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目的:在运用化学法建立小鼠狼疮样肾炎模型并对其进行生物学鉴定的基础上,探讨B7-1人-鼠嵌合抗体阻断B7/D28信号通路对小鼠狼疮样肾炎模型病理损伤的逆转效应。方法:取6周龄雌性C57BL/6J小鼠,予一次性腹腔注射Pristane 0.5ml/只,每月定期检测小鼠的尿蛋白、ANA及肾脏病理学改变。取尿蛋白含量达到++,ANA荧光强度达到++的小鼠随机分为3组,每组5只。抗体干预组用B7-1人-鼠嵌合抗体经眼眶静脉序贯给药,阳性对照组注射免疫抑制剂CTX,阴性对照组注射人同型Ig G。每月定期检测尿蛋白及ANA,干预至3个月时,处死小鼠,取肾脏进行H&E染色分析,免疫复合物(IC)检测及透射电镜观察。结果:Pristane诱导至4个月时,80%的小鼠尿蛋白含量达到+~+++,血清ANA荧光强度为++~+++,出现尿蛋白及ANA的小鼠,其肾小球炎性细胞侵润,肾小管上皮样细胞可见水肿样变性、血管充血明显,纤维组织增生。抗体干预后,尿蛋白逐渐由++~+++降为±~++,ANA由++~+++降为+~++。与阴性对照组比较具有统计学差异(P<0.01)。肾脏HE染色分析的结果显示,抗体干预组肾小球炎性细胞侵润及肾小管充血等表现均得到明显改善。免疫荧光染色可见抗体干预组抗原抗体复合物(IC)的荧光强度明显减弱。经透射电镜观察,抗体干预组与阴性对照组相比,肾小球的电子致密物沉积减少,基底膜厚度趋于均匀。结论:B7-1抗体通过抑制B7-1/CD28信号通路下调机体的免疫应答,减少自身抗体的产生,对自身免疫造成的病理损伤具有逆转作用。
OBJECTIVE: To establish a model of lupus-like nephritis in mice by chemical method and to identify the pathological model of lupus-like nephritis in mice by blocking B7 / D28 signaling pathway by chimeric antibody of B7-1 The reversal effect. Methods: Six-week-old female C57BL / 6J mice were given intraperitoneal injection of Pristane 0.5ml per mouse, and the urinary protein, ANA and pathological changes of kidney were detected regularly every month. Take urine protein content reached ++, ANA fluorescent intensity reached ++ mice were randomly divided into three groups of five. Antibody intervention group with B7-1 human-mouse chimeric antibodies by sequential orbital ophthalmic injection of positive control group injected immunosuppressive CTX, negative control group injected human homologous type G G. Urine protein and ANA were detected regularly every month. When intervention was 3 months, mice were sacrificed and kidneys were taken for H & E staining analysis, immunocomplex (IC) detection and transmission electron microscope observation. Results: After 4 months of Pristane induction, the proteinuria of urinary protein and ANA in 80% of mice reached + ~ +++, the serum ANA fluorescence intensity was ++ ~ +++, Inflammatory cells infiltration, tubular epithelial cells showed edema-like degeneration, vascular congestion was obvious, fibrous tissue hyperplasia. After antibody intervention, urinary protein gradually decreased from ++ to +++ to ± ~ ++, and ANA decreased from ++ to +++ to ++. Compared with the negative control group was statistically significant (P <0.01). The results of HE staining of kidney showed that the performance of glomerular inflammatory cell infiltration and renal tubular congestion in the antibody intervention group were significantly improved. Immunofluorescence staining showed that the antibody intensity of the antigen-antibody complex (IC) in the intervention group decreased significantly. Transmission electron microscopy showed that compared with the negative control group, the electron-dense deposits of glomeruli decreased and the thickness of basement membrane became more uniform. CONCLUSION: B7-1 antibody can down-regulate the immune response of B7-1 / CD28 signaling pathway and reduce the production of autoantibodies, reversing the pathological damage caused by autoimmunity.