OBJECTIVE: To investigate the incidence of autoantibodies directed to deproteinized transfer RNA(His) (tRNA(His)) in anti-Jo-1 positive myositis patients and to determine the major B cell epitope. METHODS: One hundred sixty-seven myositis sera were screened by immunoblotting and enzyme-linked immunosorbent assay for the presence of anti-Jo-1 antibody. Autoantibodies directed to deproteinized RNA were detected by immunoprecipitation. Ribonuclease cleavage experiments were performed to determine the tRNA(His)-specific features important for recognition. RESULTS: Approximately one-third of the anti-Jo-1 positive sera also contained autoantibodies recognizing tRNA(His). This recognition was independent of modified bases, but the presence of stabilizing Mg2+ ions appeared to be essential for efficient immunoprecipitation. Transfer RNA(His)-specific features in the anticodon loop were not protected from ribonuclease cleavage by bound antibodies, while protection of bases located in the D and T loops was observed. CONCLUSION: A significant number of anti-Jo-1 positive myositis sera contain anti-tRNA(His) activity. Formation of the major autoepitope on tRNA(His) is strongly dependent on proper folding of this molecule mediated by an interaction between D and T loops which is stabilized by either modified residues or Mg2+ ions.
OBJECTIVE: To investigate the incidence of autoantibodies directed to deproteinized transfer RNA(His) (tRNA(His)) in anti-Jo-1 positive myositispatients and to determine the major B cell epitope. METHODS: One hundred sixty-seven myositis sera were screened by immunoblotting and enzyme-linked immunosorbent assay for the presence of anti-Jo-1 antibody. Autoantibodies directed to deproteinized RNA were detected by immunoprecipitation. Ribonuclease cleavage experiments were performed to determine the tRNA(His)-specific features important for recognition. RESULTS: Approximately one-third of the anti-Jo-1 positive sera also contained autoantibodies recognizing tRNA(His). This recognition was independent of modified bases, but the presence of stabilizing Mg2+ ions appeared to be essential for efficient immunoprecipitation. Transfer RNA(His)-specific features in the anticodon loop were not protected from ribonuclease cleavage by bound antibodies, while protection of bases located in the D and T loops was observed. CONCLUSION: A significant number of anti-Jo-1 positive myositis sera contain anti-tRNA(His) activity. Formation of the major autoepitope on tRNA(His) is strongly dependent on proper folding of this molecule mediated by an interaction between D and T loops which is stabilized by either modified residues or Mg2+ ions.
Authors: R Brouwer; G J Hengstman; W Vree Egberts; H Ehrfeld; B Bozic; A Ghirardello; G Grøndal; M Hietarinta; D Isenberg; J R Kalden; I Lundberg; H Moutsopoulos; P Roux-Lombard; J Vencovsky; A Wikman; H P Seelig; B G van Engelen ; W J van Venrooij Journal: Ann Rheum Dis Date: 2001-02 Impact factor: 19.103
Authors: G J D Hengstman; L van Brenk; W T M Vree Egberts; E L van der Kooi; G F Borm; G W A M Padberg; W J van Venrooij; B G M van Engelen Journal: J Neurol Date: 2005-02-23 Impact factor: 4.849
Authors: Rick Brouwer; Wilma T M Vree Egberts; Gerald J D Hengstman; Reinout Raijmakers; Baziel G M van Engelen; Hans Peter Seelig; Manfred Renz; Rudolf Mierau; Ekkehard Genth; Ger J M Pruijn; Walther J van Venrooij Journal: Arthritis Res Date: 2001-11-12