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The role of IgG subclass in the pathogenesis of bullous pemphigoid

IgG亚类在大疱性类天疱疮发病机制中的作用

基本信息

批准号：
04670646
负责人：
SUZUKI Masayuki
金额：
$ 1.41万
依托单位：
JICHI MEDICAL SCHOOL
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670646/
关键词：
IgG subclass bullous pemphigoid igG subclass interleukin 類天疱瘡抗原

项目摘要

We examined the role of IgG subclass in the mechanism of bullous pemphigoid (BP) as follows : (1)Whether BP IgG subclass binds to the different binding site of BP antigen or not. (2)Which BP IgG subclass induces directly dermo-epidermal separation. (3)Whether BP IgG subclass bound to BP antigen passes some signal into the cell or not. (4)Whether complement activated by BP IgG1 induces cell lysis or not. (5)Whether cultured keratinocytes secrete any chemical substance (ex.protease) or not, after BP IgG subclass binds to BP antigen on the surface of cultured keratinocytes.First, we examined the binding site of BP IgG subclass on BP antigen, using blocking immunofluorescense, immunoblot and ELISA.The result suggested that BP IgG1, IgG2 and IgG4 could bind to the same epitope of BP antigen, but considerable variation occured between patients. Second, we observed vertical section of luciffer yellow loaded keratinocytes, by using confocal laser scanning microscopy. The vertical image showed that there was no difference between BP IgG and normal IgG.Third, using confocal laser scanning microscopy, we chased the time course of fluorescent intensity change of fluo-3/am, snarf-1/am and dioc-5 which indicated intracellular Ca^<2+> concentration ([Ca^<2+>]i), intracellular pH and membrane potential, respectively. The result showed BP IgG1 induced a transient increase of [Ca^<2+>]i, even though fluo-3 loaded keratinocytes were incubated in Ca removed medium. There was no significant change about membrane potential. Forth, BP IgG1 induced a transient increase of [Ca^<2+>]i, when complement was added. But at the same time, it wasn't observed the cytolysis of luciffer yellow loaded keratinocytes. Fifth, we examined the presence of urokinase receptor (UKR) in the lesional skin of various skin diseases. The result showed that UKR was observed in the intercellular space of pemphgus vulgaris, but not observed at the basement membrane zone of BP.

我们从以下几个方面探讨了IgG亚类在大疱性类天疱疮（BP）发病机制中的作用：（1）BP IgG亚类是否与BP抗原的不同结合位点结合;（2）哪种BP IgG亚类直接诱导真皮-表皮分离。（3）与BP抗原结合的BPIgG亚类是否向细胞内传递信号。（4）BPIgG_1激活的补体是否诱导细胞溶解。（5）培养的角质形成细胞是否分泌任何化学物质本文首先用阻断免疫荧光、免疫印迹和ELISA方法检测了BP IgG亚类与BP抗原的结合位点，结果表明，BP IgG亚类1、2、4能与BP抗原的同一表位结合，但患者之间存在相当大的差异。其次，利用激光共聚焦显微镜观察负载荧光黄的角质形成细胞的垂直切片。第三，采用激光共聚焦显微镜，追踪fluo-3/am、snarf-1/am和dioc-5荧光强度变化的时间过程，分别反映细胞内Ca^2+浓度（[Ca^2+]i）、细胞内pH和膜电位。结果显示，即使在去钙培养基中培养，BPIgG 1也能诱导角质形成细胞[Ca^2+]i的瞬时升高。膜电位无明显变化。第四，当加入补体时，BP IgG 1可引起[Ca^<2+>]i的一过性升高。但未观察到负载荧光黄的角质形成细胞的细胞溶解作用。第五，我们检测了各种皮肤病皮损中尿激酶受体（UKR）的存在。结果表明，寻常型天疱疮细胞间隙有UKR，而BP基底膜区无UKR。