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： To investigate the association of cyclooxygenase-2 （COX-2） expression with angiogenesis and the number and type of inflammatory cells （macrophages/Kupffer cells; mast cells） within primary hepatocellular car...AIM： To investigate the association of cyclooxygenase-2 （COX-2） expression with angiogenesis and the number and type of inflammatory cells （macrophages/Kupffer cells; mast cells） within primary hepatocellular carcinoma （HCC） tissues and adjacent non-tumorous （NT） tissues. METHODS： Immunohistochemistry for COX-2, CD34, CD68 and mast cell tryptase （MCT） was performed on 14 well-characterized series of liver-cirrhosis-associated HCC patients. COX-2 expression and the number of inflammatory cells in tumor lesions and surrounding liver tissues of each specimen were compared. Moreover, COX-2, CD34 staining and the number of inflammatory cells in areas with different histological degrees within each tumor sample were comparatively analyzed. RESULTS： The percentage of COX-2 positive cells was significantly higher in NT tissues than in tumors. COX-2 expression was higher in well-differentiated HCC than in poorly-differentiated tissues. Few mast cells were observed within the tumor mass, whereas a higher number was observed in the surrounding tissue, especially in peri-portal spaces of NT tissues. Abundant macrophages/ Kupffer cells were observed in NT tissues, whereas the number of cells was significantly lower in the tumor mass. However, a higher cell number was observed in the welldifferentiated tumor and progressively decreased in relation to the differentiation grade. Within the tumor, a positive correlation was found between COX-2 expression and the number of macrophages/Kupffer cells and mastcells. Moreover, there was a positive correlation between CD34 and COX-2 expression in tumor tissues. Comparison between well- and poorly-differentiated HCC showed that the number of CD34-positive cells decreased with dedifferentiation. However, COX-2 was the only independent variable showing a positive correlation with CD34 in a multivariate analysis. CONCLUSION： The presence of inflammatory cells and COX-2 expression in liver tumor suggests a possible relationship with tumor angiogenesis. COX-2 expr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.
基金Supported by the MIUR and Progetto Strategico Oncologia "Terapia Preclinica Moleculare Oncologia" MIUR-CNR
文摘AIM： To investigate the association of cyclooxygenase-2 （COX-2） expression with angiogenesis and the number and type of inflammatory cells （macrophages/Kupffer cells; mast cells） within primary hepatocellular carcinoma （HCC） tissues and adjacent non-tumorous （NT） tissues. METHODS： Immunohistochemistry for COX-2, CD34, CD68 and mast cell tryptase （MCT） was performed on 14 well-characterized series of liver-cirrhosis-associated HCC patients. COX-2 expression and the number of inflammatory cells in tumor lesions and surrounding liver tissues of each specimen were compared. Moreover, COX-2, CD34 staining and the number of inflammatory cells in areas with different histological degrees within each tumor sample were comparatively analyzed. RESULTS： The percentage of COX-2 positive cells was significantly higher in NT tissues than in tumors. COX-2 expression was higher in well-differentiated HCC than in poorly-differentiated tissues. Few mast cells were observed within the tumor mass, whereas a higher number was observed in the surrounding tissue, especially in peri-portal spaces of NT tissues. Abundant macrophages/ Kupffer cells were observed in NT tissues, whereas the number of cells was significantly lower in the tumor mass. However, a higher cell number was observed in the welldifferentiated tumor and progressively decreased in relation to the differentiation grade. Within the tumor, a positive correlation was found between COX-2 expression and the number of macrophages/Kupffer cells and mastcells. Moreover, there was a positive correlation between CD34 and COX-2 expression in tumor tissues. Comparison between well- and poorly-differentiated HCC showed that the number of CD34-positive cells decreased with dedifferentiation. However, COX-2 was the only independent variable showing a positive correlation with CD34 in a multivariate analysis. CONCLUSION： The presence of inflammatory cells and COX-2 expression in liver tumor suggests a possible relationship with tumor angiogenesis. COX-2 expr
