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

作者：Santer Deanna M^*; Minty Gillian E S; Mohamed Adil; Baldwin Lesley; Bhat Rakesh; Joyce Michael; Egli Adrian; Tyrrell D Lorne J; Houghton Michael^*

来源：Journal of Immunological Methods, 2017, 445: 15-22.

DOI：10.1016/j.jim.2017.03.001

• Interferon-lambda
• Interferon-lambda receptor
• Flow cytometry
• Binding assay
• Interferon-stimulated genes

Type III interferons (IFN-lambdas) are important antiviral cytokines that also modulate immune responses acting through a unique IFN-lambda R1/IL-10R2 heterodimeric receptor. Conflicting data has been reported for which cells express the IFN-lambda\R1 subunit and directly respond to IFN-lambda s. In this study we developed a novel method to measure IFN-lambda 3 binding to IFN-lambda R1/1-10R2 on the surface of cells and relate this to a functional readout of interferon stimulated gene (ISG) activity in various cell lines. We show that Huh7.5 hepatoma cells bind IFN-lambda 3 at the highest levels with the lowest K-d(app), translating to the highest induction of various ISGs. Raji and Jurkat cell lines, representing B and T cells, respectively, moderately bind IFN-lambda 3 and have lower ISG responses. U937 cells, representing monocytes, did not bind IFN-lambda 3 well and therefore, did not have any ISG induction. Importantly, knockdown of IFNLR1 in Huh7.5 cells decreased our binding signal proportionally and reduced ISG induction by up to 93%. IFN-lambda 3 responsiveness increased over time with maximal ISG responses seen at 24 h for all but one gene. These data confirm our new IFN-lambda 3 binding assay can be used to quantify IFN-X receptor surface expression on a variety of cell types and reflects IFN-lambda 3 responsiveness.

• 出版日期2017-6