摘要
目的研究趋化因子巨噬细胞炎症蛋白1α（MIP-1α）动员的树突状细胞（DC），经黑色素瘤抗原基因3（MAGE-3）腺病毒转染后对胃癌细胞的免疫效应。方法615小鼠尾静脉注射MIP-1α，分选得到B220^-CD11c^＋细胞，加入细胞因子连续培养，检测其细胞表面标志和混合淋巴细胞反应（MLR）。收集培养后的B220^-CD11c^＋细胞，加入编码MAGE-3的重组腺病毒进行转染，制备表达肿瘤抗原的DC疫苗，以荷载小鼠前胃癌（MFC）全肿瘤细胞抗原制备的DC疫苗作为阳性对照。采用二苯基溴化四氮唑蓝（MTT）法，检测活化的T淋巴细胞在体外对MFC细胞的杀伤作用。采用酶联免疫吸附试验（ELISA），检测γ干扰素（INF-γ）的分泌情况。DC疫苗皮下注射MFC荷瘤小鼠，观察小鼠瘤体生长情况和存活时间。结果MIP-1α注射后，外周血中B220-CD11c^＋细胞明显升高。新鲜分离的B220-CD11c^＋细胞在体外经过细胞因子诱导分化后具有典型的DC表面标志，并在MLR中具有极强的刺激T淋巴细胞增殖的能力。腺病毒转染MAGE-3的DC激活的T淋巴细胞表现出对MFC细胞的特异性杀伤作用，产生高水平的INF-γ[（1460．00±16．82）pg／ml]。实验组荷瘤小鼠接受DC疫苗治疗后，肿瘤生长缓慢，观察至MFC细胞接种后的第27天，其肿瘤体积仅为（3．46±1．12）cm^3；实验组小鼠的肿瘤体积与对照组之间的差异有统计学意义（P〈0．01）。实验组小鼠存活时间明显延长，与对照组之间的差异有统计学意义（P〈0．01）。结论注射趋化因子MIP—1α可快速动员并诱导分化为成熟DC。肿瘤抗原基因MAGE-3经腺病毒转染制备的DC疫苗，在体外可以诱导出针对胃癌细胞的特异性杀伤作用，在体内对MFC荷瘤小鼠有明显的免疫治疗作用。
Objective To investigate the anti-gastric carcinoma immunological efficacy of dendritic cells （DC） precursors, that were mobilized into the peripheral blood by injection of macrophage inflammation protein-1α （MIP-1α）, and induced by DC vaccine expressing melanoma antigen gene-3 （MAGE-3） ex vivo and in vivo. Methods 615 mice were injected with MIP-lc~ via the tail vein. Freshly isolated B220^- CD11c^＋ cells were cultured with cytokines and assayed by phenotype analysis and mixed lymphocyte reaction （MLR）. For adenoviral （Ad） -mediated gene transduction, cultured B220 - CD11 c^＋ cells were incubated with Ad-melanoma antigen gene-3. MIP-1α-mobilized B220 - CD11 c^＋ cells pulsed MFC cells tumor lysate were used as positive control. The stimulated DC vaccination-induced T lymphocytes, and the killing effect of the T cells on gastric carcinoma ceils were assayed by MTT. INF-3, production was determined with the INF-γ ELISA kit. To establish the solid tumor model, groups of 615 mice were injected with MFC cells subcutaneously into the abdominal wall. MIP-1α-mobilized DC vaccines expressing MAGE-3 gene were used to immunize the mice after the challenge of MFC cells, then the tumor size and the survival of mice were examined to detect the therapeutic effect of DC vaccines. Results B220^- CD11 c^＋ cells increased obviously after MIP-1α injection, and freshly isolated B220^- CD11c^＋ cells cultured with mGM- CSF, IL-4, and mTNF-α were phenotypically identical to typical DC, gained the capacity to stimulate allogeneic T cells. These MIP-1α-mobilized DCs were transduced with Ad-MAGE-3, which were prepared for DC vaccines expressing tumor antigen. T lymphocytes stimulated with DC-transduced with Ad-MAGE-3 showed specific killing effect on gastric carcinoma cells and produced high levels of INF-γ [ （ 1460.00± 16.82） pg/ml]. Five days after the MFC cells challenge, the mice were subsequently injected with DC vaccines. The tumor size of the experimental group was significantly smaller th
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2009年第5期330-334,共5页
Chinese Journal of Oncology
关键词
黑色素瘤抗原基因3
树突状细胞
巨噬细胞炎症蛋白-1Α
胃肿瘤
Melanoma antigen gene-3 （ MAGE-3 ）
Dendritic cells
Macrophage inflammation protein-1 α （ MIP-1 α）
Stomach neoplasms
