国际输血及血液学杂志
國際輸血及血液學雜誌
국제수혈급혈액학잡지
INTERNATIONAL JOURNAL OF BLOOD TRANSFUSION AND HEMATOLOGY
2013年
1期
1-6
,共6页
刘迷迷%房婷%化静%付金玉%黄玉金%吴进燕%徐开林%曾令宇
劉迷迷%房婷%化靜%付金玉%黃玉金%吳進燕%徐開林%曾令宇
류미미%방정%화정%부금옥%황옥금%오진연%서개림%증령우
骨髓移植%内皮祖细胞%免疫重建%修复
骨髓移植%內皮祖細胞%免疫重建%脩複
골수이식%내피조세포%면역중건%수복
bone marrow transplantation%EPC%immune reconstitution%repair
目的 探讨异基因骨髓移植中联合内皮祖细胞(EPC)移植对接受白消安/环磷酰胺(BU/CY)方案预处理小鼠移植后免疫重建的影响.方法 选择周龄为8～10周龄,体重为(22～24)g的健康SPF级雄性BALB/c小鼠58只作为研究对象.随机分为正常对照组(n=9):经尾静脉注射磷酸盐缓冲液(PBS)0.2 mL；BU/CY预处理组(n=9):移植前7d至前2d接受BU/CY方案预处理后,经尾静脉注射PBS 0.2 mL;单纯骨髓细胞移植组(BMT组)(n=20):BU/CY方案预处理后,经尾静脉注射供者骨髓细胞；EPC联合移植组(EPC组)(n=20):BU/CY方案预处理后,经尾静脉注射供者骨髓细胞及EPC.于移植后第14,21,28天,采用流式细胞术检测各组小鼠骨髓中B系祖(ProB)细胞(CD19+ c-kit+ IgM-)数量、胸腺中CD4+,CD8+,双阳性(DP)及双阴性(DN)T淋巴细胞比例及脾脏中CD4+,CD8+T淋巴细胞及B淋巴细胞比例,并通过苏木素-伊红染色及免疫组化染色观察中枢和外周免疫器官组织结构的恢复及损伤血管内皮的修复情况.结果 移植后BMT及EPC组小鼠骨髓中ProB细胞数量持续增高,第14天时,EPC组(0.83×106)显著高于BMT组(0.14×106)(x2=7.2,P＜0.05)；胸腺中DP细胞比例于第21天达峰值,EPC组(0.58％)较BMT组(0.39％)显著增高(x2=6.49,P＜0.05)；脾脏中CD4+T细胞比例持续增高,第28天时,EPC组(3.64％)较BMT组(2.47％)显著增高(x2=7.2,P＜0.05)；CD8+T细胞比例在移植后第21天达峰值,EPC组(3.64％)较BMT组(3.21％)显著增高(x2=5.42,P＜0.05).组织病理学检查也显示EPC组小鼠的中枢及外周免疫器官组织结构恢复较BMT组快,组织中实质细胞数量恢复较BMT组快；对脾脏血窦内皮免疫组化染色的结果也显示,EPC组小鼠脾血窦内皮完整性的恢复较BMT组快.结论 BU/CY预处理小鼠异基因骨髓移植中,联合EPC输注可促进移植受鼠免疫器官损伤血管内皮的修复、促进其组织结构的恢复,从而有效促进移植后的免疫重建.
目的 探討異基因骨髓移植中聯閤內皮祖細胞(EPC)移植對接受白消安/環燐酰胺(BU/CY)方案預處理小鼠移植後免疫重建的影響.方法 選擇週齡為8～10週齡,體重為(22～24)g的健康SPF級雄性BALB/c小鼠58隻作為研究對象.隨機分為正常對照組(n=9):經尾靜脈註射燐痠鹽緩遲液(PBS)0.2 mL；BU/CY預處理組(n=9):移植前7d至前2d接受BU/CY方案預處理後,經尾靜脈註射PBS 0.2 mL;單純骨髓細胞移植組(BMT組)(n=20):BU/CY方案預處理後,經尾靜脈註射供者骨髓細胞；EPC聯閤移植組(EPC組)(n=20):BU/CY方案預處理後,經尾靜脈註射供者骨髓細胞及EPC.于移植後第14,21,28天,採用流式細胞術檢測各組小鼠骨髓中B繫祖(ProB)細胞(CD19+ c-kit+ IgM-)數量、胸腺中CD4+,CD8+,雙暘性(DP)及雙陰性(DN)T淋巴細胞比例及脾髒中CD4+,CD8+T淋巴細胞及B淋巴細胞比例,併通過囌木素-伊紅染色及免疫組化染色觀察中樞和外週免疫器官組織結構的恢複及損傷血管內皮的脩複情況.結果 移植後BMT及EPC組小鼠骨髓中ProB細胞數量持續增高,第14天時,EPC組(0.83×106)顯著高于BMT組(0.14×106)(x2=7.2,P＜0.05)；胸腺中DP細胞比例于第21天達峰值,EPC組(0.58％)較BMT組(0.39％)顯著增高(x2=6.49,P＜0.05)；脾髒中CD4+T細胞比例持續增高,第28天時,EPC組(3.64％)較BMT組(2.47％)顯著增高(x2=7.2,P＜0.05)；CD8+T細胞比例在移植後第21天達峰值,EPC組(3.64％)較BMT組(3.21％)顯著增高(x2=5.42,P＜0.05).組織病理學檢查也顯示EPC組小鼠的中樞及外週免疫器官組織結構恢複較BMT組快,組織中實質細胞數量恢複較BMT組快；對脾髒血竇內皮免疫組化染色的結果也顯示,EPC組小鼠脾血竇內皮完整性的恢複較BMT組快.結論 BU/CY預處理小鼠異基因骨髓移植中,聯閤EPC輸註可促進移植受鼠免疫器官損傷血管內皮的脩複、促進其組織結構的恢複,從而有效促進移植後的免疫重建.
목적 탐토이기인골수이식중연합내피조세포(EPC)이식대접수백소안/배린선알(BU/CY)방안예처리소서이식후면역중건적영향.방법 선택주령위8～10주령,체중위(22～24)g적건강SPF급웅성BALB/c소서58지작위연구대상.수궤분위정상대조조(n=9):경미정맥주사린산염완충액(PBS)0.2 mL；BU/CY예처리조(n=9):이식전7d지전2d접수BU/CY방안예처리후,경미정맥주사PBS 0.2 mL;단순골수세포이식조(BMT조)(n=20):BU/CY방안예처리후,경미정맥주사공자골수세포；EPC연합이식조(EPC조)(n=20):BU/CY방안예처리후,경미정맥주사공자골수세포급EPC.우이식후제14,21,28천,채용류식세포술검측각조소서골수중B계조(ProB)세포(CD19+ c-kit+ IgM-)수량、흉선중CD4+,CD8+,쌍양성(DP)급쌍음성(DN)T림파세포비례급비장중CD4+,CD8+T림파세포급B림파세포비례,병통과소목소-이홍염색급면역조화염색관찰중추화외주면역기관조직결구적회복급손상혈관내피적수복정황.결과 이식후BMT급EPC조소서골수중ProB세포수량지속증고,제14천시,EPC조(0.83×106)현저고우BMT조(0.14×106)(x2=7.2,P＜0.05)；흉선중DP세포비례우제21천체봉치,EPC조(0.58％)교BMT조(0.39％)현저증고(x2=6.49,P＜0.05)；비장중CD4+T세포비례지속증고,제28천시,EPC조(3.64％)교BMT조(2.47％)현저증고(x2=7.2,P＜0.05)；CD8+T세포비례재이식후제21천체봉치,EPC조(3.64％)교BMT조(3.21％)현저증고(x2=5.42,P＜0.05).조직병이학검사야현시EPC조소서적중추급외주면역기관조직결구회복교BMT조쾌,조직중실질세포수량회복교BMT조쾌；대비장혈두내피면역조화염색적결과야현시,EPC조소서비혈두내피완정성적회복교BMT조쾌.결론 BU/CY예처리소서이기인골수이식중,연합EPC수주가촉진이식수서면역기관손상혈관내피적수복、촉진기조직결구적회복,종이유효촉진이식후적면역중건.
Objective To study the effects of endothelial progenitor cell (EPC) on immune reconstitution in mice undergoing allo-geneic bone marrow transplantation after busulfan/cyclophosphamide (BU/CY) pretreatment.Methods Fifty eight healthly SPF male BALB/c mice aged (8-10) weeks and weighted (22-24)g were randomly divided into control group (n=9),which injected with 0.2 mL phosphate buffer solution (PBS) ; BU/CY pretreatment group (n=9),which treated with BU/CY from day-7 to day -2 and injected with 0.2 mL PBS; bone marrow cells transplantation (BMT) group (n=20),which injected with donor derived bone marrow cells after BU/CY pretreatment; EPCs combined with BMT (EPC) group (n=20),which injected with donor derived bone marrow cells and EPCs after BU/CY pretreatment.The number of progenitor B (ProB) cells (CD19+ c-kit+ IgM-) in the bone marrow and the percentages of CD4+,CD8+,double positive (DP),double negative(DN)T cells in the thymus and the percentages of CD4+,CD8+ T cells and B cells in the spleen were detected by flow cytometry.The morphology of immune organs was observed by the light microscope to evaluate the repair of tissues and injured endothelial cells.Results The number of ProB cells in BMT group and EPC group increased continuously after transplantation,and compared with the BMT group (0.14 × 106),the EPC group (0.83×106) increased significantly on day 14 (x2 =7.2,P＜0.05).The percentage of DP cells in the thymus reached the highest peak on day 21,and increased significantly in the EPC group (0.58％) compared with the BMT group (0.39％) (x2 =6.49,P＜0.05).After transplantation,the percentage of CD4+ T cells in the spleen also had gradual increase,and the EPC group (3.64 ％) increased significantly compared with the BMT group (2.47％) (x2 =7.2,P＜0.05) on day 28;The percentage of CD8+ T cells in the spleen peaked on day 21,with the EPC group (3.64％) increased significantly compared with the BMT group (3.21％) (x2 =5.42,P＜ 0.05).The histopathological examination revealed that the repair of damaged tissue in the central and periphery immune organs and the increase of parenchymal cell number accelerated in the EPC group compared with the BMT group,the immunohistochemical staining of the spleen sinusoids also revealed a faster recovery of the integrity of endothelial cells in the EPC group than the BMT group.Conclusions EPC infusion along with bone marrow cells could repair injured blood vessels,accelerate the repair of damaged immune organs,and promote the process of immune reconstruction after allo-HSCT.
