Autoantibodies in a Subgroup of Patients with Linear IgA Disease React with the NC16A Domain of BP1801

Zillikens, Detlef; Herzele, K.; Georgi, M.; Schmidt, Enno; Chimanovitch, Iakov; Schumann, Hauke; Mascaró, J. M.; Díaz, Luis A.; Bruckner‐Tuderman, Leena; Bröcker, Eva B.; Giudice, George J.

doi:10.1046/j.1523-1747.1999.00808.x

Cited by 109 publications

(81 citation statements)

References 41 publications

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“…The ectodomain shedding of COL17 can evoke neoepitopes on the shed forms, which can be targeted by autoAbs of pemphigoid patients 16,17 . Previously, we…”

Section: Discussionmentioning

confidence: 99%

“…As is true for other transmembrane collagen family members, the extracellular domain of COL17 can be cleaved from the cell surface in vitro by a disintegrin and metalloproteases (ADAMs) 9, 10 and 17 12,13 , and interestingly, the ectodomain shedding occurs within the NC16A domain which contains major epitopes for BP autoAbs 14,15 . In addition, it is known that autoAbs from LAD 16 and certain BP patients 17 preferentially react with the shed ectodomain but not with the full-length form of COL17. This unique characteristic was also observed 3 in several monoclonal Abs (mAbs) directing the ectodomain of COL17 11,[18][19][20] , suggesting that neoepitopes appeared on the shed ectodomain of COL17.…”

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confidence: 99%

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Context-Dependent Regulation of Collagen XVII Ectodomain Shedding in Skin

Nishie

Natsuga

Iwata

et al. 2015

The American Journal of Pathology

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Abstract:Pemphigoid is a common autoimmune blistering disorder in which autoantibodies target transmembrane collagen XVII (COL17), a component of hemidesmosomes in basal keratinocytes. The ectodomain of COL17 can be cleaved from the cell surface within the juxtamembranous extracellular NC16A domain, and interestingly, certain autoantibodies of pemphigoid patients preferentially react with the shed ectodomain. These findings suggest that COL17 ectodomain shedding generates neoepitopes on the shed form; however, the regulatory mechanism of the shedding in in vivo skin and the pathogenicity of the neoepitope-targeting antibodies are still uncertain. To address these issues, we produced rabbit antibodies specifically reacting with N-terminal cleavage sites of the shed COL17 ectodomain. COL17 is a type II-oriented transmembrane glycoprotein whose N-terminus and C-terminus are in the cytoplasm and the extracellular matrix (ECM), respectively 10,11 . As is true for other transmembrane collagen family members, the extracellular domain of COL17 can be cleaved from the cell surface in vitro by a disintegrin and metalloproteases (ADAMs) 9, 10 and 17 12, 13 , and interestingly, the ectodomain shedding occurs within the NC16A domain which contains major epitopes for BP autoAbs 14, 15 . In addition, it is known that autoAbs from LAD . However, the Ab HK139 showed mottled staining at the dermal-epidermal junction (DEJ) of NHS; thus, Leu 524 may be a minor cleavage site in a physiological setting and a different cleavage site or sites may be involved in vivo. In addition, it is still uncertain whether neoepitope-specific Abs to COL17 are pathogenic for blister formation in vivo.In this study, we tried to determine whether COL17 is physiologically cleaved and whether the ectodomain shedding of COL17 produces neoepitopes in in vivo skin, and we investigated the pathogenic roles of Abs targeting the neoepitopes at the cleavage sites in blister formation. Materials and methods Cell CulturePrimary normal human epidermal keratinocytes (NHEKs) isolated from neonatal foreskin (Lonza) were cultured in keratinocyte growth medium (KGM, Lonza). To isolate primary mouse keratinocytes (NMKs), neonatal WT mice on a C57BL/6J background (Clea) were sacrificed, and whole skin sections were incubated with

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“…The ectodomain shedding of COL17 can evoke neoepitopes on the shed forms, which can be targeted by autoAbs of pemphigoid patients 16,17 . Previously, we…”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Context-Dependent Regulation of Collagen XVII Ectodomain Shedding in Skin

Nishie

Natsuga

Iwata

et al. 2015

The American Journal of Pathology

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“…Dermal extract and recombinant murine and human vWFA2 protein were fractionated by 6 and 12% SDS-PAGE, respectively, transferred to nitrocellulose, and analyzed by immunoblotting (35). For epitope mapping, patients' sera (1:100 dilution) were incubated at 4˚C overnight and were incubated Ab against human IgG-HRP (dilution 1:1000) at RT for 1 h. Circulating mouse vWFA2-specific Abs were detected by ELISA.…”

Section: If Microscopy Immunohistochemistry Immunoblot and Elisa Amentioning

confidence: 99%

B Cells, Dendritic Cells, and Macrophages Are Required To Induce an Autoreactive CD4 Helper T Cell Response in Experimental Epidermolysis Bullosa Acquisita

Iwata

Bieber

Tiburzy

et al. 2013

The Journal of Immunology

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In autoimmune bullous dermatoses (AIBD), autoantibodies induce blisters on skin or mucous membranes, or both. Mechanisms of continued autoantibody production and blistering have been well characterized using AIBD animal models. Mechanisms leading to the initial autoantibody production, however, have not been investigated in detail. Epidermolysis bullosa acquisita (EBA) is an AIBD associated with autoantibodies to type VII collagen (COL7). The majority of EBA patients’ sera recognize the noncollagenous domain 1, including the von Willebrand factor A–like domain 2 (vWFA2). In experimental EBA induced by immunization with GST-COL7, disease manifestation depended on the genetic background, a Th1 polarization, and the GST-tag. In this model, nude mice neither produced autoantibodies nor blisters. It has remained uncertain which APC and T cell subsets are required for EBA induction. We established a novel EBA model by immunization with vWFA2 fused to intein (lacking the GST-tag). All tested mouse strains developed autoantibodies, but blisters were exclusively observed in mice carrying H2s. In immunized mice, CD4 T cells specific for vWFA2 were detected, and their induction required presence of B cells, dendritic cells, and macrophages. Anti-vWFA2 autoantibodies located at the lamina densa bound to the dermal side of salt-split skin and induced blisters when transferred into healthy mice. Absence of CD8 T cells at time of immunization had no effect, whereas depletion of CD4 T cells during the same time period delayed autoantibody production and blisters. Collectively, we demonstrate the pathogenic relevance of Abs targeting the vWFA2 domain of COL7 and show the requirement of APC-induced CD4 T cells to induce experimental EBA.

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“…These results suggest that LAD sera, in contrast to the majority of BP sera, target epitopes on the entire BP180 ectodomain . 54 In another subset of patients, a combined IgA and IgG response was detected.55-57 For this disease , the term linear IgA/IgG bullous dermatosis was proposed .55 Interestingly, in addition to LAD patients , a large number of BP sera has been found to react with the 120 kD soluble ectodomain of BP180 (LAD-1) .49,58 In a recent study, we investigated the hypothesis that there is a significant overlap in the autoimmune isotype pro files associated with BP and LAD. For this study , sev eral new recombinant forms of BP180 were generated in the baculovirus expression system, including the full length protein.…”

Section: Linear Iga Diseasementioning

confidence: 99%

BP180 as the common autoantigen in blistering diseases with different clinical phenotypes.

Zillikens

2002

Keio j. med

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Abstract. Bullous pemphigoid antigen 180 (BP180, type XVII collagen) is a transmembrane hemi desmosomal glycoprotein of basal keratinocytes that spans the lamina lucida of the dermal-epidermal junction. Five autoimmune subepidermal blistering diseases are associated with an immune response to BP180, including bullous pemphigoid (BP), pemphigoid gestationis (PG), cicatricial pemphigoid (CP), lichen planes pemphigoides (LPP), and linear IgA disease (LAD). The BP180 ectodomain consists of 15 interrupted collagen domains. The largest non-collagenous (NC) 16A domain is located next to the cell membrane. In BP, autoantibodies are directed to a tightly clustered set of epitopes located at the N-terminal 45 amino acids of the NC16A domain. However, some BP sera also react with other por tions of the BP180 ectodomain or with the intracellular region of this protein. In PG, antibodies appear to exclusively recognize the immunodominant BP180 NC16A region. In CP, autoantibodies are directed to both the NC16A domain and the C-terminus of BP180 that projects into the lamina lucida/ lamina densa interface of the dermal-epidermal junction. In LPP, autoantibodies react with an epitope located at the C-terminus of NC16A, that is not targeted by BP or PG sera. Finally, the epidermal 97 kDa and the keratinoctye-derived 120 kDa autoantigens of LAD (LABD97 and LAD-1, respectively) have recently been identified as portions of the BP180 ectodomain. These observations demonstrate that different clinical phenotypes are associated with an autoinunune response to the same autoantigen yet the immunoglobulin subclass of the autoantibody and the epitope that is recognized may be dif ferent. Adhesion of basal keratinocytes to the underlying dermis is maintained by various structural proteins of hemidesmosomes, including the 230 kD bullous pem phigoid antigen (BP230), plectin, a6134 integrin, BP180, and laminin 5. Directly or through other proteins these hemidesmosomal components associate with anchor ing fibrils which originate in the lamina densa of the dermal-epidermal junction (DEJ) and extend into the subjacent connective tissue. The major component of anchoring fibrils is type VII collagen.1Mutations for the different components of this anchoring complex as well as formation of autoanti bodies against these structures result in split forma tion at the level of the DEJ. A major component of the dermal-epidermal anchoring complex and a major target of autoantibodies is BP180. BP180 is a member of the collagen protein family and is also referred to as type XVII collagen. It is a transmembrane glycoprotein with a type II orientation, i.e., the N-terminus localizes to the cytoplasm, whereas the C-terminal ectodomain consisting of about 1,000 amino acids projecting into the extracellular space.2.3 Rotary shadowing revealed that BP180 contains an intracellular globular head, an extracellular rigid rod corresponding to the 16th non collagenous (NC) A and the 15th collagenous domain spanning the lamina lucida, and a flexible tail extend ing into t...

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Autoantibodies in a Subgroup of Patients with Linear IgA Disease React with the NC16A Domain of BP1801

Cited by 109 publications

References 41 publications

Context-Dependent Regulation of Collagen XVII Ectodomain Shedding in Skin

Context-Dependent Regulation of Collagen XVII Ectodomain Shedding in Skin

B Cells, Dendritic Cells, and Macrophages Are Required To Induce an Autoreactive CD4 Helper T Cell Response in Experimental Epidermolysis Bullosa Acquisita

BP180 as the common autoantigen in blistering diseases with different clinical phenotypes.

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