Immunoreactivity of bullous pemphigoid (BP) autoantibodies against the NC16A and C-terminal domains of the 180 kDa BP antigen (BP180): immunoblot analysis and enzyme-linked immunosorbent assay using BP180 recombinant proteins

Nakatani, Chie; Muramatsu, Tsutomu; Shirai, Toshihiko

doi:10.1046/j.1365-2133.1998.02396.x

Cited by 76 publications

(82 citation statements)

References 10 publications

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“…Using rCol15 as antigen, we showed that approximately one-third of the tested autoimmune sera contained either IgG or IgA reactive with it. This is in contrast to previous studies with mainly bacterial recombinant fragments, which found most immunodominant epitopes in the NC16a domain (25,49,50), some in the distal amino-and carboxyl-terminal regions of the molecule, but none in the Col15 domain (23,28,51). By affinity adsorbing the rCol15-reactive immunoglobulins from the patient sera, we also showed that the circulating autoantibodies against rCol15 target collagen XVII at the epidermal basement membrane zone in situ.…”

Section: Discussioncontrasting

confidence: 52%

Collagen XVII Is Destabilized by a Glycine Substitution Mutation in the Cell Adhesion Domain Col15

Tasanen

Eble

Aumailley

et al. 2000

Journal of Biological Chemistry

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Collagen XVII is a hemidesmosomal transmembrane molecule important for epithelial adhesion in the skin. It exists in two forms, as a full-length protein and as a soluble ectodomain that is shed from the keratinocyte surface by furin-mediated proteolysis. To obtain information on the conformation and the functions of this unusual collagen, its largest collagenous domain, Col15, was expressed in a eukaryotic episomal expression system and purified by DEAE and fast protein liquid-Mono S chromatography. The protein was triple-helical (T m of 26.5°C) when produced in cultures containing ascorbic acid. When the vitamin supply was limited, the 4-hydroxyproline content was reduced from 74 to 9%, which, in turn, resulted in a drastic reduction of the stability of the triple helix. The glycine substitution mutation G627V associated with junctional epidermolysis bullosa, a human blistering skin disease, also had a striking effect on thermal stability of rCol15 causing partial unfolding already at 4°C. Col15 promoted cell adhesion of epithelial and fibroblastic cell lines with a ␤1 integrinmediated mechanism. In concert with this, in acquired autoimmune blistering skin diseases, circulating IgG and IgA autoantibodies were found to target rCol15r.Collagens are a family of closely related, although genetically distinct, extracellular matrix proteins. Each collagen consists of three polypeptide chains, ␣ chains, which contain a characteristic repeating "collagenous" triplet amino acid sequence -Gly-Xaa-Yaa-, where Xaa and Yaa denote amino acids other than glycine. In all collagen types, the ␣ chains also have non-collagenous domains of varying sizes (1). Typically, the three polypeptide chains are twisted around each other into a collagen triple helix; however, only suggestive data for this exist in the recently characterized transmembrane collagens, types XIII and XVII (for review, see Ref.2). Collagen XVII, also known as the 180-kDa bullous pemphigoid antigen, or BP180, is a structural component of hemidesmosomes, multiprotein complexes that mediate the adhesion of epidermal keratinocytes to the underlying basement membrane (3, 4). The cDNA sequence codes for a type II integral transmembrane protein of 1497 amino acids, with an intracellular domain of 560 amino acids, a short transmembrane stretch, and an extracellular collagenous domain of 914 amino acids with multiple interruptions. The length of the individual collagenous subdomains varies from 14 to 242 amino acid residues (5). Recently, it was established that collagen XVII exists as two molecular forms, i.e. as a full-length transmembrane homotrimer of three 180-kDa ␣1(XVII) chains and as a 120-kDa soluble form that corresponds to the extracellular domain and is presumably released from the cell surface through furin-mediated proteolytic processing (6, 7). In some situations also a shorter, approximately 90 -100-kDa fragment, has been observed (3, 4, 6). Very little is known about the molecular shape of collagen XVII under physiological conditions. Rotary shadowing ele...

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Section: Discussioncontrasting

confidence: 52%

Collagen XVII Is Destabilized by a Glycine Substitution Mutation in the Cell Adhesion Domain Col15

Tasanen

Eble

Aumailley

et al. 2000

Journal of Biological Chemistry

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“…16 ELISA analysis using recombinant COL17 NC16A demonstrated that 94-96% of BP sera reacts to COL17 NC16A peptides. 18,19 In addition, several major antigenic sites have been identified within the N-terminal 45 amino acid stretch of NC16A, while the remaining 28 amino acids of NC16A were shown to have no BP-associated epitopes. 17 Several studies have confirmed that the serum levels of anti-COL17 NC16A autoantibodies correlate with the disease severity of BP.…”

Section: Major Pathogenic Autoantigen: Type XVII Collagen (Col17)mentioning

confidence: 99%

What’s new in bullous pemphigoid

Ujiie

Shibaki

Nishie

et al. 2010

The Journal of Dermatology

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Bullous pemphigoid (BP) is the most common autoimmune blistering disease. BP patients have autoantibodies against type XVII collagen (COL17, also called BP180 or BPAG2), a type II transmembrane protein that spans the lamina lucida and projects into the lamina densa of the epidermal basement membrane. The non-collagenous 16A domain of COL17 is considered to contain pathogenic epitopes of BP. The transfer of immunoglobulin (Ig)G from BP patients fails to cause blisters on mouse skin probably due to differences between humans and mice in the amino acid sequence of NC16A pathogenic epitope of COL17. Passive transfer of rabbit IgG antibodies against the murine homolog of human COL17 NC16A triggers immune reactions to COL17 in mice, including complement activation, mast cell degranulation and neutrophilic infiltration, resulting in dermal-epidermal separation. Recent studies using COL17-humanized mice that express human COL17 but lack murine COL17 were the first to demonstrate the pathogenicity of anti-COL17 human BP IgG autoantibodies in vivo. These new findings provide a greater understanding of BP pathomechanisms and facilitate the development of novel specific and efficient therapeutic strategies for BP.

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“…16,17 Recently, it has been established that most BP serum samples react with the NC16A domain of BP180. 12,22,23 The purpose of this study was to test the hypothesis that the level of IgG reactivity to NC16A reflected disease severity in patients with BP. Fifteen consecutive patients were analyzed: 9 patients were treated with dapsone-prednisolone, and 6 patients received doxycycline-niacinamide.…”

Section: Commentmentioning

confidence: 99%

Serum Levels of Autoantibodies to BP180 Correlate With Disease Activity in Patients With Bullous Pemphigoid

Schmidt¹,

Obe²,

Bröcker³

et al. 2000

Arch Dermatol

315

257

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Immunoreactivity of bullous pemphigoid (BP) autoantibodies against the NC16A and C-terminal domains of the 180 kDa BP antigen (BP180): immunoblot analysis and enzyme-linked immunosorbent assay using BP180 recombinant proteins

Cited by 76 publications

References 10 publications

Collagen XVII Is Destabilized by a Glycine Substitution Mutation in the Cell Adhesion Domain Col15

Collagen XVII Is Destabilized by a Glycine Substitution Mutation in the Cell Adhesion Domain Col15

What’s new in bullous pemphigoid

Serum Levels of Autoantibodies to BP180 Correlate With Disease Activity in Patients With Bullous Pemphigoid

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