Identification of novel autoantigens by a triangulation approach.
High titer autoantibodies, which are often associated with specific clinical phenotypes, are useful diagnostically and prognostically in systemic autoimmune diseases. In several autoimmune rheumatic diseases (e.g. myositis and Sjogren's syndrome), 20-40% of patients are autoantibody negative as assessed by conventional assays. The recent discovery of new specificities (e.g., anti-MDA5) in a subset of these autoantibody-negative subjects demonstrates that additional specificities await identification. In this manuscript, we describe a rapid multidimensional method to identify new autoantigens. A central foundation of this rapid approach is the use of an antigen source in which a pathogenic pathway active in the disease is recapitulated. Additionally, the method involves a modified serological proteome analysis strategy which allows confirmation that the correct gel plug has been removed prior to sending for sequencing. Lastly, the approach uses multiple sources of information to enable rapid triangulation and identification of protein candidates. Possible permutations and underlying principles of this triangulation strategy are elaborated to demonstrate the broad utility of this approach for antigen discovery.
Division of Rheumatology, School of Medicine, Johns Hopkins University, 5200 Eastern Avenue, MFL-CT Suite 5300, Baltimore, MD 21224, United States. tcottre2@jhmi.edu
Tricia R
TR
Cottrell
John C
JC
Hall
Antony
A
Rosen
Livia
L
Casciola-Rosen
D001324:Autoantigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D016898:Interferon-alpha / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D032342:Muscle Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D009132:Muscles / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D018994:Myosin Light Chains / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D009220:Myositis / Q000097:blood / Q000276:immunology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D020543:Proteome / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry
D001324:Autoantigens; C116468:IFIT3 protein, human; D016898:Interferon-alpha; D047908:Intracellular Signaling Peptides and Proteins; D018994:Myosin Light Chains; D020543:Proteome