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1.
Antibodies (Basel) ; 8(2)2019 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-31544843

RESUMEN

Rheumatoid arthritis (RA) is an autoimmune disease of unknown etiology. The majority of individuals with RA are positive for the disease-specific anti-citrullinated protein antibodies (ACPAs). These antibodies are primarily of cross-reactive nature, hence, the true autoantigen to ACPA remains unidentified. In this study, we analyzed the reactivity of RA sera to several post-translationally modified epitopes, in order to further characterize the specific nature of ACPAs by immunoassays. Substituting citrulline with other amino acids, e.g., D-citrulline, homo-citrulline and methyl-arginine illustrated that ACPAs are utmost specific for citrullinated targets. Collectively, these findings support that ACPAs and citrullinated targets are specific for RA, making citrulline-containing peptide targets the most effective assays for detection of ACPAs.

2.
Scand J Clin Lab Invest ; 75(1): 51-63, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25347362

RESUMEN

Rheumatoid factors (RFs) are antibodies recognizing other antibodies usually by binding to the Fc part, while heterophilic antibodies (HAbs) are antibodies reacting with immunoglobulins (Igs) from other species. In particular, RFs have been found to cause false positive results in sandwich immunoassays. In this work, we analyzed RF-positive and RF-negative sera for content of cytokines and for heterophilic reactions by enzyme-linked immunosorbent assay and bead-based sandwich immunoassays. All sera, including those with RFs, contained insignificant amounts of cytokines and chemokines, but RF-positive sera showed large false positive values for several cytokines when analyzed by fluorescent bead-based multiplex immunoassays. This non-specific binding could be minimized by reagents designed to block HAbs, i.e. by selected animal IgGs. Furthermore, sera positive for RFs reacted with several animal IgGs, when these were immobilized on beads or coated on the polystyrene surface in enzyme-linked immunosorbent assays. This reaction could be inhibited by human IgG and by agents designed to inhibit heterophilic reactions (i.e. mixtures of IgGs from different species). In conclusion, RFs and HAbs represent an identical/overlapping set of antibodies, causing false positive reactions in sandwich and other immunoassays. Such assays must be conducted in the presence of appropriate blocking agents, e.g. HBR+, and must be carefully controlled.


Asunto(s)
Artritis Reumatoide/inmunología , Factor Reumatoide/inmunología , Animales , Artritis Reumatoide/sangre , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoglobulina G/inmunología , Factor Reumatoide/sangre , Especificidad de la Especie
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