鼻咽癌病人外周血树突状细胞的表型及功能检测  

DETECTION OF EPITOPE AND FUNCTION OF DENDRITIC CELL IN PERIPHERAL BLOOD FROM PATIENTS WITH NASOPHARYGEAL CARCINOMA

作　　者：黄光武[1] 温文胜[1] 张哲[1] 谢莹[1] 吴英鹰[1] 唐安洲[1] 

机构地区：[1]广西医科大学第一附属医院耳鼻咽喉科,南宁530021

出　　处：《广西医科大学学报》2006年第6期881-884,共4页Journal of Guangxi Medical University

基　　金：国家自然科学基金项目(No.30260115)

摘　　要：目的:观察鼻咽癌病人树突状细胞(DC)经体外分离诱导分化后表型变化和呈递功能的情况。方法:在体外分离并诱导成熟HLA-A2基因型鼻咽癌病人自身的DC,运用流式细胞仪检测DC的表型变化,然后分别负载EB病毒抗原LMP-2的HLA-A2限制型两个表位肽段CLGGLLTMV(CLG)和LTAGFIFL(LTA),诱导自身的CD8+T细胞。培养两周后,运用酶联免疫斑点法(ELISPOT)检测经DC诱导后CD8+T细胞的免疫应答情况。结果:分离鼻咽癌患者外周血DC,培养7d后用流式细胞仪检测DC的各项表型:CD80、CD86、CD83、HLA-DR等,呈现分化成熟的表型特征。上述各种表型的细胞百分率分别为(33·92±21·42)%、(90·20±12·10)%、(32·89±23·47)%、(90·37±12·82)%,并且与正常人的DC的表达无明显差异。培养成熟的DC呈递肽段CLG及LTA给自身CD8+T细胞,T细胞识别肽段并能应答的细胞数增加,呈递前分别为(2·67±1·97)2×104CD8+T细胞和(5·33±1·86)2×104CD8+T细胞,呈递后为(42·67±33·79)2×104CD8+T细胞和(25·67±18·25)2×104CD8+T细胞,呈递前、后比较差异有统计学意义(P<0·05)。同时与正常T细胞的应答能力无明显差别。结论:鼻咽癌病人外周血DC可经外周血分离培养,诱导扩增后能分化成熟并能有效呈递肿瘤抗原肽段,产生特异性CTL,对鼻咽癌病人的免疫治疗具有潜在的应用价值。Objective：To study the change of the epitope and function of dendritic cell in peripheral blood from Nasopharyngeal carcinoma patients. Methods： DCs were isolated from HLA-A2 peripheral blood of NPC patients and cultured with cytokine. Membrane marks of DCs were detected by flow cytometry after 7-day culture. Then DCs were stimulated by peptides CLGGLLTMV （CLG） and LTAGFIFL（LTA）that are the HLA-A2 restricted epitope peptides from LMP-2 protein. After 2-week culture, autologous CD8^＋ T was pulsed with the DCs, and the frequency of CD8^＋ T cells that secret IFN-garmma in response to the DCs were detected in Elispot assay. Result： The percentages of CD80^＋ DCs, CD86^＋ DCs, CD83^＋ DCs and HLA-DR^＋ DCs were （33.92±21.42）% ,（90. 20±12.10）% ,（32. 89±23.47）% and（90.37±12.82）% respectively, and there was no difference compared with the normal control group. The number of T cells that can recognize and respond to the peptides CLG and LTA are （2.67± 1.97）per 2 × 10^4 CD8^＋ T cells and （5.33±1.86） per 2× 10^4 CD8^＋ T cells before their presence, and after their presence are （42.67±33.79） per 2× 10^4 CD8^＋ T cells and （25.67±18. 25） per 2 × 10^4 CD8^＋ T cells. There are differences compared with pre-presence and pro-presence （ P 〈0.05）. But there are not any difference compared with the normal control group. The result shows the DCs have the ability to present the peptides CLG and LTA to CD8^＋T cells. Conclusion：The DCs in NPC patients can be separated and cultured from peripheral blood. After cultured in mature, DCs loaded with the epitope peptides of LMP-2 from EBV can induce the CD8^＋ T cell to cytotoxic T lymphocyte （CTL）, and this result is very useful for immunological therapy for NPC.

关 键 词：树突状细胞 鼻咽癌 细胞表型 

分 类 号：R739.63[医药卫生—肿瘤]