CAJ | 학술논문

目的探讨人源性抗大疱性类天疱疮抗原BP180单链抗体的生物学功能.方法亲和层析方法纯化大疱性类天疱疮(BP)患者血清自身抗体,通过竞争性ELISA、竞争性免疫荧光和补体活化的竞争性抑制实验来观察所制备的抗BP180单链抗体对BP-IgG自身抗体结合人BP180抗原的竞争性抑制作用.结果竞争性ELISA结果表明,在0～60 μg/ml范围内单链抗体对BP-IgG自身抗体的竞争性抑制作用呈剂量依赖关系,最大抑制率可达69.50%(与对照组相比,均有统计学意义,P＜0.01).竞争性免疫荧光实验中,单链抗体浓度增加至40μg/ml时,BP-IgG在基底膜带的沉积及其介导的补体C3活化作用均被抑制,呈阴性.结论人源性抗BP180单链抗体在体外对BP致病性自身抗体与抗原结合及后续的补体活化具有一定的抑制作用.
목적 탐토인원성항대포성류천포창항원BP180단련항체적생물학공능.방법 친화층석방법순화대포성류천포창(BP)환자혈청자신항체,통과경쟁성ELISA、경쟁성면역형광화보체활화적경쟁성억제실험래관찰소제비적항BP180단련항체대BP-IgG자신항체결합인BP180항원적경쟁성억제작용.결과 경쟁성ELISA결과표명,재0～60 μg/ml범위내단련항체대BP-IgG자신항체적경쟁성억제작용정제량의뢰관계,최대억제솔가체69.50%(여대조조상비,균유통계학의의,P＜0.01).경쟁성면역형광실험중,단련항체농도증가지40μg/ml시,BP-IgG재기저막대적침적급기개도적보체C3활화작용균피억제,정음성.결론 인원성항BP180단련항체재체외대BP치병성자신항체여항원결합급후속적보체활화구유일정적억제작용.
Objective To characterize the function of human anti-BP180 single-chain Fv antibody (scFv) in vitro. Methods The IgG autoantibodies against BP180 were purified by affinity chromatography from the sera of patients with BP. The inhibitive effect of previously constructed anti-BP180 scFv on the binding of anti-BP180 IgG autoantibodies to the recombinant NC16A domain of human BP180 antigen was observed by competitive ELISA, competitive immunofluorescence assay and competitive inhibition test for complement activation. Results As ELISA revealed, the scFv significantly inhibited the binding of anti-BP180 IgG autoantibodies to the corresponding antigen (P ＜ 0.01 ), and the inhibitive effect was dose-dependent within the concentration range from 0 to 60 μg/ml. The inhibitive rate peaked at 69.50%. The deposition of anti-BP180-IgG autoantibodies in basement membrane zone and the IgG autoantibody-mediated complement C3 activation were both suppressed by the scFv of 40 μg/ml. Conclusion The genetically engineered anti-BP180 scFv has a certain inhibitive effect on the binding of BP-IgG autoantibodies to BP180 antigen and on the subsequent complement activation in vitro.