Author: Zhou, Haixia; Zhao, Jincun; Perlman, Stanley
Title: Autocrine Interferon Priming in Macrophages but Not Dendritic Cells Results in Enhanced Cytokine and Chemokine Production after Coronavirus Infection Document date: 2010_10_19
ID: qg541qho_10
Snippet: To assess whether the upregulation of these cytokines and chemokines was IFN dependent, we analyzed only BMM, since BMDC produced very low levels of these molecules and did not express type I IFN. We also included an additional IFNresponsive gene ISG15 (IFN-stimulated gene 15) in these assays. Consistent with a previous study of lipopolysaccharide (LPS)-treated BMM and BMDC (41), we found that CXCL10, IL-10, and ISG15 expression was dependent on .....
Document: To assess whether the upregulation of these cytokines and chemokines was IFN dependent, we analyzed only BMM, since BMDC produced very low levels of these molecules and did not express type I IFN. We also included an additional IFNresponsive gene ISG15 (IFN-stimulated gene 15) in these assays. Consistent with a previous study of lipopolysaccharide (LPS)-treated BMM and BMDC (41), we found that CXCL10, IL-10, and ISG15 expression was dependent on IFNAR, but not MyD88, signaling (Fig. 5) . TNF, IL-6, and IL-12 p40 mRNA upregulation was abolished in MHV-infected MyD88 Ϫ/Ϫ cells and was variably dependent upon IFNAR signaling: the levels of IL-6 and IL-12 p40 were decreased by about 50 to 80% in IFNAR Ϫ/Ϫ BMM ( Fig. 5A and B) . These mRNA data were confirmed by measuring the levels of CXCL10, TNF, and IL-12 p70 protein in an ELISA (Fig. 5C ). As in MHV-infected cells, CXCL10 expression in SenV-infected or poly(I · C)-transfected BMM was also dependent on IFNAR, but not MyD88. However, TNF expression in these samples was not dependent on MyD88, consistent with a previous report showing that TNF expression was mediated by direct MDA5 recognition in SenV-infected cells (Fig. 5A) (42) . IL-6 and IL-12 p40 were expressed at very low levels in poly(I · C)-treated or SenV-infected BMM, so expression of these cytokines was not analyzed further.
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