Page 7 - 第10期

第41卷第10期 南京医科大学学报（自然科学版）
2021年10月 Journal of Nanjing Medical University（Natural Sciences） ·1425 ·

·基础医学·

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

任宏艳 ，康美云 ，王子妤 ，姚 琦 ，张璐瑶 ，王永韧 2*
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南京中医药大学医学院整合医学学院病理学与病理生理学系，江苏 南京 210023；南京医科大学附属儿童医院血液肿瘤
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科，江苏 南京 210008
［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（interleu⁃
kin，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⁃
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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。
［关键词］ PEI；氢氧化铝纳米颗粒；树突状细胞；抗原提呈
［中图分类号］ R392.11 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）10⁃1425⁃07
doi：10.7655/NYDXBNS20211001

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

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REN Hongyan ，KANG Meiyun ，WANG Ziyu ，YAO Qi ，ZHANG Luyao ，WANG Yongren 2*
1 Department of Pathology and Pathophysiology，School of Medicine & Holistic Integrative Medicine，Nanjing
University of Chinese Medicine，Nanjing 210023；Department of Hematology and Oncology，Children’s Hospital of
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Nanjing Medical University，Nanjing 210008，China
［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 B 3Z cells. The activation of B 3Z 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 B 3Z cells was determined by CPRG
assay after co⁃incubation with B 3Z cells. DCs loaded with HS/PEI⁃OVA were co⁃incubated with CFSE⁃labeled OT⁃Ⅰ T cells to detect
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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.
［Key words］ PEI；aluminum hydroxide nanoparticle；dendritic cell；antigen presentation
［J Nanjing Med Univ，2021，41（10）：1425⁃1431］
［基金项目］ 国家自然科学基金（81502681）
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通信作者（Corresponding author），E⁃mail：wangyongren@gmail.com