Ly49A转基因淋巴细胞对异基因骨髓移植后GVHD和GVL的影响

Effect of Ly49A transfected lymphocytes on graft versus host disease and graft versus leukemia effect after allogeneic bone marrow transplantation

目的 观察Ly49A基因转染淋巴细胞对异基因骨髓移植后移植物抗宿主病 (GVHD)和移植物抗白血病效应 (GVL)的影响。方法 经逆转录病毒介导将Ly49A基因转染至C57BL/ 6小鼠的淋巴细胞 ,以C57BL/ 6(H 2 b)小鼠为供者 ,以接种EL961 1红白血病细胞的BALB/c(H 2 d)小鼠为受者 ,进行脾淋巴细胞和骨髓细胞联合移植 ,建立异基因急性GVHD模型 ,观察Ly49A基因转染的淋巴细胞对GVHD和GVL的影响。结果 在未进行移植的单纯照射组 ,小鼠的存活时间为 (6 .50±2 .41 )d ;仅以环磷酰胺治疗的小鼠存活时间为 (2 0 .90± 2 .88)d ;非转染淋巴细胞和骨髓细胞联合移植组的存活时间为 (1 7.1 0± 4 .65)d ;空载体转染淋巴细胞和骨髓细胞联合移植组的存活时间为(1 7.40± 5 .32 )d ;Ly49A转染淋巴细胞与骨髓细胞联合移植组的存活时间为 (35 .2 0± 1 2 .52 )d ,较上述各组明显延长 (P =0 .0 0 0 )。

Objective To observe the effect of Ly49A transfected lymphocytes on graft versus host disease (GVHD) and graft versus leukemia (GVL) effect post allogeneic bone marrow transplantation. Methods Ly49A gene was transfected into lymphocytes of C57BL/6 mice by retrovirus and the expression rate of Ly49A receptor was detected by flow cytometer. The murine model of allogeneic acute GVHD was established in this way: the donor was C57BL/6(H 2 b). The recipient was BALB/c(H 2 d), which was injected EL9611 cells. After the irradiation (TBI, 60 Co 9.0 Gy) the recipient accepted the transplantation of mixed spleen cells and bone marrow cells to establish the GVHD model. The effects of Ly49A transfected spleen cells on GVHD and GVL post allogeneic bone marrow transplantation were detected. Results The expression rate of Ly49A receptor was ( 42.20 ± 4.87 )?% for lymphocytes transfected with pLXSN Ly49A; ( 18.67 ± 2.48 )?% for lymphocytes transfected with pLXSN and ( 18.73 ± 3.82 )?% for untransfected control respectively. In the case of allo BMT (C57BL/6 H 2 b→BALB/c H 2 d), survival time was ( 6.50 ± 2.41 ) days for the irradiation group; ( 20.90 ± 2.88 ) days for cyclophosphomide therapy group; ( 17.10 ± 4.65 ) days for bone marrow cells mixed with lymphocytes transplantation group; ( 17.40 ± 5.32 ) days for bone marrow cells mixed with pLXSN transfected lymphocytes transplantation group; ( 35.20 ± 12.52 ) days for bone marrow cells mixed with Ly49A transfected lymphocytes transplantation group, which was much longer than the survival time of any of the above group ( P = 0.000 ). Conclusion The Ly49A transfected lymphocytes transplantation could alleviate GVHD and retain GVL effect on acute GVHD model post allo BMT.