humidified atmosphere of 5% CO： in air. The expression of CD69 was quantified by flow cytometry with SYSTEMTMII SOFTWARE （Bechman -Coulter） on Coulter Epics XL after staining of cells with monoclonal antibodies of dual - and single - color fluorescence CD4/CD8 and CD69 （ Immunotech, France）. Results： The results showed that the expression of CD69 on PBMC from patients with oral cancers induced by Con A and PHA was significantly depressed after 4 -hour activated phase （P 〈 0.05 ）. Conclusion： This finding suggested a suppressed response of T cells to PHA in patients with oral cancers, and therefore indicated that the depressed activation function of T cells might play a role in the immunopathogenesis of oral cancer.

关 键 词：CD69 T细胞 口腔癌 流失细胞术 

分 类 号：R739.8[医药卫生—肿瘤]