Background and aims Many reports suggest that saliva could be a source of biomarkers capable of detecting certain diseases. However, very few studies conducted to profile autoantibody isotypes in the saliva of autoimmune diseases.This study was performed to establish protein microarray for saliva diagnostics and to identify distinct profiles of salivary autoantibody in patients with systemic lupus erythematosus (SLE).
Methods We constructed antigen microarrays with canonical antigens of SLE as well as cytokines to characterise autoantibodies in matched saliva and serum derived from 17 SLE patients and 13 healthy controls. The autoantibody IgG and IgA isotypes were assayed.The Axon Scanner and GenePix Pro 7.0 were used to determine median fluorescence intensities (MFI) of features and background. Data were analysed using MultiExperiment Viewer and Significance Analysis of Microarray (SAM) algorithm.
Results The dynamic range of detection on the array was 1–104 ng/mL for commercial Abs spiked into saliva. We observed a high degree of specificity for its target antigen. IgG Ab reactivity against specific antigens was found mainly in serum, while IgA Ab reactivity to given antigens was predominant in saliva. SAM identified 7 antigens including BAFF, Ro60, U1-A and Sm/RNP that were significantly more reactive to IgA Ab in the saliva of SLE patients than in healthy controls (false discovery rate <0.01). The hierarchical clustered heat-map successfully placed SLE patients into close subgroups.
Conclusions Protein microarrays facilitate detection of autoantibody in human saliva as well as serum. Saliva profiling revealed that elevated IgA autoantibody reactivity to several targets including BAFF was associated with SLE compared with controls.
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