Ly49A转基因小鼠脾细胞对半相合异基因骨髓移植后GVHD和GVL的作用

Effect of Ly49A transfected mouse spleen cells on graft versus host disease and graft versus leukemia after haploidentical allogeneic bone marrow transplantation in mice

目的 观察Ly4 9A基因转染的C5 7BL 6小鼠脾细胞对半相合异基因骨髓移植 (allo BMT)后移植物抗宿主病 (GVHD)和移植物抗白血病效应 (GVL)的影响。方法 经由逆转录病毒介导将Ly4 9A基因转染至C5 7BL 6小鼠的脾细胞 ;流式细胞仪检测Ly4 9A受体在转染后脾细胞上的表达率 ;以亲代C5 7BL 6H 2 b 小鼠为供者 ,以接种EL96 11红白血病细胞的 (BALB c×C5 7BL 6 )F1H 2 d b(CB6F1)小鼠为受者 ,预处理条件为全身照射 (TBI,6 0 Co照射 10 .5Gy) ,进行脾细胞混合骨髓细胞移植 ,建立半相合异基因急性GVHD模型并在该模型基础上观察Ly4 9A基因转染的脾细胞对GVHD和GVL的作用。结果 Ly4 9A基因转染的C5 7BL 6小鼠的脾细胞 2 4h后蛋白表达率为 (42 .2 0± 4 .87) % ,空载体转染组为(18.6 7± 2 4 8) % ,未转染对照组为 (18.73± 3.82 ) % ,在半相合异基因移植中 (C5 7BL 6 H 2b →CB6F1H 2d b) ,未进行移植的单纯照射组生存期为 (7.80± 3.36 )d ;环磷酰胺治疗组生存期为 (2 1.70±2 87)d ;脾细胞混合骨髓细胞移植组生存期为 (2 9.4 0± 6 .4 3)d ;空载体转染的脾细胞混合骨髓细胞移植组生存期为 (2 9.10± 7.39)d ;Ly4 9A转染的脾细胞混合骨髓细胞移植组生存期为 (45 .0 0± 12 .38)d ,较上述各组生存期明显延长

Objective To observe the effect of Ly49A transfected mouse spleen cells on graft versus host disease (GVHD) and graft versus leukemia (GVL) effect after haploidentical allogeneic bone marrow transplantation in mice. Methods Ly49A gene was transfected into spleen cells of C57BL/6 mice by retrovirus and the expression rate of Ly49A receptor was evaluated by flow cytometry. The murine model of haploidentical allogeneic acute GVHD was established by using C57BL/6 H-2b mouse as donor, and (BALB/c×C57BL/6)F1 H 2d/b (CB 6F 1) mouse as the recipient which was injected EL9611 cells before transplantation. After irradiation (TBI, 60 Co 10.5 Gy), the recipient received mixed graft of spleen cells and bone marrow cells to establish a GVHD model. The effects of Ly49A transfected spleen cells on GVHD and GVL post haploidentical allogeneic bone marrow transplantation were detected with this model. Results The expression rate of Ly49A receptor was (42.20±4.87)%,(18.67±2 48)% and (18 73±3.82)% for pLXSN Ly49A, pLXSN transfected and untransfected spleen cells respectively. Among haploidentical allo BMT (C57BL/6 H-2b →CB6F1 H-2d/b ) groups, the survival time was (7.80±3.36) days for irradiation group; (21.70±2.87) days for cyclophosphomide therapy group; (29.40±6.43) days for mixed bone marrow cells and spleen cells transplantation group; (29.10±7.39) days for mixed bone marrow cells and pLXSN transfected spleen cells transplantation group and (45.00±12.38) days for mixed bone marrow cells and Ly49A transfected spleen cells transplantation group, which was much longer than that of any other groups (P=0.000). Conclusion The Ly49A transfected spleen cell transplantation could alleviate GVHD and retain GVL effect in the acute GVHD model post haploidentical allo BMT.