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本研究观察异基因NK细胞在白血病小鼠单倍体相合骨髓移植中的作用。建立荷EL9611(H-2d)红白血病细胞的CB6F1H-2b/d小鼠动物模型,5天后将其随机分为7组,每组10只。CB6F1H-2b/d受鼠为移植组,9Gy(60Co)照射后以C57BL/6H-2b为供鼠,进行骨髓移植。对照组分为5组:对照1组为未治疗组;对照2组为单纯照射组;对照3组为阿糖胞苷治疗组:小鼠腹腔注射阿糖胞苷50mg/kg×6d;对照4组为单纯骨髓细胞移植组:小鼠照射后4小时移植C57BL/6H-2b小鼠骨髓细胞;对照5组为GVHD对照组:小鼠照射后4小时移植C57BL/6H-2b小鼠骨髓细胞和脾细胞。实验组分为2组:实验A组,照射后输注C57BL/6H-2b小鼠NK细胞1×106/只,4小时后移植C57BL/6H-2b小鼠骨髓细胞;实验B组:照射后同实验1组,4小时后移植C57BL/6H-2b小鼠骨髓细胞和脾细胞。以血象、体重、生存期和病理组织学变化为观察指标并作组间比较。结果表明,对照1、2、3、5组小鼠生存期分别为(10.10±0.88)天、(9.80±0.92)天、(22.70±3.23)天、(20.10±1.73)天,对照4组生存期为(30.10±15.95)天,其中2只小鼠超过30天;实验A、B组小鼠生存期分别为(39.10±18.11)天、(49.30±17.24)天,实验1组4只小鼠生存期超过30天,实验2组7只生存期超过30天,其中实验1组与对照1、2、3、5组比较均有显著性差异(p<0.01),实验2组同其他组比较均有显著性差异(p<0.05)。因白血病死亡的小鼠,可见肝脾明显肿大,结构破坏,有大量白血病细胞浸润。实验组长期存活的小鼠出现Y染色体(嵌合率80%-90%)。结论:在荷EL9611(H-2d)红白血病细胞CB6F1H-2b/d小鼠模型的单倍体相合骨髓移植中供者NK细胞具有杀灭白血病细胞并降低GVHD的双重作用。
This study was to observe the role of allogeneic NK cells in haploidentical bone marrow transplantation in leukemia mice. The animal model of CB6F1H-2b / d mice bearing EL9611 (H-2d) erythroleukemia cells was established. After 5 days, they were randomly divided into 7 groups of 10 mice. CB6F1H-2b / d mice were transplanted, and irradiated with 9Gy (60Co) irradiated C57BL / 6H-2b mice for bone marrow transplantation. The control group was divided into five groups: the control group 1 was untreated group, the control group 2 was treated with irradiation alone, and the control group 3 was cytarabine-treated group: mice were given intraperitoneal injection of cytarabine 50 mg / kg × 6 d. Control 4 C57BL / 6H-2b mouse bone marrow cells were transplanted 4 hours after irradiation in mice; control group 5 was GVHD control group: C57BL / 6H-2b mice bone marrow cells were transplanted 4 hours after irradiation and Spleen cells. The experimental groups were divided into two groups: experimental group A, irradiated with 1 × 106 NK cells of C57BL / 6H-2b mice and irradiated with C57BL / 6H-2b mouse bone marrow cells after 4 hours. The same experimental group 1, C57BL / 6H-2b mouse bone marrow cells and spleen cells were transplanted 4 hours later. Blood, weight, survival and histopathological changes were observed and compared between groups. The results showed that the survival time was (10.10 ± 0.88) days, (9.80 ± 0.92) days, (22.70 ± 3.23) days and (20.10 ± 1.73) days respectively in control group 1, (30.10 ± 15.95) days, of which two mice over 30 days; experimental group A, B mice survival was (39.10 ± 18.11) days, (49.30 ± 17.24) days, experimental group 1 4 mice Survival of more than 30 days, the experimental group 2 7 survival time of more than 30 days, including experimental group 1 and control groups 1,2,3,5 were significantly different (p <0.01), experimental group 2 compared with other groups There was significant difference (p <0.05). Mice died of leukemia, liver and spleen can be seen significantly enlarged, structural damage, a large number of leukemia cell infiltration. Mice surviving in the experimental group developed Y chromosome (chimerism rate 80% -90%). CONCLUSION: Donor NK cells have the dual effect of killing leukemia cells and reducing GVHD in haploidentical bone marrow transplantation of EL9611 (H-2d) erythroleukemia cell CB6F1H-2b / d mouse model.