KCNE1L Plays A Role In Anti-viral Innate Immune Response Against Herpes Simplex Virus-1 And Regulates TNF-? Mediated IL-8 Gene Expression

ObjectiveKCNE1L belongs to the KCNE family,and it is a potassium channel accessary protein.Previous studies on its function have been focused on its association with arrhythmia.Our previous study found that KCNE1 L gene expression was significantly decreased in peripheral blood mononuclear lymphocytes(PBMCs)upon HSV-1 stimulation in atopic dermatitis patients with a history of disseminated herpes simplex virus infection,the biological significance of this phenomenon is not clear.The purpose of this study is to investigate the role of KCNE1 L in the innate immune response against herpes simplex virus and TNF?-mediated inflammatory response.Methods The eukaryotic expression plasmids of KCNE1 L and short hairpin RNA targeting KCNE1 L were used to over-express and inhibit KCNE1 L gene expression.Real-time quantitative PCR and plaque assays were used to evaluate HSV-1 replication.Immune co-precipitation method was used to detect the interaction between KCNE1 L and other proteins that participate in innate immune signaling pathways.Immunofluorescence microscopy was used to detect the organelle location of KCNE1 L protein.Real-time quantitative PCR and ELISA were used to detect IFN?,IFN?,TNF?,IL-29 and IL-8 gene expression.Results1?Overexpression of KCNE1 L significantly inhibits HSV-1 viral replication: while inhibition of KCNE1 L gene expression using shRNA led to significantly enhanced HSV-1 replication.2?Overexpression of KCNE1 L augments HSV-1 induced gene expression of IL-29 and TNF?: while inhibition of KCNE1 L gene expression using shRNA led to reduced IL-29 and TNF? expression upon HSV-1 stimulation.3?KCNE1L and IRF7 interact with each other.Over-expression of KCNE1 L enhances IRF7-mediated IL-29 expression.4?Overexpression of KCNE1 L can enhance TNF? induced IL-8 gene expression.5?KCNE1L is located in the mitochondria and Golgi apparatus.It is translocated from cytoplasm to nucleus upon TNF? stimulation.ConclusionKCNE1L inhibits HSV-1 replication,and enhances HSV-1 induced type III interferon IL-29 gene expression.KCNE1 L also enhances TNF?-induced IL-8 gene expression,suggesting that it may be involved in TNF?-mediated inflammatory response.