不同方法制备抗原致敏的树突状细胞介导T淋巴细胞对Tca8113细胞杀伤作用的影响

Killing effect of cytotoxicty T lymphocyte primed by two different methods preparation of dendritic cells pulsed with antigen on Tca8113 cells in vitro

目的探讨树突状细胞(DCs)疫苗治疗舌癌的可行性,并选择较优的抗原负载方式。方法分别采用弱酸洗脱法、反复冻融法制备Tca8113细胞抗原,并分为3组:弱酸洗脱抗原组、反复冻融抗原组、对照组(不加肿瘤抗原)。从外周血单核细胞中诱导DCs,分离出T淋巴细胞。以不同效靶比将DCs和T淋巴细胞混合培养,MTT法测光密度值,计算刺激指数。DCs抗原负载后,与T淋巴细胞混合,以不同效靶比加入预先接种了Tca8113细胞的培养板。MTT法测光密度值,计算杀伤率。结果2种方法体外成功地构建了DCs疫苗,抗原致敏组与对照组比较差异显著,且弱酸洗脱组优于反复冻融组。DCs疫苗可诱导细胞毒性T淋巴细胞有效的杀伤Tca8113细胞,并表现明显的剂量效应。结论DCs疫苗致敏T淋巴细胞能够有效的杀伤Tca8113细胞,酸洗脱抗原冲击致敏的DCs疫苗在舌癌的免疫治疗中优于冻融抗原。

Objective To study the feasibility of dendritic cells（DCs） vaccine on the therapy of tongue carcinoma and find the better way of antigen load. Methods The antigen peptides of Tca8113 cells were obtained by acid eluted technique and repetitive freeze thaw method. Separating T cell and inducing dendritic cells were obtained from human peripheral blood monocyte. Divided into three groups： Weak acid elution method antigen group, anti-freezethaw method antigen group, and the control group（without tumor antigen）. T cells and DCs were mixed to culture by different effector-target ratio. Using MTT assay measured the quantities of absorbance and calculated stimulation index. Dendrtic cells pulsed with antigen were mixed with T cells by different effector-target ratio. MTT assay was used to measure the quantities of absorbance and calculate killing rate. Results DCs vaccine was constructed successfully. DCs vaccine can induce T lymphocytes to kill Tca8113 cells and display the dose-effect relationship. There was significant difference among the three groups. The acid eluted and repetitive freeze thaw groups were better than the control group. The acid eluted group was better than repetitive freeze thaw group. Conclusion DCs vaccine can induce T lymphocytes to kill Tca8113 cells. The antigen peptides obtained by acid eluted technique is better than repetitive freeze thaw method in immunotherapy of tongue cancer.