Autoantibody profiling for the study and treatment of autoimmune disease

Abstract

Proteomics technologies enable profiling of autoantibody responses using biological fluids derived from patients with autoimmune disease. They provide a powerful tool to characterize autoreactive B-cell responses in diseases including rheumatoid arthritis, multiple sclerosis, autoimmune diabetes, and systemic lupus erythematosus. Autoantibody profiling may serve purposes including classification of individual patients and subsets of patients based on their 'autoantibody fingerprint', examination of epitope spreading and antibody isotype usage, discovery and characterization of candidate autoantigens, and tailoring antigen-specific therapy. In the coming decades, proteomics technologies will broaden our understanding of the underlying mechanisms of and will further our ability to diagnose, prognosticate and treat autoimmune disease.

Figure 1

The 'connective tissue disease' array. A 48-feature collage derived from a 1536-feature 'connective tissue disease' array probed with serum from a patient with systemic lupus erythematosus (SLE) is presented. This array demonstrates specific detection of two representative autoantibody reactivities, against Ro52 (upper center box) and double-stranded DNA (dsDNA, lower right box). Antibodies against Candida skin test antigens (lower center box) are also detected, and serve as a positive control. This collage contains four features representing the reactive antigens (boxed) and control antigens (not boxed). Arrays were produced using a robotic microarrayer to attach putative connective tissue disease autoantigens (listed in text) to poly-L-lysine-coated microscopic slides. The depicted array was incubated with a 1:150 dilution of serum derived from a patient with SLE and with ELISA-confirmed reactivity against Ro and DNA. Antibody binding was detected by incubation with Cy-3-labeled antihuman IgG/IgM secondary antibody. Marker spots (spotted Cy-3-labeled IgG, left box) are used to orient the arrays. Detailed protocols for production, probing, and scanning antigen arrays are presented in our earlier work [17] and online [21].