title: Screening and characterization of molecules that modulate the biological activity of IFNs-I.
abstract: Type I Interferons (IFNs-I) are species-specific glycoproteins which play an important role as primary defence against viral infections and that can also modulate the adaptive immune system. In some autoimmune diseases, interferons (IFNs) are over-produced. IFNs are widely used as biopharmaceuticals for a variety of cancer indications, chronic viral diseases, and for their immuno-modulatory action in patients with multiple sclerosis; therefore, increasing their therapeutic efficiency and decreasing their side effects is of high clinical value. In this sense, it is interesting to find molecules that can modulate the activity of IFNs. In order to achieve that, it was necessary to establish a simple, fast and robust assay to analyze numerous compounds simultaneously. We developed four reporter gene assays (RGAs) to identify IFN activity modulator compounds by using WISH-Mx2/EGFP, HeLa-Mx2/EGFP, A549-Mx2/EGFP, and HEp2-Mx2/EGFP reporter cell lines (RCLs). All of them present a Z' factor higher than 0.7. By using these RGAs, natural and synthetic compounds were analyzed simultaneously. A total of 442 compounds were studied by the Low Throughput Screening (LTS) assay using the four RCLs to discriminate between their inhibitory or enhancing effects on IFN activity. Some of them were characterized and 15 leads were identified. Finally, one promising candidate with enhancing effect on IFN-α/-β activity and five compounds with inhibitory effect were described.
authors.affiliation: Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria - C.C. 242 - (S3000ZAA) Santa Fe, Provincia de Santa Fe, Argentina.
authors.forename: Milagros
authors.initials: M
authors.lastname: Bürgi
authors.affiliation: Technical University of Clausthal, Leibnizstraße 6, D-38678 Clausthal-Zellerfeld, Germany.
authors.forename: Viktor A
authors.initials: VA
authors.lastname: Zapol'skii
authors.affiliation: Department of Chemical Biology, Helmholtz Centre for Infection Research, Inhoffenstraße 7, D-38124 Braunschweig, Germany.
authors.forename: Bettina
authors.initials: B
authors.lastname: Hinkelmann
authors.affiliation: Department Gene Regulation and Differentiation, Helmholtz Centre for Infection Research, Inhoffenstraße 7, D-38124 Braunschweig, Germany.
authors.forename: Mario
authors.initials: M
authors.lastname: Köster
authors.affiliation: Technical University of Clausthal, Leibnizstraße 6, D-38678 Clausthal-Zellerfeld, Germany.
authors.forename: Dieter E
authors.initials: DE
authors.lastname: Kaufmann
authors.affiliation: Department of Chemical Biology, Helmholtz Centre for Infection Research, Inhoffenstraße 7, D-38124 Braunschweig, Germany.
authors.forename: Florenz
authors.initials: F
authors.lastname: Sasse
authors.affiliation: Department Gene Regulation and Differentiation, Helmholtz Centre for Infection Research, Inhoffenstraße 7, D-38124 Braunschweig, Germany.
authors.forename: Hansjörg
authors.initials: H
authors.lastname: Hauser
authors.affiliation: Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria - C.C. 242 - (S3000ZAA) Santa Fe, Provincia de Santa Fe, Argentina.
authors.forename: Marina
authors.initials: M
authors.lastname: Etcheverrigaray
authors.affiliation: Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria - C.C. 242 - (S3000ZAA) Santa Fe, Provincia de Santa Fe, Argentina.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Kratje
authors.affiliation: Cell Biology Unit, Institut Pasteur de Montevideo, Mataojo 2020, CP 11400, Montevideo, Uruguay. Electronic address: mbollati@pasteur.edu.uy.
authors.forename: Mariela
authors.initials: M
authors.lastname: Bollati-Fogolín
authors.affiliation: Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria - C.C. 242 - (S3000ZAA) Santa Fe, Provincia de Santa Fe, Argentina.
authors.forename: Marcos
authors.initials: M
authors.lastname: Oggero
mesh_terms: D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D055808:Drug Discovery / Q000379:methods*; D017930:Genes, Reporter / Q000235:genetics*; D005821:Genetic Techniques; D006367:HeLa Cells; D006801:Humans; D016898:Interferon-alpha / Q000187:drug effects* / Q000502:physiology*; D015203:Reproducibility of Results
chemical_list: D016898:Interferon-alpha
keywords: Cell-based RGA; Low Throughput Screening; Mx2/EGFP; Natural and synthetic compounds; hIFN-I