中文摘要：

目的：构建分泌性小鼠β-防御素2（MBD2）真核表达质粒,转染L1210淋巴细胞白血病细胞,对转染细胞进行功能及特性鉴定。方法：RT-PCR法从小鼠皮肤细胞克隆MBD2成熟部分基因片段,用overlap PCR法将小鼠Igκ信号肽与MBD2成熟片段相连,构建MBD2分泌表达载体。经测序证实序列正确后,脂质体法转染L1210细胞,筛选稳定表达细胞株。RT-PCR、Western blot检测瘤苗MBD2的表达。Transwell法检测瘤苗分泌的MBD2对不成熟树突状细胞（iDC）迁移的作用。细胞计数、PI染色及FACS观察转染MBD2对肿瘤细胞增殖、细胞周期及MHCⅠ类（H-2K^d,H-2D^d）、Ⅱ类（I-A^d）分子及共刺激分子（CD80、CD860）表达的影响。结果：转染MBD2基因的肿瘤细胞表达并分泌MBD2,并以剂量依赖方式趋化iDC,表明具有生物学活性。转染MBD2基因对肿瘤细胞生长增殖、细胞周期及细胞表面MHC分子及共刺激分子表达无明显影响。结论：本实验成功构建了MBD2白血病细胞疫苗,为进一步体内评价抗白血病效果奠定了基础。

英文摘要：

To construct eukaryotic expression plasmid which secrets murine ~-defensin 2（MBD2）,transduce L1210 lymphocytic leukemia cells, and observe the function and characteristics of the tumor vaccine. Methods：The gene for mature MBD2 was cloned from mice skin by RT-PCR. The cDNA encoding the mature MBD2 was fused with a mouse IgK signal sequence by overlap PCR amplification. After being verified by the DNA sequencing, the MBD2-expression plasmid was transduced into L1210 cells by liposome and stable transfectants were isolated. Expression of MBD2 was detected by RT-PCR and Western blot. The Transwell test was used to examine the chemoattractic ability of L1210-MBD2 to immature dendrtic cells （iDC）. The impact of MBD2 transfection on proliferation and expressions of MHC class I（H-2K^d,H-2D^d）, class 11（ I-A^d） and costimulatory molecules（ CD80 and CD860） on the L1210 cells were assessed by cell counting, cell cycle test and FACS assay. Rcsults：L1210-MBD2 cells could express and secret MBD2, which chemoattracted il）C in a dose dependent manner, demonstrating its functional activity. Transfection of MBD2 gene has little influence on cell growth and the expression of surface molecules L1210 cells. Conclusion：The MBD2 leukemic cell vaccine was constructed successfully, and this research laid a foundation for further assess its antileukemia activity in vivo.