Safety and Feasibility of CIK Cell Therapy in HCC After Tumor Resection

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and a leading cause of cancer-related mortality worldwide. Even after curative-intent surgical resection, disease recurrence occurs in approximately 60-70% of patients within five years, and no adjuvant systemic therapy has demonstrated a consistent recurrence-free survival or overall survival benefit in this setting. Cytokine-induced killer (CIK) cells are ex vivo-expanded autologous immune effector cells with a heterogeneous phenotype dominated by CD3⁺CD56⁺ cells, combining T-cell and NK-cell features and exhibiting MHC-unrestricted cytotoxicity against tumor cells. Phase II/III trials and subsequent meta-analyses conducted predominantly in East Asian populations have reported improved recurrence-free and overall survival with adjuvant CIK therapy following HCC resection, without a corresponding increase in severe (Grade ≥3) adverse events, and the South Korean Ministry of Food and Drug Safety approved a CIK-based product (Immuncell-LC) on this basis in 2022. No clinical data exist to date for an Iranian population, which differs from East Asian cohorts in HCC etiology (notably HBV prevalence and metabolic-associated steatotic liver disease burden) and genetic background.

This is a single-center, open-label, single-arm, phase I safety and feasibility study conducted at the Liver Transplant and Surgery Research Center, Imam Khomeini Hospital, Tehran University of Medical Sciences, in collaboration with Royan Institute. Full eligibility criteria are listed in the Eligibility section of this record.

For each patient, peripheral blood mononuclear cells (PBMCs) are isolated and expanded ex vivo under Good Manufacturing Practice (GMP) conditions for 14-21 days using IFN-γ, anti-CD3 antibody, and IL-2 to generate the autologous CIK cell product. Prior to release, each batch undergoes quality control testing for viability, CD3⁺CD56⁺ phenotype, antitumor cytotoxicity, sterility, and endotoxin level against pre-specified thresholds (≥85% viability, ≥60% CD3⁺CD56⁺, ≥30% cytotoxicity, endotoxin ≤5 EU/kg, negative microbial/mycoplasma testing); only batches meeting these criteria are released for infusion.

Each patient receives six intravenous infusions of autologous CIK cells administered as a slow infusion over approximately 60 minutes under sterile conditions: three weekly infusions (weeks 0, 1, and 2) followed by three biweekly infusions (weeks 4, 6, and 8). Vital signs are monitored before, during, and for two hours after each infusion. No routine premedication is given; mild fever is managed with acetaminophen. Adverse event monitoring and grading methodology are detailed under Outcome Measures. Over the 6-month follow-up period, scheduled assessments also include laboratory testing (including AFP, PIVKA-II, liver function and coagulation panels), peripheral blood lymphocyte immunophenotyping (CD56⁺CD16⁺CD3-, CD3⁺CD8⁺, CD3⁺CD56⁺CD16⁺, CD3⁺CD4⁺ populations at month 6 versus baseline), and contrast-enhanced MRI at months 3 and 6. Given the phase I descriptive design and small sample size, statistical analysis is planned to be descriptive rather than inferential.

Inclusion criteria

• Age between 18 and 80 years
• Documented HCC at BCLC stage 0-A, having undergone surgical tumor resection
• Single tumor or ≤3 nodules, each ≤3 cm
• Child-Pugh score A-B
• ECOG performance status 0-1
• Confirmed cancer-free status one month after surgery
• Written informed consent
• Leukocyte count > 3 × 10⁹/L
• Absolute neutrophil count (ANC) ≥ 1,000/µL
• Hemoglobin ≥ 8.5 g/dL
• Platelet count > 50 × 10⁹/L
• BUN and serum creatinine ≤ 1.5 × upper limit of normal No extrahepatic abdominal disease spread, confirmed by abdominal CT/MRI

Exclusion criteria

• Active infection or uncontrolled viremia (particularly HBV, HCV, or HIV)
• Any cell therapy or immunotherapy in the past 6 months, or current participation in another clinical study
• Another malignancy (prior or concurrent) differing from HCC in primary site or histology
• Clinically significant cardiovascular disease (e.g., heart failure, serious arrhythmia, symptomatic coronary artery disease)
• History of organ transplantation
• Primary or secondary immunodeficiency, or active autoimmune disease
• Severe allergic disorder or history of anaphylaxis
• Pregnancy or breastfeeding at study entry
• Women of childbearing potential intending to become pregnant