摘要
AIM: To observe the biological specialization of humanperipheral blood dendritic cells (DC) and cord blood derivedDC and its effects on effector cells killing humanhepatocarcinoma cell line BEL-7402 in vitro.METHODS: The DC biological characteristics were detectedwith immunohistochemical and MTT assay. Two antitumorexperimental groups are: peripheral blood DC and cordblood DC groups. Peripheral blood DC groups used LAKcells as the effector cells and BEL-7402 as target cells, whilecord blood DC groups used CTL induced by tumor antigentwice pulsed DC as effector cells and BEL-7402 as targetcells, additional peripheral blood DC and cord blood DC areadded to observe its stimulating activities to effector cells.The effector's cytotoxicity to tumor cells were detected withneutral red colorimetric assay at two effector/target ratios of5:1 and 10: 1.RESULTS: Peripheral blood DC and cord blood DC highlyexpressed HLA-ABC, HLA-DR, HLA-DQ, CD54 and S-100protein. The stimulating activities to lymphocyteproliferation were compared between experimental groups(DC added) and control group (no DC added). In sixexperiment subgroups, the DC/lymphocyte ratio wassequentially 0.25: 100, 0.5: 100, 1: 100, 2: 100, 4: 100 and 8:100, A values(x± s) were 0.75396± 0.009, 0.84916± 0.010,0.90894± 0.012, 0.98371 ± 0.007, 1.01299 ± 0.006 and 1.20384± 0.006 in peripheral blood DC groups and 0.77650 ± 0.005,0.83008± 0.007, 0.92725 ± 0.007, 1.05990 ± 0.010, 1.15583 ±0.011, 1.22983 ± 0.011 in cord blood DC groups. A value was0.59517 ± 0.005 in control group. The stimulating activitieswere higher in experimental groups than in control group ( P＜ 0.01 ), which were increased when the DC concentrationwas enlarged ( P ＜ 0.01 ). Two differently derived DCs hadthe same phenotypes and similar stimulating activities ( P ＞0.05). In peripheral blood DC groups, the cytotoxicity (x ±s) of the LD groups (experimental groups) and L groups(control group) was 58.16% ± 2.03% (5: 1), 46.18% ±2.25% (10:l) and 38.13% ± 1.29% (5: 1) and 65.40% ±1.56% (10: 1), respectively; in cord blood DC groups, TDgroups (experimental groups ) and T groups (controlgroups) were 69.71% ± 2.33% (5: 1 ), 77.64% ± 1.94% ( 10:1) a. nd 56.89% ± 1.82% (5:1) and 60.99% ± 1.42% (10:1),respectively. The cytotoxicity activities were enhanced withincreased effector/target ratio ( P ＜ 0.01 ). At the sameeffector/target ratio, the cytotoxicity of experimental groupswas higher than that of control groups ( P ＜ 0.01 ). Thecytotoxicity of cord blood DC groups were higher than ofperipheral blood DC groups ( P ＜ 0.01 ).CONCLUSION: Peripheral blood DC and cord blood DC aremature DCs in morphology and function, both can enhancethe effector cell killing activities to hepetocarcinoma cells.DC pulsed with tumor antigen can induce ligher specific CTLactivity than unpulsed DC.
AIM:To observe the biological specialization of human peripheral blood dendritic cells(DC)and cord blood derived DC and its effects on effector cells killing human hepatocarcinoma call line BEL-7402 in vitro. METHODS:The DC biological characteristics were detected with immunohistochemical and MTT assay.Two antitumor experimental groups are:peripheral blood DC and cord blood DC groups.Peripheral blood DC groups used LAK cells as the effector cells and BEL-7402 as target cells,while cord blood DC groups used CTL induced by tumor antigen twice pulsed DC as effector cells and BEL-7402 as target cells,additional peripheral blood DC and cord blood DC are added to observe its stimulating activities to effector cells. The effector's cytotoxicity to tumor cells were detected with neutral red colorimetric assay at two effector/target ratios of 5:1 and 10:1. RESULTS:Peripheral blood DC and cord blood DC highly expressed HLA-ABC,HLA-DR,HLA-DQ,CD54 and S-100 protein.The stimulating activities to lymphocyte proliferation were compared between experimental groups (DC added)and control group(no DC added).In six experiment subgroups,the DC/lymphocyte ratio was sequentially0.25:100,0.5:100,1:100,2:100,4:100 and 8: 100,A values(x±s)were 0.75396±0.009,0.84916±0.010, 0.90894±0.012,0.98371±0.007,1.01299±0.006 and 1.20384 ±0.006 in peripheral blood DC groups and 0.77650±0.005, 0.83008±0.007,0.92725±0.007,1.05990±0.010,1.15583± 0.011,1.22983±0.011 in cord blood DC groups.A value was 0.59517±0.005 in control group.The stimulating activities were higher in experimental groups than in control group(P <0.015,which were increased when the DC concentration was enlarged(P<0.01 5.Two differently derived DCs had the same phenotypes and similar stimulating activities(P>. 0.055.In peripheral blood DC groups,the cytotoxicity(x± s)of the LD groups(experimental groups)and L groups (control group)was 58.16%±2.03%(5:1),46.18%± 2.25%(10:15 and 38.13%±1.29%(5:1)and 65.40%± 1.56%(10:1),respectively;in cord blood DC groups,TD groups(experimental groups)and T groups(control groups)were 69.71%±2.33%(5:1),77.64%±1.94%(10: 1)and 56.89%±1.82%(5:1)and 60.99%±1.42%(10:1), respectively.The cytotoxicity activities were enhanced with increased effector/target ratio(P<0.01).At the same effector/target ratio,the cytotoxicity of experimental groups was higher than that of control groups(P<0.01).The cytotoxicity of cord blood DC groups were higher than of peripheral blood DC groups(P<0.01). CONCLUSION:Peripheral blood DC and cord blood DC are mature DCs in morphology and function,both can enhance the effector cell killing activities to hepatocarcinoma cells. DC pulsed with tumor antigen can induce ligher specific CTL activity than unpulsed DC.
基金
Natural Science Foundation of the Higher Education Office of Guangdong Province,No.9501 and No.9816
