Janssen J J, Janssen B P, van Vugt A H
Institute of Ophthalmology, University of Nijmegen, The Netherlands.
Invest Ophthalmol Vis Sci. 1994 Jan;35(1):189-98.
To purify and characterize retinal pigment epithelial proteins exhibiting uveitogenic characteristics after immunization of Lewis rats, a broad panel of monoclonal antibodies directed against retinal pigment epithelial (RPE) protein antigens, was isolated.
Bovine RPE detergent extracts were used to isolate monoclonal antibodies against RPE antigens. Species and tissue specificity within the eye was tested through immunocytochemical analysis. Western blot analysis was used to determine the molecular weight of the RPE antigens.
Several RPE-reactive antibodies were obtained. At least four monoclonal antibodies were isolated that reacted with different RPE antigen types. Against most of the antigens more than one hybridoma cell line was isolated. Two hybridoma lines were isolated producing antibodies, which on immunocytochemical analysis showed strong reactivity with the RPE and eye muscle tissue. The latter monoclonal antibody recognizes a 32 kD protein in RPE cells. Five monoclonal antibodies recognize a protein with an apparent molecular weight of 65 kD. One cell line was isolated that produced antibodies showing an irregular reaction pattern with both iris and ciliary body.
RPE cells, striated eye muscle and smooth muscle cells share a 32 kD antigenic protein. This antigen is present in almost all ocular epithelial cells. Based on reaction patterns on Western blot and immunocytochemical analysis, there are at least three different 65 kD RPE antigens, two of which are RPE-specific and one of which is also present in the kidney epithelium.
为了纯化和鉴定在免疫Lewis大鼠后表现出葡萄膜炎致病特征的视网膜色素上皮蛋白，分离出了一系列针对视网膜色素上皮（RPE）蛋白抗原的单克隆抗体。
使用牛RPE去污剂提取物分离针对RPE抗原的单克隆抗体。通过免疫细胞化学分析测试眼内的物种和组织特异性。蛋白质印迹分析用于确定RPE抗原的分子量。
获得了几种与RPE反应的抗体。至少分离出四种与不同RPE抗原类型反应的单克隆抗体。针对大多数抗原，分离出了不止一种杂交瘤细胞系。分离出两个杂交瘤细胞系，其产生的抗体在免疫细胞化学分析中显示与RPE和眼肌组织有强烈反应。后一种单克隆抗体识别RPE细胞中的一种32kD蛋白。五种单克隆抗体识别一种表观分子量为65kD的蛋白。分离出一个细胞系，其产生的抗体对虹膜和睫状体呈现不规则反应模式。
RPE细胞、眼横纹肌和平滑肌细胞共享一种32kD的抗原性蛋白。这种抗原几乎存在于所有眼上皮细胞中。基于蛋白质印迹和免疫细胞化学分析的反应模式，至少有三种不同的65kD RPE抗原，其中两种是RPE特异性的，一种也存在于肾上皮中。
