studies

1018A AASLD ABSTRACTS HEPATOLOGY, October, 2015
entially expressed Genes & miRNAs between patient groups,
key pathways & biological processes regulated were identified
by using Volcano plot, miRTarbase & GOElite tool. Differentially
expressed genes & miRNA along with pathways &
biological categories regulated, were subjected to Integrome
analysis to identify disease baseline & stage specific signatures.
Results: In AVH-B compared to controls,1178 genes,42
miRNAs were up-regulated & 1348 genes,9 miRNAs were
down regulated. In CHB-PNALT compared to controls,167
genes,1 miRNA were-up regulated & 307 genes,11 miRNAs
were down-regulated. In CHB-RALT compared to controls,82
genes,4 miRNAs were up-regulated & 58 genes,17 miRNAs
were down-regulated. In CHB-PNALT compared to AVH-B,500
genes,9 miRNAs were up regulated & 1307 genes,12 miR-
NAs were down regulated. In CHB-RALTcompared to AVH-B
1132 genes,26 miRNAs were up regulated &1694 genes,21
miRNAs were down regulated. In CHB-RALT compared to CHB-
PNALT, 183 genes, 3 miRNAs were up regulated & 70 genes,
3 miRNAs were down regulated.We identified 267 genes &
32 differentially expressed miRNAs,13 key pathways, biological
categories & gene families in one or more of the infection
stagesthat could play major role either in clearance or persistence
of HBV infection (Fig.1). Conclusion: Integrome analysis
revealed induction of unique cluster of miRNAs & repression
of their target genes that differentiates acute & chronic infection
stage, chronic infection with & without hepatic injury in DCs in
HBV infection.
Disclosures:
The following authors have nothing to disclose: Avishek K. Singh, Robert Geffers,
Sheetalnath B. Rooge, Aditi Varshney, Madavan Vasudevan, Ankit Bhardwaj,
Manoj Kumar, Pawan Malhotra, Sunil K. Mukherjee, Raj K. Bhatnagar, Nirupma
Trehanpati, Shiv K. Sarin
1660
Functional restoration of CD56 bright NK cells via IL-15
and NKG2D correlates with antiviral treatment efficacy
in chronic hepatitis B patients
Tao Chen 1 , Aichao Shi 1 , Xiaoping Zhang 1 , Lin Zhu 1 , Zeguang
Wu 1 , Weiming Yan 1 , Xiaojing Wang 1 , Hong Ma 2 , Jidong Jia 2 ,
Wei Guo 1 , Qin Ning 1 ; 1 Department and Institute of Infectious
Disease, Tongji Hospital, Tongji Medical College, Huazhong
University of Science and Technology, Wuhan, China; 2 Beijing
Friendship Hospital, Beijing, China
Hepatitis B virus is intrinsically immunogenic, and the immune
status of the host has a prognostic impact on patients with
chronic infection and influences the effects of antiviral treatment.
This current study aimed to investigate the dynamic
changes of NK cells post antiviral treatment and its potential
influence on treatment efficacy. This study involved 52 hepatitis
B e antigen (HBeAg) - positive chronic hepatitis B (CHB)
patients who received telbivudine (Ldt) for 52 weeks. Blood
samples were collected at baseline and week 12, 24, 36 and
48 of treatment, and tested for HBV DNA, HBsAg, HBeAg, ALT,
AST, and additional immunological markers. Compared with
baseline, the percentages and absolute number of peripheral
CD3 - CD56 + NK cells were significantly higher from week 36
to week 48, especially CD3 - CD56 bright NK cells. The expression
(percentage and MFI) of activating receptors NKG2D
and NKP46 was enhanced, while inhibitory receptor NKG2A
decreased. NKG2D expression was significantly enhanced on
peripheral NK cells in patients with HBeAg seroconversion,
particularly in CD3 - CD56 bright NK cell. The serum interleukin
15 (IL-15) level significantly elevated during Ldt treatment,
especially in patients with HBeAg seroconversion. Co-culture
of Ldt with purified peripheral NK cells from treatment naïve
HBeAg positive CHB patients showed significantly enhanced
expression of NKG2D and IL-15. These findings indicate that
antiviral treatment exerted by Ldt in CHB patients may play as
an “adjuvant” role capable of inducing or restoring NK cell
function via upregulated NKG2D and IL-15 expression, and
this in turn correlated with HBeAg seroconversion.
Disclosures:
Jidong Jia - Consulting: BMS, GSK, MSD, Novartis, Roche
Qin Ning - Advisory Committees or Review Panels: ROCHE, NOVARTIS, BMS,
MSD, GSK; Consulting: ROCHE, NOVARTIS, BMS, MSD, GSK; Grant/Research
Support: ROCHE, NOVARTIS, BMS; Speaking and Teaching: ROCHE, NOVAR-
TIS, BMS, MSD, GSK
The following authors have nothing to disclose: Tao Chen, Aichao Shi, Xiaoping
Zhang, Lin Zhu, Zeguang Wu, Weiming Yan, Xiaojing Wang, Hong Ma, Wei
Guo
1661
Knockdown of NTCP reduces susceptibility to HBV infection
in humanized-liver mice
Tasuku Nakabori, Hayato Hikita, Satoshi Aono, Yoshinobu
Yokoyama, Kazuhiro Murai, Takuo Yamai, Yugo Kai, Yuki Makino,
Yasutoshi Nozaki, Yoshinobu Saito, Satoshi Tanaka, Ryotaro
Sakamori, Tomohide Tatsumi, Tetsuo Takehara; Osaka University
Graduate School of Medicine, Suita, Japan
Background and Aim: Hepatitis B Virus (HBV) infects very limited
species such as humans and chimpanzees, which hamper
HBV research. Humanized-liver mice have potential to resolve
these difficulties. Recently, Na+-taurocholate cotransporting
polypeptide (NTCP) was reported as a candidate of HBV entry
receptor. However, the effect of its inhibition on HBV infection
remains unclear. In this study, we clarified it using humanized-liver
mice. Methods: TK-NOG mice were administrated
ganciclovir followed by transplantation of human primary
hepatocytes. Humanized-liver TK-NOG mice (humanized mice)
were inoculated with HBV obtained from a chronic hepatitis B
patient. The institutional ethics committee approved the study.
For in vitro study, primary hepatocytes isolated from humanized
mice were inoculated with HBV derived from the culture supernatant
of 2.2.15 cells. Results: Serum HBV-DNA were detected
in humanized mice from 2 weeks after HBV inoculation. After
8 to 10 weeks post-inoculation, HBV-DNA levels reached a
plateau (6-8 log copies/ mL) and almost all of human hepatocytes
were positive for HBc antigen by immunohistochemistry.
Administration of siRNA against human NTCP by tail vein successfully
reduced its expression level in human hepatocytes,
evidenced by immunohistochemistry and real time RT-PCR.
After 2 weeks of HBV inoculation following administration of
NTCP siRNA, the levels of serum HBV-DNA, serum HBs antigen
and intracellular covalently closed circular DNA (cccDNA) were
significantly reduced. In sharp contrast, siRNA-mediated NTCP
knockdown at 12 weeks after HBV inoculation did not affect
them. Primary hepatocytes from humanized mice were maintained
in culture over 2 months. After inoculation with HBV for
24 hours, HBV-DNA, HBs antigen and HBe antigen levels in
culture medium, gradually increased and kept detectable over
a month. cccDNA was also detected. Immunohistochemistry