Cross-reactivity of antibodies against interferon beta in multiple sclerosis patients and interference of the JAK-STAT signaling pathway

Isaac Hurtado-Guerrero,Patricia Urbaneja,Jesús Ortega-Pinazo,Pedro Serrano,Begoña Oliver-Martos,Óscar Fernández,Maria Jesus Pinto-Medel,Laura Leyva,Jose Luis Rodriguez-Bada

doi:10.1038/s41598-017-16828-x

Abstract

Interferon beta (IFNβ) therapy has immunogenic properties and induces the development of neutralizing antibodies (NAbs). From the extensive literature focused in the development of NAbs in multiple sclerosis (MS) patients, their ability to cross-react has been deficiently evaluated, despite having important consequences in the clinical practice. Here, the relation between the cross-reactivity and the NAbs titers has been evaluated in MS patients, by inhibition of the antiviral activity of IFNβ by bioassay and through the interference with the activation of the IFNß pathway (JAK-STAT), by phosphoflow. Thus, patients with intermediate-high titers of NAbs, determined by bioassay, had a 79-fold increased risk of cross-reactivity compared to patients with low titers. The cross-reactivity is also demonstrated because NAbs positive sera were able to decrease significantly the activation of pSTAT1 achieved by other different IFNβ molecules in the cells patients. Besides, a linear relationship between the STAT1 phosphorylation and NAbs titers was found. The study demonstrates that cross-reactivity increases with the titer of antibodies, which has important implications in clinical practice when switching the treatment. The direct relationship between the NAbs titer and the activation of STAT1 suggest that its determination could be an indirect method to identify the presence of NAbs.

Highlights

During last years, several drugs have been approved for the treatment of multiple sclerosis (MS), interferon beta (IFNβ) continues being a widely used treatment for newly diagnosed patients with relapsing-onset multiple sclerosis
Statistical analysis using chi-square test showed a relationship between the antibody titers and the presence of cross-reactivity (p < 0.0001), so that the number of individuals with cross-reactivity increases as higher the antibody titer is, presenting cross reactivity almost always in those individuals with neutralizing antibodies (NAbs) greater than 100 TRU/ml
Patients with intermediate and high titers were grouped and a logistic regression analysis was performed to calculate the risk of cross-reactivity according to NAbs titers

Summary

Introduction

Several drugs have been approved for the treatment of multiple sclerosis (MS), interferon beta (IFNβ) continues being a widely used treatment for newly diagnosed patients with relapsing-onset multiple sclerosis. Exogenous IFNβ is a recombinant protein available in three formulations, two as IFNβ-1a (intramuscular (i.m.) and subcutaneous (s.c.)) which are produced in mammalian cells with identical structure to the native form, and one as IFNβ-1b (s.c.), that is produced in Escherichia coli and differs from native form, mainly by lack of glycosylation These differences in the biochemical structure, as well as in the excipients, route and timing of administration, translate into different immunogenicity among preparations. The antibodies against therapeutic proteins can have serious consequences for the patient because they are able to cross react and neutralize the native proteins, with the consequent induction of adverse events This fact has been described for thromboepoetin (TPO), which induces the formation of antibodies that neutralize native TPO and cause thrombocytopenia[4] and with IFNβ, whose antibodies neutralize endogenous IFNβ5

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