分子嵌合MHC-Ⅰ基因对小鼠DC反应的研究  

The Research on Chimeric MHC-I Gene Responsing to Allogeneic Dendritic Cells

作　　者：闫永嘉[1] 付蔚华[1] 高莹[1] 朱理玮[1] 

机构地区：[1]天津医科大学总医院普通外科,300052

出　　处：《天津医药》2012年第4期378-380,I0006,共4页Tianjin Medical Journal

摘　　要：目的:构建分子嵌合主要组织相容性复合体(MHC)-Ⅰ基因小鼠骨髓造血干细胞,并探讨其诱导脾脏T细胞对异基因小鼠树突状细胞(DC)反应的机制。方法:密度梯度法分离培养BALB/c小鼠骨髓造血干细胞。构建携带C57BL/6小鼠MHC-Ⅰ基因慢病毒载体(病毒感染组),携带无意义基因慢病毒载体(阴性对照组)。分别感染BALB/c小鼠骨髓造血干细胞,构建分子嵌合细胞。分别取病毒感染组、阴性对照组及未加入病毒的空白对照组造血干细胞输注BALB/c小鼠后7d,获取脾脏T淋巴细胞,分别与C57BL/6小鼠DC进行混合淋巴细胞培养,测定刺激指数。结果:成功体外分选及培养BALB/c小鼠骨髓造血干细胞。病毒感染组C57BL/6小鼠MHC-Ⅰ蛋白表达率可达98.17%。单向混合淋巴细胞培养结果显示,C57BL/6小鼠DC对输注病毒感染组细胞后BALB/c小鼠脾脏T细胞刺激指数明显降低(P<0.01)。结论:输注分子嵌合MHC-Ⅰ基因造血干细胞后,小鼠脾脏T细胞对异基因小鼠DC反应明显减低。Objective： To construct the mouse bone marrow hematopoietic stem cells （HSCs） with chimeric MHC- I gene, and to explore the mechanism of it inducing T lymphocyte response to allogeneic dendritic ceils （DC）. Methods： The mouse （BALB/c） bone marrow HSCs were collected with the lymphocyte separation medium by density gradient separation. The identified HSCs were infected by the constructed lentiviral vectors. After infection, the molecular chimerism was success- fully constructed, called infected group. The HSC infected by lentiviral vectors without H2-K1 was counted as noninfected group. The HSC was counted as blank control group. The ability of molecular chimerism in T lymphocyte response to allogene- ic dendritic cells was observed through mixed lymphocyte culture （MLC）. Results： The BALB/c mouse bone marrow HSCs were successfully separated and cultured. Flow cytometry analysis indicated that MHC- I protein expression was 98.17% in infected group. The result of MLC demonstrated that the stimulation index of T lymphocyte was significantly decreased （P 〈 0.01）. Conclusion： The lentiviral vector carrying C57 mice＇ s MHC- I gene H2-K1 was successfully constructed. The molecular chimerism could reduce T lymphocyte response to allogeneic dendritic cells.

关 键 词：骨髓祖代细胞 慢病毒属 T淋巴细胞 免疫耐受 树突细胞 MHC-Ⅰ基因 小鼠 近交BALB C小鼠 近交C57BL 疾病模型 动物 

分 类 号：R516.7[医药卫生—内科学]