目的:建立EL9611红白血病小鼠急性移植物抗宿主病(GVHD)的动物模型。方法:同种异基因骨髓移植(allo—BMT)以C57BL/6(H-2^b)鼠为供鼠,BALB/c(H-2^d)为受鼠。设白血病组(n=10)、照射对照组(白血病鼠照射后不进行allo—BMT,n=4)、GVHD组(白血病鼠照射+allo-BMT,n=10)及正常对照组(n=4)。白血病组采用每只BALB/c鼠尾静脉输注2×10^6个EL9611红白血病细胞建立红白血病动物模型;GVHD组于接种白血病细胞7d后行总剂量为8.0Gy的1次性[60Co].γ射线全身照射(TBI),照射后5h内每只小鼠尾静脉输注C57BL/6鼠骨髓细胞2×10^6个+脾细胞1×10^7个,建立EL9611红白血病小鼠的急性GVHD动物模型。观察小鼠体位、皮毛、大便等临床表现,病理检查肝脾、皮肤、小肠、外周血和骨髓,计算生存率。结果:白血病组生存时间(14.5±2.1)d[从照射当天(第0d)算起为(7.5±0.7)d],生存时间与GVHD组相比P〈0.01,死亡率100%,无自发缓解,死亡时肝脾肿大(肝重2.40g±0.48g,脾重0.84g±0.20g,与正常对照组比P〈0.01),外周血WBC升高[死亡前(3.33±0.27)×10^10/L,与正常对照组比P〈0.05],病理检查示组织正常结构破坏,白血病细胞浸润。照射对照组生存时间为(9.0±0.7)d,生存时间与GVHD组和正常对照组相比差异显著(P〈0.01),死亡率100%,病理检查显示造血衰竭。GVHD组生存时间为(32.0±3.2)d,生存时间与其它各组相比P〈0.01,死亡率100%,allo—BMT后第10—13d出现症状,临床表现和病理检查符合Ⅰ到Ⅱ度GVHD的改变。结论:采用EL9611红白血病细胞(2×10^6/鼠)静脉输注的方式可成功建立EL9611红白血病动物模型;接种EL9611红白血病细胞第7d行TBI+allo—BMT可成功建立EL9611红白血病小鼠的急性GVHD动物模型。
AIM: To establish an acute graft- versus -host disease (GVHD) model in EL9611 erythroleukemia mice. METHODS: Using C57BL/6 (H -2b) mice as the donor and BALB/c (H -2d) mice as the recipient in allogeneic bone marrow transplantation ( allo - BMT), the acute GVHD model was established. The mice were divided into leukemia group (n = 10), radiation control group (leukemic mice given radiation without allo- BMT, n = 4), GVHD group (leukemic mice given radiation + allo - BMT, n = 10) and normal control group ( n = 4). In leukemia group, 2 ×10^6/mouse EL9611 erythroleukemic cells were transfused via tail vein into BALB/c mice to build the ervthroleukemia model. In GVHD group, 7 days after leukemic cell transfusion, the mice received total dose of 8.0 Gy [ 60Co] ? of total body irradiation(TBI), and within 5 h, 2 ×10^6 C57BL/6 bone marrow cells and 1 ×10^7 C57BL/6 spleen cells per mouse were transfused via tail vein to build the acute GVHD model in EL9611 erythroleukemia mice. The clinical manifestations of posture, fur, stool and so on were observed. Pathological examination was conducted to examine the changes of liver, spleen, skin, small intestine and peripheral blood. The survival rate was also calculated. RESULTS : ( 1 ) In leukemia group, the mean survival time (MST) was (14. 5 ±2. 1) days,or (7.5 ±0. 7) days when irradiation day was as day 0(P 〈0. 01 compared with GVHD group). The death rate was 100% with no spontaneous remission. The dead mice showed splenohepatomegalia [ liver weight (2.40 ±0. 48) g, spleen weight (0. 84 ±0. 20) g, P 〈 0. 01 compared with normal group ] and high WBC count [ (3.33 ±0. 27) ×10^10/L prior to death, P 〈0. 05, compared with normal group] Pathological examination showed disorganization of normal tissues and leukemic cell infiltration. ( 2 ) In radiation control group, MST was ( 9. 0 ± 0. 7) d, with significant difference as compared with GVHD group and normal group (P 〈 0. 01 )