The multicenter study of a new assay for simultaneous detection of multiple anti-aminoacyl-tRNA synthetases in myositis and interstitial pneumonia

Ran Nakashima, Yoshitaka Imura, Yuji Hosono, Minae Seto, Akihiro Murakami, Kizuku Watanabe, Tomohiro Handa, Michiaki Mishima, Michito Hirakata, Tsutomu Takeuchi, Keishi Fujio, Kazuhiko Yamamoto, Hitoshi Kohsaka, Yoshinari Takasaki, Noriyuki Enomoto, Takafumi Suda, Kingo Chida, Shu Hisata, Toshihiro Nukiwa, Tsuneyo Mimori

研究成果: Article

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Objective: Autoantibodies to aminoacyl-tRNA synthetases (ARSs) are useful in the diagnosis of idiopathic inflammatory myopathy (IIM) with interstitial pneumonia (IP). We developed an enzyme-linked immunosorbent assay (ELISA) system using a mixture of recombinant ARS antigens and tested its utility in a multicenter study. Methods: We prepared six recombinant ARSs: GST-Jo-1, His-PL-12, His-EJ and GST-KS expressed in Escherichia coli, and His-PL-7 and His-OJ expressed in Hi-5 cells. After confirming their antigenic activity, with the exception of His-OJ, we developed our ELISA system in which the five recombinant ARSs (without His-OJ) were mixed. Efficiency was confirmed using the sera from 526 Japanese patients with connective tissue disease (CTD) (IIM n = 250, systemic lupus erythematosus n = 91, systemic sclerosis n = 70, rheumatoid arthritis n = 75, Sjögren's syndrome n = 27 and other diseases n = 13), 168 with idiopathic interstitial pneumonia (IIP) and 30 healthy controls collected from eight institutes. IIPs were classified into two groups; idiopathic pulmonary fibrosis (IPF) (n = 38) and non-IPF (n = 130). Results were compared with those of RNA immunoprecipitation. Results: Sensitivity and specificity of the ELISA were 97.1% and 99.8%, respectively when compared with the RNA immunoprecipitation assay. Anti-ARS antibodies were detected in 30.8% of IIM, 2.5% of non-myositis CTD, and 10.7% of IIP (5.3% of IPF and 12.3% of non-IPF). Anti-ARS-positive non-IPF patients were younger and more frequently treated with glucocorticoids and/or immunosuppressants than anti-ARS-negative patients. Conclusion: A newly established ELISA detected anti-ARS antibodies as efficiently as RNA immunoprecipitation. This system will enable easier and wider use in the detection of anti-ARS antibodies in patients with IIM and IIP.

元の言語English
記事番号e85062
ジャーナルPloS one
9
発行部数1
DOI
出版物ステータスPublished - 2014 1 14

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Nakashima, R., Imura, Y., Hosono, Y., Seto, M., Murakami, A., Watanabe, K., Handa, T., Mishima, M., Hirakata, M., Takeuchi, T., Fujio, K., Yamamoto, K., Kohsaka, H., Takasaki, Y., Enomoto, N., Suda, T., Chida, K., Hisata, S., Nukiwa, T., & Mimori, T. (2014). The multicenter study of a new assay for simultaneous detection of multiple anti-aminoacyl-tRNA synthetases in myositis and interstitial pneumonia. PloS one, 9(1), [e85062]. https://doi.org/10.1371/journal.pone.0085062