摘要
目的:树突状细胞是功能最强的抗原提呈细胞,能够向T淋巴细胞有效地提呈抗原,发挥特异性的杀伤肿瘤的作用,本实验主要观察树突状细胞的体外抗癌作用。方法:实验于2002-03/2003-03在华北煤炭医学院完成。选取华北煤炭医学院健康志愿者30例,抽取外周血10mL,分离出外周血单个核细胞。联合应用重组人集落细胞刺激因子和重组人白细胞介素-4诱导外周血单个核细胞7d后获取树突状细胞。以肺癌细胞系CALU-6肿瘤细胞全抗原负载的树突状细胞和重组人白细胞介素-2联合活化自体混合T淋巴细胞,使其增殖为树突状细胞活化的杀伤细胞;体外观察树突状细胞活化的杀伤细胞及其培养上清对肿瘤细胞的杀伤活性,并检测不同的树突状细胞对其活化的杀伤细胞杀伤活性的影响。结果:①10mL健康人外周血单个核细胞经重组人集落细胞刺激因子和重组人白细胞介素-4联合培养7d收获的树突状细胞数量为(1.88±0.77)×106个。②树突状细胞活化的杀伤细胞其细胞表型CD8阳性率明显增加。③树突状细胞活化的杀伤细胞对于肿瘤细胞全抗原来源的肺癌细胞系CALU-6肿瘤细胞有明显的特异性杀伤作用。④树突状细胞浓度适中,其活化的杀伤细胞的杀伤肿瘤细胞的能力最强,树突状细胞浓度过高或过低,其活化的杀伤细胞的杀伤力均减弱。⑤树突状细胞活化的杀伤细胞上清液有一定的杀伤活性,并且随着树突状细胞浓度的降低而降低。结论:负载抗原的树突状细胞能诱导自体的混合T淋巴细胞,产生树突状细胞活化的杀伤细胞进而诱导高效而特异性的抗肿瘤免疫作用。树突状细胞诱导的抗肿瘤免疫与其浓度有关,呈双向调节作用。
AIM： The dendritic cells that are the strongest antigen presenting cell can present effectively antigen against T lymphocytes, and bring specific effect of killing tumor into full play. To see the anti-tumor effect of dendritic cell in vitro.
METHODS： The experiment was conducted at the North China Coal Medical College from March 2002 to March 2005. Thirty healthy volunteers from North China Coal Medical College were selected and 10 mL peripheral blood was extracted, and then mononuclear cells were isolated from peripheral blood. The dendritic cells were obtained after 7 days of gaining mononuclear cells with the supplement of recombination human colony stimulating factor （CSF） and interleukin （IL）-4. Tumor complete antigen （TCA） was extracted from the lysate of lung cancer cell CALU-6. Tumor-killer cells were co-stimulated by IL-2 and dendritic cell that was activated by TCA. The anti-tumor effect of dendritic cell activatied killer（DAK） cell and its supernatant were studied in vitro. The best density of dendritic cell affected the antitumor effect of DAK cells is observed.
RESULTS： ①Large amount of dendritic cells generated from mononuclear cells of 10 mL peripheral blood after cultured for 7 days with CSF and IL- 4 was （1.88±0.77）×10^6. ②Positive rate of phenotype CDs of DAK increased significantly. ③The DAK had obviously killing effects on lung cancer cell CALU-6 extracted from TCA. ④Concentration of dendritic cells that was well situated had the strongest ability of killing tumor cells. Superhigh or superlow concentration of dendritic cells could make the ability of, killing calls decrease. ⑤The supernatant of DAK had killing activity, and decreased with the decrease of the concentration of dendritic cells.
CONCLUSION： Antigen dendritic cells can induce to autologous T lymphocytes, produce DAK so as to lead to high effective and specific antitumor immunological effect. The anti-tumor effect due to dendritic cells has relation with its concentratign, which shows double regulatory function.
出处
《中国临床康复》
CSCD
北大核心
2006年第4期119-121,共3页
Chinese Journal of Clinical Rehabilitation
