【摘要】 目的:探讨用聚乳酸-羟基乙酸共聚物(PLGA)制备的纳米微粒对树突细胞的表型及功能的影响。方法:用复乳溶剂蒸发法制备纳米微粒(NP)及包含有黑色素瘤抗原肽gp100-154-162的纳米颗粒(NP-gp100-154-162),用扫描电镜、Nano Plus粒度测定仪检测NP表征。用外周血单核细胞诱导树突细胞(DC),用纳米微粒负载树突细胞,共聚焦显微镜和流式细胞仪检测DC细胞对NP的摄取情况及DC摄取NP后的表型变化。用混合淋巴细胞反应(MLR)检测负载NP的DC及单纯DC刺激同种异体淋巴细胞增殖反应;免疫斑点法(ELISPOT)检测负载NP-gp100-154-162的DC刺激效应细胞分泌IFN-γ的能力,观察纳米微粒对DC递呈抗原的影响。结果:PLGA-NP为圆形,粒径约为180～230 nm,该纳米微粒能被DC有效摄取;负载PLGA-NP后DC表面标志HLA-DR、CD80、CD86、CD83等明显增加;混合淋巴细胞反应结果显示负载纳米微粒的DC(DC/NP)能刺激T细胞增殖,刺激指数(SI)显著高于单纯DC组和单纯纳米微粒组。ELISPOT检测显示:DC细胞负载NPgp100-154-162在第4天时刺激效应细胞产生分泌IFN-γ的斑点数显著高于其在第2天时产生的斑点数,二者均显著高于DC负gp100-154-162产生的斑点数,表明纳米微粒能促进DC成熟,增强DC递呈抗原的能力。结论:PLGA纳米微粒能促进DC成熟和活化,增强并延长DC对抗原的递呈作用,提高树突状细胞免疫应答能力。更多还原

【Abstract】 Objective: To investigate the effect of nanoparticles prepared with poly( PLGA) on the phenotype and function of dendritic cells. Methods: The nanoparticles( NP)/NP-gp100-154-162 were made using double solvent evaporation method and characterized by scanning electronic microscope and Nano Plus respectively. DCs were generated from PBMC and treated with PLGA-NPs,their uptake and phenotype were evaluated by confocal laser scanning microscopy and flow cytometry. Finally,the effects of the NPs/NPgp100-154-162 on DCs mediated lymphocyte proliferation and generating IFN-γ-producing cells were investigated using mixed lymphocyte reaction( MLR) and ELISPOT respectively. Results: Nanoparticles were spherical in morphology and range in size from 180 nm to230 nm which was efficiently endocytosed when incubated with DCs. NPs promoted DC increasing phenotype expression of HLA-DR,CD80,CD86,CD83 compared with untreated DCs. Significantly proliferation was observed for effector T lymphocyte treated with DCs/NP compared with that treated with DCs or NP alone respectively. ELISPOT results indicated that NP-gp100-154-162-plusing-DCs stimulated effector cell produced more IFN-γ producing cells on day 4 than that on day 2,both stimulated effector cell produced more IFN-γ producing cells than free peptide pulsing-DCs suggested that PLGA-NPs effectively induce maturation and enhance antigen presentation of DCs. Conclusion: PLGA nanoparticle promoted DC maturation and activation,enhance antigent presentation capacity and increased immunological response of DC.更多还原