studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 707A
used for HIV infection. A coculture model of PHHs an PKCs was
also utilized. Specific RNA and DNA dependent innate and
inflammatory pathways, such as RIG-I/TLR3 and IFI16/cGAS
signaling, were studied as well as additional pathways involving
the production of interferon and chemokines such as IP10/
CXCL10, IL-1b and CCL5/Rantes that may drive subsequent
liver disease. RESULTS: We found that similar to distinct stimuli
including IFN-a and polyI:C, HCV and HIV stimulation of these
primary liver cells resulted in rapid and robust upregulation of
IP10 and other inflammatory cytokines at the mRNA and protein
level. Surprisingly, microarray analyses of HIV treated liver
cells demonstrated several innate immune pathways were activated
by this virus in the liver. siRNA experiments demonstrated
the induction of inflammatory cytokines was mediated by IRF3
and NFkB-dependent pathways. CONCLUSIONS: Overall,
our data demonstrate that the robust antiviral response that is
observed in HCV infected livers and subsequently drives liver
pathology is augmented with HIV/HCV coinfection. It is hoped
that understanding the mechanism by which coinfected patients
have more severe liver disease will enable the development of
targeted therapeutics to abrogate these poor clinical outcomes.
Disclosures:
Eugene R. Schiff - Advisory Committees or Review Panels: Bristol Myers Squibb,
Gilead, Merck, Janssen, Salix Pharmaceutical, Pfizer, Arrowhead; Consulting:
Acorda; Grant/Research Support: Bristol Myers Squibb, Abbott / AbbVee, Gilead,
Merck, Conatus, Medmira, Roche, Janssen, Orasure Technologies, Discovery
Life Sciences, Siemens, Beckman Coulter, Siemens
The following authors have nothing to disclose: Jinhee Hyun, Masato Yoneda,
Emmanuel Thomas
1018
Expression of IFNλ4 in liver is closely associated with
non-response to antiviral therapy through the regulation
of basal expression of ISGs in chronic hepatitis C
patients but not in hepatitis B patients.
Miyako Murakawa 1,3 , Yasuhiro Asahina 1 , Fukiko Kawai-Kitahata
1 , Hiroko Nagata 1 , Syun Kaneko 1 , Sayuri Nitta 1 , Takako
Watanabe 1 , Yasuhiro Itsui 1,3 , Mina Nakagawa 1 , Sei Kakinuma 1 ,
Sayuki Iijima 2 , Yasuhito Tanaka 2 , Mamoru Watanabe 1 , Yujiro
Tanaka 3,1 ; 1 Gastroenterology and Hepatology, Tokyo Medical
and Dental University, Tokyo, Japan; 2 Nagoya City University,
Nagoya, Japan; 3 Medical Education Research and Development,
Tokyo Medical and Dental University, Tokyo, Japan
Backgrounds & Aims: The innate immune system has an essential
role in host antiviral defense. IL28B SNPs are associated
with the response to anti-HCV therapy both in IFN-based and
IFN-free regimens. Recently, IFNλ4 gene was considered as
one of the mechanisms responsible for poor response influenced
by IL28B unfavorable SNPs. However, little is known
about IFNλ4 expressions and its roles in antiviral innate immunity
including intrahepatic gene expressions of IFN-stimulated
genes (ISGs). The aim of this study is to evaluate the relationship
between IFNλ4 expression and viral liver diseases, and to
investigate the effect of IFNλ4 on intrahepatic gene expressions
responsible for poor response to IFN therapy. Methods: 1)
IFNλ4 expressions were analyzed in HCVcc-infected Huh7.5.1
cells, poly (I:C)-treated Huh7, HepG2, HeLa, HEK293T cells
and B lymphocytes, and HBV-expressing HepG2.215 cells.
2) The expressions of IFNλ4 were analyzed in liver samples
obtained from 49 chronic hepatitis C (CHC), 13 chronic hepatitis
B (CHB) and 3 autoimmune hepatitis (AIH) patients. All
CHC patients were treated with PEG-IFN/ribavirin (PR) therapy
and the relationship between IFNλ4 expressions and
treatment responses. 3) Antiviral ISGs (ISG15, Mx1, RIG-I,
TLR3, SOCS2) and IFN-suppressive genes (RNF125, SOCS1,
SOCS3, A20, RNF11) were analyzed in all liver samples,
and the relationship with IFNλ4 expressions were accessed.
Results: 1) IFNλ4 expression was induced by poly (I:C) treatment
as well as HCVcc infection in IL28B-minor cell lines, but
not induced by HBV. 2) IFNλ4 mRNA was detected in 11 of
22 liver samples of CHC patients with IL28B-unfavorable SNP,
but not in CHC patients with favorable genotype, CHB and AIH
patients. IFNλ4 expression was associated with non-response
to PR therapy (p < 0.001). 3) Intrahepatic expressions of antiviral
ISGs (ISG15 and Mx1) were significantly up-regulated in
nonvirological responders (NR) compared with those in virological
responders (VR) while expressions of suppressive ISGs
(RNF125, SOCS1, SOCS3 and RNF11) were down-regulated
in NR. Expressions of antiviral ISGs (Mx1 and RIG-I) were significantly
higher in IL28B-unfavorable patients in whom IFNλ4
mRNA was detected compared with those in IFNλ4-undetected
patients. However, expressions of suppressive ISGs (RNF125,
SOCS1, SOCS3 and RNF11) were significantly lower in
IFNλ4-detected patients. Conclusions: Our data suggest IFNλ4
have a specific role only in CHC. Intrahepatic expression of
IFNλ4 is associated with higher expression of antiviral ISGs
and lower expressions of suppressive ISGs at baseline, resulting
in the poor responsiveness for IFNα-based therapy for HCV
infection.
Disclosures:
Sei Kakinuma - Grant/Research Support: The Japanese Society of Gastroenterology,
MSD Co., Ltd.
Yasuhito Tanaka - Grant/Research Support: Chugai Pharmaceutical CO., LTD.,
MSD, Bristol-Myers Squibb; Speaking and Teaching: janssen pharma, Bristol-Myers
Squibb
Mamoru Watanabe - Grant/Research Support: MSD Co., Ltd.
The following authors have nothing to disclose: Miyako Murakawa, Yasuhiro
Asahina, Fukiko Kawai-Kitahata, Hiroko Nagata, Syun Kaneko, Sayuri Nitta,
Takako Watanabe, Yasuhiro Itsui, Mina Nakagawa, Sayuki Iijima, Yujiro Tanaka
1019
Lipid-Free apolipoprotein E signaling induces an ABCG1
driven cholesterol efflux that markedly decreases HCV
replication
Emilie Crouchet 1,2 , Mathieu Lefèvre 1,3 , Eloi R. Verrier 1,2 , Thomas
F. Baumert 1,2 , Catherine Schuster 1,2 ; 1 U1110, Inserm, Strasbourg,
France; 2 Strasbourg University Hospital, Université de Strasbourg,
Strasbourg, France; 3 CNRS UPR 9002, Strasbourg, France
There is an intimate link between the hepatitis C virus (HCV)
life cycle and its host cell’s lipid metabolism. HCV is associated
with lipoproteins in blood, forming lipo-viro-particles, which
are the infectious form of the virus. Apolipoprotein E (apoE), a
key lipo-viro-particle component, plays an essential role in HCV
entry, assembly, and egress. Although both the lipoprotein-associated
and lipid-free forms of apoE can be found in blood,
the role of lipid-free apoE in the HCV life cycle is unknown. In
this study, we investigated the effect of lipid-free apoE on the
HCV life cycle using the HCVcc model system and primary
human hepatocytes (PHH). A dose-dependent decrease in HCV
replication was observed when Huh 7.5.1 cells or PHHs were
treated with increasing amounts of lipid-free apoE. We show
that lipid-free apoE acts on HCV replication independently of
previously described apoE receptors, and independently of
the lipid-free apoE recycling process. By investigating the role
of lipid-free apoE on actors of hepatic lipid metabolism we
observed that extracellular lipid-free apoE markedly increases
cholesterol efflux via the ATP binding cassette sub-family G
member 1 protein (ABCG1). Our findings highlight a new
mechanism in lipid metabolism regulation and may provide
new therapeutic strategies to fight chronic HCV infection.