B7-1人-鼠嵌合抗体对小鼠慢性移植物抗宿主病(cGVHD)狼疮样肾炎的免疫干预效应

Immune intervention effect of B7-1 chimeric antibody on lupus nephritis of chronic graft-host disease (cGVHD) mice

目的探讨在诱导c GVHD小鼠狼疮样肾炎模型并通过各项指标鉴定其造模成功的基础上,运用自行研制的B7-1人-鼠嵌合抗体阻断B7/CD28共刺激信号通路对小鼠狼疮样肾炎模型病理损伤的逆转效应。方法将6~8周龄雌性(C57BL/6×BALB/c)BCF1小鼠随机分为4组,模型组于0、3、7和11 d经眼眶静脉注射100μL雌性亲代BALB/c小鼠脾脏细胞107/只。抗体干预组在造模后第1、3、5、8、15、30及60天分别经眼眶静脉注射100μL B7-1人-鼠嵌合抗体(克隆号2B11)200μg/只,环磷酰胺(CTX)干预组在末次注射淋巴细胞后第1、3、5、8、15天分别腹腔注射100μL CTX 2.5 mg/只。按照上述时间点分析小鼠抗ds DNA抗体、ANA及尿蛋白含量,12周时处死小鼠观察肾脏组织HE染色、免疫复合物(IC)沉积等情况。结果模型组100%小鼠出现蛋白尿,尿蛋白含量为++^++++,抗体干预组12周时仅有30%的小鼠出现蛋白尿,尿蛋白含量为+^++;12周时与模型组相比,抗体干预组小鼠血清抗ds DNA抗体阳性率由50%降至0;抗体干预组ANA阳性率由90%降至40%,且荧光面积与荧光亮度均下降;HE染色镜下观察,抗体干预组肾小球囊腔大小较为均一,炎性细胞浸润减少;直接免疫荧光法可见抗体干预组肾小球血管襻IC沉积减少。结论 B7-1人-鼠嵌合抗体可通过阻断或削弱B7/CD28共刺激信号通路,减少自身抗体和IC生成,逆转狼疮肾炎的病理损伤,提示该自行研制的B7-1人-鼠嵌合抗体对狼疮肾炎具有潜在的防治作用。

Objective To study the immune intervention effect of B7-1 chimeric antibody on lupus nephritis on the basis of the use of chronic graft-host disease（ c GVHD） mice lupus nephritis model and its biological identification. Methods 6 ~ 8 weeks aged female（ C57 BL/6 × BALB/c） BCF1 mice were randomly divided into four groups.Each mouce in model group was intravenously injected by 100 μL orbital veins 107 on the 0,3 rd,7 th and 11 th day.Each mouse in antibody treatment group was injected B7-1 chimeric antibody by 100 μL orbital vein 200 μg on the1 st,3 rd,5 th,8 th,15 th,30 th and 60 th day respectively,while each mouse in CTX treatment group was injected 100 μL CTX 2. 5 mg on the 1 st,3 rd,5 th,8 th and 15 th day respectively. The urine protein,anti-ds DNA antibody and ANA content were analyzed at each time point. After the mice were sacrificed on the twelfth weeks,the HE staining of kidney tissue and immune complexes（ IC） deposition was observed. Results At the 12 th week,100% of the model mice were detected proteinuria（ ＋ ＋ ~ ＋ ＋ ＋ ＋） while only 30% of the antibody treatment group mice（ ＋ ~ ＋ ＋）. At the 12 th week,the anti-ds DNA antibody positive rate in antibody treatment group were zero while the model group were 50%,ANA positive rate were 40% while the model group were 90%; HE staining and microscopic observation,only part of the antibody treatment group appeared glomerular swelling,less lymphocyte infiltration and crescent generation; direct immunofluorescence have seen less IC deposited in glomerular vascular loops,the fluorescence area and brightness were weaker than the model group. Conclusion B7-1 chimeric antibody can decrease the generation of autoantibodies and IC,decrease the pathological damage of lupus nephritis by blocking or weakening the B7/CD28 cosignaling pathways,which suggested that self-developed B7-1 human-mouse chimeric antibody can prevent and control lupus nephritis.