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目的:探讨N-三甲基壳聚糖包衣聚乳酸-羟基乙酸共聚物[poly(lactic-co-glycolic acid),PLGA]纳米粒对诱导树突细胞交叉递呈的机制。方法:复乳法制备PLGA纳米粒并用N-三甲基壳聚糖(N-trimethyl chitosan,TMC60)进行包衣,分别包载模型抗原卵清白蛋白、异硫氰酸荧光素(fluorescein isothiocyanate,FITC)标记的卵清白蛋白(FITC-OVA)。将TMC60包衣和未包衣的纳米粒分别作用于体外培养的小鼠骨髓系树突细胞(murine bone marrow-derived dendritic cell,BMDC),用流式细胞仪检测BMDC对纳米粒子的吞噬动力学及BMDC表面分子CD83,MHCI和MHCII的表达;B3Z T细胞检测纳米粒被BMDC摄取后引起的交叉递呈反应。结果:TMC60包衣PLGA纳米粒可以促进BMDC吞噬作用;促进BMDC表达表面分子CD83和MHC I;增强BMDC对纳米粒包载抗原的交叉递呈作用。结论:TMC60包衣PLGA纳米粒可增强BMDC对外源性抗原的摄取及交叉递呈作用。
Objective: To investigate the mechanism of poly (lactic-co-glycolic acid) (PLGA) nanoparticles coated with N-trimethyl chitosan on the induction of dendritic cells cross-presentation. METHODS: PLGA nanoparticles were prepared by double emulsion method and coated with N-trimethyl chitosan (TMC60), respectively. The plasmids were packaged into model antigen ovalbumin, fluorescein isothiocyanate (FITC ) Labeled egg white albumin (FITC-OVA). TMC60-coated and uncoated nanoparticles were respectively treated with murine bone marrow-derived dendritic cells (BMDC) in vitro and the phagocytosis kinetics of BMDC nanoparticles by flow cytometry And BMDC surface molecules CD83, MHCI and MHCII expression; B3Z T cells detected by the BMDC uptake of cross-presentation reaction caused by BMDC. RESULTS: TMC60-coated PLGA nanoparticles could promote the phagocytosis of BMDC, promote the expression of surface molecules CD83 and MHC I on BMDC, and enhance the cross-presentation of BMDC on the antigen-loaded nanoparticles. CONCLUSION: TMC60-coated PLGA nanoparticles can enhance the uptake and cross-presentation of exogenous antigens by BMDC.