非清髓性单倍体骨髓移植后输注供体免疫细胞治疗白血病的研究

本研究在小鼠非清髓性单倍体骨髓移植模型上,观察移植后早期输注供体免疫细胞对促进植入及改善疗效的作用。以CB6F1雌性小鼠为受鼠,C57BL/6雄性小鼠为供鼠,移植前4 d经尾静脉注射小鼠红白血病细胞1×106个/只,移植前2 d和1 d分别通过腹腔注射阿糖胞苷0.015 g/只,移植前1d予450 cGy全身照射（TBI）,移植当天给予供鼠骨髓及脾细胞混合物6×107个/只,移植后15 d输注供体免疫淋巴细胞6×107个/只,移植后定期监测受鼠外周血供鼠CD3＋细胞嵌合状态和供鼠性别决定基因（SRY）,观察小鼠急性移植物抗宿主病（aGVHD）、疾病复发及生存情况。结果表明：经尾静脉注射小鼠红白血病细胞1×106个/只,不予治疗,小鼠平均生存期为15±1 d;混合骨髓移植组移植后供者细胞获得植入,SRY基因在移植后30和60 d时检测PCR结果均阳性,移植后14、30、60 d外周血供鼠CD3＋细胞嵌合率分别为（17.95±12.03）%、（37.34±2.78）%,（47.06±6.1）%;DLI组移植后30、45、60 d嵌合率为（69.78±12.62）%、（75±15.97）%、（83.41±16.07）%;小鼠移植后平均生存期分别为66.66±1.47 d、78.2±7.82 d。结论：移植前高肿瘤负荷影响供体细胞植入及预后,移植后早期行供体免疫淋巴细胞输注可以进一步改善治疗效果,延长生存期。

Treatment of Leukemia with Immunized Donor Cell Infusion after Nonmyeloablative Haploidentical Bone Marrow Transplantation

This study was purposed to investigate the therapeutic effects of early transfusion of immunized donor lymphocytes after haploidentical transplantation by means of mouse model of nonmyeloablative haploidentical bone marrow transplantation. CB6F1 female mouse was served as recipient and C57BL/6 male mouse was served as donor. Each CB6F1 female mouse was subjected to intravenous transfusion with 1 x10^6 erythroleukemia （EL9611） cells at day 4 before transplantation, followed with intraperitoneal injection of Ara-C （0. 015 g） respectively at day 2 and day 1, then conditioned for BMT with TB1 （450 cGy） at day 1 before transplantation. After conditioning （ day 0 ）, each of recipients was transplanted with 6 x 10^7 mixture of bone marrow and spleen cells from the C57BL/6 mice, and was infused with 6 x 10^7 immunized donor lymphocytes at day 15 after transplantation. All treated animals were evaluated for survival, development of leukemia and aGVHD. The donor CD3 ＋ cell chimerism and sex determining region Y gene （SRY） in recipients were monitored periodically after transplantation. The results showed tht all mice with only inoculation of 106 EL9611 cells survived for 15±1 days （n =4） ; all mice of other groups obtained the varying degrees of implantation. SRY could be detected at day 30 and 60 after transplantation. The chimerism of donor CD3 cells in mixed bone marrow transplantation （MT） group at day 14, 30 and 60 respectively reached 17.95% ±12.03%, 37.34% ± 2.78% and 47.06% ± 6.1%. In donor lymphocyte infusion （DLI） group it reached 69.78% ± 12.62%, 75% ± 15.97% ,83.41% ± 16.07% at day 30, 45 and 60 after transplantation. The mice of MT and DLI group survived for 66.66± 1.47 days and 78.2 ± 7.82 days. It is concluded that the high tumor burden before transplantation can affect donor cell engraftment and prognosis. Early postranslanted infusion of immunized lymphocytes from donor can help to improve the therapeutic efficacy and survival.