凋亡肿瘤细胞致敏的树突状细胞疫苗治疗肺癌的实验研究  

Dendritic cells loaded with apoptotic tumor cells in the treatment of lung cancer

作　　者：黄建安[1] 杨新静[1] 胡玉敏[2] 穆传勇[1] 张学光[2] 

机构地区：[1]苏州大学附属第一医院呼吸内科,江苏省临床免疫学重点实验室,苏州215007 [2]苏州大学生物技术研究所,苏州215007

出　　处：《肿瘤》2007年第2期88-91,共4页Tumor

基　　金：江苏省社会发展基金(编号:BS200038);江苏省医学重点人才基金(编号:RC2002032)

摘　　要：目的用凋亡肿瘤细胞致敏的树突状细胞(dendritic cells,DCs)激发肿瘤抗原特异性的细胞毒T细胞(cytotoxic Tlymphocytes,CTL)活性,观察其体内外抗肺癌的特性。方法常规方法从健康人外周血单个核细胞中诱导DCs,采用或不采用凋亡肿瘤细胞负载DCs,并利用激发型CD40单克隆抗体(CD40mAb)诱导DCs成熟;成熟DCs与自体T细胞共育,分别获得Ag-CTL及non-Ag-CTL,流式细胞仪检测Ag-CTL细胞表型的变化;3H-TdR掺入法测定DNA片段形成率;建立人肺癌细胞株A549荷瘤裸鼠模型,过继回输Ag-CTL和non-Ag-CTL,评价其在体内的抗肿瘤活性。结果CD40mAb激发可使DCs上调CD1a、CD80、CD86、CD83、HLR-DR的表达;凋亡肿瘤细胞负载联合CD40mAb激发可进一步促进DCs的成熟;成熟DCs和自体的T细胞共育活化后CD8+T细胞明显上调;Ag-CTL对A549具有高效特异的杀伤力,明显强于Ag-CTL对肝癌细胞株HepG2的作用(P<0.01),且Ag-CTL对A549的杀伤力明显强于non-Ag-CTL(P<0.01),而non-Ag-CTL对A549及HepG2细胞的杀伤力无显著性差异;体内实验表明,Ag-CTL可有效抑制裸鼠皮下移植瘤的生长,与生理盐水组(NS组)、non-Ag-CTL组相比在统计学意义上有显著差异(P<0.05),non-Ag-CTL组与NS组相比在统计学意义上有差异(P<0.05)。结论凋亡肿瘤细胞致敏的树突状细胞疫苗激发的Ag-CTL在体内外均呈现抑制肺癌细胞的特性。Objective：Using apoptotic tumor cells-loaded dendritic cells （DCs） to activate the antigen-specific cytotoxic T lymphocytes （CTL） and observe their effects against lung cancer in vivo and in vitro, nethods-DCs were induced from healthy human peripheral blood monocytes（PBMC） by rhGM-CSF and rhIL-4 and loaded with or without apoptotic tumor ceils. The immature DCs were activated by CD40mAb. The mature DCs were co-cultured with autologous T cells to obtain antigen specific CTL （Ag-CTL） and antigen non-specific CTL （ non Ag-CTL）. The phenotypes of Ag-CTL were analyzed by flow cytometry. The DNA fragment formation was detected by thymidine incorporation. The antitumor activity of Ag-CTL and non Ag-CTL in vivo was evaluated in BALB/c nude mice bearing A549 lung cancer. Results.The expressions of CD1a, CD80 CD83, CD86, HLR-DR were up-regulated after DCs were activated by CD40mAb alone. CD40mAb combined with apoptotic tumor cells further induced maturation of DCs. The CD8^＋T cells were markedly upregulated after co-culture of mature DCs and autologous T cells; Ag-CTL had specific killing effect on A549 lung cancer cells with high efficiency. The effect was stronger than that on HepG2 hepatoma carcinoma cells （P 〈0.01 ）. The tumor-killing effect of Ag-CTL on A549 lung cancer cells was stronger than non Ag-CTL （P 〈0.01 ）. The difference in the tumor killing effects of non Ag-CTL on A549 and HepG2 cells was not significant. In vivo experiments showed that Ag-CTL significantly inhibited the growth of transplanted tumors in nude mice compared with NS and non Ag-CTL （P 〈0.05 ）. The difference between NS group and non Ag-CTL group was significant（ P 〈0.05 ）. Conclusion：The apoptotic tumor cells-loaded DCs induce a strong and specific CTL response against lung cancer cells in vitro and in vivo.

关 键 词：肺肿瘤 抗原呈递细胞 树突状细胞 T淋巴细胞 细胞毒性 细胞凋亡 A549细胞 

分 类 号：R73-36[医药卫生—肿瘤]