淋巴细胞活性染色质诱导系统性红斑狼疮样小鼠模型

Active chromatin of lymphocyte induced systemic lupus erythematosus-like mouse model

目的建立活性染色质诱导系统性红斑狼疮(SLE)样小鼠模型。方法从ConA活化的BALB/c小鼠脾淋巴细胞中提取活性染色质,分别于d 0、d 14、d 21和d 28以染色质100μg在BALB/c小鼠尾根部及背部皮内注射免疫4次,诱导SLE样小鼠模型。目测半定量尿蛋白试纸法检测动物的尿蛋白变化,HE染色法检查动物的肾脏、脾脏病理改变,计算动物的胸腺和脾脏指数,MTT法检测ConA和LPS诱导的T、B淋巴细胞增殖反应,全自动生化分析仪检测血清中Crea和BUN水平,ELISA法检测小鼠血清中ANA、抗dsDNA、IgG1、IgG2a、IL-10、IFN-γ水平,流式细胞术检测脾脏T、B淋巴细胞亚群变化。结果诱导模型小鼠尿蛋白水平升高,出现肾小球肾炎、脾脏增生等病理改变;脾脏指数明显升高,LPS诱导的B淋巴细胞增殖反应增强;血清Crea、BUN、ANA、抗dsDNA、IgG1、IgG2a、IL-10和IFN-γ水平明显升高;脾脏CD19+、CD19+CD21+、CD19+CD23+、CD19+IgD+B淋巴细胞亚群百分比明显升高,CD4+CD25+T淋巴细胞百分比明显下降。结论 ConA活化淋巴细胞的染色质免疫同系BALB/c小鼠成功诱导了SLE样小鼠模型,其血清学、组织病理学及免疫学方面特征与人类SLE临床特征表现相似。

Aim To establish mouse model of systemic lupus erythematosus（SLE） induced by active chromatin in BALB/c mice.Methods Active chromatin was extracted from ConA-activated spleno-lymphocytes of BALB/c mice.BALB/c mice were immunized with 100μg active chromatin on d 0,d 14,d 21,d 28 by intradermal injection on the back and the base of the tail for 4 times to establish the SLE-like mouse model.Proteinuria was measured by Semi-quantitative Albustix paper.Histopathological changes of kidney and spleen were observed by HE staining.Thymus index and spleen index were calculated.Thymo-lymphocyte and spleno-lymphocyte proliferation stimulated by ConA and LPS was tested by MTT method.Levels of Crea and BUN were detected by automatic biochemical analyzer.Levels of ANA,anti-dsDNA,IgG1,IgG2a,IL-10 and IFN-γ in serum were tested by enzyme-linked immunosorbent assay.Splenic T,B lymphocyte subsets were analysed by flow cytometry.Results The level of proteinuria in model mice was elevated.Histopathological examination showed glomerulonephritis and spleen hyperplasia;spleen index was increased and spleno-lymphocyte proliferation stimulated by LPS was enhanced;levels of Crea,BUN,ANA,anti-dsDNA,IgG1,IgG2a,IL-10 and IFN-γ were elevated;the percentage of CD19^＋,CD19^＋CD21^＋,CD19^＋CD23^＋,CD19^＋IgD^＋ B cell subsets was increased and the percentage of CD4＋CD25＋ T cells was decreased.Conclusions SLE-like mouse model is successfully established in BALB/c mice by immunized with active chromatin isolated from ConA-activated lymphocytes.The characteristics of serology,histopathology and immunology in the mouse model are similar to the clinical features of human SLE.