Accepted Manuscript KIR2DL3+NKG2A- Natural Killer Cells are Associated with Protection from Productive Hepatitis C Virus Infection in People who Inject Drugs Christine Thoens, Christoph Berger, Martin Trippler, Holger Siemann, Melanie Lutterbeck, Ruth Broering, Jörg Schlaak, Falko M. Heinemann, Andreas Heinold, Jacob Nattermann, Norbert Scherbaum, Galit Alter, Joerg Timm PII: DOI: Reference:

S0168-8278(14)00283-9 http://dx.doi.org/10.1016/j.jhep.2014.04.020 JHEPAT 5125

To appear in:

Journal of Hepatology

Received Date: Revised Date: Accepted Date:

1 October 2013 18 March 2014 11 April 2014

Please cite this article as: Thoens, C., Berger, C., Trippler, M., Siemann, H., Lutterbeck, M., Broering, R., Schlaak, J., Heinemann, F.M., Heinold, A., Nattermann, J., Scherbaum, N., Alter, G., Timm, J., KIR2DL3+NKG2A- Natural Killer Cells are Associated with Protection from Productive Hepatitis C Virus Infection in People who Inject Drugs, Journal of Hepatology (2014), doi: http://dx.doi.org/10.1016/j.jhep.2014.04.020

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KIR2DL3+NKG2A- Natural Killer Cells are Associated with Protection from Productive Hepatitis C Virus Infection in People who Inject Drugs

Christine Thoens1, Christoph Berger2, Martin Trippler3, Holger Siemann4, Melanie Lutterbeck3, Ruth Broering3, Jörg Schlaak3, Falko. M. Heinemann5, Andreas Heinold5, Jacob Nattermann6, Norbert Scherbaum4, Galit Alter2, Joerg Timm1

1

Institute of Virology, University of Duisburg-Essen, University Hospital, Essen, Germany;

2

The Ragon Institute of MGH, MIT and Harvard, Boston, USA

3

Department for Gastroenterology and Hepatology, University of Duisburg-Essen, University

Hospital, Essen, Germany 4

Addiction Research Group, Department of Psychiatry and Psychotherapy, Rhine State

Hospital, Hospital of the University of Duisburg-Essen, Essen, Germany 5

Institute for Transfusion Medicine, University of Duisburg-Essen, University Hospital,

Essen, Germany 6

Department of Internal Medicine, University of Bonn, Bonn, Germany

corresponding author:

Jörg Timm Institute of Virology University of Duisburg-Essen, University Hospital Virchowstr. 179 45147 Essen phone: +49 201 723 2306 fax: +49 201 723 5929 [email protected]

word count: 4863 number of figures: 4 number of tables: 1

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List of abbreviations: APC: Allophycocyanin, APC-Cy7: Allophycocyanin -Cyanine7, PBMC: Peripheral blood mononuclear cell, PE: Phycoerythrin, CD: Cluster of differentiation, FITC: Fluorescein isothiocyanate, HBV: hepatitis B virus, HCV: hepatitis C virus, HLA: Human Leukocyte Antigen, IDU: injection drug user, IFN: Interferon, KIR: receptor killer cell immunoglobulin-like receptor, OMT: opioid maintenance treatment, PHH: Primary human hepatocyte, RT-PCR: Real-time polymerase chain reaction, TLR: toll-like receptor

Conflict of interest: The authors have no conflict of interest.

Financial support: The study was supported by the Federal Ministry of Education and Research (01KI1008E “Host and viral determinants for susceptibility and resistance to hepatitis C virus infection”) and by funds for the National Reference Centre for Hepatitis C. The gene expression analysis in human liver biopsies was funded by BioNRW.PROFILE, Förderkennzeichen 005-10060045. PROFILE is co-funded by the European Union (European Regional Development Fund - Investing in your future) and the German federal state North Rhine-Westphalia (NRW).

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Abstract: Background & aims: Despite continuous high-risk behavior, a subgroup among people who inject drugs (PWID) remains seronegative for hepatitis C virus (HCV) suggesting that a state of “natural resistance” to HCV Infection may exist. Homozygosity for KIR2DL3 and its ligand HLA-C1 group alleles has been associated with control of HCV infection, however, the mechanism mediating this protective effect remained unclear. Methods: Peripheral NK cells from PWID (n=104) were phenotypically and functionally characterized by multicolor flow cytometry. Expression levels of the NK cell receptor ligands were analyzed in liver biopsies and primary human hepatocytes. Results: HCV seronegative PWID (n=34) had increased levels of KIR2DL3 +NKG2A- NK cells compared to healthy controls (n=10; p