Development of an ELISA to detect anti-BP180 autoantibodies in bullous pemphigoid and herpes gestationis.
Autoantibodies associated with the subepidermal blistering disorders bullous pemphigoid and herpes gestationis react with a 180-kD transmembrane hemidesmosomal protein, designated BP180. The BP180 ectodomain is composed of a series of interrupted collagen triple helical domains. Located on one of the noncollagenous extracellular segments of this protein is an immunodominant epitope, designated MCW-1, recognized by patient autoantibodies. In this investigation we have developed an enzyme-linked immunosorbent assay system to detect antibody reactivity against the MCW-1 epitope with the use of a bacterial fusion protein containing the BP180 autoantibody-reactive site. The following sera were assayed for reactivity with this recombinant protein: bullous pemphigoid (n = 62), herpes gestationis (n = 28), endemic pemphigus foliaceus (n = 17), lupus erythematosus (n = 15), and normal human sera (n = 22). This enzyme-linked immunosorbent assay-based protocol was shown to be highly specific (98.3%) in detecting autoantibody activity in bullous pemphigoid and herpes gestationis patients. Fifty-three percent of bullous pemphigoid sera and 71% of herpes gestations sera, but none of the control sera, yielded positive results in this assay. Of the patient sera that were known to react with full-length BP180, almost all showed reactivity with the MCW-1 antigenic site of this protein. Autoantibodies detected in this assay were predominantly of the immunoglobulin G class. The results presented here lend support to the hypothesis that this well-defined antigen/antibody system may be relevant in pathogenesis.
Giudice
G J
GJ
Department of Dermatology, Medical College of Wisconsin, Milwaukee 53226.
Wilske
K C
KC
Anhalt
G J
GJ
Fairley
J A
JA
Taylor
A F
AF
Emery
D J
DJ
Hoffman
R G
RG
Diaz
L A
LA
D001323:Autoantibodies / Q000032:analysis* / Q000276:immunology; D001324:Autoantigens / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D008970:Molecular Weight; D024041:Non-Fibrillar Collagens; D006559:Pemphigoid Gestationis / Q000175:diagnosis / Q000276:immunology* / Q000378:metabolism; D010391:Pemphigoid, Bullous / Q000175:diagnosis / Q000276:immunology* / Q000378:metabolism; D011247:Pregnancy; D011993:Recombinant Fusion Proteins / Q000276:immunology; D000097805:Collagen Type XVII
D001323:Autoantibodies; D001324:Autoantigens; D000939:Epitopes; D007074:Immunoglobulin G; D024041:Non-Fibrillar Collagens; D011993:Recombinant Fusion Proteins; D000097805:Collagen Type XVII