H-2单倍体相合小鼠双供体造血干细胞移植模型的建立

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目的:建立新的H-2单倍体相合小鼠双供体造血干细胞移植模型并与经典的造血干细胞移植进行比较,以减轻移植的相关并发症。方法:建立H-2单倍体相合小鼠双供体外周血造血干细胞移植模型并与8 Gy预处理移植组进行比较。在经典移植组给予CB6F1受鼠8 Gy TBI预处理,2 h内回输经G-CSF动员的供体(雄性C57)脾单个核细胞(spMNC)3×10~7;在双供体移植(DHSCT)组给予CB6F1受鼠2 Gy TBI,2 h内回输H-2单倍体相合小鼠双供体来源的spMNC共12×10~7(每个供体各6×10~7),依据供鼠组品系和性别不同,双倍体移植组又再分为3组:A组为雄性C57+雌性BALB/c,B组为雄性C57+雄性BALB/c,C组为雄性C57+雄性C3H。观察4组的造血重建、移植物植入、GVHD及存活情况。结果:经典移植组出现严重造血抑制,WBC<1×10~9/L持续3-5d;A、B、C各组未出现造血抑制(WBC>3×10~9/L)。经典移植组快速植入,1周达到外周血完全植入;A、B、C 3组1周达混合植入,2周达完全植入。经典移植组34 d GVHD发生率及致死率均为100%。双供体移植(DHSCT)组中34 d总体GVHD发生率及致死率分别为49.6%、50%(P<0.01,P<0.05);50 d分别为60.4%和81.2%,50 d总体存活率为50.9%。A、B、C各组的造血重建、供体植入、GVHD发生率、GVHD致死率、OS等均无显著差异(P>0.05)。结论:采用2 Gy TBI预处理、双供者细胞输注、无GVHD预防,可使供体完全稳定植入、无造血抑制、GVHD发生率及死亡率明显减少;研究表明,H-2单倍体相合小鼠双供体造血干细胞移植模型成功建立。 OBJECTIVE: To establish a new model of haploidentical mouse double donor hematopoietic stem cell transplantation in H-2 and to compare with classical hematopoietic stem cell transplantation in order to reduce the complications related to transplantation. Methods: A double donor peripheral blood stem cell transplantation model of H-2 haploidentical mouse was established and compared with 8 Gy preconditioning group. In the classic transplantation group, CB6F1 was pretreated with 8 Gy TBI and donor splenic mononuclear cells (MMNCs) 3 × 10 ~ 7 mobilized by G-CSF within 2 h. DHSCT group was given 2 Gy TBI to CB6F1 mice, and 12 × 10 ~ 7 sporozoites (6 × 10 ~ 7 per donor) were transfused from H-2 haploidentical mouse double donor in 2 h. The diploid group was subdivided into three groups: male C57 + female BALB / c in group A, male C57 + male BALB / c in group B and male C57 + male C3H in group C, depending on the strain and sex of donor. The hematopoietic reconstitution, graft implantation, GVHD and survival were observed in the four groups. Results: The severe hematopoietic suppression occurred in the classic transplantation group. WBC <1 × 10 ~ 9 / L lasted for 3-5 days. No hematopoietic suppression occurred in all the three groups (WBC> 3 × 10 ~ 9 / L). The classic transplantation group was quickly implanted, and the peripheral blood was completely implanted one week later. The rats in group A, B and C were mixed for 1 week and completely implanted in 2 weeks. The incidence and mortality of GVHD in the classic transplantation group were both 100% at 34 days. The 34-day overall incidence and mortality of GVHD in DHSCT group were 49.6% and 50%, respectively (P <0.01, P <0.05). The 50-day and 60-day survival rates were 60.4% and 81.2% Is 50.9%. There was no significant difference in hematopoietic reconstitution, donor implantation, GVHD incidence, GVHD mortality and OS among groups A, B and C (P> 0.05). CONCLUSION: Pretreatment with 2 Gy TBI and double donor cell infusion without GVHD prophylaxis can completely implant the donor with no hematopoietic suppression and significantly reduce the incidence of GVHD and mortality. Studies have shown that H-2 haploidentical Body matched mice double donor hematopoietic stem cell transplantation model was successfully established.
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