低剂量氟达拉滨、环磷酰胺联合供者NK细胞作为非清髓性预处理用于小鼠单倍相合造血干细胞移植.doc

1 低剂量***达拉滨、环磷酰***联合供者 NK 细胞作为非清髓性预处理用于小鼠单倍相合造血干细胞移植作者:于津浦, 曹水, 辛宁, 安秀梅, 齐静, 任秀宝【摘要】本研究探讨低剂量***达拉滨、环磷酰***联合供者异体反应性 NK 细胞( flu+cy+allo 拟 NK ) 作为新的非清髓性单倍相合造血干细胞移植( haploidentical HSCT )预处理方案的可行性。利用免疫磁珠富集 F1 供鼠(H拟 2d/b) 脾脏 NK 细胞,检测其中 Ly49C+ 、 Ly49A+ 细胞的比例; LDH 法检测其异体反应性。建立小鼠单倍相合造血干细胞移植模型, 并比较清髓性方案( 9 Gy TBI ) 、各种非清髓性方案( Gy TBI , flu+cy ,及 flu+cy+allo 拟 NK) 的体内清髓效果、移植后供者嵌合率、 GVHD 的发生率及严重程度。结果表明: 与其他非清髓性预处理方案相比, flu+cy+allo 拟 NK 组不能增加清髓程度,但可显著提高单倍相合移植后的供者嵌合率,移植后 21 天的嵌合率在骨髓为( ± )% ,脾脏为( ± )% , 并持续 2 个月。与 flu+cy 组相比, flu+cy+allo 拟 NK 组出现的 GVHD 反应轻微,仅有 50%(5/10) 受鼠出现 2 体重减轻, flu+cy+allo 拟 NK 组小鼠的肝脏、小肠、肾脏及皮肤的病理切片均未见明显的组织损伤。结论: 供者 allo 拟 NK 具有促进单倍相合供者造血干细胞植入, 减轻 GVHD 强度的作用;低剂量 flu+cy+allo 拟 NK 方案为高龄和一般情况差的肿瘤患者开展单倍相合造血干细胞移植提供了新的途径。【关键词】非清髓性预处理 Application of Low Dose of Fludarabine and bined with Donor NK Cells asa Non 拟 myeloablative Conditio 拟 ning Regimen for the Haploidentical Hematopoietic Stem Cell Transplantation in Mice Department of Immunology, Key Laboratory of Cancer Prevention and The rapy, Tianjin Cancer Institute and Hospital, Tianjin Medical University, Tianjin 300060, China Abstract This study was aimed to investigate the feasibity of low dose of fludarabine, bined with donor derived alloreactive NK cells asa new nonmyeloablative 3 conditioning regimen in the haploidentical hematopoietic stem cell transplantation (haploidentical HSCT). F1 derived 拟 NK cells were enriched with MACS ic separation system, in which the proportions of the Ly49C+ and Ly49A+ cells were detected by flow cytometry and the alloreactivity was measured by LDH method. The haploidentical HSCT models were constructed, and the myeloablativity in vivo, donor engraftment and the intensity of GVHD pared between different myeloablative and nonmyeloablative conditioning regimens, including 9 Gy TBI, Gy TBI, flu+cy, and flu+cy+allo 拟 NK. The results showed that the flu+cy+allo 拟 NK conditioning was nonmyeloablative, but the rate of donor chimerism after haploidentical HSCT was si