CAJ | 학술논문

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C57 BL／6 J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IgG。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。
목적：재운용화학법건립소서랑창양신염모형병대기진행생물학감정적기출상，탐토B7-1인-서감합항체조단B7／D28신호통로대소서랑창양신염모형병리손상적역전효응。방법：취6주령자성C57 BL／6 J소서，여일차성복강주사Pristane 0.5ml／지，매월정기검측소서적뇨단백、ANA급신장병이학개변。취뇨단백함량체도＋＋，ANA형광강도체도＋＋적소서수궤분위3조，매조5지。항체간예조용B7-1인-서감합항체경안광정맥서관급약，양성대조조주사면역억제제CTX，음성대조조주사인동형IgG。매월정기검측뇨단백급ANA，간예지3개월시，처사소서，취신장진행H&E염색분석，면역복합물（ IC）검측급투사전경관찰。결과：Pristane유도지4개월시，80％적소서뇨단백함량체도＋～＋＋＋，혈청ANA형광강도위＋＋～＋＋＋，출현뇨단백급ANA적소서，기신소구염성세포침윤，신소관상피양세포가견수종양변성、혈관충혈명현，섬유조직증생。항체간예후，뇨단백축점유＋＋～＋＋＋강위±～＋＋， ANA유＋＋～＋＋＋강위＋～＋＋。여음성대조조비교구유통계학차이（ P＜0.01）。신장HE염색분석적결과현시，항체간예조신소구염성세포침윤급신소관충혈등표현균득도명현개선。면역형광염색가견항체간예조항원항체복합물（ IC）적형광강도명현감약。경투사전경관찰，항체간예조여음성대조조상비，신소구적전자치밀물침적감소，기저막후도추우균균。결론：B7-1항체통과억제B7-1／CD28신호통로하조궤체적면역응답，감소자신항체적산생，대자신면역조성적병리손상구유역전작용。
Objective:On the basis of the use of chemical methods to establish mouse model of lupus nephritis and its biological identification , we investigate the reverse effect of pathological lesions of B 7-1 human-mouse chimeric antibody blockade against B7/D28 signaling pathway in mice with lupus nephritis model.Methods:Pristane was injected intraperitoneally to 6-week-old female C57BL/6J mice at dose of 0.5 ml per mouse in one go,and urine protein,ANA and renal pathological changes were detected on a monthly basis.Mice whose urine protein content reached ++and ANA fluorescence intensity reached ++were randomly devided into three groups ,five each.Antibody intervention group was sequentially injected with B 7-1-mouse chimeric antibody by orbital venous , positive control group was injected with immunosuppressant CTX , negative control group was injected with isotype control IgG.Urine protein and ANA were also detected on a monthly basis.Mice were sacrificed three months after intervention was executed.Kidney was used for H&E dying , IC detection and electric microscope observation.Results: After four-month Pristane induction , urine protein content of 80%mice reached +-+++,meanwhile,serum ANA fluorescence intensity reached ++-+++.Glomerulonephritis infiltrating cells were observed Mice with urine protein and ANA , glomerular inflammatory cell infiltration , tubular epithelial cell degeneration visible edema ,vascular congestion significantly ,fibrosis.After antibody intervention ,urine protein content in antibody intervention group gradually reduced from ++-+++to ±-+++,ANA ++-+++to +-++,and were significantly different from that in the negative control group ( P<0.01 ).Analysis of kidney H&E dying showed that antibody glomerular infiltration of inflammatory cells in the intervention group and tubular congestion and other symptoms were improved significantly.Immunofluorescence staining indicated that fluorescence intensity of IC was significantly reduced in the antibody intervention group.Electron dense deposits reduction and glomerular basement membrane uniformity were observed in antibody intervention group by electric microscope when compared with the negative control group.Conclusion:B7-1 antibodies could downregulate immune response through inhibiting B 7-1/CD28 signaling pathway , reducing the production of autoantibodies and reversing pathological damage caused by autoimmune response .