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目的:探讨肝癌细胞来源分泌型自噬体，即一群表达自噬标志LC3B的细胞外囊泡(LC3Bn + extracellular vesicles, LC3Bn +EVs)对CD8n +T细胞功能耗竭的影响及其作用机制。n 方法:流式细胞术检测肝癌患者外周血和腹水中LC3Bn +EVs和PD-1n +CD8n +T细胞比例并分析二者之间的相关性；分离健康人外周血单个核细胞(peripheral blood mononuclear cells, PBMCs)，在αCD3/CD28和IL-2培养条件下加入LC3Bn +EVs或经热休克蛋白90α(heat shock protein 90α，HSP90α)封闭抗体封闭后的LC3Bn +EVs体外诱导培养72 h，流式细胞术检测PD-1n +CD8n +T细胞和IFN-γn +CD8n + T细胞比例以及上清中IL-2、TNF-α和IFN-γ的浓度。n 结果:肝癌患者血浆和腹水中LC3Bn +EVs及表达HSP90α的LC3Bn +EVs比例显著高于健康对照组和非癌性腹水组，且与PD-1n +CD8n +T细胞耗竭比例均呈显著正相关，尤其是HSP90αn +LC3Bn +EVs。另外，体外人肝癌细胞来源的LC3Bn + EVs可以诱导CD8n +T细胞耗竭，当采用HSP90α封闭抗体预封闭LC3Bn +EVs后，CD8n +T细胞耗竭比例显著降低。n 结论:肝癌细胞来源LC3Bn +EVs通过膜结合型HSP90α诱导CD8n +T细胞功能耗竭。n “,”Objective:To investigate the potential molecular mechanisms of liver cancer cell-derived secretory autophagosomes, extracellular vesicles expressing LC3B (LC3Bn + EVs), in promoting the exhaustion of CD8n + T cells.n Methods:The proportions of LC3Bn + EVs and PD-1n + CD8n + T cells in peripheral blood and ascites of liver cancer patients were measured by flow cytometry. Spearman correlation test was used to analyze the correlation between the proportions of LC3Bn + EVs and PD-1n + CD8n + T cells. Peripheral blood mononuclear cells (PBMCs) from healthy donors were treated with LC3Bn + EVs or heat shock protein 90α (HSP90α) blocking antibody-pretreated LC3Bn + EVs for 72 h in the presence of αCD3/CD28 antibodies and IL-2 n in vitro. The proportions of PD-1n + CD8n + T and IFN-γn + CD8n + T cells and the concentrations of IL-2, TNF-α and IFN-γ in the supernatants were all detected by flow cytometry.n Results:The proportions of LC3Bn + EVs and HSP90αn + LC3Bn + EVs in plasma and ascites from liver cancer patients were significantly higher than those in healthy control group and non-cancerous ascites group. The level of plasma LC3Bn + EVs, especially HSP90αn + LC3Bn + EVs, was significantly correlated with the percentage of exhausted PD-1n + CD8n + T cells. In addition, LC3Bn + EVs from human liver cancer cells up-regulated the percentage of exhausted CD8n + T cells n in vitro. However, LC3Bn + EVs pretreated with HSP90α blocking antibody could significantly inhibit LC3Bn + EVs-induced CD8n + T cell exhaustion.n Conclusions:Liver cancer cell-derived LC3Bn + EVs could effectively induce CD8n + T cell exhaustion mainly through the membrane-bound HSP90α.n