Autocrine Interferon Priming in Macrophages but Not Dendritic Cells Results in Enhanced Cytokine and Chemokine Production after Coronavirus Infection

Zhou, Haixia; Zhao, Jincun; Perlman, Stanley

Generation of BMM and BMDC. Primary bone marrow cells were isolated from the hind limbs of WT, IFNAR Ϫ/Ϫ , or MyD88 Ϫ/Ϫ C57BL/6 mice as previously described (32) . BMM were cultured in DMEM supplemented with 10% FBS, 10% L929 cell-conditioned medium (as a source for macrophage colony-stimulating factor [M-CSF]), 1 mM sodium pyruvate, and 1% penicillin-streptomycin. The cells were harvested 7 days after plating and were Ͼ90% pure (CD11b ϩ CD11c -), as determined by flow cytometry. BMDC were grown in RPMI 1640 medium supplemented with 10% FBS, 1.0 mM HEPES, 0.2 mM L-glutamine, 1% penicillinstreptomycin, 1 mM sodium pyruvate, 0.02 mM 2-mercaptoethanol, 1,000 U/ml recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) (BD Pharmingen, San Diego, CA), and 25 U/ml recombinant interleukin-4 (IL-4) (PeproTech, Rocky Hill, NJ). The cells were used after 7 days of culture and were Ͼ80% pure (CD11b ϩ CD11c ϩ ) as determined by flow cytometry.