致敏DC诱导CTL特异性杀伤HL-60细胞的实验研究

Investigation on specific Anti-HL-60 cell reaction with CTL induced by sensitive dendritic cells

目的 :探讨来自外周血单个核细胞的树突状细胞 (DC)负载冻融或弱酸洗脱的HL 6 0多肽抗原 ,体外诱导产生细胞毒性T淋巴细胞 (CTL)对HL 6 0细胞的杀伤作用。方法 :应用GM CSF、TNF α和IL 4自外周血单个核细胞诱导出DC ,使其负载两种HL 6 0多肽抗原 ,致敏同种T淋巴细胞 ,产生CTL。以胃癌细胞系SGC790 1为对照组 ,观察DC∶CTL∶HL 6 0 /SGC 790 1为 1∶10 0∶2 ,CTL对HL 6 0特异性免疫杀伤活性。结果 :弱酸洗涤法或冻融法所提取的抗原多肽致敏DC诱导的CTL对HL 6 0细胞的杀伤率分别为 6 8.2 9%± 11.37%和4 5 .95 %± 8.4 7% ,二者差异有极显著性意义 (P <0 .0 1)。对SGC 790 1细胞的杀伤率分别为 2 4 .6 0 %± 4 .4 0 %和 16 .0 9%± 4 .98% ,二者差异有极显著性意义 (P <0 .0 1)。结论 :相对于冻融HL 6 0细胞抗原多肽 ,DC可更有效地提呈酸洗HL 6 0抗原多肽 ,并通过特异性CTL产生对HL 6

Objective:To study the effects of dendritic cell (DC) pulsed with HL 60 cell peptides on stimulating the cytotoxic T lymphocyte (CTL) to get specific anti tumor activity.Method:DC was generated from normal human peripheral blood mononuclear cell (PBMC) in the presence of granulocyte/macrophage colony stimulating factor (GM CSF), interleukin 4 (IL 4) and alpha tumor necrosis factor (TNF α) in vitro.HL 60 cell was treated with citrate phosphate buffer (pH 3.3 ) or frozen thawed, and released its tumor antigen peptides (TAP) which were quantified with the Lowry's method.DC pulsed with TAP or lysates ( 40.0 ng/ml), and co culture to transfor T cell into specific CTL. DC,CTL and HL 60 cell mixed with 1∶100∶2.SGC 7901 cell line was evaluated as control group.Result:The CTL induced by DC pulsed with acid eluted TAP had more significant activity killing HL 60 cell than frozen thawed lysates (P< 0.01 ), and both of these induced CTLs were more effective to kill HL 60 cell than SGC 7901 cell (P< 0.01 ).Conclusion:The acid eluted TAP and the lysates of HL 60 cells can pulse DC to activate the CTL which might induce efficient specific anti HL 60 cell immune response.