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鼻咽癌患者放疗前后外周血EBV-DNA和细胞免疫水平的研究

Research of EB Virus Related Markers and Cell Immune Level in Peripheral Blood from Pre- radiotherapy and Post-radiotherapy Patients with Nasopharyngeal Carcinoma

  • 摘要: 目的比较鼻咽癌(NPC)患者放疗前后抗EBV特异性CTL细胞免疫功能、EBV-DNA、T细胞亚群及CD4+CD25+Tr淋巴细胞水平,为临床选择免疫治疗方案及预测预后提供理论依据及客观指标。方法收集标本105份(NPC患者放疗前35份、NPC患者放疗6月后40份和健康人30份),ELISPOT检测外周血LMP2特异性CTL;荧光定量PCR检测EBV-DNA;流式细胞术检测T细胞亚群及CD4+CD25+Tr淋巴细胞,统计分析各指标变化的特点及相互关系。结果抗EBV-LMP2特异性细胞免疫反应最大频率见于健康对照组,NPC放疗前患者中则仅少数呈阳性反应,NPC放疗后组反应率介于NPC放疗前组和健康对照组之间;NPC放疗前组血清EBV-DNA 的阳性率均高于其他两组,对EBV-LMP2特异性细胞免疫反应的阳性率与血清中EBV-DNA病毒载量的阳性率呈负相关;放疗前NPC患者外周血CD3+、CD4+T细胞百分率及CD4+/CD8+比值降低, CD8+T细胞百分率升高,CD4+CD25+Tr细胞水平升高,放疗后情况有所缓解。结论EBV-LMP2特异性CTL反应在NPC放疗前患者最低,EBV-LMP2特异性CTL反应阳性率与血清中EBV-DNA载量相关,增强CTL应答可改善EBV相关肿瘤的治疗效果,EBV-DNA载量水平一定程度上可反映NPC患者的肿瘤负荷。NPC患者外周血CD3+、CD4+T细胞降低,而CD8+T细胞升高,CD4+CD25+Tr细胞比例升高,监测患者T细胞亚群有助于对NPC患者的治疗及预后的判断。

     

    Abstract: ObjectiveThe levels of EBV-LMP2 special CTLs, EBV-DNA and T cell subsets in peripheral blood of NPC patients are analyzed to summarize the change patterns and characteristics. Methods Serum samples were obtained from 35 NPC patients waiting for radiotherapy, 40 NPC patients treated with radiotherapy and 30 volunteers without NPC.ELISPOT assay was used to evaluate the levels of EBV-LMP2 special CTLs.RT-PCR and flow cytometry were employed to detect EBV-DNA loads and T cell subsets contents,respectively. Results The positive percentages of EBV-LMP2-specific CTLs were obviously different among three groups;the healthy individuals appeared of highest response, the next was post-treated NPC patients.The positive percentages of EBV-DNA in NPC patients were higher than that in normal controls.Besides, the positive percentage of pre-treated patients was higher than that of post-treated patients.The positive percentage had a negative correlation between EBV-LMP2 special immune response and EBV-DNA.The percentages of CD3+, CD4+ T cells and the CD4+/CD8+ ratios in the pre-treated NPC group decreased;the percentage of CD8+ T cells and the CD4+CD25+ Tr cell level increased. Conclusion The special activity of CTLs in pre-treated NPC patients was lowest and correlated with the EBV-DNA load.Enhancing the response of CTLs may improve the effect on preventing and curing the EBV associated tumors.EBV-DNA load could reflect the tumor load and effectiveness of therapy in some way.The CD3+ and CD4+ T cells in peripheral blood of NPC patients decreased obviously;the levels of CD8+ T cells and CD4+CD25+Tr cell increased.The detection of the T cell subsets would be helpful to the treatment and prognosis judgement of NPC patients.

     

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