Characterization of monoclonal antibodies recognizing retinal pigment epithelial antigens.
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Janssen
J J
JJ
Institute of Ophthalmology, University of Nijmegen, The Netherlands.
Janssen
B P
BP
van Vugt
A H
AH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000918:Antibody Specificity / Q000276:immunology; D001324:Autoantigens / Q000276:immunology*; D015153:Blotting, Western; D002417:Cattle; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D005123:Eye / Q000276:immunology; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D010857:Pigment Epithelium of Eye / Q000276:immunology*; D051381:Rats; D011917:Rats, Inbred Lew; D012168:Retinal Pigments / Q000276:immunology*; D012589:Sciuridae
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D012168:Retinal Pigments