摘要
目的:探讨食管癌细胞抗原致敏、白细胞介素27(interleukin27,IL-27)基因修饰的树突状细胞(dendritic cells,DCs)疫苗的特异性抗食管癌免疫反应。方法:采用重组逆转录病毒介导IL-27基因修饰食管癌患者外周血DCs;反复冻融法提取食管癌细胞裂解产物,致敏经IL-27基因转染的DCs;ELISA法检测各组DCs和细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTLs)上清液中IL-27、IL-12和干扰素-γ(interferon-γ,IFN-γ)的分泌水平;FCM法分析各组DCs表面分子CD1a、CD83、CD80和CD86的表达水平;MTT法检测DCs刺激T细胞增殖的活性和DCs诱导CTLs杀伤食管癌细胞的效能。结果:经IL-27基因修饰和抗原致敏后的DCs高表达CD83、CD80和CD86分子,其表达量分别为(82.67±7.92)%、(78.33±7.31)%和(85.33±4.32)%;DCs上清液中IL-12、IL-27及IFN-γ分泌量明显提高,分别为(107.85±7.20)ng/L、(430.39±10.12)ng/L及(411.97±17.80)ng/L;并且,DCs明显刺激同源T细胞增殖,增强了CTLs上清液中IFN-γ水平[(751.50±31.30)ng/L]。诱导产生的CTLs对食管癌细胞有强大的杀伤作用,显著高于其他组(P<0.01)。结论:IL-27基因修饰增强了DCs在体外诱导自体T淋巴细胞产生特异性抗食管癌免疫的能力。其机制可能与IL-27基因修饰和抗原致敏活化了DCs抗原提呈第二信号,促进了DCs高分泌IL-27和IL-12因子,活化了T淋巴细胞,并致使CTLs分泌IFN-γ的能力增强等密切相关。
Objective： To investigate the anti-esophageal cancer immunity induced by antigen- sensitized and interleukin 27 （IL-27） gene-modified dendritic cells （DCs）. Methods： DCs isolated from the peripheral blood of patients with esophageal cancer were modified by recombinant retrovirus-mediated IL-27 gene, and then sensitized by the cleaved products from esophageal carcinoma Eca109 cells by using repeated freezing-thawing method. The secretion levels of IL-27, IL-12 and interferon-y （IFN-y） in culture supernatant of DCs and the level of IFN-y in cytotoxic T lymphocytes （CTLs） were examined by ELISA method. The expression levels of CDla, CD83, CD80 and CD86 molecules on DCs surface were measured by FCM. The proliferative activity of T lymphocytes and the killing efficacy of CTLs against Eca109 cells induced by DCs were evaluated by MTT method. Results： After being modified by IL-27 gene and sensitized by tumor antigen, the secretion levels of IL-12 [（107.85＋7.20） ng/LL, IL-27 [（430.39_＋10.12） ng/L] and IFN-y [（411.97±1 7.80） ng/L of DCs were increased, and the expressions of CD83 [（82.67± 7.92）%], CD80 [（78.33±7.31）%] and CD86 [（85.33±4.32）%] molecules were also elevated. Meanwhile, the proliferation capacity of autologous T lymphocytes was enhanced after stimulation by DCs, and thesecretion level of IFN-y [（751.50±31.30） ng/L] in the culture supernatant of DCs was increased. The DCs modified by IL-27 gene and sensitized by tumor antigen could significantly induce the strongest cytotoxicity of antigen-specific CTLs against Eca109 cells （P〈0.01）. Conclusion： IL-27 gene modification enhances the capacity of DCs in inducing autologous T lymphocytes to generate specific antitumor immunity against esophageal carcinoma. The mechanism of this immune response may be closely relevant to the activation of the second signal for presenting antigen cells such as DCs, the promoted capacity of DCs secreting IL-27 and IL-12, and the activation of T cells resulting in the
出处
《肿瘤》
CAS
CSCD
北大核心
2011年第7期601-607,共7页
Tumor
基金
河北省普通高等学校强势特色学科基金资助(编号:冀教高[2005]52号)
关键词
食管肿瘤
树突细胞
免疫疗法
白细胞介素-27
转染
抗原致敏
Esophageal neoplasms
Dendritic cells
Immunotherapy
Interleukin-27
Transfection
Antigen-sensitization
