小鼠非清髓性单倍体相合骨髓移植模型的建立  

Establishment of Haploidentical Bone Marrow Transplantation Model in Nonmyeloablative Conditioning

作　　者：李凤[1,2] 段连宁[2] 王喆[2] 罗渊[2] 陆承荣[2] 向培德[2] 纪树荃[2] 

机构地区：[1]第四军医大学,西安710032 [2]中国人民解放军空军总医院,北京100142

出　　处：《实验动物科学》2011年第3期6-10,共5页Laboratory Animal Science

基　　金：国家自然科学基金资助项目(No.30873014)

摘　　要：目的建立小鼠非清髓性单倍体相合骨髓移植模型,为研究移植前诱导免疫耐受或移植后输注供者细胞促进植入提供研究平台。方法以CB6F1雌性小鼠为受鼠,移植前1 d予450 cGy全身照射(TBI)后,随机分为2组,实验组移植0 d输注C57BL/6雄性小鼠骨髓有核细胞5×107/只,对照组不予移植。然后监测受鼠造血恢复、检测供鼠性别决定基因(SRY)判断植入情况,以及外周血供者细胞尤其CD3+细胞嵌合状态,同时观察小鼠急性移植物抗宿主病(aGVHD)的发生情况。结果对照组小鼠均存活,仅表现轻度aGVHD,血象移植后30 d内基本恢复正常水平。实验组小鼠SRY基因在移植后+14 d、+30 d、+60 d时检测PCR结果均阳性,供鼠外周血淋巴细胞、单核细胞、粒细胞嵌合率在移植后14 d分别为23.8%、36.9%%、19.4%;30 d分别为49.9%、53.2%、54.4%;60 d分别为67.6%、51.6%、56.9%,其中CD3+细胞嵌合率分别为4.4%、21.2%、54.4%。结论 450 cGyTBI的非清髓性预处理方案,可以诱导受鼠免疫耐受、供者骨髓细胞植入,嵌合率处于中低水平混合嵌合状态。Objective To establish the haploidentical bone marrow transplantation mouse model in nonmyeloablative conditioning,which will provide a platform for studies on inducing immune tolerance and promoting engraftment after transplantation.Methods One day before transplantation,the receptions CB6F1female mice were exposed to a dose of 450cGy total body irradiation（TBI）.Then they were divided into two groups randomly,and each mouse of the experiment group was transfused with 5×107 nucleated bone marrow cells from the donor C57BL/6 male mice.Recovery of receptions hemogarm,engraftment of the sex determining gene（SRY）,chimerism of the donor cells,especially the CD3＋ cells,and signs of aGVHD after transplantation,were examined.Results All control mice survived and the hemogram almost recovered to normal level within thirty days after transplantation.Most control mice shown slight graft-versus-host disease.For the experiment group,the sex determining gene SRY can be detected in the peripheral blood from receptions.Chimerism of the donor cells at day 14,30 and 60 after transplantation was 23.8%,49.9%,67.6% in lymphocytes,36.9%,53.2%,51.6% in monocytes,and 19.4%,54.4%,56.9% in granulocytes.In CD3＋ cells,chimerism rates were 4.4%,21.2%,and 54.4%,respectively.No significant signs of graft-versus-host disease were observed.Conclusion In the nonmyeloablative conditioning,single dose 450 cGy TBI one day before transplantation can induce immune tolerance of receptions and enable donor bone marrow cells engraftment.The chimerism of donor cells was low until the end of observation.

关 键 词：非清髓性预处理 单倍体骨髓移植 动物模型 

分 类 号：R551.3[医药卫生—血液循环系统疾病]