A novel method for detection of IFN-lambda 3 binding to cells for quantifying IFN-lambda receptor expression

Santer, Deanna M.; Minty, Gillian E. S.; Mohamed, Adil; Baldwin, Lesley; Bhat, Rakesh; Joyce, Michael; Egli, Adrian; Tyrrell, D. Lorne; Houghton, Michael

doi:10.1016/j.jim.2017.03.001

Cited by 4 publications

(4 citation statements)

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“…Neutrophils were the only cell type tested where there was higher sIFNLR1 than mIFNLR1 expression ( Fig 4A). In agreement with our data with primary T and B cells, we had previously found Jurkat T cell and Raji B cell lines also had low membrane/soluble IFNLR1 ratios of~2-3 [48]. We extended our cell line results to demonstrate that the lung BEAS-2B and A549 epithelial cell lines had a much higher membrane/soluble IFNLR1 transcript ratio of 14.9 and 6.7 respectively, while DG75 B cells had a lower ratio of 4.0 ( Fig 4A).…”

Section: Plos Pathogenssupporting

confidence: 93%

“…To date, studies on IFN-λR1 biology have been limited due to a lack of sensitive reagents to measure receptor protein expression. We recently developed a flow cytometry binding assay [48], which measures IFN-λ3 binding to the cell surface as a surrogate of IFN-λR1 expression. We quantified the binding of 6 His-tagged IFN-λ3 to immune cell subsets within peripheral blood, as well as primary liver and lung epithelial cells.…”

Section: Ifn-λ3 Binds To Both Epithelial Cells and Specific Immune Cementioning

confidence: 99%

“…HepG2 hepatocyte cell line [35]. We used our previously designed RT-qPCR assay to specifically quantify either the full length membrane form (mIFNLR1) or sIFNLR1 [48]. Using this assay, we found all immune cells tested had higher levels of sIFNLR1 relative to mIFNLR1, when compared to epithelial cells.…”

Section: Plos Pathogensmentioning

confidence: 99%

“…The IFN-λ3 binding assay was performed as published [48]. In brief, cells were cultured with or without His-tagged IFN-λ3 (R&D Systems) diluted in PBS containing 1% BSA on ice for 60 min at indicated doses.…”

Section: Flow Cytometrymentioning

confidence: 99%

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Differential expression of interferon-lambda receptor 1 splice variants determines the magnitude of the antiviral response induced by interferon-lambda 3 in human immune cells

et al. 2020

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Type III interferons (IFN-lambdas(λ)) are important cytokines that inhibit viruses and modulate immune responses by acting through a unique IFN-λR1/IL-10RB heterodimeric receptor. Until now, the primary antiviral function of IFN-λs has been proposed to be at anatomical barrier sites. Here, we examine the regulation of IFN-λR1 expression and measure the downstream effects of IFN-λ3 stimulation in primary human blood immune cells, compared with lung or liver epithelial cells. IFN-λ3 directly bound and upregulated IFN-stimulated gene (ISG) expression in freshly purified human B cells and CD8 + T cells, but not monocytes, neutrophils, natural killer cells, and CD4 + T cells. Despite similar IFNLR1 transcript levels in B cells and lung epithelial cells, lung epithelial cells bound more IFN-λ3, which resulted in a 50-fold greater ISG induction when compared to B cells. The reduced response of B cells could be explained by higher expression of the soluble variant of IFN-λR1 (sIFN-λR1), which significantly reduced ISG induction when added with IFN-λ3 to peripheral blood mononuclear cells or liver epithelial cells. T-cell receptor stimulation potently, and specifically, upregulated membrane-bound IFNLR1 expression in CD4 + T cells, leading to greater antiviral gene induction, and inhibition of human immunodeficiency virus type 1 infection. Collectively, our data demonstrate IFN-λ3 directly interacts with the human adaptive immune system, unlike what has been previously shown in published mouse models, and that type III IFNs could be potentially utilized to suppress both mucosal and blood-borne viral infections.

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Section: Plos Pathogenssupporting

confidence: 93%

Section: Ifn-λ3 Binds To Both Epithelial Cells and Specific Immune Cementioning

confidence: 99%