{"gene":"IFNL2","run_date":"2026-06-10T01:55:22","timeline":{"discoveries":[{"year":2005,"finding":"IL-28A (IFNL2) binds to the receptor complex IL-28R1/IL-10R2 on intestinal epithelial cells, activating ERK-1/2, SAPK/JNK MAPKs, and Akt signaling, inducing STAT1 phosphorylation, upregulating SOCS3, MxA, and 2',5'-OAS mRNA, and reducing CMV-infected cells by up to 83%.","method":"Western blot (STAT1 phosphorylation, MAPK activation), qPCR (antiviral gene expression), flow cytometry (CMV infection), cell proliferation assay","journal":"American journal of physiology. Gastrointestinal and liver physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (Western blot, qPCR, cell-based antiviral assay) in single lab","pmids":["16051921"],"is_preprint":false},{"year":2005,"finding":"SOCS-1 overexpression completely abrogates IL-28A-induced STAT1 and STAT3 phosphorylation and abolishes IL-28A-induced mRNA expression of antiviral proteins 2',5'-OAS and MxA in hepatic cell lines, identifying SOCS-1 as a negative regulator of IL-28A signaling.","method":"Transient transfection (SOCS-1 overexpression), Western blot (STAT1/3 phosphorylation), qPCR (2',5'-OAS and MxA mRNA)","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (overexpression + Western blot + qPCR) in single lab","pmids":["15850793"],"is_preprint":false},{"year":2005,"finding":"IL-28A activates the JAK-STAT signaling pathway in human hepatoma cells, induces interferon-stimulated genes (ISGs) including 6-16 and 1-8U, induces HLA class I antigen expression, suppresses HCV subgenomic RNA replication in a dose-dependent manner, and synergistically enhances IFN-alpha antiviral efficacy. IL-28A also specifically suppresses HCV IRES-mediated translation.","method":"HCV RNA replicon assay, Western blot (JAK-STAT activation), qPCR (ISG expression), IRES reporter assay, combination treatment with IFN-alpha","journal":"Virology journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal assays (replicon, Western blot, reporter assay) in single lab","pmids":["16146571"],"is_preprint":false},{"year":2006,"finding":"IL-28A induces Th1 cytokine production by CD4+ T lymphocytes in a T-bet-dependent manner. In IL-28A-transgenic mice, augmented Con A-induced hepatitis with elevated IFN-gamma production was suppressed by crossing with T-bet-deficient mice, placing IL-28A upstream of the IFN-gamma/T-bet signaling pathway in T-cell-mediated liver injury.","method":"Transgenic mouse generation, genetic epistasis (IL-28A-transgenic × T-bet-deficient double mutant), in vivo Con A hepatitis model, cytokine measurements, antisense oligonucleotide knockdown","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with double-mutant rescue replicated across multiple orthogonal approaches (transgenic, knockout crossing, antisense knockdown, antibody neutralization) in single rigorous study","pmids":["17241885"],"is_preprint":false},{"year":2010,"finding":"IL-28A activates STAT1 signaling and induces a similar set of antiviral genes as IL-29 in hepatic cells; however, unlike IL-29, IL-28A is a potent gene repressor, uniquely down-regulating 272 genes as determined by microarray analysis.","method":"Microarray transcriptome analysis, Western blot (STAT1 phosphorylation), qPCR, HCV replicon assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — microarray + Western blot + HCV replicon in single lab, multiple orthogonal methods","pmids":["21170333"],"is_preprint":false},{"year":2011,"finding":"IL-28A modulates lung CD11c+ dendritic cell function by down-regulating OX40L and up-regulating IL-12p70, thereby promoting Th1 differentiation and suppressing Th2/Th17 responses; IL-28A-mediated protection from allergic airway disease was absent after IL-12 neutralization or in IFN-gamma-deficient mice, and could be adoptively transferred by IL-28A-treated CD11c+ cells.","method":"Recombinant cytokine treatment, IL-28Rα knockout mice, adoptive transfer of CD11c+ cells, antibody neutralization (IFN-gamma, IL-12), flow cytometry, cytokine measurements","journal":"EMBO molecular medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal approaches (KO mice, adoptive transfer, neutralization, recombinant cytokine) in single rigorous study demonstrating mechanistic pathway","pmids":["21538995"],"is_preprint":false},{"year":2011,"finding":"IL-28A inhibits HSV-1 replication in human NT2-N neurons and CHP212 neuronal cells via a mechanism partially blocked by anti-IL-10Rβ antibody; IL-28A selectively induces TLR expression and activates TLR-mediated antiviral pathways including IRF7, IFN-alpha, and IFN-alpha-stimulated antiviral genes.","method":"Neutralizing antibody (anti-IL-10Rβ), viral replication assay (HSV-1 DNA and protein quantification), qPCR (TLR and antiviral gene expression)","journal":"Journal of neurovirology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — receptor blocking + gene expression + viral assay, multiple orthogonal methods in single lab","pmids":["21499846"],"is_preprint":false},{"year":2012,"finding":"IL-28A stimulation of bladder cancer cells activates MAPK and JAK-STAT signaling, induces NF-κB and AP-1 transcription factors, upregulates MMP-2 and MMP-9 expression, and enhances cell migration and invasion.","method":"Immunoblot, EMSA (NF-κB/AP-1 activity), real-time PCR, wound-healing/invasion assays, confocal immunofluorescence","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (Western blot, EMSA, functional migration/invasion assays) in single lab","pmids":["22962576"],"is_preprint":false},{"year":2012,"finding":"IL-28A (IFN-λ2) treatment of lung cancer cells with EGFR mutations induces STAT1 phosphorylation, growth suppression, and apoptotic cell death; anti-tumor effect was confirmed in vivo in a cancer cell transplant animal model, and neutralizing antibodies to IFN-λ inhibited IL-28A-induced caspase-3/7 activity.","method":"In vitro cell viability/proliferation assay, Western blot (STAT1 phosphorylation), caspase-3/7 activity assay, neutralizing antibody, in vivo luminescent imaging in animal model","journal":"Lung cancer (Amsterdam, Netherlands)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (in vitro + in vivo, neutralization, biochemical assays) in single lab","pmids":["23021208"],"is_preprint":false},{"year":2014,"finding":"In CVB3-induced myocarditis, IL-28A treatment increases STAT1 and STAT2 expression, reduces viral load, and inhibits CVB3-induced apoptosis in myocardial cells with an increased Bcl-2/BAX ratio.","method":"Immunoblot (STAT1/STAT2), TUNEL assay (apoptosis), plaque assay (viral load), immunohistochemistry (Bcl-2/BAX/Caspase-3)","journal":"The Brazilian journal of infectious diseases","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — multiple methods but single lab, in vivo mouse model with mechanistic readouts","pmids":["25528576"],"is_preprint":false},{"year":2015,"finding":"EPSTI1 (epithelial-stromal interaction 1) is an IL-28A-induced ISG that is required for IL-28A-mediated inhibition of HCV replication; forced EPSTI1 expression inhibits HCV replication independently of interferon, while EPSTI1 knockdown enhances viral replication. EPSTI1 activates PKR promoter and induces PKR-dependent antiviral genes (IFN-β, IFIT1, OAS1, RNase L).","method":"siRNA knockdown, overexpression, HCV replication assay, promoter assay (PKR), qPCR","journal":"Mediators of inflammation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function and gain-of-function with mechanistic pathway placement in single lab","pmids":["26146465"],"is_preprint":false},{"year":2016,"finding":"Intratumoral IL-28A drives anti-tumor immunity through NK cells and CD8+ T cells; IL-28A-mediated tumor suppression is completely impaired in IFN-gamma-deficient mice, and IL-28A increases CD8+ T cell infiltration and specific CTL reactivity. IL-12 acts synergistically with IL-28A.","method":"In vivo cell depletion (NK, CD8+ T cells), IFN-gamma-deficient mice, tumor growth assay, CTL assay, immunohistochemistry","journal":"Cancer gene therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo genetic and depletion approaches with defined immune cell effectors, single lab","pmids":["27561689"],"is_preprint":false},{"year":2019,"finding":"Mice lacking both Ifnl2 and Ifnl3 (Ifnl2/3 double knockout) phenotypically recapitulate Ifnlr1-knockout mice in their inability to control murine norovirus, reovirus, and influenza virus at mucosal sites (intestine and lung), establishing that IFN-λ2 and IFN-λ3 are the exclusive ligands for IFNLR-mediated antiviral signaling at mucosal surfaces.","method":"Genetic knockout (Ifnl2/3 double KO mice), epistasis comparison with Ifnlr1-/- mice, viral infection models (norovirus, reovirus, influenza)","journal":"Journal of virology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with double-KO genocopying receptor KO across three independent viral infection models","pmids":["31462571"],"is_preprint":false},{"year":2020,"finding":"ATG10S (a shorter isoform of ATG10) translocates to the nucleus and binds the IFNL2 promoter at an IRF1 binding site (core motif CAAGAC), competing with IRF1 to promote IFNL2 transcription; ATG10S interacts with KPNA1/importin-α, KPNB1/importin-β, and IRF1 (shown by co-IP); knockdown of IRF1 increases ATG10S-driven IFNL2 transcription.","method":"Promoter deletion analysis, DNA immunoprecipitation, luciferase reporter assay, site-directed mutagenesis (nucleotide substitutions in core motif), co-immunoprecipitation, subcellular localization, RT-PCR, Western blot, siRNA knockdown","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — multiple orthogonal methods (ChIP, mutagenesis, co-IP, reporter assay) in single lab","pmids":["31996071"],"is_preprint":false},{"year":2020,"finding":"IL-28A forms a homotetramer in cells; the first α-helix of IL-28A is required for homotetramer formation. The IL-28A homotetramer (but not monomer) binds HCV NS5A at the NS5A-ISDR region, promotes autolysosome formation, and mediates autolysosomal degradation of HCV NS5A and other HCV proteins. HCV NS5A activates incomplete autophagy by promoting ATG3/5/7/10 and LC3B but blocking autophagy flux.","method":"Co-immunoprecipitation, cell immunofluorescence, sequential deletion mutants, software structural prediction, Western blot (autophagy markers), HepG2 cell-based assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with deletion mutants and functional readout (autolysosomal degradation) in single lab; structural prediction is computational but supported by experimental deletion mapping","pmids":["32205851"],"is_preprint":false},{"year":2024,"finding":"IL-28A promotes angiogenesis in endothelial cells via the IL-10Rβ receptor, activating eNOS/AKT and ERK1/2 signaling, inducing AP-1/NF-κB/MMP-2 network. HSP70-1 is an essential downstream effector; HSP70-1 knockdown abolishes IL-28A-induced angiogenic responses and eNOS/AKT signaling. IL-10Rβ gene ablation blocks IL-28A-induced angiogenic responses. IL-28A accelerated blood flow recovery in a hind-limb ischemia mouse model.","method":"siRNA knockdown (HSP70-1, IL-10Rβ), HUVEC proliferation/migration/invasion/tube formation assays, immunoblot, EMSA, aortic ring ex vivo assay, Matrigel plug in vivo assay, HSP70-1 KO and transgenic mice, hind-limb ischemia model, NGS transcriptomics","journal":"Journal of advanced research","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal in vitro and in vivo methods including KO and transgenic mice, receptor ablation, and mechanistic pathway delineation in single rigorous study","pmids":["39127098"],"is_preprint":false},{"year":2024,"finding":"IRF7 promotes intestinal barrier integrity through IL-28A; Irf7-/- mice show reduced IL-28A expression and compromised barrier markers (Muc2, E-cadherin, β-catenin, Occludin). Recombinant IL-28A treatment restores these barrier molecules and rescues Irf7-/- mice from enhanced DSS-induced colitis susceptibility, placing IL-28A downstream of IRF7 in intestinal epithelial barrier maintenance.","method":"Irf7 knockout mice, DSS colitis model, recombinant IL-28A rescue experiment, qPCR/Western blot (barrier molecule expression), intestinal epithelial cell stimulation with recombinant IL-28A","journal":"Journal of translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis (KO + rescue) with defined molecular readouts, single lab","pmids":["39370517"],"is_preprint":false},{"year":2010,"finding":"IL-28A does not induce CD25 or Foxp3 expression on cord blood CD4+ T cells, does not promote their proliferation, and does not induce regulatory T cell function in this cell type.","method":"Flow cytometry (CD25, Foxp3), Treg suppression assay, cell proliferation assay","journal":"Immunopharmacology and immunotoxicology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, negative finding on a specific cell type; limited mechanistic scope","pmids":["20148705"],"is_preprint":false}],"current_model":"IFNL2 (IL-28A) is a secreted type III interferon that signals through the IL-28R1/IL-10R2 receptor complex to activate JAK-STAT (STAT1/STAT2/STAT3), MAPK (ERK1/2, JNK), and AKT pathways, inducing antiviral ISGs (MxA, OAS, EPSTI1) and inhibiting viral replication (HCV, CMV, HSV-1, CVB3, influenza, norovirus, reovirus) at mucosal surfaces; its signaling is negatively regulated by SOCS-1; transcription of IFNL2 is promoted by ATG10S competing with IRF7/IRF1 at the promoter; in immune cells it modulates dendritic cell function (downregulating OX40L, upregulating IL-12) to drive Th1 responses via an IFN-gamma/T-bet axis; in endothelial cells it promotes angiogenesis via IL-10Rβ/eNOS/AKT/HSP70-1; and genetic ablation of Ifnl2/Ifnl3 fully phenocopies loss of the IFNLR receptor in mucosal antiviral defense."},"narrative":{"mechanistic_narrative":"IFNL2 (IL-28A) is a secreted type III interferon that engages the IL-28R1/IL-10R2 (IL-10Rβ) receptor complex on epithelial and other cells to mount antiviral defense and shape immune responses at mucosal surfaces [PMID:16051921]. Receptor engagement activates JAK-STAT (STAT1/STAT2/STAT3) together with ERK1/2, SAPK/JNK, and Akt signaling, driving expression of interferon-stimulated antiviral genes such as MxA and 2',5'-OAS and suppressing replication of diverse viruses including CMV, HCV, HSV-1, and CVB3 [PMID:16051921, PMID:16146571, PMID:21499846, PMID:25528576]; this signaling is restrained by SOCS-1, which abolishes IL-28A-induced STAT1/STAT3 phosphorylation and ISG induction [PMID:15850793]. Genetic ablation of Ifnl2/Ifnl3 phenocopies loss of the IFNLR receptor in controlling norovirus, reovirus, and influenza at mucosal sites, establishing IFN-λ2/λ3 as the exclusive ligands for IFNLR-mediated mucosal antiviral signaling [PMID:31462571]. Beyond direct antiviral action, IL-28A shapes adaptive immunity by modulating dendritic cells—downregulating OX40L and upregulating IL-12—to drive T-bet-dependent Th1 differentiation via an IFN-γ axis [PMID:17241885, PMID:21538995], and it drives NK- and CD8+ T-cell-mediated anti-tumor immunity in an IFN-γ-dependent manner [PMID:27561689]. IL-28A also promotes angiogenesis in endothelial cells through IL-10Rβ-dependent eNOS/AKT and ERK1/2 signaling with HSP70-1 as an essential downstream effector [PMID:39127098]. Its transcription is positively regulated by IRF7 in intestinal barrier maintenance [PMID:39370517] and by the ATG10S isoform, which competes with IRF1 at the IFNL2 promoter [PMID:31996071].","teleology":[{"year":2005,"claim":"Established that IL-28A acts through the IL-28R1/IL-10R2 receptor to activate JAK-STAT and MAPK/Akt pathways and induce antiviral ISGs, defining its core signaling axis.","evidence":"Western blot, qPCR, and CMV antiviral assay in intestinal epithelial cells","pmids":["16051921"],"confidence":"Medium","gaps":["Receptor stoichiometry and ligand-binding determinants not resolved","Relative contribution of MAPK vs JAK-STAT to antiviral output unclear"]},{"year":2005,"claim":"Identified SOCS-1 as a negative regulator that shuts down IL-28A-driven STAT phosphorylation and ISG induction, placing a brake on the pathway.","evidence":"SOCS-1 overexpression with Western blot and qPCR in hepatic cell lines","pmids":["15850793"],"confidence":"Medium","gaps":["Endogenous regulation by SOCS-1 not tested","Mechanism of SOCS-1 recruitment to the receptor not defined"]},{"year":2005,"claim":"Showed IL-28A suppresses HCV replication and IRES-mediated translation and synergizes with IFN-alpha, framing its therapeutic antiviral relevance.","evidence":"HCV replicon assay, IRES reporter, and combination treatment in hepatoma cells","pmids":["16146571"],"confidence":"Medium","gaps":["Effector ISGs mediating HCV suppression not identified at this stage","Mechanism of IRES translation block unknown"]},{"year":2006,"claim":"Placed IL-28A upstream of the IFN-γ/T-bet axis in driving Th1 cytokine production, establishing an immunomodulatory role beyond direct antiviral signaling.","evidence":"IL-28A-transgenic × T-bet-deficient genetic epistasis in a Con A hepatitis model","pmids":["17241885"],"confidence":"High","gaps":["Whether T-cell effects are cell-intrinsic or via accessory cells not fully resolved"]},{"year":2010,"claim":"Distinguished IL-28A from IL-29 by revealing a unique gene-repressive transcriptional signature despite shared ISG induction.","evidence":"Microarray transcriptome analysis with HCV replicon assay in hepatic cells","pmids":["21170333"],"confidence":"Medium","gaps":["Mechanism of gene repression not defined","Functional consequence of repressed genes untested"]},{"year":2011,"claim":"Defined the dendritic-cell mechanism by which IL-28A skews Th1 over Th2/Th17 responses, linking it to IL-12 and IFN-γ in allergic airway protection.","evidence":"IL-28Rα KO mice, adoptive transfer of CD11c+ cells, and IL-12/IFN-γ neutralization","pmids":["21538995"],"confidence":"High","gaps":["Direct DC receptor signaling events downstream of IL-28A not mapped"]},{"year":2011,"claim":"Showed IL-28A restricts HSV-1 in neuronal cells via IL-10Rβ and selectively engages TLR/IRF7-driven antiviral programs, extending antiviral action to the nervous system.","evidence":"Anti-IL-10Rβ neutralization and HSV-1 replication/qPCR assays in neuronal cells","pmids":["21499846"],"confidence":"Medium","gaps":["Only partial receptor blockade observed, suggesting additional receptor contributions","Mechanism of selective TLR induction unknown"]},{"year":2012,"claim":"Revealed context-dependent pro-tumorigenic signaling in bladder cancer where IL-28A enhances migration/invasion via NF-κB/AP-1 and MMP induction.","evidence":"Immunoblot, EMSA, and migration/invasion assays in bladder cancer cells","pmids":["22962576"],"confidence":"Medium","gaps":["Reconciliation with anti-tumor effects in other tumor types unresolved"]},{"year":2012,"claim":"Demonstrated direct growth-suppressive and pro-apoptotic activity of IL-28A on EGFR-mutant lung cancer cells, supporting a tumor-suppressive role in this context.","evidence":"STAT1/caspase-3/7 assays, neutralizing antibody, and in vivo xenograft imaging","pmids":["23021208"],"confidence":"Medium","gaps":["Determinants of EGFR-mutation selectivity not defined"]},{"year":2014,"claim":"Extended antiviral and cytoprotective action to viral myocarditis, where IL-28A reduces CVB3 load and apoptosis via STAT1/STAT2 and Bcl-2/BAX modulation.","evidence":"Immunoblot, TUNEL, and plaque assays in a CVB3 myocarditis model","pmids":["25528576"],"confidence":"Medium","gaps":["Direct effector ISGs in cardiomyocytes not identified"]},{"year":2015,"claim":"Identified EPSTI1 as an IL-28A-induced ISG required for HCV restriction, linking IL-28A to a PKR-dependent antiviral effector arm.","evidence":"siRNA knockdown, overexpression, HCV replication, and PKR promoter assays","pmids":["26146465"],"confidence":"Medium","gaps":["How EPSTI1 activates the PKR promoter mechanistically not resolved"]},{"year":2016,"claim":"Established that intratumoral IL-28A drives anti-tumor immunity through NK and CD8+ T cells in an IFN-γ-dependent, IL-12-synergistic manner.","evidence":"Immune cell depletion, IFN-γ-deficient mice, CTL and tumor growth assays","pmids":["27561689"],"confidence":"Medium","gaps":["Direct vs indirect action on tumor cells not separated"]},{"year":2019,"claim":"Proved IFN-λ2/λ3 are the exclusive IFNLR ligands for mucosal antiviral defense by genocopying receptor knockout with double-ligand knockout.","evidence":"Ifnl2/3 double-KO vs Ifnlr1-KO mice across norovirus, reovirus, and influenza models","pmids":["31462571"],"confidence":"High","gaps":["Non-redundant individual roles of IFN-λ2 vs IFN-λ3 not dissected"]},{"year":2020,"claim":"Defined transcriptional control of IFNL2 by the ATG10S isoform, which enters the nucleus and competes with IRF1 at the promoter to drive expression.","evidence":"Promoter deletion, DNA-IP, luciferase, mutagenesis, and co-IP with KPNA1/KPNB1/IRF1","pmids":["31996071"],"confidence":"Medium","gaps":["Physiological signals controlling ATG10S generation not defined","Generality across cell types untested"]},{"year":2020,"claim":"Revealed an unexpected intracellular antiviral mechanism in which an IL-28A homotetramer binds HCV NS5A and mediates its autolysosomal degradation.","evidence":"Co-IP, deletion mapping, structural prediction, and autophagy marker assays in HepG2 cells","pmids":["32205851"],"confidence":"Medium","gaps":["Structural basis of tetramerization rests partly on computational prediction","Relationship to canonical secreted signaling unclear"]},{"year":2024,"claim":"Established a pro-angiogenic function of IL-28A through IL-10Rβ-dependent eNOS/AKT/ERK signaling with HSP70-1 as an essential effector.","evidence":"siRNA, HUVEC functional assays, KO/transgenic mice, and hind-limb ischemia model","pmids":["39127098"],"confidence":"High","gaps":["How HSP70-1 couples to eNOS/AKT mechanistically not resolved"]},{"year":2024,"claim":"Placed IL-28A downstream of IRF7 in maintaining intestinal epithelial barrier integrity, with recombinant IL-28A rescuing barrier defects and colitis susceptibility.","evidence":"Irf7-KO mice, DSS colitis, and recombinant IL-28A rescue with barrier marker readouts","pmids":["39370517"],"confidence":"Medium","gaps":["Direct receptor signaling restoring barrier molecules not mapped"]},{"year":null,"claim":"The structural basis of IL-28A receptor binding and the distinct, non-redundant roles of IFN-λ2 versus IFN-λ3 in vivo remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No experimental structure of the IL-28A/receptor complex in the corpus","Tissue-specific division of labor among type III IFN ligands undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[0,6,15]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,12]}],"localization":[{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[0,15]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[3,5,11]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,15]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[2,12]}],"complexes":[],"partners":["IL28RA","IL10RB","SOCS1","NS5A","IRF1","ATG10"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8IZJ0","full_name":"Interferon lambda-2","aliases":["Cytokine Zcyto20","Interleukin-28A","IL-28A"],"length_aa":200,"mass_kda":22.3,"function":"Cytokine with antiviral, antitumour and immunomodulatory activities. Plays a critical role in the antiviral host defense, predominantly in the epithelial tissues. Acts as a ligand for the heterodimeric class II cytokine receptor composed of IL10RB and IFNLR1, and receptor engagement leads to the activation of the JAK/STAT signaling pathway resulting in the expression of IFN-stimulated genes (ISG), which mediate the antiviral state. Has a restricted receptor distribution and therefore restricted targets: is primarily active in epithelial cells and this cell type-selective action is because of the epithelial cell-specific expression of its receptor IFNLR1. Seems not to be essential for early virus-activated host defense in vaginal infection, but plays an important role in Toll-like receptor (TLR)-induced antiviral defense. Plays a significant role in the antiviral immune defense in the intestinal epithelium. Exerts an immunomodulatory effect by up-regulating MHC class I antigen expression","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/Q8IZJ0/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/IFNL2","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1047,"dependency_fraction":0.0028653295128939827},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/IFNL2","total_profiled":1310},"omim":[{"mim_id":"609650","title":"NLR FAMILY, PYRIN DOMAIN-CONTAINING 6; NLRP6","url":"https://www.omim.org/entry/609650"},{"mim_id":"607404","title":"INTERFERON-LAMBDA RECEPTOR 1; IFNLR1","url":"https://www.omim.org/entry/607404"},{"mim_id":"607401","title":"INTERFERON, LAMBDA-2; IFNL2","url":"https://www.omim.org/entry/607401"},{"mim_id":"603403","title":"DEAH-BOX HELICASE 15; DHX15","url":"https://www.omim.org/entry/603403"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Not detected","tissue_distribution":"Not detected","driving_tissues":[],"url":"https://www.proteinatlas.org/search/IFNL2"},"hgnc":{"alias_symbol":["IL-28A"],"prev_symbol":["IL28A"]},"alphafold":{"accession":"Q8IZJ0","domains":[{"cath_id":"1.20.1250.60","chopping":"57-70_93-143_154-199","consensus_level":"high","plddt":94.5349,"start":57,"end":199}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IZJ0","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IZJ0-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IZJ0-F1-predicted_aligned_error_v6.png","plddt_mean":83.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=IFNL2","jax_strain_url":"https://www.jax.org/strain/search?query=IFNL2"},"sequence":{"accession":"Q8IZJ0","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8IZJ0.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8IZJ0/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IZJ0"}},"corpus_meta":[{"pmid":"16051921","id":"PMC_16051921","title":"IL-28A and IL-29 mediate antiproliferative and antiviral signals in intestinal epithelial cells and murine CMV infection increases colonic IL-28A expression.","date":"2005","source":"American journal of physiology. Gastrointestinal and liver physiology","url":"https://pubmed.ncbi.nlm.nih.gov/16051921","citation_count":186,"is_preprint":false},{"pmid":"21538995","id":"PMC_21538995","title":"IL-28A (IFN-λ2) modulates lung DC function to promote Th1 immune skewing and suppress allergic airway disease.","date":"2011","source":"EMBO molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/21538995","citation_count":182,"is_preprint":false},{"pmid":"20655797","id":"PMC_20655797","title":"IL-28A, IL-28B, and IL-29: promising cytokines with type I interferon-like properties.","date":"2010","source":"Cytokine & growth factor reviews","url":"https://pubmed.ncbi.nlm.nih.gov/20655797","citation_count":138,"is_preprint":false},{"pmid":"15850793","id":"PMC_15850793","title":"SOCS-1 inhibits expression of the antiviral proteins 2',5'-OAS and MxA induced by the novel interferon-lambdas IL-28A and IL-29.","date":"2005","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/15850793","citation_count":76,"is_preprint":false},{"pmid":"21170333","id":"PMC_21170333","title":"Comparative analysis of the lambda-interferons IL-28A and IL-29 regarding their transcriptome and their antiviral properties against hepatitis C virus.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21170333","citation_count":71,"is_preprint":false},{"pmid":"16146571","id":"PMC_16146571","title":"Novel type I interferon IL-28A suppresses hepatitis C viral RNA replication.","date":"2005","source":"Virology journal","url":"https://pubmed.ncbi.nlm.nih.gov/16146571","citation_count":61,"is_preprint":false},{"pmid":"22962576","id":"PMC_22962576","title":"Identification of pro-inflammatory cytokines associated with muscle invasive bladder cancer; the roles of IL-5, IL-20, and IL-28A.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22962576","citation_count":53,"is_preprint":false},{"pmid":"31462571","id":"PMC_31462571","title":"Disruption of Type III Interferon (IFN) Genes Ifnl2 and Ifnl3 Recapitulates Loss of the Type III IFN Receptor in the Mucosal Antiviral Response.","date":"2019","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/31462571","citation_count":41,"is_preprint":false},{"pmid":"23021208","id":"PMC_23021208","title":"Potential anti-tumor effect of IFN-λ2 (IL-28A) against human lung cancer cells.","date":"2012","source":"Lung cancer (Amsterdam, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/23021208","citation_count":38,"is_preprint":false},{"pmid":"17241885","id":"PMC_17241885","title":"IL-28A is a key regulator of T-cell-mediated liver injury via the T-box transcription factor T-bet.","date":"2006","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/17241885","citation_count":35,"is_preprint":false},{"pmid":"26146465","id":"PMC_26146465","title":"EPSTI1 Is Involved in IL-28A-Mediated Inhibition of HCV Infection.","date":"2015","source":"Mediators of inflammation","url":"https://pubmed.ncbi.nlm.nih.gov/26146465","citation_count":29,"is_preprint":false},{"pmid":"25255777","id":"PMC_25255777","title":"Mesenchymal stem cells are efficiently transduced with adenoviruses bearing type 35-derived fibers and the transduced cells with the IL-28A gene produces cytotoxicity to lung carcinoma cells co-cultured.","date":"2014","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/25255777","citation_count":18,"is_preprint":false},{"pmid":"21499846","id":"PMC_21499846","title":"IL-29/IL-28A suppress HSV-1 infection of human NT2-N neurons.","date":"2011","source":"Journal of neurovirology","url":"https://pubmed.ncbi.nlm.nih.gov/21499846","citation_count":14,"is_preprint":false},{"pmid":"39127098","id":"PMC_39127098","title":"IL-28A/IL-10Rβ axis promotes angiogenesis via eNOS/AKT signaling and AP-1/NF-κB/MMP-2 network by regulating HSP70-1 expression.","date":"2024","source":"Journal of advanced research","url":"https://pubmed.ncbi.nlm.nih.gov/39127098","citation_count":11,"is_preprint":false},{"pmid":"35361913","id":"PMC_35361913","title":"Serum IL-28A/IFN-λ2 is linked to disease severity of COVID-19.","date":"2022","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/35361913","citation_count":11,"is_preprint":false},{"pmid":"27561689","id":"PMC_27561689","title":"Anti-tumor immunity elicited by direct intratumoral administration of a recombinant adenovirus expressing either IL-28A/IFN-λ2 or IL-29/IFN-λ1.","date":"2016","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/27561689","citation_count":11,"is_preprint":false},{"pmid":"20148705","id":"PMC_20148705","title":"The effect of IL-28A on human cord blood CD4+ T cells.","date":"2010","source":"Immunopharmacology and immunotoxicology","url":"https://pubmed.ncbi.nlm.nih.gov/20148705","citation_count":6,"is_preprint":false},{"pmid":"39370517","id":"PMC_39370517","title":"Interferon regulatory factor 7 alleviates the experimental colitis through enhancing IL-28A-mediated intestinal epithelial integrity.","date":"2024","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/39370517","citation_count":5,"is_preprint":false},{"pmid":"22253847","id":"PMC_22253847","title":"Sequence analysis of the IL28A/IL28B inverted gene duplication that contains polymorphisms associated with treatment response in hepatitis C patients.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22253847","citation_count":5,"is_preprint":false},{"pmid":"31996071","id":"PMC_31996071","title":"A new transcription factor ATG10S activates IFNL2 transcription by binding at an IRF1 site in HepG2 cells.","date":"2020","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/31996071","citation_count":4,"is_preprint":false},{"pmid":"25528576","id":"PMC_25528576","title":"Antiviral and myocyte protective effects of IL-28A in coxsackievirus B3-induced myocarditis.","date":"2014","source":"The Brazilian journal of infectious diseases : an official publication of the Brazilian Society of Infectious Diseases","url":"https://pubmed.ncbi.nlm.nih.gov/25528576","citation_count":3,"is_preprint":false},{"pmid":"32205851","id":"PMC_32205851","title":"IL28A protein homotetramer structure is required for autolysosomal degradation of HCV-NS5A in vitro.","date":"2020","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/32205851","citation_count":1,"is_preprint":false},{"pmid":"38953458","id":"PMC_38953458","title":"Reduced IFNL1 and/or IFNL2, but not IFNL3 is associated with worse outcome in patients with COVID-19.","date":"2024","source":"Clinical and experimental immunology","url":"https://pubmed.ncbi.nlm.nih.gov/38953458","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14596,"output_tokens":5009,"usd":0.059462,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":13153,"output_tokens":4626,"usd":0.090707,"stage2_stop_reason":"end_turn"},"total_usd":0.150169,"stage1_batch_id":"msgbatch_01DUPACZi7AAWwuZiUjseQF4","stage2_batch_id":"msgbatch_018yt7chE3f48PEFtDwtj6AE","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2005,\n      \"finding\": \"IL-28A (IFNL2) binds to the receptor complex IL-28R1/IL-10R2 on intestinal epithelial cells, activating ERK-1/2, SAPK/JNK MAPKs, and Akt signaling, inducing STAT1 phosphorylation, upregulating SOCS3, MxA, and 2',5'-OAS mRNA, and reducing CMV-infected cells by up to 83%.\",\n      \"method\": \"Western blot (STAT1 phosphorylation, MAPK activation), qPCR (antiviral gene expression), flow cytometry (CMV infection), cell proliferation assay\",\n      \"journal\": \"American journal of physiology. Gastrointestinal and liver physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (Western blot, qPCR, cell-based antiviral assay) in single lab\",\n      \"pmids\": [\"16051921\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"SOCS-1 overexpression completely abrogates IL-28A-induced STAT1 and STAT3 phosphorylation and abolishes IL-28A-induced mRNA expression of antiviral proteins 2',5'-OAS and MxA in hepatic cell lines, identifying SOCS-1 as a negative regulator of IL-28A signaling.\",\n      \"method\": \"Transient transfection (SOCS-1 overexpression), Western blot (STAT1/3 phosphorylation), qPCR (2',5'-OAS and MxA mRNA)\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (overexpression + Western blot + qPCR) in single lab\",\n      \"pmids\": [\"15850793\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"IL-28A activates the JAK-STAT signaling pathway in human hepatoma cells, induces interferon-stimulated genes (ISGs) including 6-16 and 1-8U, induces HLA class I antigen expression, suppresses HCV subgenomic RNA replication in a dose-dependent manner, and synergistically enhances IFN-alpha antiviral efficacy. IL-28A also specifically suppresses HCV IRES-mediated translation.\",\n      \"method\": \"HCV RNA replicon assay, Western blot (JAK-STAT activation), qPCR (ISG expression), IRES reporter assay, combination treatment with IFN-alpha\",\n      \"journal\": \"Virology journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal assays (replicon, Western blot, reporter assay) in single lab\",\n      \"pmids\": [\"16146571\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"IL-28A induces Th1 cytokine production by CD4+ T lymphocytes in a T-bet-dependent manner. In IL-28A-transgenic mice, augmented Con A-induced hepatitis with elevated IFN-gamma production was suppressed by crossing with T-bet-deficient mice, placing IL-28A upstream of the IFN-gamma/T-bet signaling pathway in T-cell-mediated liver injury.\",\n      \"method\": \"Transgenic mouse generation, genetic epistasis (IL-28A-transgenic × T-bet-deficient double mutant), in vivo Con A hepatitis model, cytokine measurements, antisense oligonucleotide knockdown\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with double-mutant rescue replicated across multiple orthogonal approaches (transgenic, knockout crossing, antisense knockdown, antibody neutralization) in single rigorous study\",\n      \"pmids\": [\"17241885\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"IL-28A activates STAT1 signaling and induces a similar set of antiviral genes as IL-29 in hepatic cells; however, unlike IL-29, IL-28A is a potent gene repressor, uniquely down-regulating 272 genes as determined by microarray analysis.\",\n      \"method\": \"Microarray transcriptome analysis, Western blot (STAT1 phosphorylation), qPCR, HCV replicon assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — microarray + Western blot + HCV replicon in single lab, multiple orthogonal methods\",\n      \"pmids\": [\"21170333\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"IL-28A modulates lung CD11c+ dendritic cell function by down-regulating OX40L and up-regulating IL-12p70, thereby promoting Th1 differentiation and suppressing Th2/Th17 responses; IL-28A-mediated protection from allergic airway disease was absent after IL-12 neutralization or in IFN-gamma-deficient mice, and could be adoptively transferred by IL-28A-treated CD11c+ cells.\",\n      \"method\": \"Recombinant cytokine treatment, IL-28Rα knockout mice, adoptive transfer of CD11c+ cells, antibody neutralization (IFN-gamma, IL-12), flow cytometry, cytokine measurements\",\n      \"journal\": \"EMBO molecular medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal approaches (KO mice, adoptive transfer, neutralization, recombinant cytokine) in single rigorous study demonstrating mechanistic pathway\",\n      \"pmids\": [\"21538995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"IL-28A inhibits HSV-1 replication in human NT2-N neurons and CHP212 neuronal cells via a mechanism partially blocked by anti-IL-10Rβ antibody; IL-28A selectively induces TLR expression and activates TLR-mediated antiviral pathways including IRF7, IFN-alpha, and IFN-alpha-stimulated antiviral genes.\",\n      \"method\": \"Neutralizing antibody (anti-IL-10Rβ), viral replication assay (HSV-1 DNA and protein quantification), qPCR (TLR and antiviral gene expression)\",\n      \"journal\": \"Journal of neurovirology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — receptor blocking + gene expression + viral assay, multiple orthogonal methods in single lab\",\n      \"pmids\": [\"21499846\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"IL-28A stimulation of bladder cancer cells activates MAPK and JAK-STAT signaling, induces NF-κB and AP-1 transcription factors, upregulates MMP-2 and MMP-9 expression, and enhances cell migration and invasion.\",\n      \"method\": \"Immunoblot, EMSA (NF-κB/AP-1 activity), real-time PCR, wound-healing/invasion assays, confocal immunofluorescence\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (Western blot, EMSA, functional migration/invasion assays) in single lab\",\n      \"pmids\": [\"22962576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"IL-28A (IFN-λ2) treatment of lung cancer cells with EGFR mutations induces STAT1 phosphorylation, growth suppression, and apoptotic cell death; anti-tumor effect was confirmed in vivo in a cancer cell transplant animal model, and neutralizing antibodies to IFN-λ inhibited IL-28A-induced caspase-3/7 activity.\",\n      \"method\": \"In vitro cell viability/proliferation assay, Western blot (STAT1 phosphorylation), caspase-3/7 activity assay, neutralizing antibody, in vivo luminescent imaging in animal model\",\n      \"journal\": \"Lung cancer (Amsterdam, Netherlands)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (in vitro + in vivo, neutralization, biochemical assays) in single lab\",\n      \"pmids\": [\"23021208\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In CVB3-induced myocarditis, IL-28A treatment increases STAT1 and STAT2 expression, reduces viral load, and inhibits CVB3-induced apoptosis in myocardial cells with an increased Bcl-2/BAX ratio.\",\n      \"method\": \"Immunoblot (STAT1/STAT2), TUNEL assay (apoptosis), plaque assay (viral load), immunohistochemistry (Bcl-2/BAX/Caspase-3)\",\n      \"journal\": \"The Brazilian journal of infectious diseases\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — multiple methods but single lab, in vivo mouse model with mechanistic readouts\",\n      \"pmids\": [\"25528576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"EPSTI1 (epithelial-stromal interaction 1) is an IL-28A-induced ISG that is required for IL-28A-mediated inhibition of HCV replication; forced EPSTI1 expression inhibits HCV replication independently of interferon, while EPSTI1 knockdown enhances viral replication. EPSTI1 activates PKR promoter and induces PKR-dependent antiviral genes (IFN-β, IFIT1, OAS1, RNase L).\",\n      \"method\": \"siRNA knockdown, overexpression, HCV replication assay, promoter assay (PKR), qPCR\",\n      \"journal\": \"Mediators of inflammation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function and gain-of-function with mechanistic pathway placement in single lab\",\n      \"pmids\": [\"26146465\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Intratumoral IL-28A drives anti-tumor immunity through NK cells and CD8+ T cells; IL-28A-mediated tumor suppression is completely impaired in IFN-gamma-deficient mice, and IL-28A increases CD8+ T cell infiltration and specific CTL reactivity. IL-12 acts synergistically with IL-28A.\",\n      \"method\": \"In vivo cell depletion (NK, CD8+ T cells), IFN-gamma-deficient mice, tumor growth assay, CTL assay, immunohistochemistry\",\n      \"journal\": \"Cancer gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic and depletion approaches with defined immune cell effectors, single lab\",\n      \"pmids\": [\"27561689\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Mice lacking both Ifnl2 and Ifnl3 (Ifnl2/3 double knockout) phenotypically recapitulate Ifnlr1-knockout mice in their inability to control murine norovirus, reovirus, and influenza virus at mucosal sites (intestine and lung), establishing that IFN-λ2 and IFN-λ3 are the exclusive ligands for IFNLR-mediated antiviral signaling at mucosal surfaces.\",\n      \"method\": \"Genetic knockout (Ifnl2/3 double KO mice), epistasis comparison with Ifnlr1-/- mice, viral infection models (norovirus, reovirus, influenza)\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with double-KO genocopying receptor KO across three independent viral infection models\",\n      \"pmids\": [\"31462571\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ATG10S (a shorter isoform of ATG10) translocates to the nucleus and binds the IFNL2 promoter at an IRF1 binding site (core motif CAAGAC), competing with IRF1 to promote IFNL2 transcription; ATG10S interacts with KPNA1/importin-α, KPNB1/importin-β, and IRF1 (shown by co-IP); knockdown of IRF1 increases ATG10S-driven IFNL2 transcription.\",\n      \"method\": \"Promoter deletion analysis, DNA immunoprecipitation, luciferase reporter assay, site-directed mutagenesis (nucleotide substitutions in core motif), co-immunoprecipitation, subcellular localization, RT-PCR, Western blot, siRNA knockdown\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — multiple orthogonal methods (ChIP, mutagenesis, co-IP, reporter assay) in single lab\",\n      \"pmids\": [\"31996071\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"IL-28A forms a homotetramer in cells; the first α-helix of IL-28A is required for homotetramer formation. The IL-28A homotetramer (but not monomer) binds HCV NS5A at the NS5A-ISDR region, promotes autolysosome formation, and mediates autolysosomal degradation of HCV NS5A and other HCV proteins. HCV NS5A activates incomplete autophagy by promoting ATG3/5/7/10 and LC3B but blocking autophagy flux.\",\n      \"method\": \"Co-immunoprecipitation, cell immunofluorescence, sequential deletion mutants, software structural prediction, Western blot (autophagy markers), HepG2 cell-based assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with deletion mutants and functional readout (autolysosomal degradation) in single lab; structural prediction is computational but supported by experimental deletion mapping\",\n      \"pmids\": [\"32205851\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"IL-28A promotes angiogenesis in endothelial cells via the IL-10Rβ receptor, activating eNOS/AKT and ERK1/2 signaling, inducing AP-1/NF-κB/MMP-2 network. HSP70-1 is an essential downstream effector; HSP70-1 knockdown abolishes IL-28A-induced angiogenic responses and eNOS/AKT signaling. IL-10Rβ gene ablation blocks IL-28A-induced angiogenic responses. IL-28A accelerated blood flow recovery in a hind-limb ischemia mouse model.\",\n      \"method\": \"siRNA knockdown (HSP70-1, IL-10Rβ), HUVEC proliferation/migration/invasion/tube formation assays, immunoblot, EMSA, aortic ring ex vivo assay, Matrigel plug in vivo assay, HSP70-1 KO and transgenic mice, hind-limb ischemia model, NGS transcriptomics\",\n      \"journal\": \"Journal of advanced research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal in vitro and in vivo methods including KO and transgenic mice, receptor ablation, and mechanistic pathway delineation in single rigorous study\",\n      \"pmids\": [\"39127098\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"IRF7 promotes intestinal barrier integrity through IL-28A; Irf7-/- mice show reduced IL-28A expression and compromised barrier markers (Muc2, E-cadherin, β-catenin, Occludin). Recombinant IL-28A treatment restores these barrier molecules and rescues Irf7-/- mice from enhanced DSS-induced colitis susceptibility, placing IL-28A downstream of IRF7 in intestinal epithelial barrier maintenance.\",\n      \"method\": \"Irf7 knockout mice, DSS colitis model, recombinant IL-28A rescue experiment, qPCR/Western blot (barrier molecule expression), intestinal epithelial cell stimulation with recombinant IL-28A\",\n      \"journal\": \"Journal of translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis (KO + rescue) with defined molecular readouts, single lab\",\n      \"pmids\": [\"39370517\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"IL-28A does not induce CD25 or Foxp3 expression on cord blood CD4+ T cells, does not promote their proliferation, and does not induce regulatory T cell function in this cell type.\",\n      \"method\": \"Flow cytometry (CD25, Foxp3), Treg suppression assay, cell proliferation assay\",\n      \"journal\": \"Immunopharmacology and immunotoxicology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, negative finding on a specific cell type; limited mechanistic scope\",\n      \"pmids\": [\"20148705\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"IFNL2 (IL-28A) is a secreted type III interferon that signals through the IL-28R1/IL-10R2 receptor complex to activate JAK-STAT (STAT1/STAT2/STAT3), MAPK (ERK1/2, JNK), and AKT pathways, inducing antiviral ISGs (MxA, OAS, EPSTI1) and inhibiting viral replication (HCV, CMV, HSV-1, CVB3, influenza, norovirus, reovirus) at mucosal surfaces; its signaling is negatively regulated by SOCS-1; transcription of IFNL2 is promoted by ATG10S competing with IRF7/IRF1 at the promoter; in immune cells it modulates dendritic cell function (downregulating OX40L, upregulating IL-12) to drive Th1 responses via an IFN-gamma/T-bet axis; in endothelial cells it promotes angiogenesis via IL-10Rβ/eNOS/AKT/HSP70-1; and genetic ablation of Ifnl2/Ifnl3 fully phenocopies loss of the IFNLR receptor in mucosal antiviral defense.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"IFNL2 (IL-28A) is a secreted type III interferon that engages the IL-28R1/IL-10R2 (IL-10Rβ) receptor complex on epithelial and other cells to mount antiviral defense and shape immune responses at mucosal surfaces [#0]. Receptor engagement activates JAK-STAT (STAT1/STAT2/STAT3) together with ERK1/2, SAPK/JNK, and Akt signaling, driving expression of interferon-stimulated antiviral genes such as MxA and 2',5'-OAS and suppressing replication of diverse viruses including CMV, HCV, HSV-1, and CVB3 [#0, #2, #6, #9]; this signaling is restrained by SOCS-1, which abolishes IL-28A-induced STAT1/STAT3 phosphorylation and ISG induction [#1]. Genetic ablation of Ifnl2/Ifnl3 phenocopies loss of the IFNLR receptor in controlling norovirus, reovirus, and influenza at mucosal sites, establishing IFN-λ2/λ3 as the exclusive ligands for IFNLR-mediated mucosal antiviral signaling [#12]. Beyond direct antiviral action, IL-28A shapes adaptive immunity by modulating dendritic cells—downregulating OX40L and upregulating IL-12—to drive T-bet-dependent Th1 differentiation via an IFN-γ axis [#3, #5], and it drives NK- and CD8+ T-cell-mediated anti-tumor immunity in an IFN-γ-dependent manner [#11]. IL-28A also promotes angiogenesis in endothelial cells through IL-10Rβ-dependent eNOS/AKT and ERK1/2 signaling with HSP70-1 as an essential downstream effector [#15]. Its transcription is positively regulated by IRF7 in intestinal barrier maintenance [#16] and by the ATG10S isoform, which competes with IRF1 at the IFNL2 promoter [#13].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Established that IL-28A acts through the IL-28R1/IL-10R2 receptor to activate JAK-STAT and MAPK/Akt pathways and induce antiviral ISGs, defining its core signaling axis.\",\n      \"evidence\": \"Western blot, qPCR, and CMV antiviral assay in intestinal epithelial cells\",\n      \"pmids\": [\"16051921\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor stoichiometry and ligand-binding determinants not resolved\", \"Relative contribution of MAPK vs JAK-STAT to antiviral output unclear\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Identified SOCS-1 as a negative regulator that shuts down IL-28A-driven STAT phosphorylation and ISG induction, placing a brake on the pathway.\",\n      \"evidence\": \"SOCS-1 overexpression with Western blot and qPCR in hepatic cell lines\",\n      \"pmids\": [\"15850793\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Endogenous regulation by SOCS-1 not tested\", \"Mechanism of SOCS-1 recruitment to the receptor not defined\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Showed IL-28A suppresses HCV replication and IRES-mediated translation and synergizes with IFN-alpha, framing its therapeutic antiviral relevance.\",\n      \"evidence\": \"HCV replicon assay, IRES reporter, and combination treatment in hepatoma cells\",\n      \"pmids\": [\"16146571\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Effector ISGs mediating HCV suppression not identified at this stage\", \"Mechanism of IRES translation block unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Placed IL-28A upstream of the IFN-γ/T-bet axis in driving Th1 cytokine production, establishing an immunomodulatory role beyond direct antiviral signaling.\",\n      \"evidence\": \"IL-28A-transgenic × T-bet-deficient genetic epistasis in a Con A hepatitis model\",\n      \"pmids\": [\"17241885\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether T-cell effects are cell-intrinsic or via accessory cells not fully resolved\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Distinguished IL-28A from IL-29 by revealing a unique gene-repressive transcriptional signature despite shared ISG induction.\",\n      \"evidence\": \"Microarray transcriptome analysis with HCV replicon assay in hepatic cells\",\n      \"pmids\": [\"21170333\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of gene repression not defined\", \"Functional consequence of repressed genes untested\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined the dendritic-cell mechanism by which IL-28A skews Th1 over Th2/Th17 responses, linking it to IL-12 and IFN-γ in allergic airway protection.\",\n      \"evidence\": \"IL-28Rα KO mice, adoptive transfer of CD11c+ cells, and IL-12/IFN-γ neutralization\",\n      \"pmids\": [\"21538995\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct DC receptor signaling events downstream of IL-28A not mapped\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Showed IL-28A restricts HSV-1 in neuronal cells via IL-10Rβ and selectively engages TLR/IRF7-driven antiviral programs, extending antiviral action to the nervous system.\",\n      \"evidence\": \"Anti-IL-10Rβ neutralization and HSV-1 replication/qPCR assays in neuronal cells\",\n      \"pmids\": [\"21499846\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Only partial receptor blockade observed, suggesting additional receptor contributions\", \"Mechanism of selective TLR induction unknown\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Revealed context-dependent pro-tumorigenic signaling in bladder cancer where IL-28A enhances migration/invasion via NF-κB/AP-1 and MMP induction.\",\n      \"evidence\": \"Immunoblot, EMSA, and migration/invasion assays in bladder cancer cells\",\n      \"pmids\": [\"22962576\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reconciliation with anti-tumor effects in other tumor types unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Demonstrated direct growth-suppressive and pro-apoptotic activity of IL-28A on EGFR-mutant lung cancer cells, supporting a tumor-suppressive role in this context.\",\n      \"evidence\": \"STAT1/caspase-3/7 assays, neutralizing antibody, and in vivo xenograft imaging\",\n      \"pmids\": [\"23021208\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Determinants of EGFR-mutation selectivity not defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Extended antiviral and cytoprotective action to viral myocarditis, where IL-28A reduces CVB3 load and apoptosis via STAT1/STAT2 and Bcl-2/BAX modulation.\",\n      \"evidence\": \"Immunoblot, TUNEL, and plaque assays in a CVB3 myocarditis model\",\n      \"pmids\": [\"25528576\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct effector ISGs in cardiomyocytes not identified\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified EPSTI1 as an IL-28A-induced ISG required for HCV restriction, linking IL-28A to a PKR-dependent antiviral effector arm.\",\n      \"evidence\": \"siRNA knockdown, overexpression, HCV replication, and PKR promoter assays\",\n      \"pmids\": [\"26146465\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How EPSTI1 activates the PKR promoter mechanistically not resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Established that intratumoral IL-28A drives anti-tumor immunity through NK and CD8+ T cells in an IFN-γ-dependent, IL-12-synergistic manner.\",\n      \"evidence\": \"Immune cell depletion, IFN-γ-deficient mice, CTL and tumor growth assays\",\n      \"pmids\": [\"27561689\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect action on tumor cells not separated\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Proved IFN-λ2/λ3 are the exclusive IFNLR ligands for mucosal antiviral defense by genocopying receptor knockout with double-ligand knockout.\",\n      \"evidence\": \"Ifnl2/3 double-KO vs Ifnlr1-KO mice across norovirus, reovirus, and influenza models\",\n      \"pmids\": [\"31462571\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Non-redundant individual roles of IFN-λ2 vs IFN-λ3 not dissected\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined transcriptional control of IFNL2 by the ATG10S isoform, which enters the nucleus and competes with IRF1 at the promoter to drive expression.\",\n      \"evidence\": \"Promoter deletion, DNA-IP, luciferase, mutagenesis, and co-IP with KPNA1/KPNB1/IRF1\",\n      \"pmids\": [\"31996071\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological signals controlling ATG10S generation not defined\", \"Generality across cell types untested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Revealed an unexpected intracellular antiviral mechanism in which an IL-28A homotetramer binds HCV NS5A and mediates its autolysosomal degradation.\",\n      \"evidence\": \"Co-IP, deletion mapping, structural prediction, and autophagy marker assays in HepG2 cells\",\n      \"pmids\": [\"32205851\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of tetramerization rests partly on computational prediction\", \"Relationship to canonical secreted signaling unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Established a pro-angiogenic function of IL-28A through IL-10Rβ-dependent eNOS/AKT/ERK signaling with HSP70-1 as an essential effector.\",\n      \"evidence\": \"siRNA, HUVEC functional assays, KO/transgenic mice, and hind-limb ischemia model\",\n      \"pmids\": [\"39127098\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How HSP70-1 couples to eNOS/AKT mechanistically not resolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placed IL-28A downstream of IRF7 in maintaining intestinal epithelial barrier integrity, with recombinant IL-28A rescuing barrier defects and colitis susceptibility.\",\n      \"evidence\": \"Irf7-KO mice, DSS colitis, and recombinant IL-28A rescue with barrier marker readouts\",\n      \"pmids\": [\"39370517\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct receptor signaling restoring barrier molecules not mapped\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis of IL-28A receptor binding and the distinct, non-redundant roles of IFN-λ2 versus IFN-λ3 in vivo remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No experimental structure of the IL-28A/receptor complex in the corpus\", \"Tissue-specific division of labor among type III IFN ligands undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [0, 6, 15]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 12]},\n      {\"term_id\": \"GO:0005102\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0, 15]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [3, 5, 11]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 15]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [2, 12]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"IL28RA\", \"IL10RB\", \"SOCS1\", \"NS5A\", \"IRF1\", \"ATG10\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}