Volume 148, Issue 7, Pages 1383-1391.e6 (June 2015) Adjuvant Immunotherapy With Autologous Cytokine-Induced Killer Cells for Hepatocellular Carcinoma  Joon Hyeok Lee, Jeong-Hoon Lee, Young-Suk Lim, Jong Eun Yeon, Tae-Jin Song, Su Jong Yu, Geum-Youn Gwak, Kang Mo Kim, Yoon Jun Kim, Jae Won Lee, Jung-Hwan Yoon  Gastroenterology  Volume 148, Issue 7, Pages 1383-1391.e6 (June 2015) DOI: 10.1053/j.gastro.2015.02.055 Copyright © 2015 AGA Institute Terms and Conditions

Figure 1 Kaplan–Meier estimates of RFS, OS, and cancer-specific survival. (A) RFS was computed for all patients included in the efficacy population. Patients who had not progressed or died were censored on the data cut-off date. (B) OS was computed for all patients included in the efficacy population. Patients who had not died were censored on the data cut-off date. (C) Cancer-specific survival was computed for all patients included in the efficacy population. Patients who had not died of hepatocellular carcinoma were censored on the date of death owing to other causes or the data cut-off date. Gastroenterology 2015 148, 1383-1391.e6DOI: (10.1053/j.gastro.2015.02.055) Copyright © 2015 AGA Institute Terms and Conditions

Figure 2 Recurrence-free survival in selected subsets. The graph show the estimates of the HR for every subgroup as a square (whose size is proportional to the amount of information) and the horizontal lines depict the 95% CIs, which were calculated by means of a Cox proportional hazards model. The diamond indicates the HRs with 95% CIs for all patients enrolled. The vertical solid line at the HR of unity corresponds to the line of no effect. HR values of less than unity correspond to a reduction in the risk of recurrence or death with immunotherapy. Patients co-infected with both HBV and HCV were included in the HCV subset. HBV DNA and antiviral agent groups 1, 2, 3, and 4 indicate patients with serum HBV-DNA levels ≥ 2000 IU/mL who underwent antiviral treatment; those with serum HBV-DNA levels ≥ 2000 IU/mL who underwent no antiviral treatment; those with HBV-DNA levels < 2000 IU/mL who underwent antiviral treatment; and those with HBV-DNA levels < 2000 IU/mL who underwent no antiviral treatment, respectively. *Characteristics were post hoc subgroups. AFP, α-fetoprotein; ALP, alkaline phosphatase; ALT, alanine aminotransferase; AST, aspartate aminotransferase; ECOG, Eastern Cooperative Oncology Group; HCV, hepatitis C virus. Gastroenterology 2015 148, 1383-1391.e6DOI: (10.1053/j.gastro.2015.02.055) Copyright © 2015 AGA Institute Terms and Conditions

Supplementary Appendix CONSORT diagram. Of the enrolled patients, 15 did not meet inclusion criteria and thus were excluded from random assignment. In addition, 4 more patients were excluded from the efficacy analysis because they were found to violate inclusion criteria after randomization according to a decision from the steering committee. Gastroenterology 2015 148, 1383-1391.e6DOI: (10.1053/j.gastro.2015.02.055) Copyright © 2015 AGA Institute Terms and Conditions

Supplementary Figure 1 Manufacturing process of CIK cell agent. After collection of peripheral blood from the respective patients (day 0), mononuclear cells were separated and cultured for 12–21 days with interleukin 2 (from day 0 to the date of harvest) and anti-CD3 monoclonal antibody (from day 0 to day 5). Tests for sterility and mycoplasma were performed twice at 3 days before harvest and at the date of harvest. Tests for identification, purity, and potency were performed 1 day before harvest. At the date of harvest, CIK cell agents were administered to patients. HCV, hepatitis C virus; HIV, human immunodeficiency virus; PBMC, peripheral blood mononuclear cell. aSterility test included aerobic and anaerobic bacteria and fungi. bIdentification included proportions of CD3+, CD8+, and CD56+ cells. cPurity test included proportions of CD14+ and CD20 CD3+cells. Gastroenterology 2015 148, 1383-1391.e6DOI: (10.1053/j.gastro.2015.02.055) Copyright © 2015 AGA Institute Terms and Conditions

Supplementary Figure 2 Kaplan–Meier estimates of RFS according to the respective recurrence site. (A) Intrahepatic local recurrence (within 2 cm from surgical or ablation margin: HR, 0.85; 95% CI, 0.44–1.62; P = .30 by 1-sided log-rank test), (B) intrahepatic distant recurrence (beyond 2 cm from margin: HR, 0.63; 95% CI, 0.37–1.07; P = .04 by 1-side log-rank test), and (C) extrahepatic recurrence (HR, 0.17; 95% CI, 0.02–0.15; P = .03 by 1-side log-rank test). RFS was computed on all patients included in the efficacy population. Patients who had not progressed or died were censored on data cut-off date. Gastroenterology 2015 148, 1383-1391.e6DOI: (10.1053/j.gastro.2015.02.055) Copyright © 2015 AGA Institute Terms and Conditions