Antigen In Hepatocellular Carcinoma Research Articles

Microproteins encoded by noncanonical ORFs are a major source of tumor-specific antigens in a liver cancer patient meta-cohort.

The expression of tumor-specific antigens during cancer progression can trigger an immune response against the tumor. Here, we investigate if microproteins encoded by noncanonical open reading frames (ncORFs) are a relevant source of tumor-specific antigens. We analyze RNA sequencing data from 117 hepatocellular carcinoma (HCC) tumors and matched healthy tissue together with ribosome profiling and immunopeptidomics data. Combining human leukocyte antigen-epitope binding predictions and experimental validation experiments, we conclude that around 40% of the tumor-specific antigens in HCC are likely to be derived from ncORFs, including two peptides that can trigger an immune response in humanized mice. We identify a subset of 33 tumor-specific long noncoding RNAs expressing novel cancer antigens shared by more than 10% of the HCC samples analyzed, which, when combined, cover a large proportion of the patients. The results of the study open avenues for extending the range of anticancer vaccines.

Identification of tumor antigens and anoikis-based molecular subtypes in the hepatocellular carcinoma immune microenvironment: implications for mRNA vaccine development and precision treatment

Hepatocellular carcinoma (HCC) represents a formidable malignancy with a high lethality. Nonetheless, the development of vaccine and the establishment of prognostic models for precise and personalized treatment of HCC still encounter big challenges. Thus, the aim of this study was to develop HCC vaccines and explore anoikis-based prognostic models based on RNA sequencing data in GEO datasets (GSE10143, GSE76427) and the TCGA-LIHC cohort. Potential HCC antigens were identified using GEPIA2, cBioPortal, and TIMER2. Anoikis-related subtypes and gene clusters were defined by consensus clustering of 566 liver cancer samples based on 28 anoikis regulators, and we further analyzed their relationship with the immune microenvironment of HCC. A predictive model based on anoikis-related long noncoding RNAs (lncRNAs) was developed to accurately predict HCC prognosis. Seven overexpressed genes associated with HCC prognosis and tumor-infiltrating antigen-presenting cells were identified as potential tumor antigens for the development of HCC mRNA vaccines. Two subtypes based on anoikis-related genes (ARGs) and two gene clusters with different characteristics were identified and validated in defined cohorts. The tumor immune microenvironment between the two subtypes showed different cell infiltration and molecular characteristics. Furthermore, a prognostic score based on seven lncRNAs identified by LASSO regression was constructed, with the low-risk group having favorable prognosis, a “hot” immune microenvironment, and better response to immunotherapy. CCNB1, CDK1, DNASE1L3, KPNA2, PRC1, PTTG, and UBE2S were first identified as promising tumor antigens for mRNA vaccine development in HCC. Besides, we innovatively propose anoikis-based molecular subtypes, which not only enable personalized prognostic stratification of HCC patients but also provide a blueprint for identifying optimal candidates for tumor vaccines, enhancing immunotherapeutic strategies.

In vivo therapeutic effects of affinity-improved-TCR engineered T-cells on HBV-related hepatocellular carcinoma

BackgroundIn patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), virus-specific cytotoxic T lymphocytes (CTLs) fail to eliminate HCC cells expressing HBV antigens. As the expression of viral antigen in...

Upregulation of ubiquitin-conjugating enzyme E2Z is associated with human hepatocellular carcinoma

UBE2Z, a member of ubiquitin-conjugating enzymes, has been reported to participate in multiple biological processes. However, its roles in hepatocellular carcinoma (HCC) remain undiscovered. This study aimed at investigating the functions of UBE2Z in HCC. Firstly, we evaluated UBE2Z expression in HCC and identified associations among UBE2Z expression, clinicopathological features, copy number alterations, DNA methylation, and survival of patients using data from the Cancer Genome Atlas (TCGA). As a result, UBE2Z was remarkably overexpressed in HCC tissues relative to normal liver tissues (P < 0.05). High UBE2Z expression was significantly correlated with age, advanced TNM stage, histological grade, vascular invasion, elevated serum alpha-fetoprotein expression (AFP), worse overall survival (OS) and disease-free survival (DFS) of HCC patients (all P < 0.05). Besides, data mining in UCSC Xena Browser showed that UBE2Z DNA amplification which was significantly associated with its expression was common (108 out of 364) in HCC, and that the level of UBE2Z DNA methylation was negatively associated with its expression (Pearson’s correlation = −0.4, P < 0.0001). After analyzing the datasets from TCGA, we further confirmed the up-regulation of UBE2Z in 60 HCC tissues and several HCC cell lines. Finally, functional assays were performed and showed that knockdown UBE2Z using small interfering RNA (siRNA) could significantly restrain tumor cell proliferation and suppress cell migration and cell invasion through repressing the expression of MMP2 and MMP9. Meanwhile, UBE2Z knockdown could effectively reduce the expression of p-ERK, p-p38, p-JNK, p-Stat3 and p-JAK2, suggesting that UBE2Z might promote HCC progression by targeting ERK and stat3 signaling pathway. These findings implied that UBE2Z might be considered as a prognostic biomarker in HCC and provided a potential therapeutic tumor-associated antigen for HCC.

Biology and significance of alpha-fetoprotein in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common causes of cancer-related deaths globally due, in part, to the majority of patients being diagnosed with intermediate or advanced stage disease. Our increased understanding of the heterogeneous molecular pathogenesis of HCC has led to significant developments in novel targeted therapies. Despite these advances, there remains a high unmet need for new treatment options. HCC is a complex disease with multiple pathogenic mechanisms caused by a variety of risk factors, making it difficult to characterize with a single biomarker. In fact, numerous biomarkers have been studied in HCC, but alpha-fetoprotein (AFP) remains the most widely used and accepted serum marker since its discovery over 60years ago. This review summarizes the most relevant studies associated with the regulation of AFP at the gene and protein levels; the pathophysiology of AFP as a pro-proliferative protein; and the correlation of AFP with molecular HCC subclasses, the vascular endothelial growth factor pathway and angiogenesis. Also described are the historical and current uses of AFP for screening and surveillance, diagnosis, its utility as a prognostic and predictive biomarker and its role as a tumour antigen in HCC. Taken together, these data demonstrate the relevance of AFP for patients with HCC and identify several remaining questions that will benefit from future research.

Construction and assessment of a novel vaccine targeting hepatocellular carcinoma

IMMUNOTHERAPY is a promising and highly effective method of targeting hepatocellular carcinoma (HCC) cells and improving patient outcomes. There is a continuing need for the development and refinement of current vaccine vehicles to safely target tumors while stimulating robust cell-mediated immune responses. The facultative intracellular bacterium Listeria monocytogenes has proven to be an effective vehicle for the design of cancer vaccines that stimulate potent and long CD8+ T-cell responses. Critical facets of its effectiveness relate to the ability of gaining access to the cytosol of infected host cells and delivering tumor-associated antigens. Here we describe the development and testing of L. monocytogenes ΔprsA2 ΔhtrA strains as effective vaccine vehicles for the safe delivery of HCC antigens. Recombinant L. monocytogenes ΔprsA2 ΔhtrA was engineered to express α-fetoprotein antigens designed to elicit immunity against HCC cells, and tested in a subcutaneous mouse model together with the previously developed L. monocytogenes ΔactA prfA* strain. The results suggest that L. monocytogenes ΔprsA2 ΔhtrA strains may represent a highly attenuated yet effective vaccine vector capable of stimulating immunity against HCC cells in vivo.

Abstract 4591: Overexpression of p62/IMP2 can promote cell migration in hepatocellular carcinoma (HCC) via activating Wnt/ β-catenin pathway

Abstract There are three members in the family of insulin-like growth factor 2 mRNA binding proteins (IMP): IMP1, IMP2 and IMP3. The IMPs are oncofetal proteins, expressed during embryogenesis and lost in most tissues in adults. However, IMPs were found overexpressed in various cancers. IMPs share similar protein structure in mammals and have similar functions: to bind specific mRNAs, regulate their expression, transportation, and degradation. Although p62/IMP2 was first reported as a tumor-associated antigen in HCC around 20 years ago, we still know little about the role of p62/IMP2 in HCC progression. Our previous studies found that p62/IMP2 was not only overexpressed in HCC tissues, but also overexpressed in HCC cell lines. To explore the biological roles of p62/IMP2 in HCC progression, p62/IMP2 was knockout in two p62/IMP2 positive HCC cell lines (SNU449, HepG2). Due to the low expression level of p62/IMP2 in SNU449, we performed the transfection experiment to overexpress p62/IMP2 in this cell line. The Wound healing assay and transwell migration assay indicated that overexpressed p62/IMP2 in both cell lines showed the ability that could promote the cell migration significantly (p&amp;lt;0.05). On the contrary, the lack of p62/IMP2 expression can reduce the cell migration ability (P&amp;lt;0.05). After analyzing the HCC expression data from Gene Expression Omnibus (GEO), the high and low IMP2 expression groups were set up based on the median expression of p62/IMP2 from GSE 14520. Three genes were selected in differential gene expression analysis with a cancer-metastasis related gene profile, including CTNNB1, ACTR2, VASP (LogFC &amp;gt; 0.3, Adj. p&amp;lt;0.001). Then we performed a western blotting analysis to explore the effect of overexpressed p62/IMP2 on Wnt/β-catenin pathway-related proteins. Overexpressed p62/IMP2 significantly enhance the expression of Wnt and β-catenin, whereas inhibiting the expression of Gsk3b and E-cadherin. Moreover, the migration ability of SNU449 and HepG2 cells were significantly reduced (p&amp;lt;0.05) after cultured with 10mM Wnt/β-catenin signaling pathway inhibitor XAV939 for 24h. In summary, our data showed that overexpressed p62/IMP2 can enhance the migration ability of HCC cells via activating Wnt/β-catenin signaling pathway. Citation Format: Mengtao Xing, Jianxiang Shi, Pei Li, Jiejie Qin, Xiaojun Zhang, Yangcheng Ma, Xiao Wang, Giulio Francia, Jianying Zhang. Overexpression of p62/IMP2 can promote cell migration in hepatocellular carcinoma (HCC) via activating Wnt/ β-catenin pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4591.

Immunotherapy for hepatocellular carcinoma: recent advances and future perspectives.

The introduction of immunotherapies has been a major development in the treatment of many advanced cancers, including hepatocellular carcinoma (HCC). We are entering a new era of systemic therapy for advanced HCC associated with an explosion of clinical trial activity. Data from phase I/II studies of checkpoint inhibitors in advanced HCC have been promising, with durable objective response rates of approximately 20% seen (in both first- and second-line settings) and acceptable safety profiles (including immune-mediated hepatitis). Phase III studies evaluating anti-programmed cell death protein 1 (anti-PD-1) and anti-programmed cell death ligand 1 (anti-PD-L1) antibodies compared with sorafenib are already underway. The potential synergistic effects of anti-PD-1/anti-PD-L1 when used in combination with agents against other checkpoint molecules, systemic therapies, as well as conventional surgical and locoregional therapies are also being explored in upcoming clinical trials. Aside from this, other strategies to harness the immune system, including chimeric antigen receptor-engineered T cells, natural killer cell therapies, and peptide vaccines directed against HCC antigens have entered phase I/II studies. Current limitations of immunotherapies and areas of future research include the accurate assessment and prediction of tumor response, overcoming the immunosuppressive effects of a hypoxic microenvironment, and the management of immune-related hepatitis in patients who already have limited liver reserve.

Abstract 2027: P62/IMP2 promotes metastasis in hepatocellular carcinoma (HCC) by regulating EMT in different signaling pathway

Abstract Insulin- like growth factor 2 mRNA binding proteins (IMPs) Family contains three members: IMP1, IMP2 and IMP3. IMPs are oncofetal proteins, expressed during embryogenesis and lost in most tissues in adults. Overexpressed IMPs have been reported in various types of cancers. They share similar protein structure in mammals and have similar functions: to bind specific mRNAs, stabilize and extend their half-life. IMP2 was first reported as a tumor-associated antigen in HCC. However, only a few research was done on what a role of p62/IMP2 plays on HCC progression. Previous studies show that IMP2 is overexpressed in HCC tissues and in several human HCC cell lines. Western blotting was performed for different HCC cells has a relatively low expression in SNU449 and higher expression in HepG2 and Hep3b. Stable IMP2 overexpression SNU449 cell line was made by IMP2 cDNA plasmid transfection; stable IMP2 knockout Hep3b and HepG2 cell lines was made by IMP2 Crispr-Cas9 KO plasmid transfection. Wound healing assay has shown that overexpression of p62/IMP2 can significantly enhance cell migration ability in SNU449 cells. On the contrary, the depletion of p62/IMP2 can reduce the migration ability in HepG2 and Hep3b cells (p&amp;lt;0.05). Furthermore, we analyzed HCC gene expression data from TCGA. Median count of IMP2 was used to separate HCC expression data into high and low IMP2 expression group to conduct differential gene expression analysis. Gene Ontology over-representation analysis shows differentially expressed genes (adjusted p&amp;lt;0.05) mainly enriched in membrane part and plasma membrane. The expression of several genes involved in EMT regulation such as CTNNB1, SMAD2, SNAI2, AKT2 shows significant difference (adjust p&amp;lt;0.05). Western blotting assay was performed to show whether IMP2 is involved in EMT regulation. Wnt/beta-catenin pathway and TGF-beta/SMAD pathway were both activated and further upregulated SNAIL, which is an E-cadherin inhibitor. Western blotting results showed that E-cadherin was inhibited in IMP2 overexpressed SNU449 cells, which is supportive to our hypothesis. Citation Format: Mengtao Xing, Jianxiang Shi, Jitian Li, Beibei Chen, Giulio Francia, Jianying Zhang. P62/IMP2 promotes metastasis in hepatocellular carcinoma (HCC) by regulating EMT in different signaling pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2027.

δ-Catenin peptide vaccines repress hepatocellular carcinoma growth via CD8+ T cell activation

ABSTRACTAs classical therapy method of advanced hepatocellular carcinoma (HCC) is not effective enough, HCC immunotherapy is a hot spot for research in recent years. Although in recent years, immune checkpoint inhibitors are focused in cancer therapy, vaccines and adoptive cell therapy (ACT), as traditional immunotherapy methods for HCC are still promising. We found that δ-Catenin might be a new tumor-associated antigen for HCC, for it could be upregulated as a stress associated protein under hypoxia and irradiation treatment. δ-Catenin peptide vaccines could inhibit the growth of subcutaneous hepatocellular tumors in vivo. According to our work, δ-Catenin peptide vaccines could stimulate the activation of cytotoxic T lymphocytes (CTLs) and enhance the infiltration of CD8+ T cells into tumors. Moreover, δ-Catenin peptide vaccines could enhance the secretion of IFN-γ and the killing of tumor cells by T cells. Mechanistically, δ-Catenin peptide vaccines, presented by antigen-presenting cells to T cells, could enhance the activation of T cells via MAPK/ERK signaling and the transcriptional factors Eomes and T-bet. Our research results indicate new potential peptide vaccines, which can be applied in clinical HCC therapy.

Phase 1 studies of 3- and 6-peptide vaccine cocktail derived from GPC3, WDRPUH, and NEIL3 in patients with advanced hepatocellular carcinoma (HCC).

387 Background: Glypican 3 (GPC3), WD-Repeat-containing Protein Up-regulated in HCC (WDRPUH), and Nei Endonuclease VIII-Like 3 (NEIL3) are tumor antigens in HCC. We conducted two phase 1 multicenter open-label studies for advanced HCC: one to evaluate the HLA-A*24-restricted 3-epitope peptide vaccine cocktail named ONO-7268MX1 (MX1); the other to evaluate the HLA-A*24 and -A*02-restricted 6-epitope peptide vaccine cocktail named ONO-7268MX2 (MX2). Both vaccines contain epitope peptides derived from GPC3, WDRPUH, and NEIL3. MX1 and MX2 cover most of Japanese HCC patients (pts). Safety is the primary, and efficacy and immunological response are the exploratory objectives. Methods: Pts received SC MX1 or MX2 once weekly. In MX1 study, pts received each of 3 peptides at 0.3 mg (cohort 1), 1 mg (cohort 2), and 3 mg of GPC3, 1.1 mg of WDRPUH and 3 mg of NEIL3 (cohort 3). In MX2 study, pts received each of 6 peptides at 1 mg. Results: In MX1 study, 3, 8, and 7 pts with HLA-A*24 were enrolled to cohort 1, 2, and 3, respectively. In MX2 study, 14 pts (7 with HLA-A*24, 6 with HLA-A*02, and 1 with HLA-A*24/A*02) were enrolled. No dose limiting toxicities were observed in both studies. The most commonly related AEs were Grade 1/2 injection site reaction (erythema, induration, pain, and pruritus), 44.4% (8/18) in MX1 and 78.6% (11/14) in MX2. Grade 3-related AEs occurred only in MX2 study; neutrophil count decreased, white blood cell count decreased, and decreased appetite (each 1 patient). No grade 4-related AEs occurred in both studies. No response (CR or PR) were seen in both studies. In MX1 study, median and mean of PFS [range], median duration of SD were 2.8, 3.4 [0.5-16.8], 3.7 months respectively, in MX2 study those were 1.9 and 2.8 [0.4-8.3], 5.7 respectively. In both studies, some pts showed long disease control (≥6 months). For those pts, each peptide-specific Cytotoxic T Lymphocyte (CTL) responses were shown by the ELISPOT assay. Conclusions: MX1 and MX2 showed good tolerability. Both vaccines induced CTL responses which may be related to antitumor effect. In addition, some pts showed long disease control. These results implied the potential usefulness of these vaccines for the treatment of HCC. Clinical trial information: JapicCTI-121933 and JapicCTI-142477.

Host antitumor resistance improved by the macrophage polarization in a chimera model of patients with HCC

ABSTRACTDespite major advances in curative and palliative approaches, hepatocellular carcinoma (HCC) is still the third leading cause of cancer-related death worldwide. M1 macrophages (Mφ) play a key role in host antitumor defenses in HCC. In our study, CD14+ cells were isolated from the peripheral blood of four groups of HCC patients (group-1, patients with stage 0 HCC; group-2, patients with stage A HCC; group-3, patients with stage B HCC; and group-4, patients with stage C HCC) and characterized phenotypically. Then, CD14+ cells from group-2 and group-3 HCC patients were induced to polarize and tested for their antitumor abilities in a chimera model of HCC patients. Human HCCs (HepG2 solid tumors) grew in a chimera model of group-3 patients (group-3 HCC chimeras) but not in a chimera model of group-2 patients (group-2 HCC chimeras). In response to HCC antigens, the majority of CD14+ cells from group-2 patients (group-2 CD14+ cells) switched to the M1 phenotype (IL-12+IL-10−iNOS+cells), whereas the majority of CD14+ cells from group-3 patients (group-3 CD14+ cells) did not switch to the M1 phenotype and continued to express M2b phenotypic properties (IL-12−IL-10+CCL1+iNOS−cells). Group-3 CD14+ cells showed M1Mφ polarization after treatment with CCL1 antisense oligodeoxynucleotide (ODN). Therefore, our study indicates that anti-HCC defenses of group-3 HCC chimeras are improved after CCL1 antisense ODN treatment.

Tumor-associated autoantibodies are useful biomarkers in immunodiagnosis of α-fetoprotein-negative hepatocellular carcinoma.

AIMTo determine the prevalence and diagnostic value of autoantibodies in α-fetoprotein (AFP)-negative hepatocellular carcinoma (HCC).METHODSFifty-six serum samples from AFP-negative HCC cases, 86 from AFP-positive HCC cases, 168 from chronic liver disease cases, and 59 from normal human controls were included in this study. Autoantibodies to nucleophosmin (NPM)1, 14-3-3zeta and mouse double minute 2 homolog (MDM2) proteins in AFP-negative HCC serum were evaluated by enzyme-linked immunosorbent assay. Partially positive sera were further evaluated by western blotting. Immunohistochemistry was used to detect the expression of three tumor-associated antigens (TAAs) in AFP-negative HCC and normal control tissues.RESULTSThe frequency of autoantibodies to the three TAAs in AFP-negative HCC sera was 21.4%, 19.6% and 19.6%, which was significantly higher than in the chronic liver disease cases and normal human controls (P < 0.01) as well as AFP-positive HCC cases. The sensitivity of the three autoantibodies for diagnosis of AFP-negative HCC ranged from 19.6% to 21.4%, and the specificity was approximately 95%. When the three autoantibodies were combined, the sensitivity reached 30.4% and the specificity reached 91.6%.CONCLUSIONAutoantibodies to NPM1, 14-3-3zeta and MDM2 may be useful biomarkers for immunodiagnosis of AFP-negative HCC.

Anti-Cadherin-17 Antibody Modulates Beta-Catenin Signaling and Tumorigenicity of Hepatocellular Carcinoma

Cadherin-17 (CDH17) is an oncofetal molecule associated with poor prognostic outcomes of hepatocellular carcinoma (HCC), for which the treatment options are very limited. The present study investigates the therapeutic potential of a monoclonal antibody (Lic5) that targets the CDH17 antigen in HCC. In vitro experiments showed Lic5 could markedly reduce CDH17 expression in a dose-dependent manner, suppress β-catenin signaling, and induce cleavages of apoptotic enzymes caspase-8 and -9 in HCC cells. Treatment of animals in subcutaneous HCC xenograft model similarly demonstrated significant tumor growth inhibition (TGI) using Lic5 antibody alone (5 mg/kg, i.p., t.i.w.; ca.60–65% TGI vs. vehicle at day 28), or in combination with conventional chemotherapy regimen (cisplatin 1 mg/kg; ca. 85–90% TGI). Strikingly, lung metastasis was markedly suppressed by Lic5 treatments. Immunohistochemical and western blot analyses of xenograft explants revealed inactivation of the Wnt pathway and suppression of Wnt signaling components in HCC tissues. Collectively, anti-CDH17 antibody promises as an effective biologic agent for treating malignant HCC.

Naturally occurring, tumor-specific CD8+ T cells in patients with hepatocellular carcinoma display a clear immunodominance

Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and ranks third in mortality. This is partly due to a limited number of available therapeutic approaches. Since several studies found that the immune response directed against the tumor is associated with prognosis, immunotherapy is a potential future way of treatment. To develop such immune-based therapies, a better understanding of the natural immune response against HCC is obligatory. To address this issue, we used overlapping peptides covering the entire sequence of four prominent tumor-associated antigens (TAAs) in HCC, i.e. α-fetoprotein (AFP), Glypican–3, melanoma-associated gene-A1 (MAGE-A1) and NY-ESO–1. CD8+ T cells obtained from blood and tumor samples of a large cohort of HCC patients (n=96) as well as from the blood of controls (n=19) were stimulated with these peptides and analyzed for the production of interferon-γ. We found that TAA-specific immune responses were readily detectable in HCC patients at a significantly higher frequency than in controls (p=0.0017). We could identify a clear hierarchy between the individual antigens and several immunodominant regions within the respective TAAs. In addition, several responses were fine-mapped, leading to the identification of novel dominant tumor-specific epitopes. A further tetramer-analysis revealed that the tumor-specific CD8+ T cells could expand specifically in vitro but remained dysfunctional with respect to effector functions. In sum, our results indicate that HCC induces a broad immune response targeting several antigens and epitopes. However, the tumor-specific CD8+ T cells are clearly functionally impaired. These results indicate that insufficient immune-mediated control of HCC is not due to a lack of priming of CD8+ T cell responses but rather associated with impaired effector functions of these cells. These results have important implications for the further development of immune-based therapies for HCC.

Dendritic cells-mediated CTLs targeting hepatocellular carcinoma stem cells

Immunotherapy, especially using dendritic cells (DCs)-based vaccine, appears promising in the treatment of hepatocellular carcinoma (HCC) following surgery. However, the therapeutic efficacy of current DC vaccines loaded with HCC antigen is limited in clinical practice. One important reason might be that the DC vaccines for the treatment of HCC were not aimed at targeting the hepatocellular carcinoma cancer stem cells (HCCCSCs). Therefore, establishing an immunotherapy to kill HCC stem cells could be a novel therapeutic strategy. In this study, we have developed an immunotherapy to target CD133+ HCC cells in the treatment of HCC. This study had three main findings; (1), CD133+HCC cells RNA loaded DCs could induce special CD8+ cytotoxic T lymphocytes (CD133+Huh7-CTLs) response against CD133+ Huh7 cells in vitro. (2), Huh7 cells-induced tumor growth in vivo was effectively inhibited by CD133+Huh7-CTLs. (3), the great inhibition potential of CD133+Huh7-CTLs to Huh7-induced tumor growth might not be only associated with anti-tumor cytokines such as IFN-γ, but also to CD133+Huh7-DCs induced specific CTLs. This study shows an experimental proof that CD133+HCC cells RNA loaded DC vaccine has potential in treating HCC and may provide a new therapy for clinical post operative adjuvant therapy in future.

Dendritic cells fused with allogeneic hepatocellular carcinoma cell line compared with fused autologous tumor cells as hepatocellular carcinoma vaccines

To investigate the specific antitumor responses against autologous hepatocellular carcinoma (HCC) cells of dendritic cells (DCs) fused with allogeneic HCC cell line, and evaluated the feasibility of BEL7402 as an alternative strategy to deliver shared HCC antigens to DCs. Previous studies demonstrated fusions of patient-derived DCs and autologous tumor cells could induce T-cell responses against autologous tumors. These fusion cells require patient-derived tumor cells, which are not, however, always available. Here, we report the fusing of autologous DCs with allogeneic HCC cell line to induced cytotoxic T-lymphocyte (CTL) response against autologous HCC cells compare with autologous tumor cells. These DC/ BEL7402 fusion cells co-expressed tumor-associated antigens and DC-derived costimulatory and major histocompatibility complex molecules. Both CD4+ and CD8 T+ cells were activated by the fusion cells as demonstrated by the proliferation of T-cells, the production of cytokines and the simultaneous induction of specific CTL responses. Significantly, CTL induced by dendritic cell/allogeneic BEL7402 fusion cells were able to kill autologous HCC cells by human leukocyte antigen-A2 restricted mechanisms. The results did not show significant difference between DC fusion with autologous hepatocellular carcinoma cells and DC fusion with allogeneic hepatocellular carcinoma cell line. The fusion of allogeneic HCC cell line and autologous DCs may have applications in antitumor immunotherapy through cross-priming against shared tumor antigens and may provide a platform for adoptive immunotherapy.

AFP-specific CD4+ T-cell immunity in the blood and liver of patients with Hepatocellular Carcinoma

Background: Hepatocellular Carcinoma (HCC) is one of the most common malignancies with limited therapeutic options. Alpha-fetoprotein (AFP) is a tumor-associated antigen in HCC and is a target for the development of cancer vaccine. Until recently, most studies on HCC have analyzed AFP-specific CD8+ T-cell responses. However, less is known about AFP-specific CD4+ T-cell responses.

Patterns of Hepatoblastoma and Hepatocellular Carcinoma in Children After Universal Hepatitis B Vaccination in Taiwan

To retrospectively evaluate clinical features, treatment, and outcome of patients with hepatoblastoma (HB) and hepatocellular carcinoma (HCC). From January 1994 to December 2007, 16 patients of HB and 13 cases of HCC were reviewed. The mean age of HB patients was much younger than HCC patients (1.2 vs. 11.5 y). There was an 84.6% positive rate for hepatitis B surface antigen in HCC, but none for HB. Mean serum alpha-fetoprotein level was higher with HCC (654,158 ng/mL) than the HB patients (352,843 ng/mL), especially higher in HCC with lung metastasis. Among the HB patients, 12 (75%) had thrombocytosis and 6 (37.5%) had microcytic anemia with high or normal ferritin, whereas only 3 of 13 with HCC (23.1%) had thrombocytosis and none had microcytic anemia. All HBs were resectable either before or after chemotherapy, but only 4 (30.8%) HCCs were resectable. Five-year disease-free survival rate was significantly higher in HB (87%) than in HCC (30%, P<0.001). Hepatitis B infection was still the most important factor associated with HCC in children even after the national vaccination program against hepatitis B. Extreme thrombocytosis, anemia, alpha-fetoprotein levels are important factors associated with difference in long-term outcomes in children with HB and HCC.

Increased Immunostimulatory Activity Conferred to Antigen-presenting Cells by Exposure to Antigen Extract From Hepatocellular Carcinoma After Radiofrequency Thermal Ablation

Radiofrequency thermal ablation represents an effective treatment for hepatocellular carcinoma (HCC) and it can also exert an "adjuvant" effect on spontaneous antitumor T-cell responses, as suggested by human and animal studies. The adjuvant effect is thought to depend on the huge amount of necrotic tumor antigen made available to the immune system by HCC thermal ablation. In addition, radiofrequency thermal ablation may result in the release of local stimuli responsible for activation and maturation of antigen-presenting cells (APCs). To test this hypothesis, we studied APC maturation and function in 19 patients undergoing thermal ablation for HCC. Patients' monocytes induced to differentiate with granulocyte macrophage colony-stimulating factor (GM-CSF), or GM-CSF plus IL-4, were cocultured in vitro with tumor debris generated by radiofrequency thermal ablation. Expression of costimulatory molecules, lymphnode homing chemokine receptor, antigen presentation, and cytokine secretion were enhanced by incubation with HCC treated tissue as compared with untreated HCC and nontumor liver tissue. Moreover, HCC-specific T-cell responses could be induced by monocytes activated with GM-CSF and incubated with thermally ablated HCC tissue. HCC thermal ablation can create an antigenic source along with stimuli appropriate for maturation of APCs to induce HCC-specific T-cell responses. These results contribute to explain at least in part the adjuvant effect of HCC thermal ablation and suggest a novel strategy to induce maturation of APCs and their loading with HCC antigens for active immunotherapy protocols aimed at reducing HCC recurrence after thermal ablation.