Monoclonal antibodies to epidermal and basement membrane zone antigens have been formed and new antibodies will be formed by fusing spleen cells from immunized mice with the mouse myeloma cell line P3/NS1. Mice immunized with a human cervical carcinoma cell line (ME-180) have formed an antibody to human anchoring fibrils. The antigen in human skin and ME-180 will be completely characterized with its molecular weight, isoelectric point, peptide and cyanogen bromide map and amino acid composition. The cell of origin of the antigen in skin will be determined by biosynthetic studies and dermal/epidermal recombinations on the nude mouse. The formation of the anchoring fibril and its antigen will be sequentially followed during wound healing and during epidermal cell attachment to collagen-coated tissue culture dishes. Gene mapping of the anchoring fibril antigen will be performed by somatic cell hybridization techniques using ME-180 cells which will be fused with polyethylene glycol with mouse epidermal cells. The anchoring fibril antigen has reacted with many samples of human skin but is non-reactive with the non-blistered skin of patients with dystrophic epidermolysis bullosa, a disease in which anchoring fibrils are absent as shown by electron microscopy. Attempts to use the anchoring fibril antibody to follow therapy of epidermolysis bullosa will be made both by biopsies of patients and by following the skin biopsies of the epidermolysis bullosa skin transplanted to the nude mouse. Success with these techniques encourages us to use these techniques to form antibodies which will allow dissection of the heterogeneous population of epidermal basal cells and also which distinguishes basal from non-basal cells.
