慢病毒载体介导的CXCR4基因过表达骨髓间充质干细胞在防治小鼠移植物抗宿主病中的作用  

Effect of lentiviral vector mediated CXCR4 gene overexpressed mesenchymal stem cell on the protection of mice against graft-versus-host disease

作　　者：陈伟[1] 李淼[1] 张翠萍[1] 王祥民[1] 潘彬[1] 曾令宇[1] 李振宇[1] 徐开林[1] 

机构地区：[1]徐州医学院附属医院血液科,221002

出　　处：《中华血液学杂志》2014年第10期936-940,共5页Chinese Journal of Hematology

基　　金：国家自然科学基金（81300441）;江苏省“六大人才高峰”资助项目（2013-WSN-080）

摘　　要：目的 探讨小鼠CXC型趋化因子受体4（CXCR4）基因过表达的骨髓间充质干细胞（MSC）防治小鼠移植物抗宿主病（GVHD）作用.方法 利用慢病毒载体构建过表达小鼠CXCR4基因的重组慢病毒载体,包装病毒后转导小鼠骨髓间充质干细胞（CXCR4-MSC）.以C57BL/6小鼠为供鼠,BALB/c小鼠为受鼠,建立小鼠异基因骨髓移植（BMT）模型,同时输注骨髓MSC.分5组：全身照射（TBI）组：仅接受TBI; BMT组：TBI组输注小鼠骨髓细胞;GVHD组：BMT组输注异基因脾细胞;EGFP-MSC组：GVHD组输注EGFP-MSC; CXCR4组：GVHD组输注CXCR4-MSC.观察受鼠生存状态,统计生存率,观察受鼠组织病理学改变及炎症细胞因子变化,评价其对GVHD防治作用.结果 TBI组小鼠在照射后14d内均死于造血衰竭,BMT组受鼠可获长期存活,GVHD组、EGFP-MSC组和CXCR4-MSC组生存时间分别为（1 7.0±2.3）、（21.7±4.8）、（30.1±9.1）d,CXCR4-MSC共移植组受鼠存活时间明显长于GVHD组（P＜0.05）.各组受鼠均出现腹泻、弓背、翘毛、皮肤缺损、体重减轻等GVHD症状,共移植CXCR4-MSC组受鼠临床评分低于GVHD组（P＜0.05）.GVHD组与EGFP-MSC组受鼠肝脏、小肠与皮肤均存在典型GVHD病理改变,CXCR4-MSC组仅发生Ⅰ～Ⅱ度病理改变,受鼠组织病理评分低于以上两组.移植后第14天,各组受鼠促炎症因子水平明显升高,CXCR4-MSC组IFN-γ、IL-2、TNF-α水平明显低于GVHD组与EGFP-MSC组（P＜0.05）.结论 联合输注CXCR4过表达的骨髓MSC可通过降低受鼠移植后炎症因子分泌从而减轻GVHD.Objective To investigate the effect of the lentiviral vector mediated CXCR4 overexpressed mesenehymal stem cell （MSCs） on graft-versus-host disease （GVHD）, Methods Lentiviral vector containing CXCR4 was constructed. CXCR4 overexpressed MSC by lentiviral vector mediated were assessed. A major histocompatibility complex （MHC）-mismatched mouse model of bone marrow transplantation （BMT） from C57BL/6 donors to BALB/c recipients was constructed. Mice were divided into five groups： total body irradiation （TBI） group, mice received irradiation only; BMT group, mice were transplanted with bone marrow （BM） after TBI; GVHD group, mice were transplanted with BM and splencytes after TBI; CXCR4-MSC group, mice were transplanted with CXCR4-MSC, BM and splencytes after TBI; EGFP-MSC group, mice were transplanted with EGFP-MSC, BM and splencytes after TBI. The survival, body weight and clinical score of GVHD in transplanted mice were monitored. Liver, intestine and skin from mice in each group were obtained for histological examination. Plasma concentrations of inflammation factors such as interleukin （IL）-2, IFN-γ and TNF-α were also determined using a cytometric bead array cytokine kit. Results All mice in TBI group died within 14 days, while all of BMT group survived. The mean survival times for GVHD, EGFP-MSC and CXCR4-MSC groups were （ 17.0±2.3 ） d, （21.7±4.8） d and （30.1±9.1） d, respectively. Treatment with CXCR4 over-expressing MSCs could decrease the mortality rate, All mice in each group developed clinical signs such as hunched posture, dull fur, diarrhea and weight loss. Meanwhile, histopathological findings in target organs were confirmed the presence of GVHD. While, clinical GVHD scores and histopathological scores in CXCR4-MSC group were significantly lower than that of GVHD group. Moreover, compared with control groups, the plasma IL-2, IFN-7 and TNF-α level in recipients infused with CXCR4-MSC were significantly decreased （P〈 0.05）. Conclusions The results revea

关 键 词：慢病毒载体 骨髓移植 移植物抗宿主病 间质干细胞 CXCR4基因 

分 类 号：R457.7[医药卫生—治疗学]