大鼠异基因骨髓移植急性移植物抗宿主病模型的建立

目的建立较稳定的异基因骨髓移植急性移植物抗宿主病动物模型,为异基因骨髓移植后的急性移植物抗宿主病（aGVHD）的相关研究提供实验参照。方法以雄性SD大鼠为供鼠,雌性Wistar大鼠为受鼠,受体大鼠随机分成A、B、C、D、E 5组,移植当天所有受鼠均接受8.5 GY的全身照射（TBI）,于照射后4～6 h内,A组回输等量培养液,B组经尾静脉输注供鼠骨髓细胞（2×10^8个/kg）,C、D、E组分别回输供鼠骨髓细胞（2×10^8个/kg）＋不同比例的脾细胞。观察各组大鼠生存期、外周白细胞计数、及有无aGVHD的临床及病理表现。结果A组大鼠于15d内全部死亡,外周血白细胞计数明显减低,骨髓病理示造血组织减少,提示死于造血衰竭。B、C、D、E组大鼠外周血白细胞计数均有明显恢复,B组大鼠8只存活超过50 d,C、D、E组大鼠均于50 d观察期内死亡,并有aGVHD的临床表现及病理表现,但C组大鼠aGVHD的程度较轻且时间不集中,其中D、E组大鼠可于相对集中的时间内观察到典型aGVHD临床及病理。结论TBI预处理的方式是可行的,单纯输入异基因骨髓细胞不能引起明显的aGVHD,骨髓细胞与脾细胞1∶1及1∶1.5混合组均可作为异基因骨髓移植后理想的aGVHD动物模型。

Establishment of a Rat Model of Acute Graft-versus-Host Disease after Allogeneic Bone Marrow Transplantation

Objective The aim of this study was to establish a rat acute graft-versus-host disease （aGVHD） model after aUogeneic bone marrow transplantation （Allo-BMT） to serve experimental studies of aGVHD. Method Female Wistar rats were used as recipients and male SD rats as donors. All Wistar rats were randomized into A, B, C, D and E groups which all received total body irradiation （TBI） 8.5 Gy at 4 - 6 hours prior to transplantation. Group A received equivalent dose of RPMI 1640. Group B received donor bone marrow cells （2 ×10^8/kg） via tail vein. The other three groups received donor bone marrow cells （2 ×10^8/kg） plus spleen cells in different ratios. Their survival time, leucocyte counts of peripheral blood, clinical and pathologic aGVHD manifestation were evaluated. Result All rats in group A died within 15 days, showing significantly decreased peripheral blood leukocyte count and hematopoietic failure with bone marrow pathologic evidence. Peripheral blood leukocyte counts were increased gradually in groups B, C, D and E. 8 rats survived more than 50 days in group B. The rats in groups with cotransplantation of bone marrow cells and spleen cells died within 50 days, which showed clinical manifestation and pathological changes of aGVHD. The rats in group C showed less severe aGVHD and at a dispersed time. The rats in group D and E showed typical clinical and pathological manifestation of aGVHD at a concentrated time. Conclusion The TBI regime of pretreatment is feasible. Infusion of allogenic bone marrow ceils alone does not induce typical aGVHD. Infusion of bone marrow ceils and spleen cells in ratio of 1 ： 1 and 1 ： 1.5 is perfect as to establish aGVHD animal model after allogeneic bone marrow transplantation.