Labeling of fractured human skin with antibodies to BM 600/nicein, epiligrin, kalinin and other matrix components.
A variety of methods were used to fracture the dermal-epidermal junction (DEJ) of human skin. These included warm and hot phosphate buffered saline, trypsin, cold 1 M salt, potassium bromide and proteolytic digestion with dispase. The localization and sensitivity of basement membrane components (bullous pemphigoid antigen, BM 600/nicein, epiligrin, kalinin, laminin, collagens IV and VII (EBA antigen) and linkin) were determined after the DEJ was fractured by each method. We found that the basement membrane zone proteins, BM 600/nicein, epiligrin and kalinin remained with the dermal side of the DEJ fractured through the lamina lucida by cold salt, phosphate buffered saline and potassium bromide. BM 600/nicein, epiligrin and kalinin were not detected after treatment with trypsin. In contrast, laminin, another glycoprotein in the lamina lucida, was insensitive to all of the procedures, but co-localized to the dermal side of DEJ-fractured skin. We also found that separation of the DEJ with brief exposure of skin to 56 degrees C provided a useful substrate for testing the autoantibodies in the sera of patients with epidermolysis bullosa acquisita (EBA). Heat-separated skin can be prepared in a significantly shorter period of time than salt-separated skin.
Ceilley
E
E
Department of Dermatology, Stanford University School of Medicine, CA.
Watanabe
N
N
Shapiro
D
D
Verrando
P
P
Bauer
E A
EA
Burgeson
R
R
Briggaman
R A
RA
Woodley
D T
DT
D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D001324:Autoantigens / Q000032:analysis / Q000276:immunology; D001485:Basement Membrane / Q000737:chemistry; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules / Q000032:analysis* / Q000276:immunology; D003094:Collagen / Q000032:analysis / Q000276:immunology; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D005455:Fluorescent Antibody Technique; D006801:Humans; D007797:Laminin / Q000032:analysis / Q000276:immunology; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D012867:Skin / Q000737:chemistry*; D000097794:Kalinin; D000097805:Collagen Type XVII
D000906:Antibodies; D001324:Autoantigens; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D003094:Collagen; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D007797:Laminin; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D000097794:Kalinin; D000097805:Collagen Type XVII; C486268:DST protein, human