转染融合基因的DC抗肿瘤免疫效应

Specific Anti-tumor Immunological Effect by Dendritic Cell Transfected with GM-ΔSur Gene

目的研究转染融合基因(GM-CSF-survivin GM-ΔSur)的树突状细胞(DC)在体外诱导高效而特异的抗肿瘤免疫效应。方法用jetPEITM-Macrophage转染体系,将构建的GM-ΔSur融合基因转染入DC,用流式细胞仪检测DC的表面分子HLA-DR、CD83、CD80、CD86表达的高低;用LDH法测定转染融合基因的DC诱导特异性细胞毒性T淋巴细胞(CTLs)杀伤肿瘤细胞(HT-29和OVCAR-3)的能力。结果转染融合基因的DC细胞中可检测到GM-ΔSur融合蛋白的表达;DC表面高表达HLA-DR、CD83、CD80、CD86;PHA/rhIL-2长期培养(21d)的T细胞CD8+比例明显增加;转染融合基因的DC对HT-29肿瘤细胞的杀伤率显著高于未修饰的DC的杀伤率。结论GM-ΔSur基因转染修饰的DC能选择性诱导MHC-I类分子限制的CTL的特异性,显著提高DC的抗原提呈功能和诱导高效而特异的抗肿瘤免疫效应。

Objective To study the specific anti-tumor immunological effects induced by dendritic cells transfected with GM-CSF gene and survivin gene（GM-ASur） in vitro. Methods pcDNA 3. 1 plasmid containing GM-ASur fusion gene was constructed and was transfected into dendritic cells by jetPEI^TM- Mcrophage transfected kit; cellular surface phenotype such as CD1a, CD80, CD86, CD83 were detected by FACS; The specific cytotoxicity of induced human cytotoxic T lymphocyte （CTLs） to tumor cells was detected with LDH assay. Results GM-ΔSur expression was detected in DC cells; High-level expression of HLA-DR, CD1a, CD80, CD86, CD83 were found in transeneed DC cells; The ratio of CD8^＋ cells cultured with PHA/rhIL-2 were largely increased. The lyse rate of induced CTL cells to HT-29 cell were much higher than those of blank DC cells. Conclusion GM-ΔSur gene modified DC cells could induce MHC- Ⅰ specific cytotoxic T lymphocytes and strikingly raised DC cell＇ s antigen present function and could induce high effective and specific anti-cancer immunological effect.