ABSTRACT
A new formulation (NF) of subcutaneous (sc) interferon (IFN) ß-1a was developed in an attempt to improve injection tolerability and immunogenicity. We compared antiviral and IFNß-stimulated gene (ISG) activities of IFNß-1a sc NF with IFNß-1a sc original formulation and IFNß-1b sc. When equivalent unit amounts were compared, the IFNß formulations demonstrated similar antiviral activity and induced similar levels of ISG mRNA. However, on a weight basis (ng/mL), significantly more IFNß-1b sc was needed to equal the antiviral activity of either IFNß-1a sc formulation, and both IFNß-1a sc formulations induced significantly higher levels of ISG mRNA than IFNß-1b sc.
Subject(s)
Epithelial Cells/drug effects , Fibroblasts/drug effects , Interferon-beta/pharmacology , Viral Load/drug effects , Animals , Cell Line, Tumor , Epithelial Cells/immunology , Epithelial Cells/virology , Fibroblasts/immunology , Fibroblasts/virology , Gene Expression , Humans , Interferon beta-1a , Interferon beta-1b , Interferon-Stimulated Gene Factor 3/agonists , Interferon-Stimulated Gene Factor 3/biosynthesis , Interferon-Stimulated Gene Factor 3/immunology , Interferon-beta/immunology , Mice , Multiple Sclerosis/drug therapy , Recurrence , Vesicular stomatitis Indiana virus/immunology , Virus Replication/drug effectsABSTRACT
Nitric oxide (NO) that is produced by inducible nitric oxide synthase (iNOS) is associated with the pathophysiology of glomerulonephritis. Numerous studies have focused on the regulation of NO production by iNOS to reduce NO-mediated cytotoxicity. In the present study, we demonstrated the differential effects of two phosphatidylinositol 3-kinase (PI3K) inhibitors, LY294002 and wortmannin, on lipopolysaccharide- (LPS) and interferon (IFN)-γ-induced NO production in a glomerular mesangial cell line, MES-13 cells. At dosages without affecting cell viability of MES-13 cells, 5µM LY294002 showed a more-significant inhibitory effect on LPS/IFN-γ-induced NO production, and iNOS protein and gene expressions than did 1µM wortmannin. Akt phosphorylation in MES-13 cells declined upon the addition of wortmannin, but not upon treatment with LY294002. Suppression of PI3K expression by small interfering (si)RNA exhibited no effect on LPS/IFN-γ-stimulated NO production or iNOS protein expression in MES-13 cells. Neither LY294002 nor wortmannin reduced IFN-γ-induced STAT-1α phosphorylation. LY294002 exhibited a more-significant inhibitory effect on NF-κB luciferase activities than wortmannin in LPS/IFN-γ-stimulated MES-13 cells. Moreover, LY294002, but not wortmannin, accelerated iNOS protein degradation and reduced the iNOS dimer/monomer ratio in MES-13 cells. Although both LY294002 and wortmannin are known as PI3K inhibitors, their differential effects on iNOS expression in MES-13 cells indicate that the effects of LY294002 on inhibiting NF-κB activation and accelerating iNOS protein degradation are through a mechanism independent of PI3K.
