Expression profiles of human interferon‐alpha and interferon‐lambda subtypes are ligand‐ and cell‐dependent

Abstract

Recent genome-wide association studies suggest distinct roles for 12 human interferon-alpha (IFN-α) and 3 IFN-λ subtypes that may be elucidated by defining the expression patterns of these sets of genes. To overcome the impediment of high homology among each of the sets, we designed a quantitative real-time PCR assay that incorporates the use of molecular beacon and locked nucleic acid (LNA) probes, and in some instances, LNA oligonucleotide inhibitors. We then measured IFN subtype expression by human peripheral blood mononuclear cells and by purified monocytes, myeloid dendritic cells (mDC), plasmacytoid dendritic cells (pDC), and monocyte-derived macrophages (MDM), and –dendritic cells (MDDC) in response to poly I:C, lipopolysaccharide (LPS), imiquimod and CpG oligonucleotides. We found that in response to poly I:C and LPS, monocytes, MDM and MDDC express a subtype pattern restricted primarily to IFN-β and IFN-λ1. In addition, while CpG elicited expression of all type I IFN subtypes by pDC, imiquimod did not. Furthermore, MDM and mDC highly express IFN-λ, and the subtypes of IFN-λ are expressed hierarchically in the order IFN-λ1 followed by IFN-λ2, and then IFN-λ3. These data support a model of coordinated cell- and ligand-specific expression of types I and III IFN. Defining IFN subtype expression profiles in a variety of contexts may elucidate specific roles for IFN subtypes as protective, therapeutic or pathogenic mediators.

Different Temporal Effects of Ebola Virus VP35 and VP24 Proteins on Global Gene Expression in Human Dendritic Cells

“…To determine the effect of the mutations on the expression of IFNs over a longer period of time and to confirm the transcriptome data by an alternative method, we analyzed RNA samples from time points up to 120 h by qRT-PCR analysis (45) (Fig. 9).…”

“…Samples were subsequently treated with RNase H (New England BioLabs, Ipswich, MA) according to the manufacturer's instructions using the following run conditions: 37°C for 20 min, 65°C for 20 min, 4°C until the end of the run. The expression of types I and III IFN was measured using the qRT-PCR method described previously (45). Briefly, 384-well assay plates were prepared by adding primers and probes with the Solo automated multichannel pipettor (Hudson Robotics, Inc., Springfield, NJ), dried, and stored in the dark at 4°C until use.…”

Immune activation of human brain microvascular endothelial cells inhibits HIV replication in macrophages

“…[39][40][41] However, their numbers and ability to produce type I IFNs are significantly reduced by HIV infection. [42][43][44][45][46] Thus, an IFNmediated antiviral response mounted by nonimmune cells such as brain ECs is likely to be beneficial for CNS protection.…”

IFN‐α subtypes: distinct biological activities in anti‐viral therapy

“…A recent study by Hillyer and colleagues shows that in vitro treatment of human peripheral blood mononuclear cells, purified myeloid DCs, plasmacytoid DCs and monocytes with different TLR ligands resulted in the induction of specific IFN-a subtypes by the ligands tested (Hillyer et al, 2012). This study reveals a coordinated ligand-and cellspecific expression of type I IFN subtypes, which might be useful for future treatment of viral infections with different IFN-a subtypes or their specific induction by exogenously applied TLR ligands.…”

“…Induced Mo et al, 2007 high in poly I : C-stimulated DC Hillyer et al, 2012 High APOBEC3G induction Vazquez et al, 2011 By CpG in PBMC Puig et al, 2012 Intermediate inducer of IFIT1, CXCL10, CXCL11, ISG15 and CCL8 Moll et al, 2011 Less potent against human metapneumovirus Scagnolari et al, 2011 HSV, herpes simplex virus; IDO, indoleamine 2,3-dioxygenase; NDV, Newcastle disease virus; PBMC, peripheral blood mononuclear cells; pDC, plasmacytoid DC; RSV, respiratory syncytial virus. The nomenclature conforms to that used by van Pesch et al (2004).…”