混合骨髓移植后过继免疫治疗小鼠白血病

Adoptive immunotherapy for leukemia in mice after autologous bone marrow mixed with H-2 haploidentical allogeneic bone marrow transplantation

背景:急性白血病自体造血干细胞移植后复发率高,异基因造血干细胞移植后移植相关病死率高,混合造血干细胞移植及移植后过继免疫治疗有可能取长补短,提高疗效。目的:观察自体骨髓混合H-2半相合异体骨髓移植后供体淋巴细胞输注+白细胞介素2治疗对小鼠白血病的疗效。方法:将Balb/c小鼠经直线加速器照射3Gy后分为白血病模型组、白血病模型照射组、混合移植组、自体骨髓移植组,均尾静脉注射5×105K562(GFP+/NeoR+)或K562(GFP-/NeoR-)细胞。7d后6Gy照射,自体骨髓移植组移植自体骨髓细胞或联合白细胞介素2治疗;混合移植组移植小鼠自体骨髓细胞混合1/10的H-2半相合异体骨髓细胞后应用白细胞介素2或联合供体淋巴细胞输注治疗。4周后行小鼠外周血及骨髓细胞形态检查,外周血细胞亚群、GFP及NeoR基因测定,肝、脾匀浆细胞GFP和NeoR基因测定。结果与结论:白血病模型组小鼠因骨髓造血功能衰竭于20d内全部死亡,白血病模型照射组小鼠因造血功能衰竭于14d内全部死亡;自体骨髓移植组、混合移植组均有多少不等小鼠无白血病存活超过28d,且混合骨髓移植后及自体骨髓移植后应用白细胞介素2治疗可提高白血病小鼠长期无病生存率,在此基础上联合供体淋巴细胞输注可更进一步提高白血病小鼠长期无病生存率。

BACKGROUND：Auto hemopoietic stem cell transplantation（Auto-HSCT） has a high relapse rate in acute leukemia;allo-HSCT has a high incidence of transplant-related mortality.It may increase curative effect when leukemia patients are administered adoptive immunotherapy post mixed-HSCT.OBJECTIVE：To explore the curative effect of using donor lymphocyte infusion combined with interleukin-2（DLI＋IL-2） after autologous bone marrow mixed with H-2 haploidentical allogeneic bone marrow transplantation（MBMT） in mice with leukemia.METHODS：Leukemia models were prepared with Balb/c mice which were irradiated 3 Gy by linear accelerator and injected K562（GFP＋/NeoR＋） or K562（GFP-/NeoR-） cells 5×10^5 into caudal vein and divided into leukemia model group,irradiated leukemia model group,MBMT group,and autologous bone marrow transplantation（ABMT） group.6 Gy irradiation was performed after 7 days;the mice were treated with ABMT or MBMT respectively.Mice of MBMT group mixed with 1/10 of H-2 haploidentical allogeneic bone marrow cells underwent IL-2 or combination of DLI treatment.Peripheral blood and bone marrow cell morphous of mice were examined;cell subsets,GFP and NeoR gene in peripheral blood,and liver,spleen homogenate cells,and NeoR gene were detected after 4 weeks.RESULTS AND CONCLUSION：All of mice in leukemia model group died of bone marrow hematopoietic failure within 20 days;mice in irradiated leukemia model group died of hematopoietic failure within 14 days.Varying amounts of non-leukemic of mice survived for more than 28 days between ABMT group and MBMT group.UsingIL-2 treatment after MBMT and ABMT can promote long term disease free survival of mice with leukemia,and which combined with DLI can further improve long term disease free survival of mice with leukemia.