Safety Study of Liver Natural Killer Cell Therapy for Hepatoma Liver Transplantation

To investigate whether adoptive transfer of IL-2 stimulated NKcells extracted from cadaveric
donor liver graft exudates into recipient with HCC will lead to prevent recurrence of HCC
after liver transplantation.

The method we plan to use is to collect NK cells from the cadaveric donor liver perfusate at
the time of organ recovery. The perfusate will be condensed by centrifuging and liver
mononuclear cells (LMNC) were isolated by gradient centrifugation. LMNC will be cultured
with anti-CD3 antibody. LMNC will be separted into a CD3-/CD56+ NK cell fraction and a
non-NK cell fraction. The purity of isolated fractions will be assessed by Flow cytometric
analyses. NK cells will be cultured with human recombinant IL-2 in 10% RPMI at 37 ºC in a 5%
CO2 incubator. NK cells will be infused intravenously to the recipient with HCC who have
liver transplant from the same cadaveric donor at post operative day 4 in attempt to prevent
the metastasis and recurrence of HCC.

Inclusion Criteria:

- Primary liver transplant recipient with HCC

- Patients need to meet the liver transplant eligibility criteria

- Cardiac function; cardiac echo will indicate that ejection fraction (EF) > 35% or
right ventricular systolic pressure (RVSP) < 50mmHg. Stress echo will show no
ischemic lesion (if applicable).

- Pulmonary function; SpO2 >90% or PaO2 > 60 mmHg, or CT will show no active pulmonary
lesion.

- Complete blood count; Platelet > 20,000 /mm^3, Hematocrit > 20%, WBC > 1,000 /mm^3

- Eighteen years of age or older

- Ability to provide informed consent

- If female of childbearing potential:

Must not be lactating, must have a negative serum B-human chorionic gonadotropin (HCG)
test within 7 days prior to Day of Transplant, and must agree to practice an acceptable
and reliable form of contraception during the study Ability to provide informed consent

Exclusion Criteria:

- Living donor liver transplant; a healthy person donates part of his or her liver to
the recipient

- Multiple organ transplants

- Prior solid organ or bone marrow transplant recipients

- Fluminant hepatic failure

- The patients regularly receive the hemodialysis more than twice a week before liver
transplant

- Status 1 transplants; acute severe disease and defined as a patient with only recent
development of liver disease who is in the intensive care unit of the hospital with a
life expectancy without a liver transplant of fewer than 7 days

- ABO incompatible transplants

- Transplants utilizing livers from non-heart beating donors; the cardiac death donor

- Recipients of investigational therapy within 90 days prior to transplant procedure

- Acute viral illness

- History of malignancy within 5 years, with exception of: Adequately treated localized
squamous or basal cell carcinoma of the skin without evidence or recurrence, and/or
Hepatocellular carcinoma

- Illness other than primary liver disease (e.g., severe ischemic heart disease, left
ventricular dysfunction, or pulmonary disease), which, in the opinion of the
Investigator, may significantly increase the risk of the transplantation procedure

- Current drug or alcohol abuse or, in the opinion of the Investigator, is at risk for
poor compliance (no drug testing required)

- Serology positive donor (HCV, HBsAg, HBcAb, HTLV-1, HTLV-3, EBVIgM)

- Poor liver function donor (Total bilirubin > 3.0 mg/dl, Prothrombin time > 35 sec),

- Patients who receive intercurrent chemotherapy at the time of enrollment