Immunogenicity of An Interferon-β1a Product

Abstract

In order to determine whether Blastoferon®, a biosimilar interferon (IFN)- beta 1a formulation, shares epitopes with other known IFN-beta products, a series of neutralization bioassays were performed with a set of well-characterized anti-IFN- beta monoclonal antibodies and human sera (World Health Organization Reference Reagents). The bioassay was the interferon-induced inhibition of virus cytopathic effect on human cells in culture (EMC virus and A-549 cells). Computer-calculated results were reported as Tenfold Reduction Units (TRU)/ml. To further assess Blastoferon® immunogenicity, in vivo production of anti-IFN beta antibodies was determined in sera of patients included in the pharmacovigilance plan of Blastoferon® by the level of IFN- beta 1a binding antibodies (by enzyme immunoassay -EIA) and neutralizing antibodies (in the Wish-VSV system). The highly characterized neutralizing monoclonal antibodies A1 and A5 that bind to specific regions of the IFN- beta molecule reacted positively with the three beta 1a IFNs: Blastoferon®, Rebif®, and the IFN- beta WHO Second International Standard 00/572. As expected, the non-neutralizing monoclonal antibodies B4 and B7 did not neutralize any of the IFN- beta preparations. The commercially available monoclonal antibody B-02 reacted essentially equally with Rebif® and Blastoferon®. The WHO Reference Reagent human serum anti-IFN- beta polyclonal antibody neutralized all the IFN- beta products, whereas the WHO Reference Reagent human serum anti-IFN-alpha polyclonal antibody G037-501-572 appropriately failed to react with any of the IFN- beta products. On the basis of in vitro reactivity with known, well-characterized monoclonal and polyclonal antibody preparations, Blastoferon® shares immunological determinants with other human interferon- beta products, especially IFN- beta 1a. In vivo antibodies were detected by EIA in 72.9% of 37 chronically treated multiple sclerosis patients, whereas neutralizing antibodies were found in 8.1% of them. Blastoferon® appears to have immunological characteristics comparable to other IFN- beta 1a products.

Cell based assay identifies TLR2 and TLR4 stimulating impurities in Interferon beta

“…It is established that recombinant human IFNβ can induce binding and neutralizing antibodies that reduce their therapeutic efficacy. Among the marketed products, the immunogenicity rate appears to be higher for the IFNβ-1b products produced in E. coli than the IFNβ-1a produced in CHO cells and this has been generally ascribed to minor differences in sequence and presence of aggregates (Supplementary Table 1 ) 32 , 37 . We hypothesized that these products could also differ in the content of trace levels of IIRMI, which may contribute to the difference in immunogenicity rates.…”

“…The immunogenicity of rhuIFNβ-1b products has been ascribed to multiple factors including the absence of glycosylation, the N terminal methionine, a substitution of a cystein for a serine in position 17 31 , 52 and aggregates. Of note, the rhuIFNβ-1a products, which are more homologous to the endogenous IFN, are also immunogenic albeit to a lesser degree 32 . Several studies suggested that the difference was possibly due to the difference in aggregate and particle content between the products 40 .…”

“…Recombinant human IFNβ-1a products are produced in mammalian CHO cells, have an amino acid sequence identical to the human protein and are glycosylated. Multiple studies indicate a higher incidence of antibodies for patients treated with Betaseron and Extavia compared to those that receive Rebif and Avonex 30 – 32 . Importantly, the development of binding or neutralizing antibodies to these products correlate with a decline in bioavailability, increased frequency of relapses, and poorer clinical outcomes 32 – 35 .…”

Interferon-β lipofection II. Mechanisms involved in cell death and bystander effect induced by cationic lipid-mediated interferon-β gene transfer to human tumor cells

“…24 Wish cells were used as biological substrate and vesicular stomatitis virus was employed as challenge. IFN produced was always b type, as it was completely neutralized by a monoclonal anti-hIFNbneutralizing antibody kindly provided by BioSidus.…”

Impact of Immunity in Autism Spectrum Disorders

“…Mast cells have been implicated in inflammatory conditions that are worsened by stress (50)(51)(52). Mast cell-derived cytokines, such as IL-6, can increase bloodbrain-barrier (BBB) permeability (47,(53)(54)(55)(56). Corticotropin-releasing hormone (CRH) may have an immunomodulatory role and has been associated with intestinal inflammation.…”