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用于检测皮肌炎中抗转录中介因子-1γ和抗Mi-2自身抗体的酶联免疫吸附测定。

Enzyme-linked immunosorbent assays for detection of anti-transcriptional intermediary factor-1 gamma and anti-Mi-2 autoantibodies in dermatomyositis.

作者信息

Fujimoto Manabu, Murakami Akihiro, Kurei Shunsuke, Okiyama Naoko, Kawakami Atsushi, Mishima Michiaki, Sato Shinji, Seishima Mariko, Suda Takafumi, Mimori Tsuneyo, Takehara Kazuhiko, Kuwana Masataka

机构信息

Department of Dermatology, School of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.

Medical & Biological Laboratories Co., Ltd., 4-5-3 10F, Sakae, Naka-ku, Nagoya, Aichi 406-0008, Japan.

出版信息

J Dermatol Sci. 2016 Dec;84(3):272-281. doi: 10.1016/j.jdermsci.2016.09.013. Epub 2016 Sep 23.

Abstract

BACKGROUND

Autoantibodies against transcriptional intermediary factor 1 (TIF1) and Mi-2 are selectively detected in patients with dermatomyositis (DM). To measure these antibodies readily, the development of reliable ELISA systems has been needed.

OBJECTIVE

This study aimed to establish enzyme-linked immunosorbent assays (ELISAs) for anti-TIF1γ and anti-Mi-2β antibodies (Abs) and to assess their utility.

METHODS

Serum samples were obtained from 104 patients with classic DM, 68 with clinically amyopathic DM (CADM) and 70 with polymyositis, who were followed up at 8 medical centers across Japan. Serum samples from 190 patients with other connective tissue diseases (CTDs) and 123 healthy individuals were also assessed. Serum antibody levels were examined by ELISAs coated with full-length TIF1γ or Mi-2β proteins produced by a baculovirus expression system. To assess the cross-reactivity, partial-length Mi-2β proteins with or without mutations were produced and examined for reactivity.

RESULTS

When compared with immunoprecipitation assay, anti-TIF1γ Ab ELISA showed 100% sensitivity and 100% specificity, while anti-Mi-2β Ab ELISA showed 100% sensitivity and 99.6% specificity. Anti-TIF1γ Ab was positive in 30 (28.8%) with classic DM and 4 (5.9%) with CADM, whereas 14 (13.5%) with classic DM, but none with CADM, were positive for anti-Mi-2β Ab. Of 30 anti-TIF1γ Ab-positive DM patients, 23 (67.6%) had malignancy. Anti-Mi-2β Ab-positive serum samples exhibited modest cross-reactivity with the TIF1γ protein due to the homologous amino acid sequence containing cysteines in their plant homeodomains.

CONCLUSION

The current study demonstrates the utility of newly established ELISAs for anti-TIF1γ and anti-Mi-2β Abs, which can serve as easier detection systems for routine testing.

摘要

背景

在皮肌炎(DM)患者中可选择性检测到针对转录中介因子1(TIF1)和Mi-2的自身抗体。为了便于检测这些抗体,需要开发可靠的酶联免疫吸附测定(ELISA)系统。

目的

本研究旨在建立抗TIF1γ和抗Mi-2β抗体(Abs)的酶联免疫吸附测定(ELISA)并评估其效用。

方法

从日本8个医疗中心随访的104例典型DM患者、68例临床无肌病性DM(CADM)患者和70例多发性肌炎患者中获取血清样本。还评估了190例其他结缔组织病(CTD)患者和123名健康个体的血清样本。通过用杆状病毒表达系统产生的全长TIF1γ或Mi-2β蛋白包被的ELISA检测血清抗体水平。为评估交叉反应性,制备了有或无突变的部分长度Mi-2β蛋白并检测其反应性。

结果

与免疫沉淀法相比,抗TIF1γ抗体ELISA显示出100%的敏感性和100%的特异性,而抗Mi-2β抗体ELISA显示出100%的敏感性和99.6%的特异性。抗TIF1γ抗体在30例(28.8%)典型DM患者和4例(5.9%)CADM患者中呈阳性,而抗Mi-2β抗体在14例(13.5%)典型DM患者中呈阳性,但在CADM患者中均为阴性。在30例抗TIF1γ抗体阳性的DM患者中,23例(67.6%)患有恶性肿瘤。抗Mi-2β抗体阳性血清样本由于其植物同源结构域中含半胱氨酸的同源氨基酸序列而与TIF1γ蛋白表现出适度的交叉反应性。

结论

本研究证明了新建立的抗TIF1γ和抗Mi-2β抗体ELISA的效用,其可作为常规检测的更简便检测系统。

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