Inhibition of influenza virus-induced NF-kappaB and Raf/MEK/ERK activation can reduce both virus titers and cytokine expression simultaneously in vitro and in vivo

Expression of Raf kinase inhibitor protein is downregulated in response to Newcastle disease virus infection to promote viral replication

“…In line with others reports, our data show that IkBa/NF-kB signalling is essential for efficient virus propagation, as inhibition of IkBa phosphorylation by IKK inhibition via Bay 11-7082 results in reduced virus titres, such as Venezuelan equine encephalitis virus (Amaya et al, 2014), IV (Mazur et al, 2007;Pinto et al, 2011) and coxsackievirus (Sobotta et al, 2012). Previous studies showed that NDV replication does not require NF-kB p50 and cRel and, accordingly, embryonic fibroblasts (MEF) derived from p50 2/2 and cRel 2/2 mice revealed no defect during the early time of infection.…”

“…RKIP also works as a scaffold protein that negatively regulates the assembly of the IKK complex leading to the degradation of IkB releasing the NF-kB complex for nuclear translocation (Beshir et al, 2010;Tang et al, 2010). Furthermore, activation of Raf/MEK/ERK and the NF-kB pathway are required for normal replication of certain viruses, such as IV (Pinto et al, 2011), Kaposi's sarcomaassociated herpesvirus (KSHV) (Ford et al, 2006), human immunodeficiency virus (HIV) (Zhu et al, 2011) and others (Mathers et al, 2014), to promote viral replication, prevent virus-induced apoptosis, and mediate the immune response to the invading pathogen (Hiscott et al, 2001). However, there is little information available in the literature regarding RKIP inhibitory mechanisms in the context of NDV infection.…”

Verapamil has Antiviral Activities that Target Different Steps of the Influenza Virus Replication Cycle

“…We also observed that IAV is unable to compensate functional effects resulting from a blockade of this particular cellular pathway, for example by acquiring specific mutations [9]. Inhibition of the signalling cascade was shown to reduce IAV replication both, in vitro and in vivo, further supporting its key role in IV replication [10,11]. Interestingly, the cascade is activated at least in part by viral HA protein accumulation and the signal is transmitted via the calciumdependent PKC-alpha to Raf [12].…”

“…To determine the median cytotoxic concentration of the compounds at which 50% of the cells are still viable (CC 50 ) an MTT assay was performed [10]. Briefly, A549 cells were seeded in 96 well microtitre plates and were grown over night.…”

“…Detection of active NF-κB was performed with an ELISA based high-throughput screening system (TransAM, ActiveMotif, USA) according to the manufacturer´s instruction [10]. Briefly, in this assay, immobilized oligonucleotides containing an NF-kB (5′-GGACTTTCC-3′) consensus binding sequence detect only the active form of NF-κB that will bind to this site.…”