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Improvement of immunological tests for detecting autoantibodies in patients with lamina lucida-type linear IgA bullous dermatosis

IF 4.6

Journal of dermatological science Pub Date : 2025-01-01 DOI:10.1016/j.jdermsci.2024.12.001

Masahiro Tsutsumi , Hiroshi Koga , Kwesi Teye , Norito Ishii , Takekuni Nakama

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引用次数: 0

Abstract

Background

In the diagnosis of linear IgA bullous dermatosis (LABD), detection of IgA at the epidermal basement membrane zone and circulating IgA autoantibodies are essential. The disease has two subtypes, lamina lucida-type and sublamina densa-type, with 120 kDa LAD-1 and 97 kDa LABD97 as major autoantigens for lamina lucida-type. Normal human epidermal keratinocytes (NHEK) and HaCaT cells are widely used for immunoblotting (IB) in the diagnosis process, but they do not provide high sensitivity and semiquantitative analysis.

Objective

To develop a more sensitive and convenient method for detecting IgA antibodies in lamina lucida-type LABD patients.

Methods

The expressions of LAD-1 and LABD97 in lysates and culture supernatants from Ker-CT, HaCaT, DJM-1, and NHEK were compared. The sensitivity of IBs using concentrated culture supernatants of HaCaT and Ker-CT and ELISAs using several recombinant proteins (RPs) corresponding to BP180 ectodomain were compared using 55 sera from LABD patients.

Results

In culture supernatant, Ker-CT expressed higher amounts of LAD-1 and LABD97. IBs using concentrated culture supernatant of HaCaT and Ker-CT showed 43 % and 46 % positivity to sera from LABD patients, respectively. In ELISAs, the RP of amino acids 490–1421 of BP180 showed the highest positivity (80.0 %) among several proteins. Additionally, this ELISA showed reduced OD values in LABD and related diseases patients' sera at remission.

Conclusion

The ELISA using the RP coding amino acids 490–1421 of BP180 is useful for identifying IgA antibodies and monitoring disease activity in lamina lucida-type LABD patients.

查看原文本刊更多论文

改进免疫学试验，以检测透明层型线性 IgA 大疱性皮肤病患者的自身抗体。

背景：在线性IgA大疱性皮肤病（LABD）的诊断中，检测表皮基底膜区IgA和循环IgA自身抗体是必不可少的。该疾病有两种亚型，即透明层型和膜下致密型，其中120 kDa LAD-1和97 kDa LABD97是透明层型的主要自身抗原。正常人表皮角质形成细胞（NHEK）和HaCaT细胞在诊断过程中广泛用于免疫印迹（IB），但它们不能提供高灵敏度和半定量分析。目的：建立一种更灵敏、简便的检测透明层型LABD患者IgA抗体的方法。方法：比较Ker-CT、HaCaT、DJM-1和NHEK的裂解液和培养上清液中LAD-1和LABD97的表达。用55例LABD患者的血清比较HaCaT和Ker-CT的浓培养上清和BP180外结构域对应的重组蛋白（RPs）的elisa的敏感性。结果：培养上清中，Ker-CT表达较高的lad1和LABD97。使用HaCaT和Ker-CT浓缩培养上清的IBs对LABD患者血清的阳性率分别为43% %和46% %。在酶联免疫吸附试验中，BP180的490 ~ 1421氨基酸的RP在几种蛋白中阳性率最高（80.0 %）。此外，该ELISA显示，缓解期LABD及相关疾病患者血清中的OD值降低。结论：利用RP编码BP180蛋白490 ~ 1421氨基酸的酶联免疫吸附试验（ELISA）可用于荧光层型LABD患者的IgA抗体鉴定和疾病活动性监测。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of dermatological science

Journal of dermatological science Dermatology

CiteScore

7.60

自引率

0.00%

发文量

0

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