Betaferon

Inclusion criteria were: (1) confirmed diagnosis of RRMS, based on medical records; (2) taking first-line drugs such as Avonex (Biogen Manufacturing ApS, Hillerød, Denmark), Rebif (Merck Serono S.p.A., Bari, Italy), Betaferon (Bayer AG, Berlin, Germany), Extavia (Novartis Pharma GmbH, Nürnberg, Germany), Copaxone (Teva Pharmaceuticals Europe B.V., Haarlem, the Netherlands), or Tecfidera (Biogen Manufacturing ApS, Hillerød, Denmark); (3) treatment for at least six months prior to participation in the study; (4) age over 18; and (5) written informed consent prior to participation in the study.
Exclusion criteria were: (1) progressive forms of MS, (2) confirmed diagnosis of RRMS but not taking the above mentioned first-line DMTs, (3) treatment initiated less than six months before participation in the study, (4) severe cognitive impairment making the patient unable to follow the test instructions, (5) treatment due to the presence of mood or anxiety disorders, and (6) lack of written consent to participate in the study.

The study enrolled patients with RRMS treated with IFNβ for at least 3 months and affected by FLS despite receiving the standard of therapy for FLS [ie, acetaminophen or non-steroidal anti-inflammatory drugs (NSAIDs)]. Patients were using following one of the IFNβ products: subcutaneous IFNβ-1b (Betaferon^®, Bayer), subcutaneous IFNβ-1a (Rebif^®, Merck Serono), and intramuscular IFNβ-1a (Avonex^®, Biogen). Inclusion criteria were: subjects of both sexes; age ≥18 years and ≤55 years; diagnosis of RRMS; therapy with IFNβ for at least 3 months; negative pregnancy test performed no more than 30 days from the baseline visit; FLS-score (FLS-S) ≥2 despite standard therapy for FLS; ability to provide written informed consent for participation in the study; absence of clinically relevant conditions or situations which in the opinion of the investigator would interfere with the study evaluation or with the participation in the study; use of effective birth control methods, or condition of menopause for at least 6 months. Exclusion criteria were: subjects (male or female) potentially fertile not using contraception; pregnant or breastfeeding women; intolerance or known contraindications to the use of cetirizine; hereditary problems of galactose intolerance, Lapp lactase deficiency, glucose-galactose malabsorption; contemporary participation in other studies.

Sera of IFNβ-treated patients were tested for the presence of NAbs against the three commercial forms of IFN-β (IFN-β 1a (Avonex, Biogen, Inc.), IFN-β 1b (Betaferon, Bayer Pharma AG) or IFN-β 1a (Rebif, Merck Serono Ltd) by a calibrated antiviral CPE test, as previously described by 29 (link). Briefly, A549 cells were seeded in 96 well plates in 100 μl of DMEM medium supplemented with 2% FCS. After 24 h, serial dilutions of the patient sera were incubated for 1 h with IFN-β, and without IFN-β to control the presence of endogenous IFN-β in the patient serum. These dilutions were added to the A549 cell culture, seeded the day before. Each plate included a viral control which did not contain IFN-β, a cell control which did not contain virus, and a standard of IFN-β. After 24 h incubation, cells were infected with Encephalomyocarditis virus (EMCV) (except for the cell control). Twenty-four hours later, the cells were stained with crystal violet and the absorbance was read at 630 lambdas in a spectrophotometer. The neutralization titre was calculated according to 30 (link) and expressed in 10-fold reduction units per millilitre (TRU/mL). Titres ≥20 TRU/mL were considered as positive.