Subepidermal blistering induced by human autoantibodies to BP180 requires innate immune players in a humanized bullous pemphigoid mouse model.

Bullous pemphigoid (BP) is a cutaneous autoimmune inflammatory disease associated with subepidermal blistering and autoantibodies against BP180, a transmembrane collagen and major component of the hemidesmosome. Numerous inflammatory cells infiltrate the upper dermis in BP. IgG autoantibodies in BP fix complement and target multiple BP180 epitopes that are highly clustered within a non-collagen linker domain, termed NC16A. Anti-BP180 antibodies induce BP in mice. In this study, we generated a humanized mouse strain, in which the murine BP180NC14A is replaced with the homologous human BP180NC16A epitope cluster region. We show that the humanized NC16A (NC16A+/+) mice injected with anti-BP180NC16A autoantibodies develop BP-like subepidermal blisters. The F(ab')(2) fragments of pathogenic IgG fail to activate the complement cascade and are no longer pathogenic. The NC16A+/+ mice pretreated with mast cell activation blocker or depleted of complement or neutrophils become resistant to BP. These findings suggest that the humoral response in BP critically depends on innate immune system players.

[1]  E. Bröcker,et al.  IgG4 and IgE are the major immunoglobulins targeting the NC16A domain of BP180 in Bullous pemphigoid: serum levels of these immunoglobulins reflect disease activity. , 2000, Journal of the American Academy of Dermatology.

[2]  G. Till,et al.  The role of complement in experimental bullous pemphigoid. , 1995, The Journal of clinical investigation.

[3]  S. Katz,et al.  Herpes gestationis. Immunopathology and characterization of the HG factor. , 1976, The Journal of clinical investigation.

[4]  M. Kramer,et al.  The autoimmune blistering skin disease bullous pemphigoid. The presence of plasmin/alpha 2-antiplasmin complexes in skin blister fluid indicates plasmin generation in lesional skin. , 1993, The Journal of clinical investigation.

[5]  K. Nishioka,et al.  High histamine level in the blister fluid of bullous pemphigoid , 2004, Archives of Dermatological Research.

[6]  D M Lewis,et al.  An in vitro model of immune complex-mediated basement membrane zone separation caused by pemphigoid antibodies, leukocytes, and complement. , 1982, The Journal of investigative dermatology.

[7]  J. Uitto,et al.  Demonstration of collagenase and elastase activities in the blister fluids from bullous skin diseases. Comparison between dermatitis herpetiformis and bullous pemphigoid. , 1983, The Journal of investigative dermatology.

[8]  J. Uitto,et al.  Human bullous pemphigoid antigen (BPAG1). Amino acid sequences deduced from cloned cDNAs predict biologically important peptide segments and protein domains. , 1991, The Journal of biological chemistry.

[9]  M. Kramer,et al.  Enhanced association of plasminogen/plasmin with lesional epidermis of bullous pemphigoid , 1992, The British journal of dermatology.

[10]  A. Sonnenberg,et al.  Current insights into the formation and breakdown of hemidesmosomes. , 2006, Trends in cell biology.

[11]  K. Hara,et al.  Lichen planus pemphigoides: identification of 180 kd hemidesmosome antigen. , 1995, Journal of the American Academy of Dermatology.

[12]  L. Diaz,et al.  Cloning and primary structural analysis of the bullous pemphigoid autoantigen BP180. , 1992, The Journal of investigative dermatology.

[13]  J. Usukura,et al.  Demonstration of the Molecular Shape of BP180, a 180-kDa Bullous Pemphigoid Antigen and Its Potential for Trimer Formation* , 1996, The Journal of Biological Chemistry.

[14]  D. Roop,et al.  Isolation of complementary DNA for bullous pemphigoid antigen by use of patients' autoantibodies. , 1988, The Journal of clinical investigation.

[15]  K. Owaribe,et al.  HD4, a 180 kDa bullous pemphigoid antigen, is a major transmembrane glycoprotein of the hemidesmosome. , 1993, Journal of biochemistry.

[16]  E. Bröcker,et al.  Elevated levels of interleukin-8 in blister fluid of bullous pemphigoid compared with suction blisters of healthy control subjects. , 1996, Journal of the American Academy of Dermatology.

[17]  T. Tomasi,et al.  Evidence for complement activation via the alternate pathway in skin diseases, I. Herpes gestationis, systemic lupus erythematosus, and bullous pemphigoid. , 1973, The Journal of clinical investigation.

[18]  L. Diaz,et al.  A recombinant form of the human BP180 ectodomain forms a collagen-like homotrimeric complex. , 1997, Biochemistry.

[19]  M. Polette,et al.  Respective contribution of neutrophil elastase and matrix metalloproteinase 9 in the degradation of BP180 (type XVII collagen) in human bullous pemphigoid. , 2001, The Journal of investigative dermatology.

[20]  E. Shevach,et al.  Characterization of bullous pemphigoid antigen: A unique basement membrane protein of stratified squamous epithelia , 1981, Cell.

[21]  J. C. Jones,et al.  Cytoplasmic domain of the 180-kD bullous pemphigoid antigen, a hemidesmosomal component: molecular and cell biologic characterization. , 1992, The Journal of investigative dermatology.

[22]  M. Mihm,et al.  Bullous pemphigoid, an ultrastructural study of the inflammatory response: eosinophil, basophil and mast cell granule changes in multiple biopsies from one patient. , 1982, The Journal of investigative dermatology.

[23]  D. Priebat,et al.  Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. , 1982, The Journal of investigative dermatology.

[24]  Enno Schmidt,et al.  Autoantibodies to bullous pemphigoid antigen 180 induce dermal-epidermal separation in cryosections of human skin. , 2002, The Journal of investigative dermatology.

[25]  K. Hashimoto,et al.  Eosinophilic spongiosis in bullous pemphigoid. , 1984, Archives of dermatology.

[26]  J. Saurat,et al.  Autoantibodies to the extracellular and intracellular domain of bullous pemphigoid 180, the putative key autoantigen in bullous pemphigoid, belong predominantly to the IgG1 and IgG4 subclasses , 2001, The British journal of dermatology.

[27]  J. Saurat,et al.  The major cicatricial pemphigoid antigen is a 180-kD protein that shows immunologic cross-reactivities with the bullous pemphigoid antigen. , 1992, The Journal of investigative dermatology.

[28]  R. Dummer,et al.  Detection of elevated levels of IL-4, IL-6, and IL-10 in blister fluid of bullous pemphigoid , 1996, Archives of Dermatological Research.

[29]  J. Zone,et al.  The 97 kDa linear IgA bullous disease antigen is identical to a portion of the extracellular domain of the 180 kDa bullous pemphigoid antigen, BPAg2. , 1998, The Journal of investigative dermatology.

[30]  E. Bröcker,et al.  Serum levels of autoantibodies to BP180 correlate with disease activity in patients with bullous pemphigoid. , 2000, Archives of dermatology.

[31]  R. Jordon,et al.  The complement system in bullous pemphigoid. III. Fixation of C1q and C4 by pemphigoid antibody. , 1975, The Journal of laboratory and clinical medicine.

[32]  A. Cochran,et al.  Mononuclear cell infiltrates in bullous disease. , 1987, The Journal of investigative dermatology.

[33]  J. R. McMillan,et al.  Humanization of autoantigen , 2007, Nature Medicine.

[34]  S. Grando,et al.  Mediators of inflammation in blister fluids from patients with pemphigus vulgaris and bullous pemphigoid. , 1989, Archives of dermatology.

[35]  E. Bröcker,et al.  Granulocyte‐derived elastase and gelatinase B are required for dermal–epidermal separation induced by autoantibodies from patients with epidermolysis bullosa acquisita and bullous pemphigoid , 2004, The Journal of pathology.

[36]  H. Tagami,et al.  Induction of leukocyte adherence at the basement membrane zone with subsequent activation of their metabolic pathway by pemphigoid antibodies and complement. , 1983, Acta dermato-venereologica.

[37]  W. Parks,et al.  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. , 1994, The Journal of clinical investigation.

[38]  I. Iwamoto,et al.  Increased immunoreactive interleukin-5 levels in blister fluids of bullous pemphigoid , 2004, Archives of Dermatological Research.

[39]  E. Beutner,et al.  Basement zone antibodies in bullous pemphigoid. , 1967, JAMA.

[40]  J. Troy,et al.  A passive transfer model of the organ-specific autoimmune disease, bullous pemphigoid, using antibodies generated against the hemidesmosomal antigen, BP180. , 1993, The Journal of clinical investigation.

[41]  S Grover,et al.  Fitzpatrick's Dermatology in General Medicine , 2008 .

[42]  D. Broide,et al.  Inhibition of eosinophilic inflammation in allergen-challenged, IL-1 receptor type 1-deficient mice is associated with reduced eosinophil rolling and adhesion on vascular endothelium. , 2000, Blood.

[43]  N. Day,et al.  The complement system in bullous pemphigoid. II. Immunofluorescent evidence for both classical and alternate-pathway activation. , 1975, Clinical immunology and immunopathology.

[44]  Zhi Liu.  Bullous pemphigoid: using animal models to study the immunopathology. , 2004, The journal of investigative dermatology. Symposium proceedings.

[45]  D. Parry,et al.  Comparison of molecularly cloned bullous pemphigoid antigen to desmoplakin I confirms that they define a new family of cell adhesion junction plaque proteins. , 1991, The Journal of biological chemistry.

[46]  H. Sonozaki,et al.  An eosinophil chemotactic factor present in blister fluids of bullous pemphigoid patients. , 1976, Journal of immunology.

[47]  W. Saunders,et al.  Isolation of a human epidermal cDNA corresponding to the 180-kD autoantigen recognized by bullous pemphigoid and herpes gestationis sera. Immunolocalization of this protein to the hemidesmosome. , 1990, The Journal of clinical investigation.

[48]  S. Kawana,et al.  Increased levels of immunoreactive leukotriene B4 in blister fluids of bullous pemphigoid patients and effects of a selective 5-lipoxygenase inhibitor on experimental skin lesions. , 1990, Acta dermato-venereologica.

[49]  M. Mihm,et al.  Morphologic and functional evidence for release of mast-cell products in bullous pemphigoid. , 1978, The New England journal of medicine.

[50]  D. Zillikens,et al.  Tight clustering of extracellular BP180 epitopes recognized by bullous pemphigoid autoantibodies. , 1997, The Journal of investigative dermatology.

[51]  Z. S. Wang,et al.  Recruitment of neutrophils during IgE-dependent cutaneous late phase reactions in the mouse is mast cell-dependent. Partial inhibition of the reaction with antiserum against tumor necrosis factor-alpha. , 1991, The Journal of clinical investigation.

[52]  M. G. Fleming,et al.  Mast cells play a key role in neutrophil recruitment in experimental bullous pemphigoid. , 2001, The Journal of clinical investigation.

[53]  K. Yancey,et al.  Detection of IgG autoantibodies in the sera of patients with bullous and gestational pemphigoid: ELISA studies utilizing a baculovirus-encoded form of bullous pemphigoid antigen 2. , 1998, The Journal of investigative dermatology.

[54]  B. Zelickson,et al.  Bullous pemphigoid and herpes gestationis autoantibodies recognize a common non-collagenous site on the BP180 ectodomain. , 1993, Journal of immunology.

[55]  G. Till,et al.  A major role for neutrophils in experimental bullous pemphigoid. , 1997, The Journal of clinical investigation.

[56]  J. Bonnetblanc,et al.  Immunoblot analysis of IgG subclasses of circulating antibodies in bullous pemphigoid. , 1990, Clinical immunology and immunopathology.