【摘要】： 目的:本研究構(gòu)建肝癌抗原肽-熱休克蛋白復(fù)合物修飾樹突狀細胞(DC),觀察這種修飾的DC細胞表型及其分泌細胞因子的變化,進一步觀察其在體外的抗腫瘤作用。 方法:從H22肝癌細胞中提取腫瘤抗原肽,并在體外與熱休克蛋白70(Hsp70)進行結(jié)合;采用細胞培養(yǎng)技術(shù),培養(yǎng)rmGM-CSF、rmIL-4誘導(dǎo)的小鼠骨髓細胞,體外獲取大量的DC,用HSP-70抗原肽復(fù)合物刺激DC,取上清液,測細胞因子IL-10,IL-12含量,并用流式細胞儀分析DC表面分子。 結(jié)果:DC體外誘導(dǎo)培養(yǎng)成功,HSP70-H22復(fù)合物可使DC成熟,DC表面分子CD40,CD80在HSP70-H22復(fù)合物修飾DC組表達較單純DC組有顯著性增高(P0.01);10ug/ml, 20ug/ml組之間相比無顯著性差異(P0.05);三組之間CD11c無差異性(P0.05)。Hsp70-H22抗原肽(10ug)修飾的DC組的細胞因子IL-12明顯高于單純DC組(P0.01),Hsp70-H22抗原肽(20ug)修飾的DC組的細胞因子IL-12也明顯高于單純DC組(P0.01),Hsp70-H22抗原肽(10ug)和Hsp70-H22抗原肽(20ug)相比具有顯著性差異(P0.01);Hsp70-H22抗原肽修飾的DC組上清中,IL-10的含量明顯下降,以Hsp70-H22抗原肽修飾的DC組(20ug)上清中含量最少與單純DC組相比有顯著性差異(P0.01),Hsp70-H22抗原肽修飾的DC組(10ug)與單純DC組相比差異也有顯著性(P0.05)。 結(jié)論:HSP70-H22復(fù)合物能提高DC表面分子CD40,CD80等表達,誘導(dǎo)DC的成熟,并能誘導(dǎo)DC分泌細胞因子IL-12明顯增高,IL-10降低,達到抗腫瘤效應(yīng)。
[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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R392;R735.7

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