血管内皮生长因子对成熟树突状细胞蛋白质组的影响

Proteomics Study on the Effect of Vascular Endothelial Growth Factor on Mature Dendritic Cells

目的:探索血管内皮生长因子(vascular endothelial growth factor,VEGF)对成熟树突状细胞(maturedendritic cells,mDCs)蛋白质表达谱的影响,以进一步理解肿瘤的免疫逃逸机制,为改善基于DCs的抗肿瘤免疫的临床治疗效率寻找新的线索。方法:用免疫磁珠从人外周血分离CD14+单核细胞,加入粒-巨噬细胞集落刺激因子(Recombinant human granulocyte-macrophage CSF,rhGM-CSF)、白介素4(Recombinant human interleukins 4,IL-4)将单核细胞诱导分化为未成熟DCs(immature DCs,imDCs),利用肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)将imDCs诱导为成熟DCs(mature DCs,mDCs),VEGF作用于mDCs,用基于质谱的蛋白质组技术研究VEGF对mDCs的蛋白质表达谱的影响。结果:VEGF可上调或下调mDCs的细胞骨架及其相关蛋白、迁移相关蛋白、免疫功能相关蛋白和抗氧化相关蛋白的表达。结论:VEGF可能通过重组mDCs的细胞骨架来损伤其运动能力和免疫学功能,这对进一步理解DCs的生物学功能和肿瘤的免疫逃逸机制具有重要意义。

Objective： To investigate the effect of vascular endothelial growth factor（VEGF） on protein expression profile of mature dendritic cells（mDCs）,and so as to further understanding of the immune escape mechanism of tumor and looking for the new clue for improvement of clinical efficiency of DCs-based immune therapy against cancer.Methods： CD14＋ monocytes were isolated from peripheral blood by immunomagnetic beads.The monocytes were then cultured with recombinant human granulocyte-macrophage colony stimulating factor（rhGM-CSF） and recombinant human interleukins 4（rhIL-4） for 7 days to develop into immature DCs（imDCs）.Tumor necrosis factor-α（rhTNF-α） was used to induce imDCs into mCDs.The mDCs were treated with VEGF.The protein expression profiles of mCDs were investigated by mass spectrometry-based proteomics technique.Results： Some cytoskeletal proteins and their related proteins,migration-associated proteins,immune function-related proteins and antioxidation-associated proteins in mCDs were up or down-regulated by VEGF.Conclusions： The motility and immune function of mDCs might be impaired by VEGF through cytoskeletal reorganization,which is significant for further understanding of biological function of mDCs and immune escape mechanism of tumor.