异基因造血干细胞移植后MHC单倍体相合脾脏抗白血病疗效分析  

The anti-leukemia effect of major histocompatibility complex haploidentical spleen in hematopoietic stem cell transplantation

作　　者：谢媚 陈伟红[1] 杜新[1] 蔡云[1] 张巧霞[2] 李明[2] 罗畅如[1] 蔡力生[1] 卓家才[1] 冯春锐[1] 李玉珠[2] 汪鹏程[1] 赵江丽[1] 

机构地区：[1]深圳市第二人民医院深圳大学第一附属医院血液内科,深圳518035 [2]深圳市第二人民医院深圳大学第一附属医院深圳市血液病研究所,深圳518035

出　　处：《广西医科大学学报》2017年第10期1406-1410,共5页Journal of Guangxi Medical University

基　　金：深圳市知识创新计划基础研究项目(No.JCYJ20140414170821270);深圳骨髓移植技术临床应用公共服务平台项目(No.s2015003610009)

摘　　要：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 evaluate the anti-leukemia effect of major histocompatibility complex （MHC）haploi- dentical spleen in nonmyeloablative allogeneic hematopoietic stem cell transplantation （allo-HSCT）. Methods： Take C57/BL/6 inbred mice （B6 mice）of 8- 12 weeks as donor rats, and their bone marrow and spleen cells were collected. P815 cells were leukemia cells. The first generation B6D2F1 mice （F1 mice）were as recipients who were hybridized by C57/BL/6 mice and DBA/2 mice. The recipients were intraperitoneally injected with cy- clophosphamide 300 mg/（m2· d） at day -5 to -3 before transplantation. On day 0, F1 mice were divided into group A, B, C and D （n=12 per group）. The mice in group A were injected with mixture of P815 cells （1×10^4 cells）, bone marrow cells（5× 10^6 cells）and spleen cells（2 ×10^2cells）via caudal vein. Mice in group B were injected with mixture of P815 cells （ 1 × 10^4 cells） and bone marrow cells （5 × 10^6 cells）. Mice in group C were injected with P815 cells （1× 10^4 cells） , and those in group D were injected with RPMI 1640 cell culture medium. The status of the mice blood, cute graft versus host disease （GVHD）score and the pathological leukemia infiltration of skin, liver and colon were observed and compared. Results： （ 1 ） The GVHD score in group A was 2～4 on the 14th day after transplantation. Three mice died on day 15. The GVHD score of 2 mice was 6 on day 26, and the rest were scored at 0～3. A mouse died on day 28, and the rest survived mice were scored at 1-6. In group B, the mice began to die on the 6th day, eleven mice died till day 17 and only one survived on day 30. Mice in group C were all dead on day 18. （2）Day 7 after transplantation, the white blood cell （WBC） in group A, B and C were significantly increased. There was significant difference among groups （P〈 0. 05 ）. Day 14 after transplantation, WBC count of all mice were significantly decreased. Compared with group B, C and D, the difference of

关 键 词：异基因造血干细胞移植 脾脏 P815细胞 白血病 

分 类 号：R733.7[医药卫生—肿瘤]