Background:To date, no adjuvant treatment has been shown to have a clear benefit in patients with hepatocellular carcinoma (HCC). In this prospective phase I/IIa study, we evaluated the safety and efficacy of adjuvant dendritic cell (DC) therapy in HCC patients who received primary treatment for HCC.Methods:Twelve HCC patients who had no viable tumour after primary treatments were included. Dendritic cell vaccines pulsed with cytoplasmic transduction peptide-attached alpha-fetoprotein, glypican-3 and melanoma-associated antigen 1 recombinant fusion proteins were injected subcutaneously near to inguinal lymph nodes. Adverse effects, time to progression (TTP), and associated immune responses were evaluated after DC vaccination.Results:Nine of 12 patients had no tumour recurrence up to 24 weeks after DC vaccination. Among a total of 144 adverse events, 129 events (89.6%) were regarded as adverse drug reactions, all of which were grade 1 or 2. The majority of patients showed enhanced anti-tumour immune responses after DC vaccination. Recurrence-free patients exhibited relatively stronger anti-tumour immune responses than patients who developed recurrence after DC vaccination, as evidenced by lymphocyte proliferation and IFN-γ ELISPOT assays. The median time of TTP was 36.6 months in the DC-vaccination group and 11.8 months in the control group (hazard ratio, 0.41; 95% confidence interval, 0.18–0.95; P=0.0031 by log-rank test).Conclusions:Adjuvant DC vaccine for HCC was safe and well tolerated in phase I/IIa study, and preliminary efficacy data are encouraging to warrant further clinical study in patients with HCC after primary treatments.

The POZ domain is a highly conserved protein-protein interaction motif found in many regulatory proteins. Nuclear factor-B (NF-B) plays a key role in the expression of a variety of genes in response to infection, inflammation, and stressful conditions. We found that the POZ domain of FBI-1 (factor that binds to the inducer of short transcripts of human immunodeficiency virus-1) interacted with the Rel homology domain of the p65 subunit of NF-B in both in vivo and in vitro proteinprotein interaction assays. FBI-1 enhanced NF-B-mediated transcription of E-selectin genes in HeLa cells upon phorbol 12-myristate 13-acetate stimulation and overcame gene repression by IB␣ or IB␤. In contrast, the POZ domain of FBI-1, which is a dominant-negative form of FBI-1, repressed NF-B-mediated transcription, and the repression was cooperative with IB␣ or IB␤. In contrast, the POZ domain tagged with a nuclear localization sequence polypeptide of FBI-1 enhanced NF-B-responsive gene transcription, suggesting that the molecular interaction between the POZ domain and the Rel homology domain of p65 and the nuclear localization by the nuclear localization sequence are important in the transcription enhancement mediated by FBI-1. Confocal microscopy showed that FBI-1 increased NF-B movement into the nucleus and increased the stability of NF-B in the nucleus, which enhanced NF-B-mediated transcription of the E-selectin gene. FBI-1 also interacted with IB␣ and IB␤.The BTB/POZ (broad complex, Tramtrack, and bric-a-brac/ poxvirus and zinc finger) domain is an evolutionarily conserved protein-protein interaction domain that is found at the N terminus of various cellular and viral regulatory proteins. The proteins containing the BTB/POZ domain have several C-terminal structures important in their biological functions, such as the zinc finger, actin-binding repeats, and ion channel motifs (1-3). The POZ domains of PLZF (promyelocytic leukemia zinc finger) and Bcl-6 (B-cell lymphoma-6) have been shown to interact with SMRT (silencing mediator for retinoid and thyroid hormone receptors)/N-CoR (nuclear receptor co-repressor), mSin3A, and histone deacetylases (4, 5).FBI-1 (factor that binds to the inducer of short transcripts of human immunodeficiency virus-1) was purified as a cellular factor that binds specifically to the wild-type IST (inducer of short transcripts) elements of human immunodeficiency virus-1 long terminal repeats and the proximal promoter of the ADH5/FDH gene, and its cDNA was cloned (6 -9). FBI-1 is a ubiquitous transcription factor that contains a BTB/POZ domain at its N terminus and Krü ppel-like zinc fingers at its C terminus. There have been several recent reports on the function of FBI-1. FBI-1 stimulates Tat (transactivator of transcription) activity on the human immunodeficiency virus-1 long terminal repeat (8) and represses human ADH5/FDH gene expression by interacting with Sp1 zinc fingers (9). The mouse counterpart of FBI-1, LRF (leukemia/lymphoma-related factor), is co-immunoprecipitated and co-localized with Bcl-6 (...

Using messenger RNA (mRNA) in situ hybridization, we investigated estrogen receptor-beta (ERbeta) mRNA levels in normal mammary, benign breast tumor (BBT), breast cancer (BC), and metastatic lymph node tissues to verify the role of ERbeta in BC development and progression. ERbeta expression was significantly decreased in BC and metastatic lymph node tissues compared with normal mammary and BBT tissues (p < 0.01). The intensity and extent of ERbeta mRNA signals were also significantly lower in BC and metastatic lymph node tissues than in the normal mammary and BBT tissues (p < 0.01). An inverse relationship was found between ERbeta mRNA level and both histologic grade (p = 0.091) and progesterone receptor expression (p = 0.052) with marginal significance, but no significant association was noted between ERbeta expression in cancer tissues and the other clinico-pathologic data. The 3-year distant relapse-free survival probability was found to be independent of ERbeta expression. Collectively, ERbeta mRNA decreases in the process of BC development, but seems to be associated with poor differentiation.

Our previous phase I/IIA study showed that autologous dendritic cells (DCs) pulsed with tumor-associated antigens are well tolerated in patients with hepatocellular carcinoma (HCC). In this randomized, multicenter, open-label, phase II trial, we investigated the efficacy and safety of this DC-based adjuvant immunotherapy with 156 patients, who treated for HCC with no evidence of residual tumor after standard treatment modalities. Patients were randomly assigned to immunotherapy ( = 77; injection of 3 × 10 DC cells, six times over 14 weeks) or control ( = 79; no treatment). The primary end point was recurrence-free survival (RFS), and the secondary endpoints were immune response and safety. The RFS between the immunotherapy and control groups was not significantly different (hazard ratio [HR], 0.97; 95% confidence interval [CI], 0.60-1.56; = 0.90). However, post-hoc subgroup analyses revealed that DC immunotherapy significantly reduced the risk of tumor recurrence of non-radiofrequency ablation (non-RFA) group patients ( = 83, HR, 0.49; 95% CI, 0.26-0.94; = 0.03), whereas unexpectedly increased the risk of recurrence in RFA group ( = 61, = 0.01). Tumor-specific immune responses were significantly enhanced (both < 0.01) in the immunotherapy group. Baseline serum interleukin (IL)-15 was statistically correlated with RFS prolongation (HR, 0.16; 95% CI, 0.03-1.58; = 0.001) within the immunotherapy groups. Overall adverse events were more frequent in the immunotherapy group ( < 0.001) but were mainly mild to moderate in severity. In conclusion, adjuvant immunotherapy with DC vaccine reduces the risk of tumor recurrence in HCC patients who underwent standard treatment modalities other than RFA. Baseline IL-15 might be a candidate biomarker for DC-based HCC immunotherapy.