studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 1067A
liver failure was induced in C57BL/6J mice using thioacetamide
(TAA) (200mg/kg, i.p) once a day for 2 days. In phase
I, animals were divided in three groups: 1) TAA+saline via tail
vein; 2) TAA+MSCs via tail vein; 3) TAA+Scaffold loaded with
MSCs to evaluate the mortality and changes in liver function.
PLGA Poly (lactic-co-glycolic acid) scaffold alone, and loaded
with 1x10 6 human MSCs were implanted on dorsum of animals.
In phase II, animals were divided into three groups: 1)
TAA; 2) TAA+Scaffold; 3) TAA+Scaffold loaded with MSCs
to evaluate liver function and mortality. Animals were sacrificed
3 days after implantation, serum and liver samples were
collected. Liver tissue immunohistochemistry for proliferation
marker (Ki67) was performed. Results: In phase I, the mortality
rate of TAA+Scaffold group was 36% as compared to
TAA+PBS tail vein (66%) and TAA+Scaffold only groups (54%).
In phase II, TAA+Scaffold loaded with MSCs group had only
11.8% mortality as compared to TAA (30.4%) and TAA+Scafold
only (52.6%) (p=0.014) groups. In phase II, serum AST
levels of TAA+Scaffold loaded with MSCs was lower than other
two groups, while serum ALT levels of TAA+Scaffold group
were the lowest. Furthermore, TAA+Scaffold loaded with MSCs
group showed increased liver tissue staining for Ki67. Later, we
performed PCR using human specific primer, to evaluate homing
of MSCs in mouse liver. There was no detection of human
cells residing in mouse liver. Conclusion: Scaffold loaded MSCs
showed protective effects via paracrine signaling on acute liver
failure model.
Disclosures:
The following authors have nothing to disclose: Yong Kyun Cho, Dae Won Jun,
Eun Chul Jang, Seung Min Lee, Joo Hee Kwak
1761
Indeterminate Etiology of Acute Liver Failure: Fact or
Fiction?
Jody A. Rule 1 , Jorge Rakela 2 , Norman L. Sussman 3 , Nathan M.
Bass 4 , Holly Tillman 5 , Jaime L. Speiser 5 , R. Todd Stravitz 6 , Ryan
Taylor 7 , William M. Lee 1 , Daniel Ganger 8 ; 1 Internal Medicine,
University of Texas Southwestern Medical Center, Dallas, TX; 2 Gastroenterology
and Hepatology, Mayo Clinic Hospital, Phoenix,
AZ; 3 Division of Abdominal Transplantation, Baylor College of
Medicine, Houston, TX; 4 Department of Medicine, University of
California San Francisco, San Francisco, CA; 5 Data Coordination
Unit, Department of Public Health Sciences, Medical University
of South Carolina, Charleston, SC; 6 Internal Medicine, Virginia
Commonwealth University, Richmond, VA; 7 Gastroenterology and
Hepatology, University of Kansas Medical Center, Kansas City,
KS; 8 Gastroenterology and Hepatology, Northwestern Memorial
Hospital, Chicago, IL
Background: Approximately 11% of acute liver failure or
acute liver injury (ALF/ALI) cases are initially listed as “indeterminate”
by attending hepatologists at academic transplant
centers; this has been attributed in the past to an unidentified
virus or toxic injury. Although nearly 20% of these cases
demonstrate a positive acetaminophen (APAP)-adduct assay
(Hepatology 2011;53:567), the remainder still are considered
indeterminate. Methods: We examined 302 indeterminate
cases from 2713 consecutively enrolled patients in the
ALFSG Registry using pre-defined characteristics developed by
an expert committee, to identify each etiology as either Highly
Likely/Definite (HL/D: >75% certainty) or Probable (>50% certainty).
Examples: Criteria for autoimmune ALF (AI-ALF): calculated
globulins elevated >ULN or serum IgG >ULN; ANA,
ASMA or LKMA-positive titer >1:40; biopsy features: plasma
cells, centrilobular necrosis or interface hepatitis. Thus, Highly
Likely/Definite (HL/D) AI-ALF: All 3 criteria met, Probable AIH:
Any 2 of 3 met. For APAP, HL/D: Positive APAP-adduct assay
or history of APAP ingestion or APAP level (or both) and AST
or ALT >1000/Bili 1 etiology. Among the remaining
86 cases, 51 had at least 1 primary diagnostic test missing
(APAP-adducts, ANA, aHAVIgM, HBsAg, aHBcIgM, or aHCV).
Only 35 cases had a complete set of medical history and laboratory
tests, and no etiology defined, fulfilling our diagnosis of
“indeterminate” ALF. Among patients initially diagnosed with
indeterminate ALF or ALI, 216 (72%) could be assigned to a
single defined etiology with additional testingand standardized
criteria. Dual etiology (or uncertainty with two possibilities)
will need further adjudication. Incomplete history or diagnostic
testing precluded assignment of etiology in 17%. Conclusions:
Only 11.5% of “indeterminates”, 1.2% of the overall ALF registry,
fulfilled stringent criteria for “indeterminate etiology” (data
complete, no cause discernible). With a defined/adjudicated
approach to causality, true “indeterminate” ALF is much less
common than previously estimated.
Disclosures:
Norman L. Sussman - Advisory Committees or Review Panels: BMS, Merck;
Board Membership: HepaHope; Grant/Research Support: AbbVie, BMS, Biotest,
Conatus, Esai, Genfit, Gilead, Hyperion, Immuron, Intercept, Lumena, Merck,
Novartis, Ocera; Speaking and Teaching: AbbVie, Gilead, Jannsen
Nathan M. Bass - Advisory Committees or Review Panels: Quest Diagnostics
William M. Lee - Consulting: Eli Lilly, Sanofi; Grant/Research Support: Gilead,
BMS, Vertex, Merck
Daniel Ganger - Advisory Committees or Review Panels: Bristol Myers, Abbvie;
Grant/Research Support: Merck, Ocera; Speaking and Teaching: Gilead
The following authors have nothing to disclose: Jody A. Rule, Jorge Rakela, Holly
Tillman, Jaime L. Speiser, R. Todd Stravitz, Ryan Taylor
1762
High CXCR-1 and CXCR-2 Expressing Neutrophils
Induce Early Hepatocyte Death and promote Liver Injury
in Acute-on-Chronic Liver Failure
Arshi Khanam 1 , Nirupma Trehanpati 1 , Peggy Riese 2 , Archana Rastogi
1 , Carlos A. Guzman 2 , Shiv K. Sarin 1 ; 1 research, Institute of
liver and biliary sciences, New Delhi, India; 2 Helmholtz Centre for
Infection Research, Braunschweig, Germany
Background and aims: Acute-on-chronic liver failure (ACLF) is a
serious and often fatal liver disease. Mechanisms of liver injury
and development of sepsis in these patients are unclear. Neutrophils
contribute to disease pathogenesis in acute liver failure
and serve as a biomarker of organ dysfunction. We investigated
the mechanisms of CXCR-1 and CXCR-2 expressing
neutrophils mediated hepatic injury in ACLF patients (alcohol
and hepatitis B related) and compared with chronic hepatitis
B (CHB) and healthy controls (HC). Methods: CXCR-1 and
CXCR-2 expressions on neutrophils were measured by flowcytometry,
immunohistochemistry and RT-PCR. In vitro co-culture
assays were performed to determine the mechanisms of hepatic
injury and cell death. Mechanism of contact dependant cell
death was investigated by co-culture of neutrophils with HepG2
and HepG2.2.15 cells. Contact independent cell death was
checked by culture of HepG2 and HepG2.2.15 cells with LPS
stimulated neutrophil’s supernatant. In patients, cell death was
determined by intrahepatic caspase-3 (apoptosis) and recep-