Native Autoantigen Complex Detects Pemphigoid Autoantibodies
Pemphigoid diseases are a group of autoimmune disorders characterized by subepidermal blistering in the skin and mucosa. Among them, mucous membrane pemphigoid (MMP) autoantibodies are characterized by targeting multiple molecules in the hemidesmosomes, including collagen XVII, laminin-332, and inte...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-05-01
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| Series: | JID Innovations |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667026723000152 |
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| author | Shoko Mai Kentaro Izumi Yosuke Mai Ken Natsuga Norito Ishii Daisuke Sawamura Franziska Schauer Dimitra Kiritsi Wataru Nishie Hideyuki Ujiie |
| author_facet | Shoko Mai Kentaro Izumi Yosuke Mai Ken Natsuga Norito Ishii Daisuke Sawamura Franziska Schauer Dimitra Kiritsi Wataru Nishie Hideyuki Ujiie |
| author_sort | Shoko Mai |
| collection | DOAJ |
| description | Pemphigoid diseases are a group of autoimmune disorders characterized by subepidermal blistering in the skin and mucosa. Among them, mucous membrane pemphigoid (MMP) autoantibodies are characterized by targeting multiple molecules in the hemidesmosomes, including collagen XVII, laminin-332, and integrin a6/β4. Traditionally, recombinant proteins of the autoantigens have been employed to identify circulating autoantibodies by immune assays. However, developing an efficient detection system for MMP autoantibodies has been challenging because the autoantibodies have heterogeneous profiles and the antibody titers are typically low. In this study, we introduce an ELISA that takes advantage of a native autoantigen complex rather than simple recombinant proteins. We generated HaCaT keratinocytes with a DDDDK-tag knocked in at the COL17A1 locus by CRISPR/Cas9-mediated gene editing. Immunoprecipitation using the DDDDK-tag isolated a native complex that contained full-length and processed collagen XVII and integrin α6/β4. Then, we used the complex proteins to prepare an ELISA system and enrolled 55 MMP cases to validate its diagnostic performance. The sensitivity and specificity of the ELISA for detecting MMP autoantibodies were 70.9% and 86.7%, respectively, far superior to those of conventional assays. In autoimmune diseases such as MMP, in which autoantibodies target various molecules, isolating the antigen-protein complexes can help establish a diagnostic system. |
| first_indexed | 2024-03-13T10:56:27Z |
| format | Article |
| id | doaj.art-946c79ccc7d547afbe03c3072a90a596 |
| institution | Directory Open Access Journal |
| issn | 2667-0267 |
| language | English |
| last_indexed | 2024-03-13T10:56:27Z |
| publishDate | 2023-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | JID Innovations |
| spelling | doaj.art-946c79ccc7d547afbe03c3072a90a5962023-05-17T04:23:39ZengElsevierJID Innovations2667-02672023-05-0133100193Native Autoantigen Complex Detects Pemphigoid AutoantibodiesShoko Mai0Kentaro Izumi1Yosuke Mai2Ken Natsuga3Norito Ishii4Daisuke Sawamura5Franziska Schauer6Dimitra Kiritsi7Wataru Nishie8Hideyuki Ujiie9Department of Dermatology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, JapanDepartment of Dermatology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, Japan; Correspondence: Kentaro Izumi, Department of Dermatology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, North 15 West 7, Kita-Ku, Sapporo 060-8638, Japan.Department of Dermatology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, JapanDepartment of Dermatology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, JapanDepartment of Dermatology, Kurume University School of Medicine, Kurume, JapanDepartment of Dermatology, Hirosaki University Graduate School of Medicine and School of Medicine, Hirosaki, JapanDepartment of Dermatology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, GermanyDepartment of Dermatology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, GermanyDepartment of Dermatology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, JapanDepartment of Dermatology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, JapanPemphigoid diseases are a group of autoimmune disorders characterized by subepidermal blistering in the skin and mucosa. Among them, mucous membrane pemphigoid (MMP) autoantibodies are characterized by targeting multiple molecules in the hemidesmosomes, including collagen XVII, laminin-332, and integrin a6/β4. Traditionally, recombinant proteins of the autoantigens have been employed to identify circulating autoantibodies by immune assays. However, developing an efficient detection system for MMP autoantibodies has been challenging because the autoantibodies have heterogeneous profiles and the antibody titers are typically low. In this study, we introduce an ELISA that takes advantage of a native autoantigen complex rather than simple recombinant proteins. We generated HaCaT keratinocytes with a DDDDK-tag knocked in at the COL17A1 locus by CRISPR/Cas9-mediated gene editing. Immunoprecipitation using the DDDDK-tag isolated a native complex that contained full-length and processed collagen XVII and integrin α6/β4. Then, we used the complex proteins to prepare an ELISA system and enrolled 55 MMP cases to validate its diagnostic performance. The sensitivity and specificity of the ELISA for detecting MMP autoantibodies were 70.9% and 86.7%, respectively, far superior to those of conventional assays. In autoimmune diseases such as MMP, in which autoantibodies target various molecules, isolating the antigen-protein complexes can help establish a diagnostic system.http://www.sciencedirect.com/science/article/pii/S2667026723000152 |
| spellingShingle | Shoko Mai Kentaro Izumi Yosuke Mai Ken Natsuga Norito Ishii Daisuke Sawamura Franziska Schauer Dimitra Kiritsi Wataru Nishie Hideyuki Ujiie Native Autoantigen Complex Detects Pemphigoid Autoantibodies JID Innovations |
| title | Native Autoantigen Complex Detects Pemphigoid Autoantibodies |
| title_full | Native Autoantigen Complex Detects Pemphigoid Autoantibodies |
| title_fullStr | Native Autoantigen Complex Detects Pemphigoid Autoantibodies |
| title_full_unstemmed | Native Autoantigen Complex Detects Pemphigoid Autoantibodies |
| title_short | Native Autoantigen Complex Detects Pemphigoid Autoantibodies |
| title_sort | native autoantigen complex detects pemphigoid autoantibodies |
| url | http://www.sciencedirect.com/science/article/pii/S2667026723000152 |
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