中国病理生理杂志2025，Vol.41Issue(10)：1938-1945,8.DOI:10.3969/j.issn.1000-4718.2025.10.008
负载H22-MPs的树突状细胞可增强CD8+T细胞对肝癌细胞的杀伤作用
Dendritic cells loaded with H22-MPs enhance killing effect of CD8+T cells on hepatocellular carcinoma cells
摘要
Abstract
AIM:To establish an optimal protocol for isolating and extracting mouse hepatocellular carcinoma H22 cell-detrived microparticles(H22-MPs),evaluate their stimulating effect on dendritic cells(DC),and investigate the cytotoxic capacity of CD8+T cells induced by H22-MPs on hepatocellular carcinoma cells.METHODS:The H22-MPs were extracted by high-speed differential centrifugation,and the morphology of H22-MPs was observed by transmis-sion electron microscopy.The particle size distribution of H22-MPs was detected by dynamic light scattering,and the ex-pression of vesicle surface markers was detected by Western blot.The protein components and pathways involved by MPs were analyzed by labeling free quantitative proteomics combining with bioinformatics GO and KEGG enrichment methods.The effect of MPs antigen on the expression of CD11c,a mature marker of DC,was detected by laser confocal microscopy.Next,CD8+T cells were sorted by magnetic beads,and the secretion of tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and interleukin-2(IL-2)in the supernatant of CD8+T cells stimulated by DC loaded with H22-MPs was detected by ELISA.Lactate dehydrogenase(LDH)test was conducted to evaluate the specific killing effect of CD8+T cells induced by DC loaded with H22-MPs on hepatocellular carcinoma cells.RESULTS:The results showed that the morphology,size and vesicle markers of H22-MPs met the requirements.Most of the H22-MPs protein antigens were derived from its parent cell H22 and coordinated various signal transduction pathways in the cells.Subsequently,it was detected that H22-MPs increased the expression of CD11c on the surface of DC,and H22-MPs were mainly localized in the lysosomes of DC.In addition,DC loaded with H22-MPs stimulated CD8+T to release high levels of TNF-α,IFN-γ and IL-2,and it also pro-moted CD8+T to overexpress CD69,thus inducing CD8+T to killhepatocellular carcinoma cells specifically.CONCLU-SION:H22-MPs promote the maturation of DC,and the mature DC may induce CD8+T cells to play an anti-hepatocellu-lar carcinoma immune response by promoting the activation of CD8+T cells.关键词
肝细胞癌/树突状细胞/微粒/蛋白质组/肿瘤疫苗/CD8+T细胞Key words
hepatocellular carcinoma/dendritic cells/microparticles/proteome/tumor vaccine/CD8+T cells分类
临床医学引用本文复制引用
湛钊,伍嘉豪,黄蓉,卓伟彬,杨萍兰,陈定宇..负载H22-MPs的树突状细胞可增强CD8+T细胞对肝癌细胞的杀伤作用[J].中国病理生理杂志,2025,41(10):1938-1945,8.基金项目
福建医科大学启航基金研究生创新项目(No.2021QH2002) （No.2021QH2002）
荆州市科技计划项目(No.2025HD157) （No.2025HD157）
