Der f 1/IGF-1纳米颗粒促进调节性T细胞生成的实验研究

Mechanism of Der f 1/IGF-1 nanoparticle promoting the production of regulatory T cell

目的:旨在制备负载Der f 1/IGF-1的聚乳酸-乙醇酸共聚物(poly lactide-co-glycolide,PLGA)纳米颗粒(Der f 1/IGF-1 NPs),探讨其促进Treg细胞生成的作用。方法:通过复乳法制备包裹Der f 1/IGF-1的NPs并分析其理化性质及体外累计释放率;预处理BMDC将细胞分为Saline组、Blank NPs组、Der f 1/IGF-1组以及Der f 1/IGF-1 NPs组;利用ELISA实验测定IL-10及TGF-β的表达;采用流式细胞术检测Treg细胞的数量变化。结果:Der f 1/IGF-1 NPs为球形结构,具有良好的分散性,粒径均<200 nm,Zeta电位为负电荷且具有稳定的缓释效果;BMDC预处理后,相比较Blank NPs组,Der f 1/IGF-1 NPs组,BMDC细胞TGF-β、IL-10的表达水平明显增高,差异有统计学意义(P<0.001);且与CD4^(+)T细胞共培养后,Der f 1/IGF-1 NPs组Treg细胞生成的比例明显提高,差异有统计学意义(P<0.001)。结论:Der f 1/IGF-1 NPs在体外实验中可诱导Treg细胞生成。本研究为重建免疫耐受功能失常提供了一种新的、更有效的方法。

Objective: To prepare PLGA nanoparticles loaded with Der f 1/IGF-1(Der f 1/IGF-1 NPs)and investigate their role in promoting the formation of Treg cells.Methods: NPs coated with Der f 1/IGF-1 were prepared by double emulsion method and their physicochemical properties and cumulative release rate in vitro were analyzed.After pretreatment,BMDC was divided into Saline group,Blank NPs group,Der f 1/IGF-1 group and Der f 1/IGF-1 NPs group.Determination of the expression of IL-10 and TGF-βin BMDC by ELISA.The number of Treg cells was detected by flow cytometry.Results: The results showed that Der f 1/IGF-1 NPs were spherical structures,with good dispersion,particle size less than 200 nm,negative charge and stable slow-release effect of Zeta potential.After BMDC pretreatment,the expression levels of TGF-βand IL-10 in BMDC cells in the Der f 1/IGF-1 NPs group were significantly increased compared with the Blank NPs group,and the difference was statistically significant(P<0.001).After co-culture with CD4^(+)T cells,the proportion of Treg cells produced in the Der f 1/IGF-1 NPs group was significantly increased,and the difference was statistically significant(P<0.001).Conclusion: Der f 1/IGF-1 NPs can induce Treg cell generation in vitro.This study provides a new and more effective method for the reconstruction of immune tolerance dysfunction.