肝癌抗原肽-HSP70复合物修饰树突状细胞增强抗肝癌作用初步研究
发布时间：2018-08-06 08:47
【摘要】： 目的:本研究构建肝癌抗原肽-热休克蛋白复合物修饰树突状细胞(DC),观察这种修饰的DC细胞表型及其分泌细胞因子的变化,进一步观察其在体外的抗肿瘤作用。 方法:从H22肝癌细胞中提取肿瘤抗原肽,并在体外与热休克蛋白70(Hsp70)进行结合;采用细胞培养技术,培养rmGM-CSF、rmIL-4诱导的小鼠骨髓细胞,体外获取大量的DC,用HSP-70抗原肽复合物刺激DC,取上清液,测细胞因子IL-10,IL-12含量,并用流式细胞仪分析DC表面分子。 结果:DC体外诱导培养成功,HSP70-H22复合物可使DC成熟,DC表面分子CD40,CD80在HSP70-H22复合物修饰DC组表达较单纯DC组有显著性增高(P0.01);10ug/ml, 20ug/ml组之间相比无显著性差异(P0.05);三组之间CD11c无差异性(P0.05)。Hsp70-H22抗原肽(10ug)修饰的DC组的细胞因子IL-12明显高于单纯DC组(P0.01),Hsp70-H22抗原肽(20ug)修饰的DC组的细胞因子IL-12也明显高于单纯DC组(P0.01),Hsp70-H22抗原肽(10ug)和Hsp70-H22抗原肽(20ug)相比具有显著性差异(P0.01);Hsp70-H22抗原肽修饰的DC组上清中,IL-10的含量明显下降,以Hsp70-H22抗原肽修饰的DC组(20ug)上清中含量最少与单纯DC组相比有显著性差异(P0.01),Hsp70-H22抗原肽修饰的DC组(10ug)与单纯DC组相比差异也有显著性(P0.05)。 结论:HSP70-H22复合物能提高DC表面分子CD40,CD80等表达,诱导DC的成熟,并能诱导DC分泌细胞因子IL-12明显增高,IL-10降低,达到抗肿瘤效应。
[Abstract]:Aim: to investigate the phenotype and cytokine secretion of dendritic cells modified with hepatoma antigen peptide-heat shock protein complex (DC),). Methods: tumor antigen peptides were extracted from H22 hepatoma cells and combined with heat shock protein 70 (Hsp70) in vitro. A large number of DCs were obtained in vitro. The supernatant of DCS was stimulated with HSP-70 antigen peptide complex. The content of IL-10 IL-12 was measured and the surface molecules of DC were analyzed by flow cytometry. Results the expression of CD40T CD80 on the surface of mature DC cells was significantly increased in HSP70-H22 modified DC group (P0.01) than that in DC treated with HSP70-H22 complex (P0.01). There was no significant difference between 20ug/ml group and 20ug/ml group (P0.05), but there was no significant difference among the three groups (P0.05). The cytokine IL-12 of DC group modified with Hsp70-H22 antigen peptide (10ug) was significantly higher than that of DC group modified with Hsp70-H22 antigen peptide (P0.01). The cytokine IL-12 of DC group modified with Hsp70-H22 antigen peptide (20ug) was significantly higher than that of DC group (P0.01) with Hsp70-H22 antigen peptide (10ug) and Hsp70-H22 antigen peptide (20ug). (P0.01) the content of IL-10 in the supernatant of DC group modified with Hsp70-H22 antigen peptide was significantly decreased. The content of supernatant in DC group (20ug) modified with Hsp70-H22 antigen peptide was significantly lower than that in DC group (P0.01). There was also significant difference between DC group (10ug) modified with Hsp70-H22 peptide and DC group (P0.05). Conclusion the expression of CD40, CD80, CD40 and CD80 on DC surface can be increased, the mature of DC can be induced, and the level of IL-12 secreting by DC can be significantly increased and IL-10 is decreased, which can achieve the anti-tumor effect.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392;R735.7
本文编号：2167182
[Abstract]:Aim: to investigate the phenotype and cytokine secretion of dendritic cells modified with hepatoma antigen peptide-heat shock protein complex (DC),). Methods: tumor antigen peptides were extracted from H22 hepatoma cells and combined with heat shock protein 70 (Hsp70) in vitro. A large number of DCs were obtained in vitro. The supernatant of DCS was stimulated with HSP-70 antigen peptide complex. The content of IL-10 IL-12 was measured and the surface molecules of DC were analyzed by flow cytometry. Results the expression of CD40T CD80 on the surface of mature DC cells was significantly increased in HSP70-H22 modified DC group (P0.01) than that in DC treated with HSP70-H22 complex (P0.01). There was no significant difference between 20ug/ml group and 20ug/ml group (P0.05), but there was no significant difference among the three groups (P0.05). The cytokine IL-12 of DC group modified with Hsp70-H22 antigen peptide (10ug) was significantly higher than that of DC group modified with Hsp70-H22 antigen peptide (P0.01). The cytokine IL-12 of DC group modified with Hsp70-H22 antigen peptide (20ug) was significantly higher than that of DC group (P0.01) with Hsp70-H22 antigen peptide (10ug) and Hsp70-H22 antigen peptide (20ug). (P0.01) the content of IL-10 in the supernatant of DC group modified with Hsp70-H22 antigen peptide was significantly decreased. The content of supernatant in DC group (20ug) modified with Hsp70-H22 antigen peptide was significantly lower than that in DC group (P0.01). There was also significant difference between DC group (10ug) modified with Hsp70-H22 peptide and DC group (P0.05). Conclusion the expression of CD40, CD80, CD40 and CD80 on DC surface can be increased, the mature of DC can be induced, and the level of IL-12 secreting by DC can be significantly increased and IL-10 is decreased, which can achieve the anti-tumor effect.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392;R735.7
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