Major controversies exist in the literature on the presence of blood group antigens on the endothelial and stromal layers of the cornea, and the importance of major histocompatibility typing for keratoplasty. Antibodies were raised in BALB/C mice against water soluble proteins of corneal epithelium. Following fusion of spleen cells with myeloma cells (Sp2/0-Ag14) hybrid colonies were maintained in HAT selective medium. The supernates of each colony were measured and screened for antibody production by radioimmunoassay. Gel electrophoresis of the antigen showed nine major bands. The antibodies were partially characterized by cross reaction against other soluble corneal fractions.
Human corneal epithelial water insoluble proteins were used to immunize mice. Immune splenocytes were fused with Sp 2/0-Ag14 mouse myeloma cells by 40% PEG. Hybridoma antibodies obtained by somatic cell fusion were tested by radioimmunoassay. Supernatants from antibody positive hybrid colonies were used in immunofluorescence and crossreaction assays to locate and characterize corneal epithelial antigens. At least three distinct epithelial cell antigens were detected, one of which cross-reacts with rabbit corneal epithelial cells.
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