TBI与环磷酰胺选择性去除同种异基因反应 供者淋巴细胞的研究.docVIP

TBI和环磷酰胺选择性去除同种异基因反应 供者淋巴细胞的研究 作者：陈宝安，董伟民, 丁家华，孙雪梅，邓晓静, 张琰，毕延智 赵刚, 高冲，孙耘玉，王骏，程坚，Schmitt M2 ，Schmitt A2 【摘要】 　　本 研究 用全身照射(TBI)和环磷酰胺（CY）选择性去除同种异基因反应的供者淋巴细胞,为预防移植物抗宿主病(GVHD)的发生探索新的手段。以(BALB/c×C57BL/6)F1雌性小鼠（H2d/b）为受鼠，于第0天接受亚致死量的60Coγ射线全身照射，总剂量为4 Gy，第1天接种P388D1白血病细胞，第2天输注由C57BL／6雄性小鼠（H2b）为供鼠提供的MHC不匹配的供者脾淋巴细胞，在造血干细胞移植前诱导移植物抗白血病效应（GVL）。第6天腹腔注射环磷酰胺（CY）200 mg/kg或再次TBI 9 Gy，选择性去除同种异基因反应供者淋巴细胞，第7天输注(BALB/c×C57BL/6)F1雄性小鼠（H2d/b）提供的骨髓造血干细胞。结果显示：以CY和TBI选择性去除同种异基因反应供者淋巴细胞组的小鼠无白血病和GVHD的发生，生存期超过了210天，于移植后第21天出现完全供者嵌合，然后嵌合率下降，第90天表现为混合嵌合体（MC）。对照组的小鼠出现了白血病和GVHD，出血、感染明显，生存期短，为20-36天（P0.01）。结论：同基因骨髓移植前输注不相匹配的供者脾细胞诱导GVL效应,然后再通过TBI和CY选择性去除同种异基因反应的供者淋巴细胞来预防移植物抗宿主病(GVHD)的发生是可能的。 【关键词】 同种异基因造血干细胞移植；同种异基因反应供者淋巴细胞；移植物抗白血病效应；移植物抗宿主病；全身照射；环磷酰胺 　　Selective Elimination of Alloreactive Donor Lymphocytes by Using TBI and Cyclophosphamide Abstract This study was aimed to investigate a new method of avoiding graftvshost disease (GVHD) through selective elimination of alloreactive donor lymphocytes by using total body irradiation (TBI) and cyclophosphamide (Cy). Female (BALB/c×C57BL/6)F1 mice (H2d/b) as recipients recEived 60Co γray sublethal TBI of 4 Gy on day 0 followed by bEIng inoculated with P388D1 leukemia cell line on day 1,injection of allogeneic splenocytes from C57BL／6 male mice（H2b） was carried out for induction of graftvsleukemia (GVL) effect prior to stem cell transplantation (SCT) , intraperitoneally injection of cyclophosphamide (Cy) (200 mg/kg) and TBI (9 Gy) was given on day 6. One day later, treated mice were rescued with bone marrow hematopoietic stem cells from (BALB/c×C57BL/6)F1 male mice（H2d/b）. The results showed that recipients had no occurrence of leukemia and GVHD through selective elimination of alloreactive donor lymphocytes by Cy and TBI, survived more than 210 days，the completedonor chimerism occured on day 21 after transplantation. The ratio of chimerism descended subsequently, but still displayed mixedchimerism at 90 days. Con