Presentation is loading. Please wait.

Presentation is loading. Please wait.

C-Terminal Processing of Collagen XVII Induces Neoepitopes for Linear IgA Dermatosis Autoantibodies  Ellen Toyonaga, Wataru Nishie, Kentaro Izumi, Ken.

Published byMárton Török Modified over 5 years ago

Similar presentations

Presentation on theme: "C-Terminal Processing of Collagen XVII Induces Neoepitopes for Linear IgA Dermatosis Autoantibodies  Ellen Toyonaga, Wataru Nishie, Kentaro Izumi, Ken."— Presentation transcript:

1 C-Terminal Processing of Collagen XVII Induces Neoepitopes for Linear IgA Dermatosis Autoantibodies 
Ellen Toyonaga, Wataru Nishie, Kentaro Izumi, Ken Natsuga, Hideyuki Ujiie, Hiroaki Iwata, Jun Yamagami, Yoshiaki Hirako, Daisuke Sawamura, Wataru Fujimoto, Hiroshi Shimizu  Journal of Investigative Dermatology  Volume 137, Issue 12, Pages (December 2017) DOI: /j.jid Copyright © 2017 The Authors Terms and Conditions

2 Figure 1 Indirect immunofluorescence and Western blotting detected by LAD autoantibodies and mAbs. (a) Schematic of full-length human COL17 (COL17), LAD-1, LABD97, and recombinant C-terminal–deleted COL17. The epitope of each antibody is shown at the bottom. (b) Western blotting using a mixture of full-length COL17, LAD-1, and LABD97 was analyzed using LAD (numbers 1, 2, 5, 7, and 10) and BP sera (numbers 3 and 4) followed by incubation with horseradish peroxidase-conjugated anti-human IgA and IgG, respectively. (c) Schematic of recombinant mouse COL17 and partially humanized mouse COL17. (d) Indirect immunofluorescence staining of normal human skin using hC17-ect15. Scale bar = 100 μm. (e) Western blotting using full-length COL17 with versus without digestion by plasmin. The right panel shows Coomassie blue staining. BP, bullous pemphigoid; C-terminal, carboxyl-terminal; COL17, collagen XVII; LAD, linear IgA dermatosis; N, normal control sera. Journal of Investigative Dermatology  , DOI: ( /j.jid ) Copyright © 2017 The Authors Terms and Conditions

3 Figure 2 Immunoprecipitation and Western blotting using five different recombinant COL17 proteins detected by LAD autoantibodies and different mAbs. (a) Western blotting using recombinant human full-length COL17, COL17 (Δ1–2), COL17 (Δ1–6), COL17 (Δ1–10), and COL17 (Δ1–14) proteins were detected by the mAbs anti-DDDDK-tag, hC17-ect15, and BMM62. (b) Western blotting using mouse COL17 (upper panels) and partially humanized mouse COL17 (lower panels) detected by the mAb hC17-ect15 and the polyclonal antibody Mo-NC14A as a positive control. (c) The full-length COL17 and COL17 (Δ1–2), COL17 (Δ1–6), COL17 (Δ1–10), and COL17 (Δ1–14) were immunoprecipitated by the mAbs TS39-3, hC17-ect15, and control mIgG1, followed by Western blotting using anti-DDDDK-tag mAbs. (d) Western blotting using COL17, COL17 (Δ1–2), COL17 (Δ1–6), COL17 (Δ1–10), and COL17 (Δ1–14) detected by the mAb anti-DDDDK-tag, normal human sera, and LAD sera. The mAb anti-DDDDK-tag shows that the amounts loaded for the five proteins were equal. COL17, collagen XVII; LAD, linear IgA dermatosis. Journal of Investigative Dermatology  , DOI: ( /j.jid ) Copyright © 2017 The Authors Terms and Conditions

4 Figure 3 ELISA using full-length COL17 and COL17 (Δ1–6). Sera from normal control subjects (n = 10) and LAD patients (n = 9, numbers 1–9 in Figure 1b and Supplementary Figure S1a) were assayed in an ELISA using (left) full-length COL17 (COL17) and (middle) COL17 (Δ1–6) as substrates without normalization using the mAb anti-DDDDK-tag. Note that COL17 (Δ1–6) and full-length COL17 (COL17) can detect LAD autoantibodies. LAD autoantibodies have significantly greater binding affinity to COL17 (Δ1–6) than to (right) COL17. OD was normalized by the mAb anti-DDDDK-tag. Dotted lines indicate +2 standard deviations from the normal control. COL17, collagen XVII; LAD, linear IgA dermatosis; OD, optical density. Journal of Investigative Dermatology  , DOI: ( /j.jid ) Copyright © 2017 The Authors Terms and Conditions

5 Figure 4 In vivo and in vitro pathogenicity of the mAb hC17-ect15. (a) Normal control mouse IgG1, the mAbs TS39-3, and hC17-ect15 were injected into the ears of adult COL17-humanized mice (n = 4/group) 48 hours before rubbing. TS39-3 causes skin detachment, whereas hC17-ect15 fails to induce skin fragility. (b) Direct immunofluorescence shows linear deposition at the DEJ in all the injected mice (yellow arrows). Scale bar = 100 μm. (c) NHEKs were treated with mouse IgG1 or the mAbs TS39-3 or hC17-ect15, for 1.5, 3, and 6 hours, respectively. Total cell lysates were subjected to Western blotting detected with anti-COL17 (mAb NC16A-3) and β-tubulin. The expression level of collagen XII is reduced in the mAb TS39-3–treated NHEKs. However, control mouse IgG1 and the mAb hC17-ect15 fail to deplete the expression of COL17. DEJ, dermal-epidermal junction; Epi, epidermis; h, hour; NHEK, normal human epidermal keratinocyte. Journal of Investigative Dermatology  , DOI: ( /j.jid ) Copyright © 2017 The Authors Terms and Conditions

Download ppt "C-Terminal Processing of Collagen XVII Induces Neoepitopes for Linear IgA Dermatosis Autoantibodies  Ellen Toyonaga, Wataru Nishie, Kentaro Izumi, Ken."

Similar presentations

Gene-Corrected Fibroblast Therapy for Recessive Dystrophic Epidermolysis Bullosa using a Self-Inactivating COL7A1 Retroviral Vector  Joanna Jacków, Matthias.

Gene-Corrected Fibroblast Therapy for Recessive Dystrophic Epidermolysis Bullosa using a Self-Inactivating COL7A1 Retroviral Vector  Joanna Jacków, Matthias.
Autoantibodies to Multiple Epitopes on the Non-Collagenous-1 Domain of Type VII Collagen Induce Blisters  Artem Vorobyev, Hideyuki Ujiie, Andreas Recke,

Autoantibodies to Multiple Epitopes on the Non-Collagenous-1 Domain of Type VII Collagen Induce Blisters  Artem Vorobyev, Hideyuki Ujiie, Andreas Recke,
Targeted Disruption of the Lama3 Gene in Adult Mice Is Sufficient to Induce Skin Inflammation and Fibrosis  Monika Pesch, Sabrina König, Monique Aumailley 

Targeted Disruption of the Lama3 Gene in Adult Mice Is Sufficient to Induce Skin Inflammation and Fibrosis  Monika Pesch, Sabrina König, Monique Aumailley 
Characterization of the Human Hair Shaft Cuticle–Specific Keratin-Associated Protein 10 Family  Hiroki Fujikawa, Atsushi Fujimoto, Muhammad Farooq, Masaaki.

Characterization of the Human Hair Shaft Cuticle–Specific Keratin-Associated Protein 10 Family  Hiroki Fujikawa, Atsushi Fujimoto, Muhammad Farooq, Masaaki.
Autoantibodies in Scurfy Mice and IPEX Patients Recognize Keratin 14

Autoantibodies in Scurfy Mice and IPEX Patients Recognize Keratin 14
TWEAK/Fn14 Activation Contributes to the Pathogenesis of Bullous Pemphigoid  Yale Liu, Lingling Peng, Liang Li, Chengfei Liu, Xiao Hu, Shengxiang Xiao,

TWEAK/Fn14 Activation Contributes to the Pathogenesis of Bullous Pemphigoid  Yale Liu, Lingling Peng, Liang Li, Chengfei Liu, Xiao Hu, Shengxiang Xiao,
IgG from Patients with Bullous Pemphigoid Depletes Cultured Keratinocytes of the 180- kDa Bullous Pemphigoid Antigen (Type XVII Collagen) and Weakens Cell.

IgG from Patients with Bullous Pemphigoid Depletes Cultured Keratinocytes of the 180- kDa Bullous Pemphigoid Antigen (Type XVII Collagen) and Weakens Cell.
S100A12 Induced in the Epidermis by Reduced Hydration Activates Dermal Fibroblasts and Causes Dermal Fibrosis  Jingling Zhao, Aimei Zhong, Emily E. Friedrich,

S100A12 Induced in the Epidermis by Reduced Hydration Activates Dermal Fibroblasts and Causes Dermal Fibrosis  Jingling Zhao, Aimei Zhong, Emily E. Friedrich,
TSLP Down-Regulates S100A7 and ß-Defensin 2 Via the JAK2/STAT3-Dependent Mechanism  Hana Lee, Woo-In Ryu, Hee Joo Kim, Hyun Cheol Bae, Hwa Jung Ryu, Jung.

TSLP Down-Regulates S100A7 and ß-Defensin 2 Via the JAK2/STAT3-Dependent Mechanism  Hana Lee, Woo-In Ryu, Hee Joo Kim, Hyun Cheol Bae, Hwa Jung Ryu, Jung.
Overexpression of Laminin-8 in Human Dermal Microvascular Endothelial Cells Promotes Angiogenesis-Related Functions  Jie Li, Lisa Zhou, Hoang T. Tran,

Overexpression of Laminin-8 in Human Dermal Microvascular Endothelial Cells Promotes Angiogenesis-Related Functions  Jie Li, Lisa Zhou, Hoang T. Tran,
The Antifibrotic Effect of α2AP Neutralization in Systemic Sclerosis Dermal Fibroblasts and Mouse Models of Systemic Sclerosis  Yosuke Kanno, En Shu,

The Antifibrotic Effect of α2AP Neutralization in Systemic Sclerosis Dermal Fibroblasts and Mouse Models of Systemic Sclerosis  Yosuke Kanno, En Shu,
Inhibition of UVB-Induced Skin Tumor Development by Drinking Green Tea Polyphenols Is Mediated Through DNA Repair and Subsequent Inhibition of Inflammation 

Inhibition of UVB-Induced Skin Tumor Development by Drinking Green Tea Polyphenols Is Mediated Through DNA Repair and Subsequent Inhibition of Inflammation 
The Autoantigen in Anti-p200 Pemphigoid Is Synthesized by Keratinocytes and Fibroblasts and Is Distinct from Nidogen-2  Silke C. Hofmann, Ursula Voith,

The Autoantigen in Anti-p200 Pemphigoid Is Synthesized by Keratinocytes and Fibroblasts and Is Distinct from Nidogen-2  Silke C. Hofmann, Ursula Voith,
The Autoantigen of Anti-p200 Pemphigoid Is an Acidic Noncollagenous N-Linked Glycoprotein of the Cutaneous Basement Membrane  Iakov Shimanovich, Yoshiaki.

The Autoantigen of Anti-p200 Pemphigoid Is an Acidic Noncollagenous N-Linked Glycoprotein of the Cutaneous Basement Membrane  Iakov Shimanovich, Yoshiaki.
Bullous Pemphigoid: Using Animal Models to Study the Immunopathology

Bullous Pemphigoid: Using Animal Models to Study the Immunopathology
Deletion of the Major Bullous Pemphigoid Epitope Region of Collagen XVII Induces Blistering, Autoimmunization, and Itching in Mice  Tiina Hurskainen,

Deletion of the Major Bullous Pemphigoid Epitope Region of Collagen XVII Induces Blistering, Autoimmunization, and Itching in Mice  Tiina Hurskainen,
Epidermolysis Bullosa Acquisita Develops in Dominant Dystrophic Epidermolysis Bullosa  Ryota Hayashi, Ken Natsuga, Mika Watanabe, Hiroaki Iwata, Satoru.

Epidermolysis Bullosa Acquisita Develops in Dominant Dystrophic Epidermolysis Bullosa  Ryota Hayashi, Ken Natsuga, Mika Watanabe, Hiroaki Iwata, Satoru.
Bullous Pemphigoid Sera that Contain Antibodies to BPAg2 also Contain Antibodies to LABD97 that Recognize Epitopes Distal to the NC16A Domain  Conleth.

Bullous Pemphigoid Sera that Contain Antibodies to BPAg2 also Contain Antibodies to LABD97 that Recognize Epitopes Distal to the NC16A Domain  Conleth.
Pathogenic Epitopes of Autoantibodies in Pemphigus Reside in the Amino-Terminal Adhesive Region of Desmogleins Which Are Unmasked by Proteolytic Processing.

Pathogenic Epitopes of Autoantibodies in Pemphigus Reside in the Amino-Terminal Adhesive Region of Desmogleins Which Are Unmasked by Proteolytic Processing.
Refractory oral ulcers with multiple immunoglobulin G/immunoglobulin A autoantibodies without skin lesions  Teruki Dainichi, MD, Bungo Ohyama, MD, Norito.

Refractory oral ulcers with multiple immunoglobulin G/immunoglobulin A autoantibodies without skin lesions  Teruki Dainichi, MD, Bungo Ohyama, MD, Norito.

Similar presentations

Ads by Google