慢性移植物抗宿主病狼疮样肾炎小鼠模型的建立及免疫病理学鉴定  

Development of,and pathological evaluation of murine lupus nephritis model of chronic graft-versus-host disease

作　　者：韩莲花[1] 蔡磊[1] 朱莹[1] 朱华亭[1] 夏永洁[1] 邱玉华[1] 

机构地区：[1]苏州大学医学部免疫学系,苏州215123

出　　处：《现代免疫学》2014年第2期115-119,共5页Current Immunology

基　　金：国家自然科学基金项目(81373236)

摘　　要：对慢性移植物抗宿主病狼疮样肾炎小鼠模型进行免疫学、血清学及肾脏病理损伤鉴定。以考察该模型作为系统性红斑狼疮(SLE)疾病模型的可靠性。将亲代雌性BALB/c小鼠脾脏细胞悬液经尾静脉注射(C57BL/J6×BALB/c)F1代小鼠,每隔3d注射1次,共4次。建模第7d,采用免疫荧光及流式细胞仪分析小鼠脾脏中抗原提呈细胞(APC)及抗体形成细胞的百分率。每隔2周检测血清中anti-dsDNA抗体及ANA;每隔4周检测尿蛋白含量的动态变化;建模12周终止实验,取肾脏组织作HE染色观察病理改变,直接免疫荧光法检测免疫复合物(IC)沉积以及透射电镜观察肾小球超微结构变化。流式细胞术的分析结果显示,造模后7d脾脏中膜型CD11b+、CD11c+及Gr1+细胞的百分率较对照组明显升高(P<0.05),同时抗体形成细胞B220+表面CD21、CD23、CD80及CD86分子的表达均上调(P<0.05);建模2周时,血清anti-dsDNA抗体出现阳性,4周时ANA出现阳性;12周时80%的小鼠存在蛋白尿,蛋白含量++^++++;肾小球体积增大并有大量炎性细胞浸润;肾脏IC沉积;透射电镜下可见肾基底膜节段性增厚,脏层上皮细胞和基底膜之间存在大量的电子致密物。慢性移植物抗宿主病小鼠狼疮样肾炎模型具有人类SLE相似的免疫细胞异常活化及肾脏病理损伤的特征性表现,用于相关研究具有可靠性。A model of murine lupus nephritis induced by chronic graft-versus-host disease was estavusneo anu examined by im-munological, serological and nephrological methods to test the validity for lupus nephritis in SLE. （C57BL/J6 ~ BALB/c） F1 hybrids were used as recipients of BALB/c donor. Single-cell suspensions from donor＇s spleen were injected intravenously 4 times at intervals of 3 days. Seven days after the last injection, immunofluorescence staining and flow cytometry were carried out to analyze percentage of APCs and antibody producing cells among the recipients＇ spleen cells. Anti-double-stranded DNA antibody and anti-nuclear antibodies in the serum were detected at intervals of 2 weeks ; Dynamic variation of proteinuria was tested every 4 weeks; 12 weeks after the last injection, the pathology of kidneys was examined to observe glomerulonephritis, and deposition of immunoglobulins by direct immunofluoreseence staining on kidney sections. Electron microscopy was used to observe ultrastructure of glomerulus＇s change. The results showed that seven days after the induction of the model, populations of cells with membranous CD11b,CDllc, Grl were increased significantly（P^0.05）, in the meanwhile , the expressions of CD21 ,CD23, CDS0,CD86 on B220＋ cells were also increased（P^0.05）. Anti-dsDNA antibody, occurred after 2 weeks and ANA was observed after 4 weeks. After 12 weeks, 80~ experimental mice had proteinuria（＋ ＋ - ＋ ＋ -[- -t- ）, both optical microscopy and immunoflorescence test indicated typical evidence of glomerulonephritis. Electron micrographs showed dense e- lectron deposits localized in subepithelia[ lesions of glomeru｜ar basement membrane. Taken together, this murine lupus nephri- tis model of chronic graft-versus-host disease has characteristic manifestations similar to human SLE and maybe a receivable model for related research.

关 键 词：慢性移植物抗宿主病 狼疮样肾炎 动物模型 

分 类 号：R593.24[医药卫生—内科学]