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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。
Immunological, serological and renal pathological lesions were identified in a mouse model of chronic graft versus host disease lupus-like nephritis. To examine the reliability of this model as a model of systemic lupus erythematosus (SLE) disease. Spleen cell suspension of parental female BALB / c mice were injected via the tail vein (C57BL / J6 × BALB / c) F1 mice, once every 4 days for 4 times. At the 7th day after modeling, the percentage of antigen presenting cells (APCs) and antibody-forming cells in spleen of mice was analyzed by immunofluorescence and flow cytometry. Serum anti-dsDNA antibody and ANA were detected every 2 weeks. Urine protein content was detected every 4 weeks. The model was terminated at 12 weeks. The kidneys were taken for HE staining to observe the pathological changes. The immunofluorescence assay (IC) deposition and transmission electron microscopy glomerular ultrastructure changes. The results of flow cytometry showed that the percentages of CD11b +, CD11c + and Gr1 + cells in the spleen were significantly higher than those in the control group 7 d after modeling (P <0.05). Meanwhile, CD21, CD23, CD80 and CD86 on the surface of antibody- The expression of anti-dsDNA antibody was positive at 2 weeks after modeling, and positive at 4 weeks. Proteinuria was found in 80% of the mice at 12 weeks +++; glomerular volume increased with a large number of inflammatory cell infiltration; renal IC deposition; transmission electron microscopy showed segmental renal basement membrane thickening, visceral epithelial cells and the basement membrane between a large number of electron-dense material. Chronic graft-versus-host disease in mice with lupus-like nephritis model with human SLE similar abnormal activation of immune cells and kidney pathological features of the performance of the injury for the relevant studies with reliability.