不同照射剂量建立非清髓性移植物抗宿主病模型小鼠外周血T细胞消长性研究  

Growth and decline changes of T cells in peripheral blood of non-myeloablative graft versus host disease with different irradiation doses in murine model

作　　者：胡晓丽[1] 李志强[1] 

机构地区：[1]上海交通大学附属第六人民医院输血科,上海200233

出　　处：《中国输血杂志》2017年第3期262-266,共5页Chinese Journal of Blood Transfusion

基　　金：上海市科学技术委员会科研计划项目基金(13140902903);第四轮(2015-2017年)上海市公共卫生重点建设项目<输血医学>(15GWZK0501)

摘　　要：目的探讨不同照射剂量建立非清髓性移植物抗宿主病(GVHD)模型小鼠外周血T细胞消长性变化,了解GVHD发病中T细胞的作用。方法 50只BALB/C小鼠随机分成5组(10只/组),包括对照组:不作任何处理;8Gy单纯照射(IRD)组:仅接受8Gy全身照射(TBI);7.5 GyIRD组:仅接受7.5 GyTBI;8 GyGVHD组:接受8 GyTBI后<24 h移植供鼠(10只C57BL/6)1×107个骨髓细胞和2.5×107个脾细胞;7.5 GyGVHD组:除接受7.5 GyTBI,其他实验条件同8 GyGVHD组。观察2个GVHD组小鼠移植后GVHD临床症状,肝、肺与肠病理变化,嵌合率以及T细胞亚群变化。结果 8与7.5 GyGVHD组小鼠移植后均为供鼠H-2Kb阳性细胞>90%。10、17、24 d的GVHD评分,8与7.5 GyGVHD组分别为7.25±0.29 vs 5.63±0.25(P<0.01),5.00±0.41 vs 4.00±0.41(P<0.05),6.50±0.41 vs 8.00±0.41;GVHD评分组内两两比较,除8 GyGVHD组10与24 d比较有较大变化(P<0.05)外,其余变化更明显(P<0.01)。CD4+/CD8+比值,8与7.5 GyGVHD组分别为0.42±0.03 vs 0.31±0.03,0.27±0.02 vs 0.19±0.01,0.43±0.03vs 0.32±0.03(P<0.01);CD4^+/CD8^+比值组内两两比较,除10与24 d相近外,10较17 d和17较24 d都变化明显(P<0.01)。CD4^+CD25^+T比例,8与7.5 GyGVHD组分别为0.028 4±0.009 1 vs 0.020 9±0.004 9(P>0.05),0.014 0±0.004 3 vs 0.006 6±0.001 2(P<0.05),0.009 4±0.000 8 vs 0.005 1±0.000 7(P<0.01);组内两两比较,均为17与24d相近,10较17 d和10较24 d变化明显(P<0.01)。结论非清髓性移植物抗宿主病模型小鼠发病过程中CD4+T细胞起更重要的作用;随着小鼠移植后生存时间的延长,其体内的CD4^+CD25^+T细胞呈阶梯式下降,下降幅度与照射剂量呈负相关。Objective To investigate the possible role of T cells in peripheral blood of non-myeloablative graft versus host disease with different irradiation doses in murine model by testing their growth and decline changes.Methods 50 SPF grade BALB/C mice were randomly devided into 5 groups（10 mice/group）.Control group：were with no treatment;8 GyIRD group：only received 8 Gy Xray TBI（total body irradiation）;7.5 GyIRD group：only received 7.5 Gy Xray TBI;8 GyGVHD group：were injected with bone marrow cells （1 ＆#215; 107） and splenocytes （2.5 ＆#215; 107） of donors （C57 BL/6） via tail vein within 24 hours after receiving 8 Gy Xray TBI;7.5 GyGVHD group：treatment is the same with 8 GyGVHD group except the 7.5 Gy irradiation dose.The clinical symptoms,histopathological analysis of the liver,lung and intestinal,the chimeric rate and growth and decline changes of T cell subsets were observed in both two GVHD groups after transplantation.Results Allogeneic H-2Kb positive cells proportion was higher than 90% in both 7.5 GyGVHD group and 8 GyGVHD group after transplantation.The GVHD score in 8 GyGVHD group is higher than 7.5 GyGVHD group on the 10th,17th,24th day,7.25±0.29 vs 5.63±0.25,5.00±0.41 vs 4.00±0.41,8.00±0.41 vs 6.50±0.41 respectively;Compared within the 7.5 Gy and 8 Gy GVHD group on the 10th,17th,24th day,differences are all P〈0.01 except difference on 10 d vs 24 d within 8 GyGVHD group is P〈0.05.Value of CD4/CD8 is compared between 8Gy and 7.5Gy GVHD group on the 10th,17th,24th day,0.42 ±0.03 vs 0.31±0.03,0.27±0.02 vs 0.19±0.01,0.43±0.03 vs 0.32±0.03（P〈0.01）;Value of CD4/CD8 within the 7.5 Gy and 8 GyGVHD group,10 d vs 24 d（P〉0.05）,10 d vs 17 d and 17 d vs 24 d（P〈0.01）.The propotion of the CD4＋ CD25＋ between 8 Gy and 7.5 Gy GVHD group on the 10th,17th,24th day,0.028 4±0.009 1 vs 0.020 9±0.004 9,（P〉0.05）,0.014 0±0.0043 vs 0.006 6±0.001 2,（P〈0.05）,0.009 4±0.000 8 vs 0.005 1±0.000 7,（P〈0.01） respectively;the proportion of CD4＋ CD25

关 键 词：移植物抗宿主病 非清髓性 干细胞移植 骨髓细胞 脾细胞 动物模型 小鼠 T细胞 照射剂量 移植时间 

分 类 号：R457.12[医药卫生—治疗学] R457.7[医药卫生—临床医学]