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目的:观察主要组织相容性复合体(MHC)单倍体相合脾脏对非清髓异基因造血干细胞移植(allo-HSCT)抗白血病疗效的分析。方法:用8~12周C57/BL/6近交系小鼠(B6鼠)作为供鼠,取其骨髓细胞和脾细胞;P815细胞即白血病细胞,近交系DBA/2小鼠与近交系C57/BL/6小鼠杂交的第一代小鼠B6D2F1(F1鼠)为受鼠,受鼠骨髓移植d-5~d-3,用环磷酰胺300mg/(m~2·d)腹腔注射预处理,受鼠移植当天(d0),将F1鼠随机分为A、B、C、D 4组,每组12只,其中A组经尾静脉注射P815细胞(1×10~4个)、骨髓细胞(5×10~6个)和脾细胞(2×10~7个)的混合细胞悬液,B组经尾静脉注射P815细胞(1×10~4个)和骨髓细胞(5×10~6个)的混合细胞悬液,C组经尾静脉注射P815细胞(1×10~4个)的细胞悬液,D组经尾静脉注射RPMI 1640细胞培养基。比较4组小鼠血象、急性移植物抗宿主病(GVHD)评分以及皮肤、肝、结肠的病理白血病细胞浸润情况。结果:(1)移植后第14天,A组GVHD评分2~4分,第15天3只小鼠死亡,第26天2只小鼠GVHD评分6分,其它7只小鼠GVHD评分0~3分,第28天死亡1只,存活的8只小鼠GVHD临床评分1~6分。B组小鼠第6天开始出现死亡,至第17天死亡11只,第30天时仅存活1只。C组第18天时全部死亡。(2)4组小鼠移植后第7天时,A、B、C组小鼠白细胞数量明显增高,组间比较有明显差异(P<0.05),与D组比较差异亦有统计学意义(P<0.05)。移植后第14天时,4组小鼠白细胞数均明显下降,A组分别与B、C、D组比较差异均有统计学意义(P<0.05)。(3)移植后第28天,A、D组白细胞数均回升,组间比较无明显差异(P>0.05)。A组小鼠的肝脏可见炎性细胞浸润,汇管区肝细胞萎缩、坏死,中央静脉区少量白血病细胞浸润。B组可见肝细胞数量减少,肝窦内白血病细胞弥散性浸润。结论:非清髓allo-HSCT MHC单倍体相合脾脏可以提高抗白血病的疗效。
OBJECTIVE: To investigate the efficacy of haploidentical haploidentical splenic lymphocyte (MHC) haploidentical spleen transplantation in the treatment of leukemia with allo-HSCT for allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: C57 / BL / 6 inbred mice (B6 mice) were used as donors for 8 ~ 12 weeks, and their bone marrow cells and splenocytes were harvested. P815 cells, leukemia cells and inbred DBA / 2 mice The first generation mice B6D2F1 (F1 mice) crossed with C57 / BL / 6 mice were transplanted with d-5-d-3 bone marrow and treated with cyclophosphamide 300mg / (m ~ 2 · d) The rats were randomly divided into 4 groups (A, B, C and D), 12 rats in each group. Group A received injection of P815 cells (1 × 10 ~ 4) , Bone marrow cells (5 × 10 ~ 6) and spleen cells (2 × 10 ~ 7) were mixed with cell suspensions. Group B received intravenous injection of P815 cells (1 × 10 ~ 4 cells) and bone marrow cells 10 to 6). The cells in group C were injected with the cell suspension of P815 cells (1 × 10 ~ 4 cells) through the tail vein, and the cells in group D were injected into the tail vein with RPMI 1640 cells. Blood samples were collected from four groups of mice, acute graft-versus-host disease (GVHD) scores and pathological leukemic cell infiltration in the skin, liver and colon. On the 14th day after transplantation, the GVHD score of group A was 2 to 4, and the 3 mice died on the 15th day. The GVHD score of 2 mice on the 26th day was 6 and that of the other 7 mice was 0 ~ 3 points, one death on the 28th day, and the surviving 8 mice had clinical scores of 1-6 points for GVHD. The mice in group B started to die on the 6th day, died on the 17th day and only survived on the 30th day. Group C all died on the 18th day. (2) The number of white blood cells in group A, B and C increased significantly on the 7th day after transplantation (P <0.05), and the difference was also statistically significant compared with group D (P <0.05). On the 14th day after transplantation, the number of white blood cells in the four groups of mice was significantly decreased. The difference between group A and group B, C and D was statistically significant (P <0.05). (3) On the 28th day after transplantation, the number of white blood cells in group A and group D rose back with no significant difference between the two groups (P> 0.05). A group of mice showed inflammatory cell infiltration of the liver, portal area hepatocyte shrinkage, necrosis, central venous area a small amount of leukemia cell infiltration. Group B showed a decrease in the number of liver cells, sinusoidal leukemia cells diffuse infiltration. Conclusion: Nonmyelo-allo-HSCT MHC haploidentical spleen can improve the anti-leukemia efficacy.