PEI修饰的氢氧化铝纳米颗粒增强树突状细胞抗原提呈作用的初步研究
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国家自然科学基金(81502681)


The study on the enhancement of antigen presentation in dendritic cells by polyethylene⁃imine modification of aluminum hydroxide nanoparticle
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    摘要:

    目的:探索聚乙烯亚胺(polyethylenimine,PEI)修饰的氢氧化铝纳米颗粒(HS)增强树突状细胞(dendritic cell,DC)抗原提呈的作用及其机制。方法:DC负载PEI修饰前后的HS联合卵清蛋白(ovalbum,OVA)抗原后与B3Z细胞共孵育,CPRG(chlorophenol red-β-d-galactopyranoside)方法检测DC对OVA的抗原提呈能力,Western blot检测DC中泛素化OVA蛋白的表达;以不同刺激剂[干扰素(interferon,IFN)-α、R848]与负载HS/PEI-OVA的DC混合后,收集培养上清ELISA检测白介素(interleukin,IL)-12的表达,与B3Z共孵育后CPRG方法检测不同刺激剂对HS/PEI增强DC抗原提呈的作用;DC负载HS/PEI-OVA后与OT-Ⅰ T细胞共孵育,检测OT-Ⅰ T细胞增殖;皮下接种小鼠黑色素瘤细胞B16-OVA构建小鼠黑色素瘤模型,探讨负载HS/PEI-OVA的DC疫苗体内诱导CD8+IFN-γ+T细胞比率上升及特异性T细胞分泌IFN-γ的能力。结果:与未经PEI修饰的HS相比,HS/PEI能显著增强DC对OVA抗原的提呈能力,并增加DC中泛素化OVA蛋白的表达及IL-12的分泌;IFN-α、R848均可进一步增强DC对HS/PEI-OVA的提呈作用;DC负载HS/PEI-OVA可显著促进OT-Ⅰ T细胞的增殖;DC-HS/PEI-OVA疫苗小鼠体内能诱导高效的T细胞应答。结论:HS/PEI能显著增强DC对抗原的交叉提呈,诱导高效的T细胞应答。

    Abstract:

    Objective:To investigate the effect and mechanism of polyethylenimine (PEI) modified alumina hydroxide nanoparticles(HS/PEI) on enhancing dendritic cell (DC) antigen presentation. Methods:DCs were loaded with HS combined with ovalbum (OVA) antigen in the presence or absence of PEI and co-incubated with B3Z cells. The activation of B3Z cells was determined by CPRG assay. The expression of ubiquitinated OVA protein in DCs was detected by Western blot. After incubated with HS/PEI-OVA and different stimulants(IFN-α,R848),IL-12 expression of DCs was detected by ELISA,and the activation of B3Z cells was determined by CPRG assay after co-incubation with B3Z cells. DCs loaded with HS/PEI-OVA were co-incubated with CFSE-labeled OT-Ⅰ T cells to detect the proliferation of OT-Ⅰ T cells. The increased percentage of CD8+IFN-γ+T cells and IFN-γ secretion of specific T cells induced by DC-HS/PEI-OVA vaccine were examined in B16-OVA tumor-bearing mice. Results:Compared with HS without PEI,HS/PEI significantly enhanced the OVA antigen presentation ability of DCs,increased the ubiquitinated OVA protein expression and IL-12 secretion of DCs. IFN-α and R848 could further enhance the presentation of HS/PEI-OVA by DCs. HS/PEI-OVA could significantly promote OT-Ⅰ T cells proliferation in vitro and induce efficient T-cell responses in vivo. Conclusion:HS/PEI could enhance antigen cross-presentation of DCs and induce efficient T-cell responses.

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任宏艳,康美云,王子妤,姚 琦,张璐瑶,王永韧. PEI修饰的氢氧化铝纳米颗粒增强树突状细胞抗原提呈作用的初步研究[J].南京医科大学学报(自然科学版),2021,(10):1425-1431

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  • 收稿日期:2021-05-11
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  • 在线发布日期: 2021-11-05
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