studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 851A
Disclosures:
Holger J. Møller - Grant/Research Support: Danish Council for Strategic
Research; Independent Contractor: IQ-Products, NL; Patent Held/Filed: Aarhus
University; Stock Shareholder: Affinicon Aps
Rajiv Jalan - Consulting: Ocera Therapeutics, Conatus; Grant/Research Support:
Grifols, Gambro; Patent Held/Filed: Yaqrit, Cyberliver
The following authors have nothing to disclose: Francesco Figorilli, Karen Louise
Thomsen, Jane MacNaughtan, I.Jane Cox, Alba Moratalla, Isidora Ranchal,
Rajeshwar Mookerjee, Nathan Davies
1301
Hepatitis B X-interacting protein (HBXIP) negatively
regulates IFN-β production and antiviral response by
promoting proteasomal degradation of TANK-binding
kinase 1
Hang Zhang; Basic medical, Hangzhou Normal University, Hangzhou,
China
TANK-binding kinase 1 (TBK1) plays an essential role in Tolllike
receptor (TLR)– and retinoic acid–inducible gene I (RIG-I)–
mediated induction of type I interferon (IFN; IFN-α/β) and host
antiviral responses. How TBK1 activity is negatively regulated
remains largely unknown. We report that Hepatitis B virus X-interacting
protein (HBXIP) promotes proteasomal degradation of
TBK1 and inhibits RIG-I-induced IFN-β signaling. HBXIP knockdown
resulted in augmented activation of IFN regulatory factor
3 (IRF3) and enhanced expression of IFN-β in RIG-I-activated
primary hepatocyte cells, whereas overexpression of HBXIP
had opposite effects. Consistently, HBXIP impaired vesicular
stomatitis virus (VSV) infection–induced IRF3 activation and
IFN-β production and promoted VSV replication. HBXIP negatively
regulated the cellular levels of TBK1 by directly binding
to and promoting K48-linked polyubiquitination of TBK1. Therefore,
we identified HBXIP as a negative regulator in RIG-I–triggered
antiviral responses and suggested HBXIP as a potential
target for the intervention of diseases with uncontrolled IFN-β
production.
Disclosures:
The following authors have nothing to disclose: Hang Zhang
1302
Toll-like receptor 9 and B-cell activating factor signals
induce impaired immunological memory in cirrhosis
Hiroyoshi Doi 1 , David E. Kaplan 3 , Eiichi Hayashi 1 , Jun Arai 1 , Risa
Omori 1 , Manabu Uchikoshi 1 , Kenichi Morikawa 4 , Junichi Eguchi 2 ,
Takayoshi Ito 2 , Hitoshi Yoshida 1 ; 1 Gastroenterology, Showa University,
Shinagawa, Japan; 2 Digestive Disease Center, Showa University
Koto Toyosu Hospital, Toyosu, Japan; 3 Gastroenterology,
University of Pennsylvania, Philadelphia, PA; 4 Gastroenterology
and Hepatology, Hokkaido University, Sapporo, Japan
[Background] Cirrhosis is an immunocompromised state and
their immune cells are thought to be impaired. We previously
reported peripheral CD27+ memory B-cells in cirrhotic patients
are decreased and dysfunctional compared to those in even
non-cirrhotic chronic liver disease patients. We also found Tolllike
receptor (TLR) 4 and TLR 9 signals play important roles for
B-cell activation and survival. (Doi et al. Hepatology 2012).
Clinically, we confirmed elevated immunoglobulin levels in cirrhosis
regardless of the etiology. The abnormal humoral immunity
could be caused by antigen non-specific way such as TLR
signals and results in polyclonal B-cell activation. The purpose
of this study is to investigate the impact of antigen non-specific
signals on immunological change and immunodeficiency
in cirrhosis. [Methods] We isolated peripheral blood mononuclear
cells (PBMC) and serum from whole blood. Then we
measured memory B-cell and memory T-cell defined by CCR7
and CD45RA expressions using PBMC by flow cytometry. As
we previously reported, B-cell stimulants such as TLR ligands
increased in cirrhotic plasma probably due to bacterial translocation
and we investigated TLR 4 and TLR 9 expressions on
B-cell. B-cell activating factors (BAFF and APRIL) in sera were
measured by ELISA. To analyze B-cell signaling, we negatively
isolated CD19+ B-cell from healthy donor PBMC and cultured
the B-cell for 72 hours with LPS (TLR4 agonist), CpG-ODN
(TLR9 agonist), anti-IgG/A/M (B-cell receptor agonist) and
combinations. Then we measured activation marker expressions
and apoptosis by flow cytometry. [Results] In accordance
with our previous study, cirrhotic patients (CIR) had decreased
peripheral memory B-cell compared to non-cirrhotic chronic
liver disease patients (CLD). (CLD vs CIR; p=0.021) They also
showed lower frequency of both CD4+ and CD8+ effector
memory T-cells. (p= 0.003, 0.027, respectively) TLR9 but not
TLR4 expression on memory B-cell was increased in cirrhotic
patients. (p=0.043) We also found serum BAFF but not APRIL
level was elevated as well. (p=0.003) Memory B-cell frequency
tended to be associated with the TLR 9 expression and BAFF
levels. (p=0.016, 0.025, respectively) These findings are more
apparent in advanced cirrhosis. In vitro, B-cell stimulants shown
above didn’t make much impact on CD27 expression but
CD27- B-cells, which relatively dominant in cirrhosis, tended
to be apoptotic by TLR 4 and TLR 9 stimulations. [Conclusion]
Cirrhotic patients especially in advanced stage have elevated
BAFF and TLR9 expression on memory B-cell. Those antigen
non-specific signals have an impact on dysfunctional immunological
memory observed in cirrhosis.
Disclosures:
David E. Kaplan - Grant/Research Support: Bayer Pharmaceuticals, Inovio Pharmaceuticals
The following authors have nothing to disclose: Hiroyoshi Doi, Eiichi Hayashi,
Jun Arai, Risa Omori, Manabu Uchikoshi, Kenichi Morikawa, Junichi Eguchi,
Takayoshi Ito, Hitoshi Yoshida
1303
Low Dose Zinc Sulfate (220mg) Supplementation for
Three Months Normalizes Zinc Levels, Endotoxeima,
Pro-Inflammatory/Fibrotic Biomarkers & Improves Clinical
Parameters in Alcoholic Cirrhosis– A Double-Blind
Placebo Controlled – (ZAC) Clinical Trial
Mohammad K. Mohammad 1,2 , Keith C. Falkner 1 , Ming Song 1 ,
Craig J. McClain 1,2 , Matthew C. Cave 1,2 ; 1 Hepatology, gastroenterology
and nutrition, university of louisville, Louisville, KY; 2 Hepatology,
Robely Rex Veteran Hospital, Louisville, KY
Purpose: Zinc deficiency occurs in human subjects with alcoholic
cirrhosis (AC), and zinc supplementation attenuates liver
injury/inflammation in murine models of alcoholic liver disease.
The aim of this NIH-funded clinical trial was to determine
if zinc sulfate therapy improves serologic biomarkers of liver
injury/inflammation & clinical status in AC Methods: 22 consented
subjects with Child-Pugh class A-B alcoholic cirrhosis
were randomized to placebo (n=11) or zinc sulfate 220 mg
daily (n=11) in the single center, IRB approved, double-blind,
placebo-controlled clinical trial. 10 non-drinking, healthy volunteers
were recruited as controls (HC). Serum cytokines (IL-1β,
IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IFN-γ, PAI-1, MCP-1 &
TNF-α) and whole blood ex-vivo unstimulated and lipopolysacharide-stimulated
(LPS) cytokine production were measured
by Luminex, while TGF-β & LPS were measured by ELISA. Differences
between means were evaluated by t-test, p < 0.05
was considered significant Results: Of the 22 AC subjects,
20 subjects completed 3 months treatment: placebo (n=10);