Toll-like receptor dual-acting agonists are potent inducers of PBMC-produced cytokines that inhibit hepatitis B virus production in primary human hepatocytes
Vaclav Janovec 1 2 , Jan Hodek 2 , Kamila Clarova 2 , Tomas Hofman 1 2 , Pavel Dostalik 1 , Jiri Fronek 3 4 , Jaroslav Chlupac 3 4 , Laurence Chaperot 5 , Sarah Durand 6 , Thomas F Baumert 6 7 , Iva Pichova 2 , Barbora Lubyova 2 , Ivan Hirsch 8 9 10 , Jan Weber 11
Affiliations
Affiliations
1
Department of Genetics and Microbiology, Faculty of Science, Charles University, BIOCEV, 25150, Vestec, Czech Republic.
2
IOCB & Gilead Research Center, Institute of Organic Chemistry and Biochemistry of the Czech Academy of Science, 16610, Prague, Czech Republic.
3
Transplantation Surgery Department, Institute for Clinical and Experimental Medicine, 14021, Prague, Czech Republic.
4
Department of Anatomy, Second Faculty of Medicine, Charles University, 15006, Prague, Czech Republic.
5
CNRS UMR5309, Inserm U1209, CHU Grenoble Alpes, IAB, EFS, Université Grenoble Alpes, 38000, Grenoble, France.
6
Inserm, Institut de Recherche Sur Les Maladies Virales Et Hepatiques UMRS 1110, Universite de Strasbourg, 67000, Strasbourg, France.
7
Pole Hepato-Digestif, Institut Hospitalo-Universitaire, Hopitaux Universitaires de Strasbourg, 67000, Strasbourg, France.
8
Department of Genetics and Microbiology, Faculty of Science, Charles University, BIOCEV, 25150, Vestec, Czech Republic. [email protected].
9
IOCB & Gilead Research Center, Institute of Organic Chemistry and Biochemistry of the Czech Academy of Science, 16610, Prague, Czech Republic. [email protected].
10
Institute of Molecular Genetics of the Czech Academy of Sciences, 14220, Prague, Czech Republic. [email protected].
11
IOCB & Gilead Research Center, Institute of Organic Chemistry and Biochemistry of the Czech Academy of Science, 16610, Prague, Czech Republic. [email protected].
PMID: 32728070 DOI: 10.1038/s41598-020-69614-7
Abstract
Recombinant interferon-α (IFN-α) treatment functionally cures chronic hepatitis B virus (HBV) infection in some individuals and suppresses virus replication in hepatocytes infected in vitro. We studied the antiviral effect of conditioned media (CM) from peripheral blood mononuclear cells (PBMCs) stimulated with agonists of Toll-like receptors (TLRs) 2, 7, 8 and 9. We found that CM from PBMCs stimulated with dual-acting TLR7/8 (R848) and TLR2/7 (CL413) agonists were more potent drivers of inhibition of HBe and HBs antigen secretion from HBV-infected primary human hepatocytes (PHH) than CM from PBMCs stimulated with single-acting TLR7 (CL264) or TLR9 (CpG-B) agonists. Inhibition of HBV in PHH did not correlate with the quantity of PBMC-produced IFN-α, but it was a complex function of multiple secreted cytokines. More importantly, we found that the CM that efficiently inhibited HBV production in freshly isolated PHH via various cytokine repertoires and mechanisms did not reduce covalently closed circular (ccc)DNA levels. We confirmed our data with a cell culture model based on HepG2-NTCP cells and the plasmacytoid dendritic cell line GEN2.2. Collectively, our data show the importance of dual-acting TLR agonists inducing broad cytokine repertoires. The development of poly-specific TLR agonists provides novel opportunities towards functional HBV cure.
Grant support
GA17-15422S/Grantová Agentura České Republiky
SVV 260426/Grantová Agentura, Univerzita Karlova
CZ.1.05/1.1.00/02.0109/European Regional Development Fund
LQ1604/Ministry of Education, Youth and Sports of the Czech Republic
ANR-10-LABX-0028_HEPSYS/Agence Nationale de la Recherche