miHA不合异基因骨髓移植后慢性移植物抗宿主病小鼠模型的建立  

Establishment of a Murine Model of Chronic Graft-Versus-Host Disease After Minor Histocompatibility Antigen Mismatched Bone Marrow Transplantation

作　　者：黄欣[1] 翁建宇[1] 吴萍[1] 童嘉琦[1] 杜欣[1] 

机构地区：[1]广东省医学科学院广东省人民医院肿瘤中心血液科,广州510080

出　　处：《国际输血及血液学杂志》2014年第2期97-101,共5页International Journal of Blood Transfusion and Hematology

基　　金：国家自然科学基金资助项目（81270648,81100384,81370665）;广东省自然科学基金资助项目（S2012010009560）;卫生公益性行业科研专项经费项目（201202017）

摘　　要：目的 通过主要组织相容性复合物（MHC）相合,次要组织相容性抗原（miHA）不合的异基因骨髓移植,建立慢性移植物抗宿主病（cGVHD）小鼠模型,为cGVHD研究提供活体模型.方法 选择20只BALB/c（H2d）雌鼠为研究对象.将其分为空白对照组、放射对照组、移植对照组和cGVHD实验组,每组5只.除空白对照组外,其余各组小鼠均经直线加速器700 cGy的全身照射（TBI）.放射对照组照射后,经尾静脉输注RPMI 1640培养液0.2 mL;移植对照组输注miHA不合的B10.D2（Hc1 H2d H2-T18c）雄鼠骨髓细胞8×106个;cGVHD组输注骨髓细胞与脾细胞各8×106个.观察小鼠移植后造血重建、植入及一般状态.小鼠移植14d后每3d进行一次临床评分,50 d后处死,评估靶器官病理变化（实验过程中对动物的处置符合动物伦理学标准）.结果 移植对照组小鼠在移植后7d均获得造血重建,至实验结束（移植后50 d）全部存活,且染色体均为供者型.cGVHD实验组小鼠移植后20 d受累皮肤临床评分均＞0.6分;50 d时,皮肤、肝脏、肺的病理改变典型,病理评分明显高于对照组（F=88.02,P＜0.05）.结论 照射剂量为700 cGy,输注骨髓细胞数为8×106个、脾细胞数为8×106个的小鼠能成功诱导出cGVHD（造模成功）,可作为临床研究骨髓移植后cGVHD的理想模型.Objective To establish a murine model of chronic graft-versus-host disease （cGVHD） after major histocompatibility complex （MHC） matched and minor histocompatibility antigen （miHA） mismatched allogeneic bone marrow transplantation,and to provide in vivo research tools for cGVHD.Methods 20 recipients BALB/c （H2d） female mice were included in this study.They were divided into blank control group （n =5）,irradiation control group （n =5）,transplantation group （n =5） and the cGVHD group （n=5）.In addition to the blank control group,the rest of the three groups of mice were irradiated with 700 cGy dose of linear accelerator.Then the irradiation control group were injected with 0.2 ml PPMI 1640; the trarsplantation group were injected with 8 × 106 bone-marrow cells,and cGVHD group were injected with bone-marrow cells and spleen cells （8× 106,1 ∶ 1） from B10.D2（Hc1 H2d H2-T18c） male mice.The observed items post-transplantation included hematopoietic reconstruction,implant,and general condition.Clinical scores were assessed every 3 days after ＋14 d.At ＋ 50 d,mice were put to death for evaluation target organ pathological changes.Animal Intervention met animal ethical standard.Results Mice in transplantation groups were in hematopoietic reconstruction at ＋7 d,and all survived to the end point （＋ 50 d）.Chromosomes of recipient mice were in donor form.Clinical scores of cGVHD group have been more than 0.6 since ＋20 d.Pathological changes of skin,liver and lung were obviously,and pathological scores were significant higher than those of transplantation control groups （F=88.02,P＜ 0.05）.Conclusions Irradiation dose for 700 cGy,8× 106 of bone marrow and spleen cell number infusion induce a successful cGVHD murine model,which might be an ideal study model of clinical cGVHD after bone marrow transplantation.

关 键 词：骨髓移植 次要组织相容性抗原 移植物抗宿主病 小鼠 

分 类 号：R457.7[医药卫生—治疗学]