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The identification of autoantigens in mucous membrane pemphigoid using immortalized oral mucosal keratinocytes
- Source :
- Journal of Oral Pathology & Medicine. 48:60-67
- Publication Year :
- 2018
- Publisher :
- Wiley, 2018.
-
Abstract
- Background Mucous membrane pemphigoid (MMP) is a rare chronic autoimmune subepithelial blistering disorder, targeting multiple basement membrane zone (BMZ) proteins including collagen XVII (COL17). Circulating autoantibodies of MMP are often undetected due to their lower titers. The oral mucosa is a valuable substrate for the detection of autoantibodies in MMP patients. However, obtaining normal human oral mucosa is more difficult than obtaining normal human skin. We established immortalized normal human oral mucosal keratinocytes (OMKs) and performed immunoblotting using immortalized OMK lysate for detecting autoantigens in MMP. Methods Immortalized OMKs were generated from primary OMKs using E6/E7 proteins of HPV. We compared the protein expression levels of major BMZ proteins between primary OMKs and immortalized OMKs. We performed immunoblotting to detect autoantigens using cell lysates from immortalized OMKs in 30 MMP patients. Results There were no significant differences between primary OMKs and immortalized OMKs in terms of protein expression levels of the BMZ proteins, including COL17, laminin 332, integrin α6/β4, collagen VII, and collagen IV. Cell lysates of immortalized OMKs effectively identified MMP autoantigens in 60% (18/30) of MMP sera. We found an interesting case of MMP whose autoantibodies preferentially reacted to the 120-kD protein that is an ectodomain of COL17. Conclusion We demonstrated that a cell lysate of immortalized OMKs is a reliable substrate for the detection of MMP autoantigens. This newly developed immunoblotting analysis method promises to contribute to the diagnosis of MMP.
- Subjects :
- Adult
Keratinocytes
Male
Cancer Research
Lysis
Immunoblotting
Pemphigoid, Benign Mucous Membrane
Human skin
Matrix metalloproteinase
Autoantigens
Pathology and Forensic Medicine
03 medical and health sciences
0302 clinical medicine
Laminin
medicine
Humans
Oral mucosa
Aged
Aged, 80 and over
biology
Chemistry
Mouth Mucosa
Autoantibody
030206 dentistry
Middle Aged
Molecular biology
Titer
medicine.anatomical_structure
Otorhinolaryngology
Ectodomain
030220 oncology & carcinogenesis
biology.protein
Periodontics
Female
Oral Surgery
Biomarkers
Subjects
Details
- ISSN :
- 09042512
- Volume :
- 48
- Database :
- OpenAIRE
- Journal :
- Journal of Oral Pathology & Medicine
- Accession number :
- edsair.doi.dedup.....b2f7e5481cea44259ed149c441bad410
- Full Text :
- https://doi.org/10.1111/jop.12780