N-linked glycosylation on laminin γ1 influences recognition of anti-laminin γ1 pemphigoid autoantibodies

Li, Xiaoguang; Qian, Hua; Takizawa, Mamoru; Koga, Hiroshi; Tsuchisaka, Atsunari; Ishii, Norito; Hayakawa, Taihei; Ohara, Koji; Sitaru, Cassian; Zillikens, Detlef; Sekiguchi, Kiyotoshi; Hirako, Yoshiaki; Hashimoto, Takashi

doi:10.1016/j.jdermsci.2014.12.003

Cited by 10 publications

(12 citation statements)

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“…Binding of specific autoantibodies to their target may be influenced by their posttranslational modifications of the antigen, as shown in previous studies also for collagen XVII/BP180 and pemphigoid autoantibodies [ 36 ]. Posttranslational modifications of laminin 332 could therefore be relevant for the recognition by MMP autoantibodies and thus for the development of an immunoassay.…”

Section: Discussionmentioning

confidence: 83%

“…Previous studies showed that posttranslational modifications of autoantigens such as phosphorylation and deglycosylation may alter the binding of autoantibodies in pemphigoid diseases [ 35 , 36 ]. It has also been shown that glycosylation might influence the laminin 332 functions in tumorigenesis such as tumour invasion or tumour suppression [ 37 ].…”

Section: Resultsmentioning

confidence: 99%

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Molecular diagnosis of anti-laminin 332 (epiligrin) mucous membrane pemphigoid

Chiorean

Dănescu

Virtic

et al. 2018

Orphanet J Rare Dis

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BackgroundMucous membrane pemphigoid is a group of chronic subepithelial autoimmune blistering diseases that mainly affect mucous membranes. Laminin 332-specific autoantibodies are present in approximately 1/3 of the patients, being associated with an increased risk of malignancy. Because of the severe complications, an early recognition of the disease allowing a timely therapy is essential. The gold standard methods for detection of laminin 332-specific autoantibodies, including the immunoprecipitation and immunoblotting are non-quantitative, laborious and restricted to a few specialized laboratories worldwide. In addition, the use of radioimmunoassays, although highly sensitive and specific, are laborious, expensive and tightly regulated. Therefore, there is a stringent need for a quantitative immunoassay for the routine detection of laminin 332-specific autoantibodies more broadly available to diagnostic laboratories. The aim of this study was to compare different antigenic substrates, including native, recombinant laminin 332 and laminin 332-rich keratinocyte extracellular matrix, for development of an ELISA to detect autoantibodies in mucous membrane pemphigoid.ResultsUsing a relatively large number of sera from MMP patients with well-characterized autoantibody reactivity we show the suitability of ELISA systems using laminin 332 preparations as adjunct diagnostic tools in MMP. While glycosylation of laminin 332 does not appear to influence its recognition by MMP autoantibodies, ELISA systems using both purified, native and recombinant laminin 332 demonstrated a high sensitivity and good correlation with the detection of autoantibodies by immunoblotting. ELISA systems using different laminin 332 preparations represent a feasible and more accessible alternative for a broad range of laboratories.ConclusionsOur findings qualify the use of immunoassays with the laminin 332-rich preparations as an ancillary diagnostic tool in mucous membrane pemphigoid.Electronic supplementary materialThe online version of this article (10.1186/s13023-018-0855-x) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Molecular diagnosis of anti-laminin 332 (epiligrin) mucous membrane pemphigoid

Chiorean

Dănescu

Virtic

et al. 2018

Orphanet J Rare Dis

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“…Five known LMα chains, three known LMβ chains and three known LMγ chains in HLE B-3 cell lysate, HLE B-3 cell culture supernatant and HLE B-3 BMs harvested at days 0, 3, 6, 9 and 12, and human ALC lysate were analyzed by western blotting, as described previously ( 28 ). Briefly, antigen sources inclding HLE B-3 cell lysate, culture supernatant or protein lysate of human ALCs were mixed with 2X sample buffer, boiled for 2 min and separated by SDS-PAGE.…”

Section: Methodsmentioning

confidence: 99%

Laminins in an inï¿½vitro anterior lens capsule model established using HLE B-3 cells

Qian

Jiang

et al. 2018

Mol Med Report

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Cataracts are the most common eye disease to cause blindness in patients. The abnormal deposition of laminins (LMs) in the lens capsule and the disruption of capsular epithelium contribute to cataract development, although the mechanism by which this occurs is currently unclear. The present study aimed to reproduce HLE B-3 basement membranes (BMs) using HLE B-3 cells and to analyze the similarities of LM expression between HLE B-3 BMs and human anterior lens capsule (ALC). Immunohistochemistry (IHC), ELISA, western blot analysis and immunoprecipitation (IP)-western blot analysis were used to detect total LMs, LM trimers and 11 LM subunits in HLE B-3 cells, HLE B-3 BMs and human ALCs. In IHC staining, HLE B-3 cells and human ALCs were positive for LMs. In LM ELISA, all samples analyzed were positive for LMs. Western blot analysis detected all LM subunits except for LMγ3 in HLE B-3 cell lysate, 4 subunits (LMα4, LMα2, LMα1 and LMγ1) in HLE B-3 cell culture supernatant, 5 subunits (LMα4, LMα2, LMα1, LMβ3 and LMγ1) in HLE B-3 BMs, and 3 subunits (LMα4, LMγ2 and LMγ1) in human ALCs. The results of IP-western blot analysis revealed that the LM411 trimer was detected in HLE B-3 cell culture supernatant. These results indicated that HLE B-3 BMs were similar to human ALCs in terms of LM expression. Therefore, HLE B-3 BMs could be used as an in vitro ALC model to determine the role of LMs in ALC in the pathogenesis of cataracts and to select potential anti-cataract drugs.

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“…All sera were kept at −80 °C for long durations and at 4 °C with 0·1% sodium azide as a preservative during experiments. Studies of the considerably large number of patients used in this study have previously been reported . All previously reported studies for smaller numbers of patients used in this study are listed in Appendix S2 (see Supporting Information).…”

Section: Methodsmentioning

confidence: 99%