LMPs特异性T淋巴细胞对鼻咽癌细胞的杀伤作用
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江苏省临床医学科技专项(BL2013038); 江苏省普通高校研究生科研创新计划(CXZZ12_0563)


The cytotoxic effect of LMPs specific T lymphocytes on nasopharyngeal carcinoma cells
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    摘要:

    目的:研究树突状细胞负载EB病毒潜伏膜蛋白(latent membrane proteins,LMPs)介导生成的LMPs特异性细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)LMPs-CTL的生物学特性,观察其对LMPs阳性鼻咽癌细胞SUNE的杀伤作用-方法:用淋巴细胞分离液分离人外周血单核细胞,采用贴壁分离及白细胞介素(interleukin,IL)-4-粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)等细胞因子诱导成熟树突状细胞(dendritic cell,DC),通过DC细胞负载LMPs多肽抗原并递呈给同源T淋巴细胞,制备LMPs-CTL-CCK8法检测LMPs-CTL对SUNE细胞的杀伤作用;ELISA和羟基荧光素二醋酸盐琥珀酰亚胺脂(carboxy fluoroscein succinimidyl ester,CFSE)染色法检测LMPs-CTL分泌IFN-γ及其增殖-结果:LMPs-CTL对SUNE细胞的杀伤率在12 h和24 h分别为(43.47 ± 1.93) %和(77.15 ± 3.18) %,显著高于对照组的(11.45 ± 3.06) % 和(24.27 ± 13.20)%(P < 0.05)-SUNE细胞刺激后,LMPs-CTL增殖能力较对照组有明显增强,IFN-γ分泌量达到(613.40 ± 121.77) pg/ml,显著高于对照组(86.90 ± 3.70) pg/ml(P < 0.05)-结论:通过DC细胞负载LMPs混合多肽可诱导生成LMPs-CTL,对LMPs阳性鼻咽癌细胞有较强的杀伤作用-

    Abstract:

    Objective:To study the biological characteristics of cytotoxic T lymphocyte (CTL) originated from dendritic cells (DC) loaded with the Epstein-Birr virus latent membrane proteins (LMPs),and to detect the cytotoxic effect of LMPs-CTL on the LMPs positive nasopharyngeal carcinoma cell SUNE. Methods:The mononuclear cells were isolated from human peripheral blood by lymphocyte separation medium. The adherent method was performed. Cytokines of IL-4 and GM-CSF were performed to induce mature DC. Loaded with LMPs polypeptide antigen,the mature DC presented antigen to autologous T lymphocyte for preparation of LMPs-CTL. The cytotoxic effect of LMPs-CTL on SUNE cells was detected by the CCK8 method. The secretion of interferon-γ (IFN-γ) of LMPs-CTL and cell proliferation was detected by ELISA and carboxy fluoroscein succinimidyl ester (CFSE),respectively. Results:The killing efficiency of LMPs-CTL for SUNE was (43.47 ± 1.93)% in 12h and (77.15 ± 3.18)% in 24 h,which were higher than those in the control group[(11.45 ± 3.06)% and (24.27 ± 13.2)% (P < 0.05)]. The ability of LMPs-CTL cell proliferation simulated with SUNE cells was stronger than that in the control group. The IFN-γ secretion level secreted by LMPs-CTL was (613.40 ± 121.77) pg/ml,which was significantly higher than that secreted by the control group (86.90 ± 3.70) pg/ml (P < 0.05). Conclusion:LMPs specific CTL can be successfully induced by mature DC and possess preferable specific killing activity on LMPs positive nasopharyngeal carcinoma cells.

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李文杰,唐小军,熊四平,陈 志,蒋 帅,冯振卿,朱 进,陈仁杰. LMPs特异性T淋巴细胞对鼻咽癌细胞的杀伤作用[J].南京医科大学学报(自然科学版),2014,(6):705-710

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  • 收稿日期:2014-01-14
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  • 在线发布日期: 2014-06-19
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