AIM:To investigate the influence of autologous cytokine- induced killer (CIK) cells on the phenotypes of CIK effector cells,peripheral T lymphocyte subsets and dendritic cell subsets in patients with primary hepatocel...AIM:To investigate the influence of autologous cytokine- induced killer (CIK) cells on the phenotypes of CIK effector cells,peripheral T lymphocyte subsets and dendritic cell subsets in patients with primary hepatocellular carcinoma (HCC). METHODS:Peripheral blood mononuclear cells (PBMC) were collected by a blood cell separator from 13 patients with HCC,then expanded by priming them with interferon- gamma (IFN-γ) followed by monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2) the next day.The phenotypic patterns of CIK cells were characterized by flow cytometry on d 0,4,7,10,13 and 15 of incubation, respectively.Then,5 mL of venous blood was obtained from HCC patients before or 8-10 d after CIK cells were transfused into patients to assess the influence of CIK cells on the percentages of effector cells,and proportions of DC1 or DC2 in peripheral blood by flow cytometry. RESULTS:After two weeks of in vitro incubation,the percentages of CD3^+CD8^+,CD3^+CD56^+,and CD25^+ cells increased significantly from 33.5±10.1%,7.7±2.8%,and 12.3±4.5% to 36.6±9.0% (P<0.05),18.9±6.9% (P<0.01), and 16.4±5.9% (P<0.05),respectively.However,the percentages of CD3^+CD4^+ and NK cells had no significant difference.The percentages of CD3^+ and CD3^+CD8^+ cells were kept at high levels during the whole incubation period,but those of CD25^+,and CD3^+CD56^+ cells began to decrease on d 7 and 13,respectively.The proportions of type Ⅰ dendritic cell (DC1) and type Ⅱ dendritic cell (DC2) subsets increased from 0.59±0.23% and 0.26±0.12% before CIK cell therapy to 0.85±0.27% and 0.43±0.19% (all P<0.01) after CIK cell transfusion,respectively.The symptoms and characteristics of HCC patients were relieved without major side effects. CONCLUSION:Our results indicated that autologous CIK cells can efficiently improve the immunological status in HCC patients,and may provide a potent approach for HCC patients as the adoptive immunotherapy.展开更多
AIM: This study investigated the anti-cancer effect ofcombined quercetin and a recombinant adenovirus vectorexpressing the human p53, GM-CSF and B7-1 genes(designated BB-102) on human hepatocellular carcinoma(HCC) cel...AIM: This study investigated the anti-cancer effect ofcombined quercetin and a recombinant adenovirus vectorexpressing the human p53, GM-CSF and B7-1 genes(designated BB-102) on human hepatocellular carcinoma(HCC) cell lines in vitro.METHODS: The sensitivity of HCC cells to anticancer agentswas evaluated by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The viability of cells infectedwith BB-102 was determined by trypan blue exclusion. Theexpression levels of human wild-type p53, GM-CSF and B7-1genes were determined by Western blot, enzyme-linkedimmunosorbent assay (ELISA) and flow cytometric analysis,respectively. The apoptosis of BB-102-infected or quercetin-treated HCC cells was detected by terminal deoxynucleotidyltransferase (TdT) assay or DNA ladder electrophoresis.RESULTS: Quercetin was found to suppress proliferation ofhuman HCC cell lines BEL-7402, HUH-7 and HLE, with peaksuppression at 50 μmol/L quercetin. BB-102 infection wasalso found to significantly suppress proliferation of HCC celllines. The apoptosis of BB-102-infected HCC cells was greaterin HLE and HUH-7 cells than in BEL-7402 cells. Quercetin didnot affect the expression of the three exogenous genes inBB-102-infected HCC cells (P＞0.05), but it was found to furtherdecrease proliferation and promote apoptosis of BB-102-infected HCC cells.CONCLUSION: BB-102 and quercetin synergeticallysuppress HCC cell proliferation and induce HCC cell apoptosis,suggesting a possible use as a combined anti-cancer agent.展开更多
文摘AIM:To investigate the influence of autologous cytokine- induced killer (CIK) cells on the phenotypes of CIK effector cells,peripheral T lymphocyte subsets and dendritic cell subsets in patients with primary hepatocellular carcinoma (HCC). METHODS:Peripheral blood mononuclear cells (PBMC) were collected by a blood cell separator from 13 patients with HCC,then expanded by priming them with interferon- gamma (IFN-γ) followed by monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2) the next day.The phenotypic patterns of CIK cells were characterized by flow cytometry on d 0,4,7,10,13 and 15 of incubation, respectively.Then,5 mL of venous blood was obtained from HCC patients before or 8-10 d after CIK cells were transfused into patients to assess the influence of CIK cells on the percentages of effector cells,and proportions of DC1 or DC2 in peripheral blood by flow cytometry. RESULTS:After two weeks of in vitro incubation,the percentages of CD3^+CD8^+,CD3^+CD56^+,and CD25^+ cells increased significantly from 33.5±10.1%,7.7±2.8%,and 12.3±4.5% to 36.6±9.0% (P<0.05),18.9±6.9% (P<0.01), and 16.4±5.9% (P<0.05),respectively.However,the percentages of CD3^+CD4^+ and NK cells had no significant difference.The percentages of CD3^+ and CD3^+CD8^+ cells were kept at high levels during the whole incubation period,but those of CD25^+,and CD3^+CD56^+ cells began to decrease on d 7 and 13,respectively.The proportions of type Ⅰ dendritic cell (DC1) and type Ⅱ dendritic cell (DC2) subsets increased from 0.59±0.23% and 0.26±0.12% before CIK cell therapy to 0.85±0.27% and 0.43±0.19% (all P<0.01) after CIK cell transfusion,respectively.The symptoms and characteristics of HCC patients were relieved without major side effects. CONCLUSION:Our results indicated that autologous CIK cells can efficiently improve the immunological status in HCC patients,and may provide a potent approach for HCC patients as the adoptive immunotherapy.
文摘AIM: This study investigated the anti-cancer effect ofcombined quercetin and a recombinant adenovirus vectorexpressing the human p53, GM-CSF and B7-1 genes(designated BB-102) on human hepatocellular carcinoma(HCC) cell lines in vitro.METHODS: The sensitivity of HCC cells to anticancer agentswas evaluated by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The viability of cells infectedwith BB-102 was determined by trypan blue exclusion. Theexpression levels of human wild-type p53, GM-CSF and B7-1genes were determined by Western blot, enzyme-linkedimmunosorbent assay (ELISA) and flow cytometric analysis,respectively. The apoptosis of BB-102-infected or quercetin-treated HCC cells was detected by terminal deoxynucleotidyltransferase (TdT) assay or DNA ladder electrophoresis.RESULTS: Quercetin was found to suppress proliferation ofhuman HCC cell lines BEL-7402, HUH-7 and HLE, with peaksuppression at 50 μmol/L quercetin. BB-102 infection wasalso found to significantly suppress proliferation of HCC celllines. The apoptosis of BB-102-infected HCC cells was greaterin HLE and HUH-7 cells than in BEL-7402 cells. Quercetin didnot affect the expression of the three exogenous genes inBB-102-infected HCC cells (P＞0.05), but it was found to furtherdecrease proliferation and promote apoptosis of BB-102-infected HCC cells.CONCLUSION: BB-102 and quercetin synergeticallysuppress HCC cell proliferation and induce HCC cell apoptosis,suggesting a possible use as a combined anti-cancer agent.
