4148 Glucose dependent transcriptional regulation of GLUT-3 mRNA in human retinal endothelial cells

Repare In order to purify and characterize retinal pigment epithelial proteins exhibiting uveitogenic characteristics upon immunization of Lewis rats, a broad panel of monoclonal antibodies directed against retinal pigment epithelial (RPE) protein antigens, was isolated. Merh&s Bovine RPE detergent extracts were used to isolate monoclonal antibodies against RIPE antigens. Species and tissue specificity within the eye was ter.ted through immunocytochemical analysis. Western blot analysis was used to determine the molecular weight of the RPE antigens. Rest&. 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 upon immunocytochemical analysis showed strong reactivity with the RPE and eye muscle tissue. The latter monoclonal antibody recognizes a 32 kD protein in retinal pigment epithelium 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. ConcLLpionc Retinal pigment epitlmlium cells, striated eye muscle and smooth muscle cells share a 32 kE antigenic protein. This antigen is present in almost all ocular epithelial cells. Based on reaction patterns upon western blot and immunocytochemical analysis, there are at least three different 65 kD KPE antigens, two of which are RPE specific and one of which is also present in the kidney epithelium.