Heat shock protein 90 is required for ex vivo neutrophil-driven autoantibody-induced tissue damage in experimental epidermolysis bullosa acquisita.
A broad range of immunosuppressive and immunomodulatory effects of heat shock protein 90 (Hsp90) blockade has been described in models of autoimmune bullous diseases, but the direct contribution of this chaperone to neutrophil effector pathways in the context of autoantibody-driven blistering is generally unknown. Therefore, this has been addressed in the current study on the basis of the subepidermal blistering disease epidermolysis bullosa acquisita (EBA) characterized by autoantibodies against type VII collagen, in which a crucial role of neutrophils and both their reactive oxygen species and matrix metalloproteinases in mediating tissue injury has been established. First, the Hsp90 antagonist 17-DMAG dose-dependently inhibited dermal-epidermal separation ex vivo in cryosections of human skin induced by co-incubation of EBA patient autoantibodies with neutrophils from healthy blood donors. Next, 17-DMAG dose-dependently suppressed production and release of reactive oxygen species by human neutrophils induced by both fMLP ± LPS and EBA-specific immune complexes. In addition, co-immunoprecipitation studies revealed that extracellular Hsp90 interacted with secreted matrix metalloproteinases 2 and 12 in sera of EBA patients, suggesting that these basement membrane-degrading proteolytic enzymes are client proteins of Hsp90 and dependent on its chaperone function. Our findings add to the knowledge of the multimodal anti-inflammatory effects of Hsp90 blockade and implicate that Hsp90 is closely involved in the effector mechanisms of neutrophil-driven autoantibody-induced tissue damage, thus being a relevant therapeutic target in patients with neutrophil-mediated autoimmune diseases such as inflammatory types of EBA.
Department of Dermatology, University of Lübeck, Lübeck, Germany.
Stefan
S
Tukaj
Institute of Medical Microbiology and Hygiene, University of Lübeck, Lübeck, Germany.
Lars
L
Hellberg
Institute of Medical Microbiology and Hygiene, University of Lübeck, Lübeck, Germany.
Christopher
C
Ueck
Department of Dermatology, University of Lübeck, Lübeck, Germany.
Martin
M
Hänsel
Department of Dermatology, University of Lübeck, Lübeck, Germany.
Unni
U
Samavedam
Department of Dermatology, University of Lübeck, Lübeck, Germany.
Detlef
D
Zillikens
Department of Dermatology, University of Lübeck, Lübeck, Germany.
Ralf J
RJ
Ludwig
Institute of Medical Microbiology and Hygiene, University of Lübeck, Lübeck, Germany.
Tamás
T
Laskay
Department of Dermatology, University of Lübeck, Lübeck, Germany.
Michael
M
Kasperkiewicz
D001323:Autoantibodies / Q000276:immunology / Q000502:physiology*; D016227:Benzoquinones / Q000494:pharmacology; D016022:Case-Control Studies; D002478:Cells, Cultured; D024143:Collagen Type VII / Q000276:immunology; D004305:Dose-Response Relationship, Drug; D016107:Epidermolysis Bullosa Acquisita / Q000276:immunology / Q000473:pathology* / Q000503:physiopathology*; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D047468:Immunoprecipitation; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D053508:Matrix Metalloproteinase 12 / Q000502:physiology; D020778:Matrix Metalloproteinase 2 / Q000502:physiology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012867:Skin / Q000187:drug effects / Q000473:pathology* / Q000503:physiopathology
D001323:Autoantibodies; D016227:Benzoquinones; D024143:Collagen Type VII; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D017382:Reactive Oxygen Species; C448659:17-(dimethylaminoethylamino)-17-demethoxygeldanamycin; D020778:Matrix Metalloproteinase 2; D053508:Matrix Metalloproteinase 12
autoantibody; heat shock protein 90; inflammation; neutrophil