外周血干细胞骨髓腔内输注对移植物抗宿主病影响.docVIP

外周血干细胞骨髓腔内输注对移植物抗宿主病影响 　 作者：毕延智,曾冬香,盛桂凤,张琰,陈宝安 【摘要】 本研究建立髓腔内外周造血干细胞输注(IBMPBSCT)动物移植模型并观察该方法对移植物抗宿主病(GVHD)的影响。选取雌性C57BL/6小鼠为移植受鼠，于第0天接受全身照射(TBI) 4Gy后，输注经rhGCSF动员的雄性BABL/c小鼠外周血造血干细胞(1×107)，2天后腹腔注射环磷酰胺(CTX)，观察各组小鼠GVHD发生情况。结果显示：髓腔内PBSCT组的受鼠GVHD发生率较尾静脉输注组明显减低(plt;0.05)，移植后在受鼠体内观察到供鼠来源的Y染色体存在。 结论：与尾静脉输注比较，髓腔内输注外周血造血干细胞不仅能减少GVHD的发生，而且更有利于干细胞植入。 【关键词】 移植物抗宿主病 Influence of Intrabone Marrow Infusion of Peripheral Hematopoietic Stem Cells on GraftVersusHost Disease Abstract This study was aimed to explore whether the graftversushost disease (GVHD) could be alleviated by intrabone marrow (IBM) infusion of allogeneic hematopoietic stem cells. Female C57BL/6 mice as recipients received total body irradiation (TBI) 4 Gy on day 0, followed by injection of peripheral hematopoietic stem cells (1×107) from mobilized male BALB/c with granulocytecolony stimulating factor (rhGCSF), and cyclophosphamide (200 mg/kg) was injected intraperitoneally two days later. The results showed that the incidence and severity of GVHD were more low and alleviative in group IBMPBSCT than that in group TVPBSCT (plt;0.05). Y chromosome of doenor mice could be detected in the bone marrow of recipient mice. It is concluded that the method of intrabone marrow infusion is superior to injection via the tail vein in the engraftment of hematopoietic stem cells in terms of stem cell homing while the frequency and severity of GVHD in allogeneic mice decrease. Key words hematopoietic stem cell transplantation; intrabone marrow infusion; graftversushost disease 外周血来源的造血干细胞移植因其干细胞采集方便、供者痛苦少、移植后造血和免疫功能重建快等优势已成为目前治疗某些恶性血液病和自身免疫性疾病的重要方法，但移植后严重的移植物抗宿主病(GVHD)的发生在一定程度上限制了其应用。有证据表明，大量造血干细胞移植可以克服主要组织相容性复合物(MHC)屏障，促进造血干细胞的植入(大剂量效应)［1］。我们将供者骨髓细胞直接注入到受者骨髓腔内(IBMBMT)，成功建立了稳定的免疫耐受模型，且未观察到GVHD的发生［2］。在此基础上，我们进一步实验证实了重组人粒细胞集落刺激因子(rhGCSF)动员的外周血干细胞髓腔内输注较尾静脉输注更有利于促进嵌合体形成，减少GVHD发生，现报道如下。 材料和方法 实验动物 供鼠为雄性BALB/c小鼠(H2d)，8-10周龄，体重18-20 g；受鼠为雌性C57BL/6小鼠(H2b,B6)，8-10周龄，体重18-20 g，均为清洁级，购自上海SLAC实验动物有限责任公司。 主要试剂 环磷酰胺（