studies

1244A AASLD ABSTRACTS HEPATOLOGY, October, 2015
hepatocyte supernatants induced HIF-1a DNA binding and
up-regulated VEGF in RAW 264.7 macrophages. Conclusion:
Modified western diet accelerated carcinogen initiated hepatocellular
carcinoma, involving hepatocytes derived signals promoting
M2 macrophage polarization.
Disclosures:
The following authors have nothing to disclose: Aditya Ambade, Abhishek Satishchandran,
Banishree Saha, Karen Kodys, Gyongyi Szabo
2128
CD8 + T-cells drive liver injury and hepatocellular carcinoma
in chronic liver disease
Jessica Endig 1 , Laura E. Buitrago 1 , Silke Marhenke 1 , Anna Saborowski
1 , jutta Schütt 2 , Florian Reisinger 3 , Florian Limbourg 1 ,
Christian Koenecke 1 , Alina Schreder 1 , Robert Geffers 4 , Michael
P. Manns 1 , Mathias Heikenwälder 5 , Thomas Longerich 6 , Arndt
Vogel 1 ; 1 Hannover Medical School, Hannover, Germany;
2 Philipps University Marburg, Marburg, Germany; 3 Technische
Universität München, Munich, Germany; 4 Helmholtz Centre for
Infection Research, Braunschweig, Germany; 5 Helmholtz Zentrum
München, Munich, Germany; 6 Aachen University Hospital,
Aachen, Germany
Tumors frequently arise at sites of inflammation and chronic
inflammation is increasingly recognized as a key factor in
the pathogenesis of many malignancies. Hepatocellular carcinoma
(HCC), one of the most lethal and prevalent cancers
worldwide, represents a classic example of inflammation-linked
cancer. Although activation of different cytokines has been
reported in chronic liver disease, the critical components linking
inflammation and hepatocarcinogenesis remain elusive. To
characterize the role of the immune system in hepatic injury
and tumor development, we comparatively studied the extent
of liver disease and hepatocarcinogenesis in immunocompromised
vs. immunocompetent Fah-deficient mice. Flares of liver
injury were repeatedly induced which led to HCC formation
within 3-4 months. Strikingly, chronic liver injury and tumor
development were markedly suppressed in alymphoid Fah -/-
mice despite an overall increased mortality. Mechanistically,
we show that CD8 + Tcell-derived lymphotoxin-β (LTβ) is a
central mediator of HCC formation in Fah -/- mice. Pharmacological
inhibition of the lymphotoxin-β receptor (LTβR) indeed
delays tumors development in mice with chronic liver injury
without preventing liver damage. We here demonstrate that
lymphocytes play a decisive, yet ambiguous role in chronic
liver disease: while infiltrating lymphocytes mediate hepatocyte
damage, liver fibrosis and tumor development, they also protect
mice from acute on chronic liver failure and are required
for activation of liver progenitor cells during regeneration. Our
data illustrate that the immune system needs to be tightly regulated
in a context-specific fashion, in order to balance immune
surveillance and cancer risk. We propose that targeting specifically
the tumor-promoting pathways such as LTβ might be an
attractive chemopreventive strategy for patients at risk.
Disclosures:
Michael P. Manns - Consulting: Roche, BMS, Gilead, Boehringer Ingelheim,
Novartis, Idenix, Achillion, GSK, Merck/MSD, Janssen, Medgenics; Grant/
Research Support: Merck/MSD, Roche, Gilead, Novartis, Boehringer Ingelheim,
BMS; Speaking and Teaching: Merck/MSD, Roche, BMS, Gilead, Janssen, GSK,
Novartis
The following authors have nothing to disclose: Jessica Endig, Laura E. Buitrago,
Silke Marhenke, Anna Saborowski, jutta Schütt, Florian Reisinger, Florian
Limbourg, Christian Koenecke, Alina Schreder, Robert Geffers, Mathias Heikenwälder,
Thomas Longerich, Arndt Vogel
2129
Combined bone marrow plus liver transplantation in
cynomolgus monkeys
Joshua Weiner, Yojiro Kato, Sulemon Chaudhry, Raimon Duran-
Struuck, Mercedes Martinez, Paula Alonso-Guallart, Jonah Zitsman,
Jay H. Lefkowitch, Tomoaki Kato, Megan Sykes, Adam D.
Griesemer; Columbia Center for Translational Immunology, New
York Presbyterian-Columbia University Medical Center, New York,
NY
Introduction Combined kidney/bone marrow transplantation
(CKBMT) induces tolerance in both primates and humans.
Since liver is thought to be more tolerogenic than kidney, we
hypothesized that combined liver/bone marrow transplantation
(CLBMT) should be successful. We performed this pre-clinical
study in cynomolgus monkeys. Methods Five pairs of animals
were used. All were mismatched for class I, and the first 2 pairs
were mismatched for class II. Liver and ~3 x 10^8 donor bone
marrow cells/kg were transplanted on day 0. The induction
regimen consisted of 1.5Gy total body irradiation on day-6,
ATGAM (Pfizer) (50mg/kg) on days -2, -1, and 0, rituximab
(Genentech) (20mg/kg) on day -6 (Recipients 2-5, also day
6 for Recipients 4 and 5), 7Gy thymic irradiation on day -1,
splenectomy on day 0, and 28 days of either IM cyclosporine
(Novartis) or continuous IV tacrolimus (Astellas) followed by
4-week taper (tacrolimus only). Recipients 4 and 5 additionally
received LoCD2 (Mass Biologics) and anti-CD40 mAb (Mass
Biologics). Anti-donor MLR was assessed by plating recipient
and irradiated donor cells (1 x 10^6 each) for 5 days and
pulsing with tritiated thymidine for 24 hours. Results Recipients
1-3 survived 42, 69, and 57 days with multilineage mixed
chimerism (MC) up to days 36, 40, and 23 respectively. They
had normal to increased anti-donor cellular responses, except
Recipient 1, and severe histological cellular rejection. Flow
cytometry showed high percentages of CD8 T effector memory
cells (TEM) in the blood, graft, and draining node but not
peripheral nodes. Therefore, we added costimulatory blockade
(anti-CD40) and depletion of CD2+ memory cells (LoCD2b)
for Recipients 4 and 5, and subsequently this cell population
did not expand. In contrast to the first three recipients, the
fourth recipient survived 61 days (final 3 weeks with negligible
immunosuppression levels) and died of diarrhea/electrolyte
abnormalities on day 61 with normal LFTs, no histological
rejection or GVHD, significant MC, and decreased anti-donor
cellular responses. Recipient 5 is now 40 days post-transplantation
with 70-80% chimerism in all lineages and no signs
of rejection despite weaning immunosuppression. Conclusion
CLBMT induces transient MC, but rejection eventually occurs.
High levels of CD8 T effector memory cells likely contribute.
However, when additional costimulatory blockade and CD2
depletion are given, these cells are significantly depleted, preventing
rejection, prolonging MC, decreasing anti-donor cellular
responses, and possibly permitting tolerance.
Disclosures:
Tomoaki Kato - Grant/Research Support: Novartis
The following authors have nothing to disclose: Joshua Weiner, Yojiro Kato,
Sulemon Chaudhry, Raimon Duran-Struuck, Mercedes Martinez, Paula Alonso-Guallart,
Jonah Zitsman, Jay H. Lefkowitch, Megan Sykes, Adam D. Griesemer