CAJ | 학술논문

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.
목적 탐토소서CXC형추화인자수체4(CXCR4)기인과표체적골수간충질간세포(MSC)방치소서이식물항숙주병(GVHD)작용.방법 이용만병독재체구건과표체소서CXCR4기인적중조만병독재체,포장병독후전도소서골수간충질간세포(CXCR4-MSC).이C57BL/6소서위공서,BALB/c소서위수서,건립소서이기인골수이식(BMT)모형,동시수주골수MSC.분5조:전신조사(TBI)조:부접수TBI; BMT조:TBI조수주소서골수세포;GVHD조:BMT조수주이기인비세포;EGFP-MSC조:GVHD조수주EGFP-MSC; CXCR4조:GVHD조수주CXCR4-MSC.관찰수서생존상태,통계생존솔,관찰수서조직병이학개변급염증세포인자변화,평개기대GVHD방치작용.결과 TBI조소서재조사후14d내균사우조혈쇠갈,BMT조수서가획장기존활,GVHD조、EGFP-MSC조화CXCR4-MSC조생존시간분별위(1 7.0±2.3)、(21.7±4.8)、(30.1±9.1)d,CXCR4-MSC공이식조수서존활시간명현장우GVHD조(P＜0.05).각조수서균출현복사、궁배、교모、피부결손、체중감경등GVHD증상,공이식CXCR4-MSC조수서림상평분저우GVHD조(P＜0.05).GVHD조여EGFP-MSC조수서간장、소장여피부균존재전형GVHD병리개변,CXCR4-MSC조부발생Ⅰ～Ⅱ도병리개변,수서조직병리평분저우이상량조.이식후제14천,각조수서촉염증인자수평명현승고,CXCR4-MSC조IFN-γ、IL-2、TNF-α수평명현저우GVHD조여EGFP-MSC조(P＜0.05).결론 연합수주CXCR4과표체적골수MSC가통과강저수서이식후염증인자분비종이감경GVHD.
Objective To investigate the effect of the lentiviral vector mediated CXCR4 overexpressed mesenchymal 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-γ and TNF-α level in recipients infused with CXCR4-MSC were significantly decreased (P＜0.05).Conclusions The results revealed that CXCR4-transduced MSCs could effectively control the occurrence of mouse GVHD following allogeneic BM transplantation.