POSTERS

Table (abstract P163): T cell responses to HBV peptides

HBV peptide Proliferation (SI) IFNg SFU/M PBL IL-10 SFU/M PBL

Vacca cHBVb p-value Vacca cHBVb p-value Vacca cHBVb p-value

HBV-S 4.4 1.3 <0.0001 68 12 <0.0001 168 12 0.0001HBV-PreS 1.2 1.3 0.024 15 8 0.024 20 7 0.0065HBV-PreCore 1.5 1.1 0.005 30 12 0.005 102 13 0.0004HBV-Core 1.3 1.1 0.91 43 18 0.007 102 18 0.0001HBV-RT1 1.4 1.4 0.2930 25 15 0.0281 92 18 0.0016HBV-RT2 2.2 1.4 0.0284 28 15 0.0007 287 17 0.0001

*Vacc: 17 vaccinees; cHBV: 168 chronic hepatitis B participants in HBRN.

P163

IMMUNE CORRELATES OF VACCINE-MEDIATED PROTECTIVE

IMMUNITY VERSUS VIRAL PERSISTENCE IN HEPATITIS B VIRUS

INFECTION

J.-J. Park1,2, D.K. Wong3, A.S. Wahed4, W.M. Lee5, J.J. Feld3,

N. Terrault6, M. Khalili7, K.V. Kowdley8, D.T. Lau9, R.K. Sterling10,

W.R. Kim11, C. Smith12,13, R. Carithers14, D.L. Levine1,2, J. Keith1,2,

M.E. Valiga1,2, A.S.F. Lok15, K.-M. Chang1,2. 1University of Pennsylvania,2Philadelphia VA Medical Center, Philadelphia, PA, United States;3University of Toronto, Toronto, ON, Canada; 4University of Pittsburgh,

Pittsburgh, PA, 5Division of Digestive and Liver Diseases, University

of Texas Southwestern Medical Center, Dallas, TX, 6Division of

Gastroenterology, 7University of California San Francisco, San

Francisco, CA, 8Virginia Mason Medical Center, Seattle, WA, 9Division

of Gastroenterology, Harvard University, Boston, MA, 10Virginia

Commonwealth University, Richmond, VA, 11Mayo Clinic Rochester,

Rochester, 12Mayo Clinic, 13University of Minnesota, Plymouth, MN,14University of Washington Medical Center, Seattle, WA, 15Division of

Gastroenterology, University of Michigan Health System, Ann Arbor,

MI, United States

E-mail: kmchang@mail.med.upenn.edu

Background and Aims: Hepatitis B virus (HBV) is cleared with

long-lasting antiviral T cell memory whereas HBV persists with

dysfunctional antiviral T cells. Recently, memory T cell responses

to HBV core and reverse transcriptase (RT) in vaccinated anti-HBc-

negative healthcare workers (Werner, Gastro 2013) suggested that

HBV vaccination is protective but not sterilizing. Here, we looked

for immune correlates of vaccine-mediated immunity relative to

immune dysregulation in chronic hepatitis B (cHBV).

Methods: Peripheral lymphocytes from 17 HBV vaccinees and 168

cHBV participants of the NIDDK-sponsored Hepatitis B Research

Network (HBRN) were analyzed for lymphoproliferation and

IFNg/IL10 production following stimulation with HBV peptide pools

(S, preS, preC, Core, RT) and immune phenotype by multi-color

FACS. Statistical analyses included Mann Whitney U and Fisher’s

Exact or Chi-square.

Results: HBV-vaccinees displayed greater proliferative, IFNg+

and IL10+ T cell responses to S (included in HBV vaccine)

and non-S antigens (not included in HBV vaccine) as shown

below. HBV-vaccinees displayed both CD4 and CD8 T cell

responses to HBV-RT suggestive of immune induction during

HBV infection in-vivo. Compared to vaccinees, cHBV participants

displayed increased %Foxp3+/CD4+ Tregs (p = 0.017), reduced

%CD56+CD16− NK-cells (p = 0.0001) and reduced %mDC (p =0.004)

with differential expression in activating, inhibitory and/or

costimulatory molecules.

Conclusions: HBV-specific T cells in HBV-vaccinees show greater

virus-specific proliferation and IFNg as well as IL-10 production

compared to cHBV participants who display differential immune

regulatory phenotype. T cell response to HBV non-S antigens in

HBV vaccinees also suggest protective but non-sterilizing vaccine-

mediated immunity.

P164

MONOCYTE EXHAUSTION: A MECHANISTIC EXPLANATION FOR

SUSCEPTIBILITY TO INFECTION IN LIVER FAILURE

C. Bernsmeier1, V. Patel1, R. Tidswell1, E. Triantafyllou1,

A. Singanayagam1, W. Khamri2, K. Agarwal1, M. Heneghan1,

C. Willars1, W. Bernal1, G. Auzinger1, Y. Ma1, M. Thursz2,

J. Wendon1, C.G. Antoniades1,2. 1Institute of Liver Studies, King’s

College Hospital, 2Section of Hepatology, St. Mary’s Hospital, Imperial

College London, London, United Kingdom

E-mail: christine.bernsmeier@kcl.ac.uk

Background and Aims: Monocyte dysfunction is postulated to

account for the marked predisposition to infection and high

mortality in acute (ALF) and acute-on-chronic (ACLF) liver failure.

Defective fractalkine (CX3CR1–CX3CL1) signaling is associated with

impaired monocyte responses to microbial challenge, migration and

survival. We sought to determine monocyte function in relation to

CX3CR1/CX3CL1 expression in patients with liver failure.

Methods: Using flow cytometry, immunophenotyping (CD32,

CD64, CD86, CD163, CD206, HLA-DR, CX3CR1, CCR2/5) and

functional responses (phagocytosis [E. coli/S. aureus]; oxidative

burst [E. coli], LPS-induced TNF-a/IL-6 secretion) of circulating

monocytes (CD14+CD16−; CD14++CD16+; CD14−CD16+) were assessed

in patients with ACLF (n =35), ALF (n =24) cirrhosis (n = 8) and

healthy-controls (n = 23). CX3CL1 serum levels were measured

using ELISA.

Results: Monocytes in ALF and ACLF patients display an anti-

inflammatory phenotype: CD163high/HLA-DRlow/CX3CR1low. CX3CR1

expression is significantly down-regulated (ALF, 1746 vs. 549.5

mean fluorescence intensity (MFI), p = 0.0001 / ACLF, 1746 vs. 243

MFI, p < 0.0001) affecting all monocyte subsets with concomitantly

elevated circulating CX3CL1 levels (p = 0.0004/p =0.0010). In

ALF/ACLF patients, CX3CR1low monocytes exhibited an array of

innate functional defects: (i) significant reductions in phagocytosis

of E. coli and S. aureus (p = 0.0009/ p =0.0083 and p=0.0257/

p =0.0373) and (ii) oxidative burst responses to phagocytosed

E. coli (p = 0.0018/0.0835), (iii) impaired dynamic pathogen uptake

of E. coli (AUC, p = 0.0129/ p =0.0091) and (iv) attenuated TNF-a and

IL-6 secretion in response to LPS (p =0.0041/ p =0.0001; p =0.0053/

p =0.0187) (see figure).

Conclusions: In liver failure, we demonstrate that circulating

monocytes exhibit functional “exhaustion” leading to impaired

pathogen clearance and responses to microbial challenge. Further

studies are required to evaluate the role of defective CX3CR1–

CX3CL1 interaction in monocyte homing to tissue sites of infection.

S120 Journal of Hepatology 2014 vol. 60 | S67–S214

POSTERS

0 30 600

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ALF ACLFE CX3CR1 CX3CR1

Figure (abstract P164): Monocytes from patients with acute (ALF) and acute-on-chronic liver failure (ACLF) exhibit diverse functional defects and lowCX3CR1 expression. (A) Phagocytosis capacity of E. coli and S. aureus; (B) dynamics of phagocytosis (E. coli); (C) oxidative burst; (D) LPS-induced cytokinesecretion (TNF-a/IL-6); (E) CX3CR1 expression.

P165

CD4+ T CELLS INDUCE LIVER DAMAGE AFTER ADENOVIRAL

INFECTION OF HEPATOCYTES

M. Wittlich1, V. Staudt2, T. Bopp2, E. Schmitt2, P.A. Knolle1,3,

D. Wohlleber1. 1Institutes for Molecular Medicine and Experimental

Immunology (IMMEI), University Hospital Bonn, Bonn, 2Institute for

Immunology, Johannes Gutenberg University Mainz, Mainz, 3Institute

of Molecular Immunology, Technical University of Munich, Munich,

Germany

E-mail: michaela.wittlich@uni-bonn.de

Background and Aims: CD8+ cytotoxic T cells (CTL) are capable of

releasing TNF and inducing apoptosis of infected hepatocytes after

cross-presentation of viral antigen by liver sinusoidal endothelial

cells (LSEC). This novel non-canonical CTL effector function

accounts for about 40% of the antiviral effector function. CD4+ T

cells in turn are reported to be necessary for an efficient licensing

of dendritic cells and priming of CD8+ T cells. But since especially

Th1 cells are a source of inflammatory cytokines such as TNF, they

could be able to induce liver damage in viral infection. To investigate

if CD4+ Th1 cells possess a non-canonical effector function, we used

a similar model.

Methods: Mice were infected with a recombinant hepatotropic

adenovirus, which primarily infects hepatocytes and expresses

ovalbumin, followed by adoptive transfer of in vitro differentiated

CD4+ T cells. Protein fragments of ovalbumin can be presented to

the OVA-specific H2-Ab1 (I-Ab)-restricted T helper cells.

Results: Our findings support the ability of Th1 differentiated cells

to induce liver damage, whereas Th2 differentiated cells, incapable

of producing TNF, could not induce a comparable ALT elevation.

This effect of Th1 cells is furthermore dependent on the amount of

transferred cells.

Conclusions: Th1 cells have a new non-canonical effector function

that is based on the secretion of TNF, since Th2 cells didn’t induce

comparable effects. Future experiments will elucidate if other T

helper cells with the potential to produce inflammatory cytokines

such as TNF, e.g. Th17 cells, are also capable of inducing liver injury

and possess an antiviral activity.

P166

A COACH, A PLAYMAKER, A DEFENDER AND STRIKERS:

AN EPIGENETIC-TARGET INTERPLAY IN HCV/HCC

M.M. Fouad1, N.M. Elemam1, H.B. Sherif1, S.A. El Sobky1,

R.A. Yacoub1, A.K. Abelhamid1, T. Elbaz2, M.A. Mohey El Din2,

I.O. Fawzy1, H.M. El Tayebi1, N. El-Ekiaby1, R.Y. Mekky1, G. Esmat2,

A.I. Abdelaziz1. 1The Molecular Pathology Research Group, German

University in Cairo, 2Department of Endemic Medicine and Hepatology,

Cairo University, Cairo, Egypt

E-mail: noha.mousaad@guc.edu.eg

Background and Aims: PU.1 is a key regulator modulating the

expression of numerous cell surface receptors in hematopoietic

cells. NKG2A is an inhibitory receptor, while NKG2D is an activating

Journal of Hepatology 2014 vol. 60 | S67–S214 S121