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目的:本研究通过非清髓性预处理方案联合髓腔内骨髓移植建立异基因小鼠免疫耐受模型,并探讨其诱导耐受的机理。方法:受鼠C57BL/6(B6)于第0天接受60Coγ射线全身照射(TBI,550 cGy),4 h内输注雄性BALB/C(H-2d)小鼠来源的骨髓细胞,2 d后腹腔注射环磷酰胺(CTX,200 mg.kg-1)。通过皮肤移植、混合淋巴细胞反应(mixed lymphocyte reaction,MLR)检测耐受状态,并应用体外过继转移实验I、L-2逆转实验等探讨免疫耐受机制。结果:经骨髓移植的B6小鼠对BALB/C小鼠的皮肤移植物平均存活时间超过300 d,较空白组明显延长(P<0.001);MLR结果证明B6小鼠获得供体特异性耐受,该耐受可以被IL-2逆转且可被过继转移;所有受鼠均未出现GVHD表现。结论:非清髓预处理联合髓腔内骨髓移植可以有效诱导异基因小鼠的免疫耐受。
OBJECTIVE: To establish a model of immune tolerance in allogeneic mice by non-myeloablative preconditioning combined with intramedullary bone marrow transplantation and to explore its mechanism of induction tolerance. METHODS: Bone marrow cells from male BALB / C (H-2d) mice were infused within 4 h after C57BL / 6 (B6) exposure to 60Co γ-rays (TBI, 550 cGy) on day 0 Cyclophosphamide (CTX, 200 mg.kg-1) was injected intraperitoneally. Tolerance status was tested by skin graft and mixed lymphocyte reaction (MLR), and adoptive transfer assay I and L-2 reversal assay were used to investigate the mechanism of immune tolerance. Results: The average survival time of BALB / C mice skin grafts was longer than 300 days in bone marrow-transplanted B6 mice, which was significantly longer than that in blank control group (P <0.001). MLR results showed that donor-specific tolerance , This resistance can be reversed by IL-2 and can be adoptively transferred; no manifestation of GVHD occurs in all mice. Conclusion: Non-myeloablative preconditioning combined with intramedullary bone marrow transplantation can effectively induce immune tolerance in allogeneic mice.