Isolation and immunologic identification of basement membrane zone antigens from human skin
Scaletta, L.J.; Occhino, J.C.; MacCallum, D.K.; Lillie, J.H.
Laboratory Investigation; A Journal of Technical Methods and Pathology 39(1): 1-9
1978
ISSN/ISBN: 0023-6837 PMID: 682587 Document Number: 133393
Antigens isolated from the basement membrane zone (BMZ) of human skin following reduction and alkylation were used to stimulate antibody formation in rabbits. Antibodies to BMZ antigens isolated after 2 h of reduction produced BMZ and connective tissue fluorescence in skin and oral mucosa when examined by indirect immunofluorescence. Antibodies to antigens isolated after a 30-min reaction period localized primarily in the BMZ with minimal connective tissue localization. Antisera to subsequent fractions isolated between 30-60 min and 60-120 min demonstrated progressively weaker BMZ staining and increased connective tissue localization. Adsorption of these antisera with an extract of dermis eliminated connective tissue localization without affecting anti-BMZ activity. The adsorbed BMZ antibody reacted only with basement membranes of stratified squamous epithelia. Other basement membranes were nonreactive. Strips of epidermis-free skin were treated with the reducing mixture for times ranging from 30 min to 24 h to determine the rate of loss of BMZ antigens from the dermal surface and to correlate these findings with ultrastructural changes in the basal lamina and associated structures. A gradual loss of BMZ antigens during the first 2 h of reduction were demonstrated, after which time antigens could no longer be detected on the dermal surface by immunofluorescence. EM examination of skin basal lamina treated for periods up to 24 h showed progressive structural changes in the lamina, associated structures and dermal collagen during the first 8 h. Although altered, a recognizable basal lamina remained after 24 h in the solubilizing solution.