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抗原基因修饰的树突状细胞诱导机体产生特异性免疫应答的实验研究 |
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唐华 曹雪涛 于益芝 朱学军 银平章 张明徽
摘 要 目的：探讨腺病毒介导的肿瘤抗原基因修饰的树突状细胞(DC)体内免疫后对抗原特异性抗肿瘤免疫反应的诱导作用。方法：以β-gal为模拟抗原，以转染有LacZ基因并稳定表达β-gal的淋巴瘤细胞E22为肿瘤细胞模型，用携带编码β-gal的LacZ基因的重组腺病毒载体(AdLacZ)转染小鼠骨髓DC，检测转染的效率及LacZ基因修饰DC刺激T细胞增殖的能力，观察皮下免疫LacZ基因修饰DC后小鼠引流区淋巴结细胞数量和组分的变化以及诱导产生CTL和抵抗E22细胞再攻击的能力。结果：LacZ基因修饰后24、48、72 h，均能检测到80％以上的DC表达β-gal，此基因修饰的DC可有效刺激同基因型小鼠脾脏T淋巴细胞增殖反应；将其皮下免疫小鼠1 w 后再接种E22细胞，小鼠的存活期较其他DC免疫小鼠显著延长，但对B16黑色素瘤细胞的攻击无免疫保护作用。此外，LacZ基因修饰的DC免疫小鼠的引流淋巴结细胞数量显著增加，且产生了针对E22的而非EL-4或B16的特异性CTL。结论：腺病毒介导的肿瘤抗原基因修饰的DC能有效诱导机体产生特异的抗肿瘤免疫反应。
关键词 树突状细胞 抗原 基因修饰 腺病毒 CTL 疫苗
中国图书分类号 R392.1
Induction of specific antitumor immune responses by vaccination
with tumor antigen gene-modified dendritic cells
TANG Hua,CAO Xue-Tao,YU Yi-Zhi et al.
Department of Immunology,Second Military Medical University,Shanghai 200433
Abstract Objective:To investigate whether vaccination with antigen gene-modified dendritic cells(DC) could induce specific antitumor immunity,using β-gal as a model tumor antigen.Methods:DC were generated from bone marrow in the presence of mGM-CSF(3.3 ng/ml) and mIL-4(1 ng/ml)，and then,transfected with a recombinant adenovirus encoding LacZ gene(AdLacZ). The efficacy of LacZ gene transfection and T cell stimulating activity of DC were detected.The number and cell subsets of draining lymph nodes,CTL activity of the mice s.c vaccinated with DC were observed.Results:Visual X-gal staining showed that more than 80％ DCs were transfected with LacZ gene.Vaccination with those genetically modified DC could increase the cell number of draining lymph node and induce β-gal specific CTL most significantly. After vaccination with the β-gal gene-modified DC,the survival time of the mice challenged with wild-type E22 cells (a clone of EL-4 lymphoma expressing β-gal)1 week later was prolonged more potently than that of the mice vaccinated with other DCs.No proteckive effect was observed in the m[1] [2] 下一页
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