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

PLoS One. 2014 Jan 14;9(1):e85062. doi: 10.1371/journal.pone.0085062. eCollection 2014.

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.

Publication types

  • Multicenter Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acyl-tRNA Synthetases / immunology*
  • Autoantibodies / blood*
  • Autoantibodies / immunology*
  • Autoantigens / immunology
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Humans
  • Lung Diseases, Interstitial / blood
  • Lung Diseases, Interstitial / diagnosis*
  • Myositis / diagnosis*

Substances

  • Autoantibodies
  • Autoantigens
  • Amino Acyl-tRNA Synthetases

Grants and funding

This work was supported by Grants-in-Aid for Scientific Research and for Challenging Exploratory Research from the Japan Society for the Promotion of Science, and grants for intractable diseases from the Ministry of Health, Labour and Welfare in Japan.The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.