军事医学2025，Vol.49Issue(4)：265-272,8.DOI:10.7644/j.issn.1674-9960.2025.04.004
CD8+T细胞体外耗竭模型的建立
Establishment of a CD8+T cell exhaustion model in vitro
摘要
Abstract
Objective To establish a stable in vitro model of CD8+T cell exhaustion.Methods CD8+T cells were isolated and purified from the spleens of ovalbumin-specific CD8+T cell receptor(OT-I)transgenic mice and subjected to chronic antigen stimulation to induce exhaustion in vitro.Flow cytometry was employed to evaluate the expressions of exhaustion markers,secretion of effector cytokines,and transcription factor profiles in CD8+T cells.Exhausted and effector(non-exhausted)CD8+T cells were co-cultured with tumor cells before tumor cell viability was measured to assess the cytotoxic potential of CD8+T cells.Additionally,N-acetyl-L-cysteine(N-AC)was used as a positive control during exhaustion induction to validate the model.Results Chronic stimulation resulted in a significant upregulation of inhibitory receptors,including programmed cell death protein 1(PD-1),T cell immunoglobulin and mucin domain-containing protein 3(TIM-3),T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motifdomain(TIGIT),and lymphocyte activation gene 3(LAG-3).Concurrently,the secretion of key effector cytokines such as tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)was markedly reduced.Exhausted CD8+T cells exhibited diminished cytotoxicity against tumor cells compared to effector CD8+T cells.Notably,treatment with N-AC effectively restored the function of exhausted CD8+T cells and enhanced their anti-tumor activity.Conclusion This study has established an effective in vitro model for CD8+T cell exhaustion.The use of N-AC demonstrates its potential to restore functionality in exhausted CD8+T cells,underscoring the reliability and utility of this model for investigating the anti-tumor potential of exhausted T cells.关键词
CD8+T细胞/T细胞耗竭/耗竭体系/免疫治疗Key words
CD8+T cells/T-cell exhaustion/exhaustion system/immunotherapy分类
生物科学引用本文复制引用
曾令敏,路丁乙,陈佳意,张昊仟,高俊,韩秋影,潘欣..CD8+T细胞体外耗竭模型的建立[J].军事医学,2025,49(4):265-272,8.基金项目
国家杰出青年科学基金项目(8202500506) （8202500506）
