Monocytes differentiated into macrophages and dendritic cells in the presence of human IFN-λ3 or IFN-λ4 show distinct phenotypes

Abstract

Human IFN-4 is expressed by only a subset of individuals who possess the ΔG variant allele at the dinucleotide polymorphism rs368234815. Recent genetic studies have shown an association between rs368234815 and different infectious and inflammatory disorders. It is not known if IFN-4 has immunomodulatory activity. The expression of another type III IFN, IFN-3, is also controlled by genetic polymorphisms that are strongly linked to rs368234815. Therefore, it is of interest to compare these two IFNs for their effects on immune cells. Herein, using THP-1 cells, it was confirmed that IFN-4 could affect the differentiation status of macrophage-like cells and dendritic cells (DCs). The global gene expression changes induced by IFN-4 were also characterized in in vitro generated primary macrophages. Next, human PBMC-derived CD14 + monocytes were used to obtain M1 and M2 macrophages and DCs in the presence of IFN-3 or IFN-4. These DCs were cocultured with CD4 + Th cells derived from allogenic donors and their in vitro cytokine responses were measured. The specific activity of recombinant IFN-4 was much lower than that of IFN-3, as shown by induction of IFN-stimulated genes. M1 macrophages differentiated in the presence of IFN-4 showed higher IL-10 secretion than those differentiated in IFN-3. Coculture experiments suggested that IFN-4 could confer a Th2-biased phenotype to allogenic Th cells, wherein IFN-3, under similar circumstances, did not induce a significant bias toward either a Th1 or Th2 phenotype. This study shows for the first time that IFN-4 may influence immune responses by immunomodulation.

Distinct molecular phenotypes involving several human diseases are induced by IFN-λ3 and IFN-λ4 in monocyte-derived macrophages

“…Human recombinant IFN-λ4 (catalog #9165-IF; carrier-free form; E. coli ­-derived) and IFN-λ3 (catalog #5259-IL/CF; Chinese Hamster Ovary cell line, CHO-derived), containing <0.1 EU/µg of endotoxin purchased from R&D Systems (Minneapolis, MN, USA) were included in the differentiation medium at 0.05 µg/ml and 6 µg/ml for IFN-λ3 and IFN-λ4 respectively. These concentrations gave comparable interferon stimulated gene (ISG) induction by the two IFNs tested on different cell types as was determined in our earlier study [ 22 ]. After six days of differentiation, lipopolysaccharides (LPS) at 100 ng/ml (Sigma-Aldrich, St. Louis, MO, USA) was used to stimulate the MDMs before processing them for RNA isolation.…”

“…The protocol for generation of M1 and M2-MDMs (monocyte-derived macrophages) in presence of recombinant IFN-λ3/4 has been described in our earlier study [ 22 ]. Briefly, 10 million CD14 + monocytes (PromoCell, Heidelberg, Germany) each, obtained from four unrelated Caucasian donors were grown in RPMI-1640 medium supplemented with antibiotics and 10% FBS (all from Gibco, Waltham, MA, USA).…”

Interferon lambda signals in maternal tissues to exert protective and pathologic effects in a gestational-stage dependent manner

“…Multiple distinct subsets of macrophages have been identified in the maternal decidua, and imbalance between macrophages subtypes is associated with adverse pregnancy outcomes (39,40). IFN-λ changes the transcriptional profile and increases pro-inflammatory phenotypes of monocytes differentiated into macrophages in culture (41,42). Macrophages skew towards a M2 phenotype as pregnancy progresses and IFN-λ could increase proportions of M1 macrophages, potentially leading to inflammation and fetal rejection.…”

An Unbiased Flow Cytometry-Based Approach to Assess Subset-Specific Circulating Monocyte Activation and Cytokine Profile in Whole Blood

“…As classical monocytes are mainly involved in phagocytosis, they can be recruited quickly to sites of infection. The remaining 15% consists of CD14 dim and CD14 ++ CD16 + monocytes, which predominantly unfold pro-inflammatory characteristics (5)(6)(7)(8)(9). The number of CD16-positive monocytes can increase or decrease strongly under various conditions.…”