92-kD gelatinase is produced by eosinophils at the site of blister formation in bullous pemphigoid and cleaves the extracellular domain of recombinant 180-kD bullous pemphigoid autoantigen.
Eosinophils are prominent in bullous pemphigoid (BP), and proteases secreted from these and other inflammatory cells may induce disruption of the basement membrane. We used in situ hybridization and immunohistochemistry to localize the sites of 92-kD gelatinase expression in BP lesions. In all samples (20/20), a strong signal for gelatinase mRNA was detected only in eosinophils and was most pronounced where these cells accumulated at the floor of forming blisters. No other cells were positive for enzyme mRNA. Both eosinophils and neutrophils, however, contained immunoreactive 92-kD gelatinase indicating that active expression occurred only in eosinophils. Degranulated eosinophils were also seen near blisters, and as demonstrated by gelatin zymography, immunoblotting, and ELISA, 92-kD gelatinase protein was prominent in BP blister fluid. No other gelatinolytic activity was specifically detected in BP fluid, and only small amounts of 92-kD gelatinase were present in suction blister fluids. As demonstrated in vitro, 92-kD gelatinase cleaved the extracellular, collagenous domain of recombinant 180-kD BP autoantigen (BP180, BPAG2, HD4, type XVII collagen), a transmembrane molecule of the epidermal hemidesmosome. Our results suggest that production and release 92-kD gelatinase by eosinophils contributes significantly to tissue damage in BP.
Ståhle-Bäckdahl
M
M
Department of Dermatology, Karolinska Hospital, Stockholm, Sweden.
Inoue
M
M
Guidice
G J
GJ
Parks
W C
WC
D001324:Autoantigens / Q000378:metabolism*; D001768:Blister / Q000201:enzymology* / Q000209:etiology / Q000473:pathology; D002352:Carrier Proteins; D003094:Collagen; D017364:Collagenases / Q000235:genetics / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D004804:Eosinophils / Q000201:enzymology*; D006801:Humans; D017403:In Situ Hybridization; D020780:Matrix Metalloproteinase 9; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D010391:Pemphigoid, Bullous / Q000201:enzymology* / Q000473:pathology; D011487:Protein Conformation; D015347:RNA Probes; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000302:isolation & purification; D000097805:Collagen Type XVII
D001324:Autoantigens; D002352:Carrier Proteins; D003094:Collagen; D017364:Collagenases; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D020780:Matrix Metalloproteinase 9; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D015347:RNA Probes; D016372:RNA, Antisense; D012333:RNA, Messenger; D000097805:Collagen Type XVII; C486268:DST protein, human