CD26基因敲除小鼠异基因骨髓移植的研究  

Investigation on allogenic bone marrow transplantation with CD26 gene knockout mice

作　　者：薛兴奎 韩新华 蔡思齐 陈晓玲 Melody Y.Zeng XUE Xing-kui;HANXin-hua;CAI Shi-qi;CHEN Xiao-ling(Central Laboratory, the People’s Hospital of ShenzhenLonghua District, Shenzhen 518109, Guangdong, CHINA;Department of Encrinology, Shekou People’s Hospital ofShenzhen Nanshan District, Shenzhen 518109, Guangdong, CHINA;Graduate School of South Medical School,Guangzhou 501505, Guangdong, CHINA)

机构地区：[1]深圳市龙华区人民医院中心实验室,广东深圳518109 [2]深圳市南山区蛇口人民医院内分泌科,广东深圳518109 [3]南方医科大学研究生院,广东广州501505

出　　处：《海南医学》2019年第10期1230-1233,共4页Hainan Medical Journal

基　　金：国家自然科学基金(编号:31650110474);广东省深圳市科创委项目(编号:JCYJ20160426094752965);广东省深圳市卫生系统科研项目(编号:201601059);广东省深圳市龙华区科技创新项目(编号:20160831A1030180;20160831A0410020)

摘　　要：目的研究CD26基因敲除对建立异基因骨髓移植模型的影响。方法分别提取CD26基因敲除C57BL/6小鼠(CD26^(-/-))骨髓细胞和脾T细胞,输注经过900 cGy剂量照射的BALB/C小鼠,进行异基因小鼠骨髓移植。根据回输的细胞不同分为骨髓细胞组(BM组)、小鼠骨髓加脾脏T细胞组(BM+SP组)。采用流式细胞术检测异基因小鼠移植的嵌合体。移植后监测小鼠体质量、活动度、弓背情况、毛发等,统计移植物抗宿主病(GVHD)指数。取小鼠肝脏、皮肤、肠道组织,制备病理切片,镜下观察病理变化。结果敲除CD26的造血干细胞顺利植入受者小鼠体内,BM组、BM+SP组在第8周嵌合率[(71.16±7.34)%vs (66.72±6.02)%]比较差异无统计学意义(P>0.05);植入的供者造血干细胞能在受者体内引起GVHD的发生,在移植后第35天,BM组、BM+SP组小鼠体质量分别平均下降2.2%与28.5%,GVHD分数分别为0.30±0.44与5.2±1.01,差异均有统计学意义(P<0.05);提取小鼠肝脏、皮肤、肠道组织,病理切片评估,结果显示,BM组和BM+SP组均有明显的病理损害,提示有GVHD的发生。结论 CD26基因敲除小鼠造血干细胞能够顺利植入受者小鼠并构建异基因骨髓移植模型,导致GVHD发病,为研究CD26在异基因骨髓移植中的作用机制提供有效方法。Objective To investigate the model of allogeneic bone marrow transplantation of CD26 gene knockout mice. Methods The CD26 gene knockout C57 BL/6 mice(CD26-/-) were used as donors and the bone marrow and spleen T cells were harvested from the euthanized mice. BALB/C mice were used as recipient, and the bone marrow and/or spleen T cells from donors were infused through the tail vein after the recipients were irradiated at 900 cGy. The mice infused with bone marrow were enrolled as BM group, and the mice infused with bone marrow and spleen T cells were enrolled as BM+SP group. Flow cytometry was used to monitor the chimerism after transplantation. Graft versus host disease(GVHD) indexes were evaluated based on monitoring the body weight, fur, activity, and hunched-back of the mice. The liver, intestine, and skin of mice were examined with pathological slides. Results Hematopoietic stem cells from CD26-/-mice were successfully engrafted into the recipient and there were no statistically significance in the chimerism between the BM group and BM+SP group 8 weeks post transplantion:(71.16±7.34)% vs(66.72±6.02)%, P>0.05. Moderate GVHD were determined after transplantation in BM+SP group. On day 35 after transplantation, the body weight of mice in the BM group and BM+SP group decreased 2.2% and 28.5% respectively, while the GVHD score were0.30 ± 0.44 and 5.2 ± 1.01(P<0.05), respectively. Pathological evaluation with the liver, skin, and intestine samples showed significant pathological damage in BM group and BM + SP group, which indicates the occurrence of GVHD.Conclusion The bone marrow of CD26 gene knockout mice were engrafted into allogeneic mice and caused GVHD,which provides effective approach for the study on the role of CD26 in the bone marrow transplantation.

关 键 词：CD26 移植物抗宿主疾病 异基因骨髓移植 嵌合体 基因敲除 

分 类 号：R-332[医药卫生]