studies

700A AASLD ABSTRACTS HEPATOLOGY, October, 2015
of HCV-expression in Huh7.5 cells confirmed by anti-NS5A Ab
and HCV-specific T cell activation examined by MHC peptide
tetramer, intracellular cytokine staining and CD107a mobilization
using FACS. Result: Jc1-1073-1a and Jc1-2594-1a with
genotype 1a-derived HCV epitopes could infect over 90% of
Huh7.5 cells and support endogenous processing/presentation
of 1a-based CD8 T cell epitopes to activate HCV-specific CD8
T cells in-vitro. Co-culture with HCV-specific CD8 T cells and
HCV-replicating Huh7.5 cells resulted in reduced number of
total and HCV-infected Huh7.5 cells. PD-1 blockade enhanced
HCV-specific CD8 T cell activation with reduced HCV expression
as well as viability of Huh7.5 cells. Conclusion: PD-1
blockade may enhance virus control through cytopathic pathway
in an infectious HCV co-culture system, highlighting the
need for caution in clinical application of checkpoint inhibitors.
*acknowledgment: HCV and T-cell reagents were generously
provided by Drs. Takaji Wakita, Charles Rice, Brett Lindenbach,
Stanley Lemon, Jim Riley, Annelise Vuidepot, Peter Molloy
and Alan Bennett.
Disclosures:
Kyong-Mi Chang - Consulting: Genentech, Tekmira, Alnylam; Stock Shareholder:
BMS (spouse employment)
The following authors have nothing to disclose: Keisuke Ojiro, Xiaowang Qu,
Jang-June Park, Hyosun Cho
1002
HCV RNA and HCV core antigen are frequently detectable
in stool of men chronically infected with HCV: Is
feces a potential source of infection?
Benjamin Heidrich 1,3 , Eike Steinmann 4 , Iris Plumeier 2 , Janina
Kirschner 1 , Lisa Sollik 1 , Szilvia Ziegert 1 , Patrick Lehmann 1 ,
Michael Engelmann 4 , Tim Lankisch 1 , Michael P. Manns 1,3 , Dietmar
H. Pieper 2,3 , Heiner Wedemeyer 1,3 ; 1 Dept. of Gastroenterology,
Hepatology and Endocrinology, Hannover Medical School, Hannover,
Germany; 2 Microbial Interactions and Processes Research
Group, Centre for Infection Research, Braunschweig, Germany;
3 Partner site Hannover-Braunschweig, German Center for Infection
Research (DZIF), Hannover-Braunschweig, Germany; 4 Institute for
Experimental Virology, TWINCORE Centre for Experimental and
Clinical Infection Research, Hannover, Germany
Background and Aims: HCV is one of the leading causes of
liver cirrhosis and hepatocellular carcinoma worldwide. Transmission
of HCV is thought to be mainly parenteral. However,
unprotected anal intercourse seems to be a risk factor for acquisition
of HCV in men having sex with men (MSM) as well as in
heterosexual partnerships. HCV has been detected in blood,
saliva, and bile. Surprisingly, there are no studies investigating
the presence of HCV in stool. Methods: Stool samples of 98
patients were prospectively collected, mixed with RNAlater
and stored at -20°C. RNA was isolated using TRIzol reagent
and transcribed in cDNA by using the SuperScript III kit with
random hexamers. Specific HCV primers were used to identify
samples positive for HCV RNA. PCR results were confirmed by
Sanger sequencing. HCV RNA positive samples were tested for
occult blood using the hemoCARE guajak test and were tested
for HCVcoreAg using the Architect HCVAg from Abbott. Additionally,
viral stability of recombinant HCV particles was investigated
in vitro by incubation of the genotype 2a chimeric virus
Jc1 with different bile and stool suspensions. Results: Overall,
68/98 (69%) stool samples tested positive for HCV RNA. Men
tested more often positive for HCV RNA in stool than women
(83% vs. 52%; p