studies

708A AASLD ABSTRACTS HEPATOLOGY, October, 2015
Disclosures:
The following authors have nothing to disclose: Emilie Crouchet, Mathieu Lefèvre,
Eloi R. Verrier, Thomas F. Baumert, Catherine Schuster
1020
Substitution in amino acid 70 of hepatitis C virus core
protein changes the adipokine profile via toll-like receptor
2/4 signaling
Masahiko Tameda, Kazushi Sugimoto, Yoshiyuki Takei; Mie University,
Tsu, Japan
Background & Aims: It has been suggested that amino acid
(aa) substitution at position 70 from arginine (70R) to glutamine
(70Q) in the genotype 1b hepatitis C virus (HCV) core protein
is associated with insulin resistance and worse prognosis.
However, the precise mechanism is still unclear. The aim of
this study was to investigate the impact of the substitution at
position 70 in HCV core protein on adipokine production by
murine and human adipocytes. Methods: The influence of treatment
with HCV core protein (70R or 70Q) on adipokine production
by both 3T3-L1 and human adipocytes were examined
with real-time PCR and enzyme-linked immunosorbent assay
(ELISA), and triglyceride content was also analyzed. The effects
of toll-like receptor (TLR)2/4 inhibition on IL-6 production by
3T3-L1 induced by HCV core protein were examined. Results:
IL-6 production was significantly increased and adiponectin
production was reduced without a change in triglyceride content
by treatment with 70Q compared to 70R core protein in
both murine and human adipocytes. IL-6 induction of 3T3-L1
cells treated by 70Q HCV core protein was significantly inhibited
with anti-TLR2 antibody by 42%, and by TLR4 inhibitor
by 40%. Conclusions: Our study suggests that extracellular
HCV core protein with substitution at position 70 enhanced IL-6
production and reduced adiponectin production from visceral
adipose tissue, which can cause insulin resistance, hepatic
steatosis, and ultimately development of HCC.
Disclosures:
The following authors have nothing to disclose: Masahiko Tameda, Kazushi
Sugimoto, Yoshiyuki Takei
1021
Variability of the hepatitis delta virus quasispecies in
chronic infection is high and similar to that of the hepatitis
C virus quasispecies
Maria Homs 1,2 , Josep Gregori 3 , Hadi Karimzadeh 4 , Damir G.
Cehic 3 , Maria Blasi 1,2 , Rosario Casillas 3 , David Tabernero 1,2 ,
Josep Quer 1,3 , Michael Roggendorf 4 , Mar Riveiro-Barciela 5 , Rafael
Esteban 1,5 , Francisco Rodriguez-Frias 1,2 , Maria Buti 1,5 ; 1 Centro de
Investigacion Biomedica en Red de Enfermedades Hepaticas y
Digestivas, CIBERehd, Barcelona, Spain; 2 Microbiology, Hospital
Vall d’Hebron, Barcelona, Spain; 3 Liver Diseases, Research Institute
Vall d’Hebron, Barcelona, Spain; 4 Institut für Virologie, Technische
Universität München, München, Germany; 5 Hepatology,
Hospital Vall d’Hebron, Barcelona, Spain
Background Hepatitis D virus (HDV) is a satellite RNA virus with
a small 1.7-kb genome that uses the hepatitis B virus envelope
to infect hepatocytes. The complexity of the hepatitis D viral
population has not been described. Hepatitis C virus (HCV)
is another RNA virus that circulates in infected individuals as
a quasispecies; the NS5A region is one of the most complex
of the non-structural encoding HCV genes. Aim To study the
complexity of the HDV quasispecies in patients with chronic
hepatitis D and compare it with that of the HCV quasispecies.
Samples and methods Fifteen serum samples from 10
chronic HCV- and 5 chronic HDV-infected treatment-naïve
patients were analyzed. Samples with similar viremia levels
were selected (mean HCV-RNA 6.51 log copies/mL, SD 0.57
and HDV-RNA 6.31 log copies/mL, SD 0.61 p=0.84). HCV-
RNA was quantified by COBAS-AmpliPrep (Roche) (detection
limits 1.63-7.83 log IU/mL) and HDV-RNA, by an in-house
method (detection limits 3.5-8.5 log copies/mL). HCV-RNA
values were converted to log copies/mL. One essential region
in each virus was deep-sequenced: the NS5A of HCV (positions
6299-6735, 436 bp) and the C-terminal region of HDV
antigen (positions 912-1298, 386 bp). Abundance filters for
deep-sequencing were set at 0.25%. Quasispecies complexity
was evaluated by determining the mutation frequency (Mf)
and nucleotide diversity (Pi). Both parameters take into account
amplicon length, and represent the number of differences/site
with respect to the most represented haplotye (Mf) or between
each pair of genomes (Pi). Results are presented as mean and
standard deviation (SD). Results A total of 187,606 sequences
were analyzed. Mf and Pi showed a normal distribution and
significantly correlated (R=0.985, p