Antigens, Neoplasm ; immunology ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; immunology ; therapy ; Humans ; Immunotherapy ; Liver Neoplasms ; immunology ; therapy

This study investigated the changes of CD4(+) CD25(+) regulatory T cells (Tregs) in peripheral blood of patients with hepatocellular carcinoma before and after transcatheter arterial chemoembolization (TACE). The proportion of CD4(+) CD25(+) Tregs among CD4(+) T lymphocytes in peripheral blood of 33 patients with hepatocellular carcinoma was determined by flow cytometry before, 1 week and 1 month after TACE. And 25 healthy volunteers served as control. One month after TACE, the patients were divided into two groups: 22 in group A, who were in stable condition or getting better; and 10 in group B, who were deteriorating. One patient died and was excluded. The results showed that the percentage of CD4(+)CD25(+) Tregs among CD4(+) T lymphocytes did not significantly change in the 33 patients 1 week after TACE as compared with that before TACE, however, the difference was significant (P<0.01) between the patients with hepatocellular carcinoma and the healthy subjects. The percentage of CD4(+) CD25(+) Tregs among CD4(+) T lymphocytes in group A 1 month after TACE was decreased significantly in comparison with that before and 1 week after TACE (P<0.01), whereas, that in group B was increased significantly 1 month after TACE (P<0.01). It was concluded that patients with hepatocellular carcinoma had a higher proportion of CD4(+)CD25(+) Tregs in peripheral blood. TACE did not significantly affect the level of CD4(+) CD25(+) Tregs within short time (such as 1 week). The proportion of CD4(+)CD25(+) Tregs in peripheral blood 1 month after TACE was related to the prognosis of hepatocellular carcinoma.
Antineoplastic Agents/administration & dosage ; Carcinoma, Hepatocellular/*immunology ; Carcinoma, Hepatocellular/therapy ; Chemoembolization, Therapeutic/*methods ; Liver Neoplasms/*immunology ; Liver Neoplasms/*therapy ; T-Lymphocytes, Regulatory/*immunology

CD4-Positive T-Lymphocytes ; immunology ; CD56 Antigen ; analysis ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; immunology ; Dendritic Cells ; immunology ; Humans ; Liver Neoplasms ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

B7-1 Antigen ; genetics ; immunology ; Carcinoma, Hepatocellular ; genetics ; immunology ; Enterotoxins ; genetics ; immunology ; Genetic Vectors ; Humans ; Liver Neoplasms ; genetics ; immunology ; T-Lymphocytes ; immunology ; Transfection ; Tumor Cells, Cultured

OBJECTIVESTo obtain a single-chain antibody with high affinity against hepatocellular carcinoma (HCC).

METHODSA second single-chain antibody mutant library was established using an error-prone PCR and a phage display. Single-chain antibodies with high affinity for hepatocellular carcinoma were selected using ELISA.

RESULTSThe content of the second single-chain antibody mutant library was about 4.5 x 10(7). Two selected mutants, M90 and M116, were obtained after 3 rounds of panning and ELISA. Immunoassay showed that both M90 and M116 could bind to human HCC cells. The relative affinity of M90 was 1.7-fold higher than that of the original antibody, and M116 was 2-fold higher than that of the original antibody.

CONCLUSIONError-prone PCR is an effective and simple method for affinity maturation of antibodies isolated from a phage antibody library.

Antibodies, Neoplasm ; immunology ; Antibody Affinity ; Antibody Specificity ; Carcinoma, Hepatocellular ; immunology ; pathology ; Humans ; Immunoglobulin Fragments ; immunology ; Immunoglobulin Variable Region ; genetics ; immunology ; Liver Neoplasms ; immunology ; pathology ; Mutation ; Peptide Library

OBJECTIVETo investigate the ultrastructure of small artery wall in patients with spontaneous rupture of hepatocellular carcinoma (HCC).

METHODSTransmission electron microscopy was used to study 11 specimens from ruptured HCC and 11 cases with non-ruptured HCC.

RESULTSThe phenomenon of activated phagocytosis in macrophage could be found in 3 cases with ruptured HCC and 10 cases with non-ruptured HCC, respectively (P < 0.05). In 9 specimens with ruptured HCC, the evidence of vascular injury characterized as less cell junctions and larger fenestrae in endothelial cells, broken elastic lamina, proliferated and fragmented elastin and damaged structure of collagen was found in small arteries. The phenomenon of electron-dense deposit in the elastic lamina, and signs of more protein synthesis in endothelial cells were also present in these specimens. In the patients with non-ruptured HCC, the evidence of vascular injury can be found only in 2 cases (P < 0.01). Less cell junctions and larger fenestrae could increase the permeability of vascular wall. The electron-dense deposition in elastic lamina may represent the deposition of antigen-antibody complex in elastic membrane which had been found in our previous study. The vascular injury was postulated to be caused by the deposition of antigen-antibody complex in vascular wall which was identified by our previous study. The vascular wall in the patient with ruptured HCC could become stiff and weak due to the proliferated fragment elastin and damaged collagen which would make the blood vessels more prone to splitting and result in hemorrhage and the rupture of HCC.

CONCLUSIONSThe vascular injury caused by antigen-antibody complex deposition might related to the spontaneous rupture of HCC.

Carcinoma, Hepatocellular ; pathology ; ultrastructure ; Humans ; Liver Neoplasms ; pathology ; ultrastructure ; Macrophages ; immunology ; Microscopy, Electron ; Rupture, Spontaneous ; pathology

As a functionally unique subset of T cells, regulatory T cells (Treg) suppress tumor immune responses effectively through a variety of mechanisms and play an important role in tumorigenesis and tumor progression. There is growing evidence to suggest that Treg participates in the formation and development of hepatic tumor, especially the HCC. Elucidation of the mechanisms for involvement of Treg in HCC progression may provide new ideas for liver cancer therapy through a point of view regarding immunology.
Carcinoma, Hepatocellular ; immunology ; Disease Progression ; Humans ; Liver Neoplasms ; T-Lymphocytes, Regulatory

OBJECTIVETo explore the clinical significances of dendritic cells and lymphocytes infiltration in hepatocellular carcinoma (HCC) tissue.

METHODSClinicopathological data were collected from 44 patients with HCC who had under/gone curative tumor resection in our hospital. Immunohistochemical staining was used to detect the infiltration of dendritic cells in the tumor tissue, and lymphocytes infiltration was assessed simultaneously. The correlation between the infiltration of dendritic cells and lymphocytes and postoperative tumor recurrence and survival rate was analyzed.

RESULTSTumor recurrence was markedly late in patients with dendritic cells count >/= 20 and positive lymphocytes infiltration (group A, n = 17) as compared with those who did not meet both criteria simultaneously (group B, n = 27), with a median interval of 21.6 months for group A and 4.1 months for group B (U value = 105.5, P = 0.009). The 1-, 3-, 4-year survival rates were significantly greater in group A than in group B; they were 83.5% vs. 42.2%, 61.8% vs. 28.4% and 48.7% vs. 23.0%, respectively (Log rank = 7.68, P < 0.01).

CONCLUSIONThe infiltration of dendritic cells and lymphocytes in HCC tissue, as an independent prognostic factor, was closely related to postoperative prognosis.

Adult ; Aged ; Carcinoma, Hepatocellular ; immunology ; Cell Count ; Dendritic Cells ; immunology ; Female ; Humans ; Liver Neoplasms ; immunology ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Middle Aged ; Neutrophil Infiltration ; Prognosis ; Recurrence ; Secondary Prevention

This study was aimed to investigate the proliferation activities and phenotype changes of DC, CIK and DC-CIK, and their cytotoxicity against hepatocarcinoma cells in co-culture of DC with CIK. Peripheral blood mononuclear cells (PBMNC) were isolated from healthy adult donors. After incubation of PBMNC for 2 hours, DCs were induced from adherent cells by some cytokines and CIKs were generated from non-adherent cells. Mature DCs were harvested after incubation for 9 days, and then were co-cultured with CIK at ratio of 1:5 for 3 days. The cytotoxicity activity against SMMC-7721 hepatocellular carcinoma cell line was detected by MTT assay. The results showed that CIK cells were able to lyse SMMC-7721 hepatocellular carcinoma cells at low ratios of effector to target. This effect was significantly enhanced by co-culture with DCs. It is concluded that CIK cells have high lytic activity against 7721 hepatocellular carcinoma cell line, which can be enhanced by co-culture with DC. DC-CIK cells are highly effective immune cells.
Carcinoma, Hepatocellular ; immunology ; pathology ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Humans ; Killer Cells, Lymphokine-Activated ; cytology ; immunology ; Liver Neoplasms ; immunology ; pathology

OBJECTIVETo separate and identify the exosomes derived from a mouse hepatoma carcinoma cell line (H22) and to detect their protein composition, and to investigate the possibility of using these exosomes as a kind of tumor vaccine.

METHODSExosomes were purified by serial ultracentrifugation and sugar density ultracentrifugation, and then they were observed and identified by electron microscopy. Exosomes underwent peptide mass fingerprint and Western blot analyses.

RESULTSH22 cell-derived exosomes were 20-90 nm round or oval vesicles. The exosomes expressed HSP70, ICAM-1, EF-G2, DLC-A, C-myc protein and Vav-2 protein.

CONCLUSIONSerial ultracentrifugation and sugar density ultracentrifugation can be used to purify H22 cell-derived exosomes. H22 cell-derived exosomes express a distinct set of proteins involving in and/or relating to antigen presentation (HSP70, ICAM-1), migration (DLC-A), adhesion (ICAM-1), cytoskeleton (EF-G2) and tumour antigens (C-myc, Vav-2). The exosomes have immunogenicity.

Animals ; Carcinoma, Hepatocellular ; immunology ; metabolism ; Cell Line, Tumor ; Exosomes ; immunology ; secretion ; Histocompatibility Antigens Class II ; immunology ; Liver Neoplasms ; immunology ; metabolism ; Mice ; Peptide Mapping