Differential Responses of Plasmacytoid Dendritic Cells to Influenza and Distinct Viral Pathogens. Homo sapiens

Plasmacytoid dendritic cells (pDCs) are key components of the innate immune response that are capable of synthesizing and rapidly releasing vast amounts of type I interferons (IFNs), particularly IFN-alpha. Here we investigated whether pDCs, often regarded as a mere source of IFN, discriminate between various functionally discrete stimuli and to what extent this reflects differences in pDC responses other than IFN-alpha release. To examine the ability of pDCs to differentially respond to various doses of intact and infectious HIV, hepatitis C virus, and H1N1 influenza virus, whole-genome gene expression analysis, enzyme-linked immunosorbent assays, and flow cytometry were used to investigate pDC responses at the transcriptional, protein, and cellular levels. Our data demonstrate that pDCs respond differentially to various viral stimuli with significant changes in gene expression, including those involved in pDC activation, migration, viral endocytosis, survival, or apoptosis. In some cases, the expression of these genes was induced even at levels comparable to that of IFN-alpha. Interestingly, we also found that depending on the viral entity and the viral titer used for stimulation, induction of IFN-alpha gene expression and the actual release of IFN-alpha are not necessarily temporally coordinated. In addition, our data suggest that high-titer influenza A (H1N1) virus infection can stimulate rapid pDC apoptosis. Overall design: pDCs from 3 separate donors (biological replicates) were stimulated with PBS (control 1) (n=3), conditioned medium (control 2) (n=3), or HCV JFH-1 at 1.00x10^4 (n=3), 1.00x10^5 (n=3), or 1.00x10^6 (n=3) RNA copies/ml. pDCs from 3 separate donors (biological replicates) were stimulated with PBS (control) (n=3) or influenza virus A/PR/8/34 (H1N1; Charles River Laboratories) at 4 (n=3), 32 (n=3), 256 (n=3), or 2,048 (n=3) HAU/ml. pDCs from 3 separate donors (biological replicates) were stimulated with PBS (control) (n=3) or HIV-1 IIIB (Advanced Biotechnologies) at 1.00x10^2 (n=3), 1.00x10^3 (n=3), 1.00x10^4 (n=3), or 1.00x10^5 (n=3) 50% tissue culture infective doses (TCID50)/ml. pDCs from 10 separate donors (biological replicates) were treated with PBS (control) (n=10), HCV at 1.00x10^6 RNA copies/ml (n=10), HIV-1 BaL at 1.00x10^5 TCID50/ml (Advanced Biotechnologies) (n=10), HIV-1 IIIB at 1.00x10^5 TCID50/ml (Advanced Biotechnologies) (n=10), or influenza virus A/PR/8/34 (H1N1; Charles River Laboratories) at 4 HAU/ml (n=10).