摘要
目的探讨培养时间对人单核细胞来源树突状细胞(DC)及其外泌体(DEX)免疫膜蛋白的影响。方法采用重组粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素4(IL-4)诱导人单核细胞分化成DC,并经肿瘤坏死因子α(TNF-α)促使其成熟;超速离心法分离DEX,透射电子显微镜及Amnis量化成像流式细胞术鉴定DEX并检测DC及DEX的免疫膜蛋白表达情况。结果体外培养至第10天,DC表面即高表达CD11c、CD80、CD86、主要组织相容性复合体Ⅰ(MHC-Ⅰ)及MHC-Ⅱ分子,至第18天达峰值,表达率分别为CD11c(78.66±20.33)%、CD80(76.41±10.02)%、CD86(96.43±0.43)%、MHC-Ⅰ(84.71±2.96)%、MHC-Ⅱ(80.01±7.03)%,第24天后表达总体呈下降趋势(除CD80和MHC-Ⅱ外,差异均具有统计学意义);培养至第30天,仍有80%的DC表达CD80、CD86、MHC-Ⅱ分子。DEX表面免疫膜蛋白表达亦于第18天最高,随后随培养时间延长总体下降(除CD80外,差异均具有统计学意义)。相关性分析显示,DC与DEX表面的免疫膜蛋白表达水平呈显著正相关(CD11c:r=0.98;CD80:r=0.65;CD86:r=0.82;MHC-Ⅰ:r=0.86;MHC-Ⅱ:r=0.93)。结论人单核细胞来源的DC在体外培养中可高效表达免疫膜蛋白,并在一定时间内保持稳定;其分泌的DEX表面同样高表达免疫膜蛋白,且二者表达变化趋势基本一致。
Objective To investigate how culture duration affects the expression of immune membrane proteins in human monocyte-derived dendritic cells(DCs)and their exosomes(DEXs).Methods Human monocytes were induced with recombinant granulocyte-macrophage colony-stimulating factor(GM-CSF)and interleukin 4(IL-4)to differentiate into DCs and were subsequently matured with tumor necrosis factorα(TNF-α).Exosomes were isolated by ultracentrifugation,and DEXs were identified by transmission electron microscopy and Amnis imaging flow cytometry,which were also used to quantify the expression of immune membrane proteins on DCs and DEXs.Results On the 10th day of culture,DCs displayed high surface expression of CD11c,CD80,CD86,major histocompatibility complex class I(MHC-I),and MHC-II.Expression peaked at day 18(CD11c:78.66%±20.33%,CD80:76.41%±10.02%,CD86:96.43%±0.43%,MHC-I:84.71%±2.96%,MHC-II:80.01%±7.03%).After day 24,the overall expression showed a declining trend,with statistically significant differences observed for all markers except CD80 and MHC-II.By day 30,80%of the DCs still expressed CD80,CD86,and MHC-II.The expression of immune membrane proteins on DEX surfaces also reached its peak on day 18,followed by an overall decline with prolonged culture time,with statistically significant differences observed for all markers except CD80.Correlation analysis revealed a significant positive relationship between the expression levels of immune membrane proteins on DC and DEX surfaces(CD11c:r=0.98;CD80:r=0.65;CD86:r=0.82;MHC-I:r=0.86;MHC-II:r=0.93).Conclusion Human monocyte-derived DCs in vitro express high expression of immune membrane proteins and maintain stable expression over a specific period.The exosomes secreted by these cells similarly demonstrate high surface expression of immune membrane proteins,with temporal trends aligned with those of the parent DCs.
作者
罗淑敏
徐芳
路鹏鹏
王一越
李传云
李伟华
LUO Shumin;XU Fang;LU Pengpeng;WANG Yiyue;LI Chuanyun;LI Weihua(Beijing Institute of Infectious Diseases Integrated with Traditional Chinese and Western Medicine,Beijing Youan Hospital,Capital Medical University,Beijing Youan Hospital,Capital Medical University,Beijing 100069,China;Beijing Institute of Hepatology,Beijing Youan Hospital,Capital Medical University,Beijing Youan Hospital,Capital Medical University,Beijing 100069,China;Department of Surgery,Beijing Youan Hospital,Capital Medical University,Beijing 100069,China)
出处
《细胞与分子免疫学杂志》
北大核心
2025年第11期971-977,共7页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(82274447)
北京市高层次公共卫生技术人才建设资助项目(2022-2-024)
首都卫生发展科研专项(2024-2-1152)
北京肝病研究所开放性项目(2025Y-KF-C02)内蒙古自治区人才开发基金高层次人才个人项目
北京市属医学科研院所公益发展改革试点项目(2019-6,2021-10)。
