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

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Article numbere85062
JournalPLoS One
Volume9
Issue number1
DOIs
Publication statusPublished - 2014 Jan 14

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aminoacyl tRNA ligases
myositis
Amino Acyl-tRNA Synthetases
Myositis
Interstitial Lung Diseases
pneumonia
Multicenter Studies
Assays
fibrosis
Immunosorbents
assays
muscular diseases
Idiopathic Interstitial Pneumonias
lungs
Pulmonary Fibrosis
Enzyme-Linked Immunosorbent Assay
Immunoprecipitation
enzyme-linked immunosorbent assay
Idiopathic Pulmonary Fibrosis
Connective Tissue Diseases

ASJC Scopus subject areas

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

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The multicenter study of a new assay for simultaneous detection of multiple anti-aminoacyl-tRNA synthetases in myositis and interstitial pneumonia. / Nakashima, Ran; Imura, Yoshitaka; Hosono, Yuji; Seto, Minae; Murakami, Akihiro; Watanabe, Kizuku; Handa, Tomohiro; Mishima, Michiaki; Hirakata, Michito; Takeuchi, Tsutomu; Fujio, Keishi; Yamamoto, Kazuhiko; Kohsaka, Hitoshi; Takasaki, Yoshinari; Enomoto, Noriyuki; Suda, Takafumi; Chida, Kingo; Hisata, Shu; Nukiwa, Toshihiro; Mimori, Tsuneyo.

In: PLoS One, Vol. 9, No. 1, e85062, 14.01.2014.

Research output: Contribution to journalArticle

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, vol. 9, no. 1, e85062. https://doi.org/10.1371/journal.pone.0085062
Nakashima, Ran ; Imura, Yoshitaka ; Hosono, Yuji ; Seto, Minae ; Murakami, Akihiro ; Watanabe, Kizuku ; Handa, Tomohiro ; Mishima, Michiaki ; Hirakata, Michito ; Takeuchi, Tsutomu ; Fujio, Keishi ; Yamamoto, Kazuhiko ; Kohsaka, Hitoshi ; Takasaki, Yoshinari ; Enomoto, Noriyuki ; Suda, Takafumi ; Chida, Kingo ; Hisata, Shu ; Nukiwa, Toshihiro ; Mimori, Tsuneyo. / The multicenter study of a new assay for simultaneous detection of multiple anti-aminoacyl-tRNA synthetases in myositis and interstitial pneumonia. In: PLoS One. 2014 ; Vol. 9, No. 1.
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abstract = "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, Sj{\"o}gren'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.",
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T1 - The multicenter study of a new assay for simultaneous detection of multiple anti-aminoacyl-tRNA synthetases in myositis and interstitial pneumonia

AU - Nakashima, Ran

AU - Imura, Yoshitaka

AU - Hosono, Yuji

AU - Seto, Minae

AU - Murakami, Akihiro

AU - Watanabe, Kizuku

AU - Handa, Tomohiro

AU - Mishima, Michiaki

AU - Hirakata, Michito

AU - Takeuchi, Tsutomu

AU - Fujio, Keishi

AU - Yamamoto, Kazuhiko

AU - Kohsaka, Hitoshi

AU - Takasaki, Yoshinari

AU - Enomoto, Noriyuki

AU - Suda, Takafumi

AU - Chida, Kingo

AU - Hisata, Shu

AU - Nukiwa, Toshihiro

AU - Mimori, Tsuneyo

PY - 2014/1/14

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N2 - 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.

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