PMC:3291650 / 15209-17622 JSONTXT

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In accordance with these data, cell culture supernatant collected from these cells contained higher levels of TNF and IFNβ (figure 2B).\n10.1371/journal.ppat.1002570.g002 Figure 2 A20 negatively regulates IAV-induced gene expression in BMDM and alveolar macrophages.\n(A–B) BMDM isolated from A20myel-WT and A20myel-KO animals were infected with IAV (moi 1). At different hours post infection (hours p.i.) cells were lysed and IL-6, IFNβ and A20 mRNA expression was analyzed by qPCR (A). 18 hours post infection cell culture supernatant was analyzed for TNF and IFNβ protein levels by ELISA and multiplexing technologies (B). (C–D) Alveolar macrophages were mock treated or infected with IAV (moi 1) for 18 hours. IL-6 and IFNβ mRNA expression was determined by qPCR (C). Cell culture supernatant was analyzed for IL-6 and IFNβ protein levels by ELISA and multiplexing technologies (D). Error bars represent mean values (+/− SEM) of 2–3 samples. Results are representative for 2 independent experiments. ***p\u003c0,001. Upon host infection with IAV, alveolar macrophages are an important source of cytokines and chemokines, attracting innate immune cells to the lung during the primary phase of infection. To test whether A20 directly controls IAV-induced gene expression in alveolar macrophages, we isolated these cells from lungs of A20myel-KO and control littermates and infected them in vitro with IAV X-47. Expression and secretion of the proinflammatory cytokines IL-6 and TNF, the type-I IFN IFNβ and IFNα4 and the chemokines MCP-1 (ccl2) and KC (cxcl1) was significantly higher in IAV infected cells lacking A20 compared to infected wild type cells (figure 2C and figure S1). Taken together these results demonstrate that A20 negatively regulates IAV-induced proinflammatory and antiviral gene expression in alveolar macrophages, consistent with the inhibitory effect of A20 seen on RIG-I-induced NF-κB and IRF3 activation.","divisions":[{"label":"title","span":{"begin":0,"end":60}},{"label":"p","span":{"begin":61,"end":622}},{"label":"figure 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