Distinct transcriptional isoforms of the interferon lambda receptor 1 (IFNLR1) are expressed in hepatocytes, but whether corresponding full-length and truncated IFNLR1 protein variants have discrete function is unclear. We quantitated IFNLR1 isoforms in liver and blood from individuals with chronic hepatitis C virus (HCV) infection before and after antiviral treatment, hypothesizing their relative expression may differentially change during resolution of virus-induced inflammation. We also expressed FLAG-tagged IFNLR1 variants in stem cell-derived hepatocytes (iHeps) with abrogated endogenous IFNLR1 to evaluate their function. IFNLR1 isoforms decreased in liver and blood during treatment of HCV, but no distinct pattern of decline was observed for any individual isoform. Expression of full-length IFNLR1 enabled lambda interferon (IFNL)-induced expression of antiviral and proinflammatory genes and augmented inhibition of hepatitis B virus (HBV) replication relative to wild-type (WT) iHeps. A noncanonical IFNLR1 variant missing part of the JAK1 binding domain enabled IFNLs to induce antiviral genes but could not support induction of proinflammatory genes or augmented HBV inhibition beyond that observed in WT iHeps with intact endogenous IFNLR1. A secreted IFNLR1 variant had no identified function in iHeps lacking endogenous IFNLR1. Although relative expression of individual IFNLR1 isoforms did not distinctly change during HCV treatment, functional studies in iHeps suggest IFNLR1 variants could function to titrate antiviral versus proinflammatory responses in hepatocytes in the context of viral hepatitis.
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References
1.
AdasMA, AlveynE, CookE, et al.The infection risks of JAK inhibition. Expert Rev Clin Immunol, 2022; 18(3):253–261; doi: 10.1080/1744666X.2022.2014323
2.
AntosD, ParksOB, DurayAM, et al.Cell-intrinsic regulation of phagocyte function by interferon lambda during pulmonary viral, bacterial super-infection. PLoS Pathog, 2024; 20(8):e1012498; doi: 10.1371/journal.ppat.1012498
3.
BhushalS, WolfsmullerM, SelvakumarTA, et al.Cell polarization and epigenetic status shape the heterogeneous response to type III interferons in intestinal epithelial cells. Front Immunol, 2017; 8:671; doi: 10.3389/fimmu.2017.00671
4.
BlumerT, Coto-LlerenaM, DuongFHT, et al.SOCS1 is an inducible negative regulator of interferon lambda (IFN-lambda)-induced gene expression in vivo. J Biol Chem, 2017; 292(43):17928–17938; doi: 10.1074/jbc.M117.788877
5.
BroggiA, TanY, GranucciF, et al.IFN-lambda suppresses intestinal inflammation by non-translational regulation of neutrophil function. Nat Immunol, 2017; 18(10):1084–1093; doi: 10.1038/ni.3821
6.
CaiD, WangX, YanR, et al.Establishment of an inducible HBV stable cell line that expresses cccDNA-dependent epitope-tagged HBeAg for screening of cccDNA modulators. Antiviral Res, 2016; 132:26–37; doi: 10.1016/j.antiviral.2016.05.005
7.
ChanHLY, AhnSH, ChangTT, et al;LIRA-B Study Team. Peginterferon lambda for the treatment of HBeAg-positive chronic hepatitis B: A randomized phase 2b study (LIRA-B). J Hepatol, 2016; 64(5):1011–1019; doi: 10.1016/j.jhep.2015.12.018
8.
ChangMY, RheeYH, YiSH, et al.Doxycycline enhances survival and self-renewal of human pluripotent stem cells. Stem Cell Rep, 2014; 3(2):353–364; doi: 10.1016/j.stemcr.2014.06.013
9.
DavidsonS, McCabeTM, CrottaS, et al.IFNlambda is a potent anti-influenza therapeutic without the inflammatory side effects of IFNalpha treatment. EMBO Mol Med, 2016; 8(9):1099–1112; doi: 10.15252/emmm.201606413
10.
de WeerdNA, NguyenT. The interferons and their receptors–distribution and regulation. Immunol Cell Biol, 2012; 90(5):483–491; doi: 10.1038/icb.2012.9
11.
DowlingJW, ForeroA. Beyond good and evil: Molecular mechanisms of type I and III IFN functions. J Immunol, 2022; 208(2):247–256; doi: 10.4049/jimmunol.2100707
12.
DoyleSE, SchreckhiseH, Khuu-DuongK, et al.Interleukin-29 uses a type 1 interferon-like program to promote antiviral responses in human hepatocytes. Hepatology, 2006; 44(4):896–906; doi: 10.1002/hep.21312
13.
DumoutierL, LejeuneD, HorS, et al.Cloning of a new type II cytokine receptor activating signal transducer and activator of transcription (STAT)1, STAT2 and STAT3. Biochem J, 2003; 370(Pt 2):391–396; doi: 10.1042/BJ20021935
14.
DuongFH, TrincucciG, BoldanovaT, et al.IFN-lambda receptor 1 expression is induced in chronic hepatitis C and correlates with the IFN-lambda3 genotype and with nonresponsiveness to IFN-alpha therapies. J Exp Med, 2014; 211(5):857–868; doi: 10.1084/jem.20131557
15.
EvansJG, NovotnyLA, MeissnerEG. Influence of canonical and non-canonical IFNLR1 isoform expression on interferon lambda signaling. Viruses, 2023; 15(3); doi: 10.3390/v15030632
16.
FerraoR, WallweberHJ, HoH, et al.The structural basis for class II cytokine receptor recognition by JAK1. Structure, 2016; 24(6):897–905; doi: 10.1016/j.str.2016.03.023
17.
ForeroA, OzarkarS, LiH, et al.Differential activation of the transcription factor IRF1 underlies the distinct immune responses elicited by type I and type III interferons. Immunity, 2019; 51(3):451–464.e456; doi: 10.1016/j.immuni.2019.07.007
18.
FuchsS, Kaiser-LabuschP, BankJ, et al.Tyrosine kinase 2 is not limiting human antiviral type III interferon responses. Eur J Immunol, 2016; 46(11):2639–2649; doi: 10.1002/eji.201646519
19.
GalaniIE, TriantafylliaV, EleminiadouEE, et al.Interferon-lambda mediates non-redundant front-line antiviral protection against influenza virus infection without compromising host fitness. Immunity, 2017; 46(5):875–890.e876; doi: 10.1016/j.immuni.2017.04.025
20.
GlassmanCR, TsutsumiN, SaxtonRA, et al.Structure of a Janus kinase cytokine receptor complex reveals the basis for dimeric activation. Science, 2022; 376(6589):163–169; doi: 10.1126/science.abn8933
21.
HuangH, ZhouW, ZhuH, et al.Baicalin benefits the anti-HBV therapy via inhibiting HBV viral RNAs. Toxicol Appl Pharmacol, 2017; 323:36–43; doi: 10.1016/j.taap.2017.03.016
22.
JilgN, LinW, HongJ, et al.Kinetic differences in the induction of interferon stimulated genes by interferon-alpha and interleukin 28B are altered by infection with hepatitis C virus. Hepatology, 2014; 59(4):1250–1261; doi: 10.1002/hep.26653
23.
JohnsonD, CarbonettiN. Roles and effects of interferon lambda signaling in the context of bacterial infections. J Interferon Cytokine Res, 2023; 43(9):363–369; doi: 10.1089/jir.2023.0037
24.
KotenkoSV, GallagherG, BaurinVV, et al.IFN-lambdas mediate antiviral protection through a distinct class II cytokine receptor complex. Nat Immunol, 2003; 4(1):69–77; doi: 10.1038/ni875
25.
KreinsAY, CiancanelliMJ, OkadaS, et al.Human TYK2 deficiency: Mycobacterial and viral infections without hyper-IgE syndrome. J Exp Med, 2015; 212(10):1641–1662; doi: 10.1084/jem.20140280
26.
KumarA, JohnsonD, BukowskiA, et al.IFNlambda signaling in neutrophils enhances the pathogenesis of Bordetella pertussis infection. J Leukoc Biol, 2024; doi: 10.1093/jleuko/qiae202
27.
LauberC, VieyresG, Terczyńska-DylaE, et al.Transcriptome analysis reveals a classical interferon signature induced by IFNlambda4 in human primary cells. Genes Immun, 2015; 16(6):414–421; doi: 10.1038/gene.2015.23
28.
LevineRL, HubbardSR. Unlocking the secrets to Janus kinase activation. Science, 2022; 376(6589):139–140; doi: 10.1126/science.abo7788
29.
LinAE, MesevEV, ToettcherJE, et al.Engineered chimeric receptors for dissecting interferon signaling. J Virol, 2024; 98(10):e0168023; doi: 10.1128/jvi.01680-23
30.
LudwigTE, BergendahlV, LevensteinME, et al.Feeder-independent culture of human embryonic stem cells. Nat Methods, 2006a;3(8):637–646; doi: 10.1038/nmeth902
31.
LudwigTE, LevensteinME, JonesJM, et al.Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol, 2006b;24(2):185–187; doi: 10.1038/nbt1177
32.
MajorJ, CrottaS, LlorianM, et al.Type I and III interferons disrupt lung epithelial repair during recovery from viral infection. Science, 2020; 369(6504):712–717; doi: 10.1126/science.abc2061
33.
MeissnerEG, WuD, OsinusiA, et al.Endogenous intrahepatic IFNs and association with IFN-free HCV treatment outcome. J Clin Invest, 2014; 124(8):3352–3363; doi: 10.1172/JCI75938
34.
MesevEV, GuareEG, PlossA, et al.Synthetic heterodimers of type III interferon receptors require TYK2 for STAT activation. J Interferon Cytokine Res, 2023; 43(9):414–426; doi: 10.1089/jir.2023.0039
35.
Metz-ZumaranC, UckeleyZM, DoldanP, et al.The population context is a driver of the heterogeneous response of epithelial cells to interferons. Mol Syst Biol, 2024; 20(3):242–275; doi: 10.1038/s44320-024-00011-2
36.
NovotnyLA, EvansJG, GuoH, et al.Interferon lambda receptor-1 isoforms differentially influence gene expression and HBV replication in stem cell-derived hepatocytes. Antiviral Res, 2023; 221:105779; doi: 10.1016/j.antiviral.2023.105779
37.
NovotnyLA, EvansJG, SuL, et al.Review of lambda interferons in hepatitis B virus infection: Outcomes and therapeutic strategies. Viruses, 2021; 13(6); doi: 10.3390/v13061090
38.
NovotnyLA, MeissnerEG. Expression and function of interferon lambda receptor 1 variants. FEBS Lett, 2024. doi: 10.1002/1873-3468.15041
39.
OrrC, XuW, MasurH, et al.Peripheral blood correlates of virologic relapse after Sofosbuvir and Ribavirin treatment of Genotype-1 HCV infection. BMC Infect Dis, 2020; 20(1):929; doi: 10.1186/s12879-020-05657-5
40.
OsinusiA, MeissnerEG, LeeYJ, et al.Sofosbuvir and ribavirin for hepatitis C genotype 1 in patients with unfavorable treatment characteristics: A randomized clinical trial. JAMA, 2013; 310(8):804–811; doi: 10.1001/jama.2013.109309
41.
PeasleeC, Esteva-FontC, SuT, et al.Doxycycline significantly enhances induction of induced pluripotent stem cells to endoderm by enhancing survival through protein Kinase B phosphorylation. Hepatology, 2021; 74(4):2102–2117; doi: 10.1002/hep.31898
42.
PervolarakiK, Rastgou TalemiS, AlbrechtD, et al.Differential induction of interferon stimulated genes between type I and type III interferons is independent of interferon receptor abundance. PLoS Pathog, 2018; 14(11):e1007420; doi: 10.1371/journal.ppat.1007420
43.
PhillipsS, MistryS, RivaA, et al.Peg-interferon lambda treatment induces robust innate and adaptive immunity in chronic hepatitis B patients. Front Immunol, 2017; 8:621; doi: 10.3389/fimmu.2017.00621
44.
PiperakisA, GalaniIE, AndreakosE. Type III interferons in innate and adaptive immunity in the respiratory tract. Curr Opin Immunol, 2024; 87:102430; doi: 10.1016/j.coi.2024.102430
45.
RanFA, HsuPD, WrightJ, et al.Genome engineering using the CRISPR-Cas9 system. Nat Protoc, 2013; 8(11):2281–2308; doi: 10.1038/nprot.2013.143
46.
RusinolL, PuigL. Tyk2 targeting in immune-mediated inflammatory diseases. Int J Mol Sci, 2023; 24(4); doi: 10.3390/ijms24043391
47.
SanterDM, MintyGES, GolecDP, et al.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. PLoS Pathog, 2020; 16(4):e1008515; doi: 10.1371/journal.ppat.1008515
SheppardP, KindsvogelW, XuW, et al.IL-28, IL-29 and their class II cytokine receptor IL-28R. Nat Immunol, 2003; 4(1):63–68; doi: 10.1038/ni873
50.
Si-TayebK, NotoFK, NagaokaM, et al.Highly efficient generation of human hepatocyte-like cells from induced pluripotent stem cells. Hepatology, 2010; 51(1):297–305; doi: 10.1002/hep.23354
51.
SommereynsC, PaulS, StaeheliP, et al.IFN-lambda (IFN-lambda) is expressed in a tissue-dependent fashion and primarily acts on epithelial cells in vivo. PLoS Pathog, 2008; 4(3):e1000017; doi: 10.1371/journal.ppat.1000017
52.
StaniferML, BoulantS. Differential signaling by type-I and type-III interferons in mucosa. Curr Opin Immunol, 2024; 86:102400; doi: 10.1016/j.coi.2023.102400
53.
StaniferML, GuoC, DoldanP, et al.Importance of Type I and III interferons at respiratory and intestinal barrier surfaces. Front Immunol, 2020; 11:608645; doi: 10.3389/fimmu.2020.608645
54.
ThomasE, GonzalezVD, LiQ, et al.HCV infection induces a unique hepatic innate immune response associated with robust production of type III interferons. Gastroenterology, 2012; 142(4):978–988; doi: 10.1053/j.gastro.2011.12.055
55.
WitteK, GruetzG, VolkHD, et al.Despite IFN-lambda receptor expression, blood immune cells, but not keratinocytes or melanocytes, have an impaired response to type III interferons: Implications for therapeutic applications of these cytokines. Genes Immun, 2009; 10(8):702–714; doi: 10.1038/gene.2009.72
ZhangD, WlodawerA, LubkowskiJ. Crystal structure of a complex of the Intracellular Domain of Interferon lambda Receptor 1 (IFNLR1) and the FERM/SH2 domains of human JAK1. J Mol Biol, 2016; 428(23):4651–4668; doi: 10.1016/j.jmb.2016.10.005
58.
ZhouZ, HammingOJ, AnkN, et al.Type III interferon (IFN) induces a type I IFN-like response in a restricted subset of cells through signaling pathways involving both the Jak-STAT pathway and the mitogen-activated protein kinases. J Virol, 2007; 81(14):7749–7758; doi: 10.1128/JVI.02438-06
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