【摘要】 目的探讨磷脂酰肌醇蛋白聚糖3（GPC3）抗原表位肽致敏的树突状细胞（DC）对T淋巴细胞活化的影响。方法选用基因型人类白细胞抗原A2阳性的剖宫产妇的胎盘端脐带血50 m L,分离培养DC及T细胞,用人工合成的GPC3抗原表位肽(GPC3_144-152、GPC3_298-306)、冻融Hep G2细胞抗原致敏DC (DC-GPC3_144-152组、DCGPC3_298-306组、DC-HepG2组、DC组),经致敏DC处理的T细胞随机分为DC-GPC3_144-152-T组、DC-GPC3_298-306-T组、DC-HepG2-T组、DC-T组,以未经活化的T细胞作对照（T细胞组）。用流式细胞术检测DC表型、T细胞分类及表面趋化因子的表达。结果 GPC3_144-152、GPC3_298-306抗原表位肽和冻融Hep G2细胞抗原三种致敏方式均能诱导DC呈典型成熟DC的形态,表面均高表达CD83、CD80和CD86抗原,且不同致敏方式之间CD8⁺T、CD4⁺T淋巴细胞比例差异无统计学意义（P均> 0. 05）;与T细胞组比较,GPC3_144-152-T组和DC-GPC3_298-306-T组CD4+T细胞中Th1细胞比例高（P均<0. 05）;与DC-T组比较,DC-GPC3_144-152-T组、DC-GPC3_298-306-T组、DC-HepG2-T组CCR6表达高（P均<0. 05）。结论 GPC3_144-152和GPC3_298-306表位抗原肽均能够有效致敏DC,并促进与DC共培养的T淋巴细胞活化。更多还原

【Abstract】 Objective To explore the role of glypican 3（ GPC3） epitope peptide-sensitized dendritic cells in activating T lymphocytes. Methods The placental umbilical cord blood（ 50 m L） of cesarean section with genotype HLA-A2-positive was selected,and dendritic cells（ DCs） and T-lymphocytes（ T cells） were isolated and cultured. Sensitized DCs were prepared by artificially synthesized GPC3 epitope peptides( GPC3_144-152,GPC3_298-306) and freeze-thawing Hep G2 cell antigens( DC-GPC3_144-152 group,DC-GPC3_298-306 group,DC-HepG2 group,DC group). Then the T cells treated by sensitized DCs were successively taken as the DC-GPC3_144-152-T group,DC-GPC3_298-306-T group,DC-HepG2-T group,and DC-T group,and meanwhile,the unactivated T cells were taken as the control group（ T cell group）,respectively. Flow cytometry was used to detect DC phenotype and T cell classification and surface chemokine expression. Results GPC3_144-152,GPC3_298-306 epitope peptide and freeze-thaw Hep G2 cell antigen all induced sensitization of DCs with typical mature DC morphology,and the surface expressed CD83,CD80 and CD86 antigens. There was no significant difference in the proportion of CD8⁺T and CD4⁺T lymphocytes between different sensitization methods（ all P > 0. 05）. Compared with the T cell group,the proportion of Th1 cells in CD4+T cells increased significantly in the DC-GPC3_144-152-T group and DCGPC3_298-306-T group（ both P < 0. 05）; compared with the DC-T group,the expression of T cell surface chemokine receptor CCR6 increased in the DC-GPC3_144-152-T group,DC-GPC3_298-306-T group,and DC-HepG2-T group（ all P < 0. 05）. Conclusion GPC3_144-152 and GPC3_298-306 epitope peptide is capable of sensitizing DC and then activating T lymphocytes.更多还原