Pemphigus foliaceus antigen: characterization of a keratinocyte envelope associated pool and preparation of a soluble immunoreactive fragment.
In both the endemic and sporadic forms of pemphigus foliaceus (PF), antiepidermal autoantibodies against desmoglein I are present. Desmoglein I is a highly insoluble 160-kD transmembrane glycoprotein of the desmosomal core. The detailed immunochemical characterization of the epitope(s) recognized by the PF autoantibodies is hampered by its large molecular weight and the insolubility of desmoglein I in nondenaturing buffers. This study was designed to identify alternative methods that could yield soluble immunoreactive PF antigen (Ag) from normal human epidermis. The presence of PF Ag in human epidermis and in its soluble or insoluble fractions was monitored by indirect immunofluorescence, immunoadsorption of PF sera, and immunoprecipitation of radiolabeled fractions. The PF Ag from trypsin-resistant, radiolabeled cell envelope preparations was cleaved by papain and immunoprecipitated by PF sera. A 50-kD peptide, isoelectric at pH 5.5-5.8, was immunoprecipitated by sera from all patients with endemic PF (n = 15) or idiopathic PF (n = 4), and by two of four pemphigus vulgaris sera, but by no control sera (n = 7). This study shows that a significant fraction of the PF Ag is insoluble, trypsin-resistant, and is associated with the cornified cell envelope fraction, but an Ag fragment can be obtained in a small molecular weight, soluble, and immunoreactive form by papain digestion. This 50-kD papain fragment is more amenable to detailed chemical and immunologic characterization than the native molecule.
Labib
R S
RS
Department of Dermatology, Johns Hopkins University, Baltimore, Maryland.
Camargo
S
S
Futamura
S
S
Martins
C R
CR
Rock
B
B
Anhalt
G J
GJ
Diaz
L A
LA
D000941:Antigens / Q000276:immunology*; D002454:Cell Differentiation; D004591:Electrophoresis, Polyacrylamide Gel; D000078404:Epidermal Cells; D004817:Epidermis / Q000276:immunology*; D006801:Humans; D007633:Keratins; D010206:Papain / Q000494:pharmacology; D010392:Pemphigus / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology / Q000302:isolation & purification*; D012995:Solubility; D014357:Trypsin / Q000494:pharmacology
D000941:Antigens; D010446:Peptide Fragments; D007633:Keratins; D014357:Trypsin; D010206:Papain