Dendritic cells loaded with H22-MPs enhance killing effect of CD8+T cells on hepatocellular carcinoma cells
10.3969/j.issn.1000-4718.2025.10.008
- VernacularTitle: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
- Author:
Zhao ZHAN
1
;
Jiahao WU
;
Rong HUANG
;
Weibin ZHUO
;
Pinglan YANG
;
Dingyu CHEN
Author Information
1. 长江大学附属第一医院检验科,湖北 荆州 434000;福建医科大学基础医学院,福建 福州 350122
- Publication Type:Journal Article
- Keywords:
hepatocellular carcinoma;
dendritic cells;
microparticles;
proteome;
tumor vaccine;
CD8+T cells
- From:
Chinese Journal of Pathophysiology
2025;41(10):1938-1945
- CountryChina
- Language:Chinese
-
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.
