1. Viral infection triggers rapid differentiation of human blood monocytes into dendritic cells
by Wanqiu Hou, James S. Gibbs, Xiuju Lu, Christopher B. Brooke, Devika Roy, Robert L. Modlin, Jack R. Bennink, and Jonathan W. Yewdell
Blood
Volume 119(13):
March 29, 2012
©2012 by American Society of Hematology

2. Virus infection of human monocytes induces differentiation into DCs
Virus infection of human monocytes induces differentiation into DCs. (A) Human PBMCs were infected for 6 hours with the indicated virus at a multiplicity of infection (MOI) of 10, and surface expression of IAV hemagglutinin or virus-encoded enhanced green f...
Virus infection of human monocytes induces differentiation into DCs. (A) Human PBMCs were infected for 6 hours with the indicated virus at a multiplicity of infection (MOI) of 10, and surface expression of IAV hemagglutinin or virus-encoded enhanced green fluorescent protein on CD3+ T cells, CD14+ monocytes, CD19+ B cells, CD56+ NK cells, lineage-negative (CD3/14/19/56)−HLA-DR+CD11c+CD123− cDCs, or lineage-negative HLA-DR+CD11c−CD123+ pDCs was determined by flow cytometry. The results represent 1 of 2 separate experiments with similar results. (B) Flow cytometric analysis of T/monocyte/B/NK cells 6 or 18 hours after IAV infection at indicated MOI. % Max indicates percent of maximum. (C) Effect of IAV infection at indicated MOI 6 or 18 hours after infection on lineage-negative HLA-DR+ DC population (red box; left panel) or on cDC (circled CD123lowCD11c+ population)/pDC (circled CD123+CD11c− population) populations (middle panel), and the cell number of monocytes, cDCs, and pDCs in 5 × 105 PBMCs 18 hours after IAV infection (right panel). (D) CD14+ monocyte-depleted PBMCs were infected for 18 hours with IAV and then stained with lineage markers and HLA-DR. The results in panels B through D are representative examples of 5 independent experiments. (E) Purified CD14+ monocytes were infected for 6 and 18 hours with IAV and then stained with anti-hemagglutinin antibody. The results are representative examples of 3 independent experiments.
Wanqiu Hou et al. Blood 2012;119:
©2012 by American Society of Hematology

3. Characterizing virus-induced DCs
Characterizing virus-induced DCs. (A) Monocytes irradiated with 2000 rad of γ-radiation or nonirradiated cells were infected with live or UV-inactivated IAV. At 18 hours after infection, CD11c+CD123− cDCs instead of CD123+ monocytes or pDCs among the CD14−H...
Characterizing virus-induced DCs. (A) Monocytes irradiated with 2000 rad of γ-radiation or nonirradiated cells were infected with live or UV-inactivated IAV. At 18 hours after infection, CD11c+CD123− cDCs instead of CD123+ monocytes or pDCs among the CD14−HLA-DR+ DC population were assessed by flow cytometry. (B) Surface CD16 or CD83 expression on the cDCs in panel A from infected and uninfected samples at 18 hours after IAV infection. The results represent 1 of 3 separate experiments. (C) Quantitative real-time PCR analysis of monocyte/DC-related markers in monocytes after 6 hours after IAV infection at a MOI of 10 vs uninfected sample. The results represent the average of 5 subject samples. (D) Intracellular TNF production by IAV-infected PBMCs or monocytes at 6 hours after infection. (E) Levels of surface CD1c, CD141, CLEC4C, and CLEC9A, as well as intracellular CLEC9A, by IAV-infected monocytes at 18 hours after infection. Shaded and open histograms, respectively, represent isotype and specific antibody staining. (F) Monocytes infected for 6 hours with different doses of VSV were cocultured for 7 days with purified CD4+ T cells from the same donor at a monocyte/T-cell ratio of 1:5 in the presence of varied concentrations of UV-inactivated IAV antigen. IFN-γ levels in supernatants of the cocultures were determined by ELISA. Data in panels D and F are representative of 3 separate experiments. % Max indicates percent of maximum.
Wanqiu Hou et al. Blood 2012;119:
©2012 by American Society of Hematology