四烯甲萘醌促进CD8^(+)T细胞激活和抗肿瘤效应  

Menaquinone-4 enhances CD8^(+) T cell activation and anti-tumor function

作　　者：王佳荣 梁丽云 樊思淼 张学敏[1] 李慧艳[1] 胡怀斌 宋增庆 WANG Jiarong;LIANG Liyun;FAN Simiao;ZHANG Xuemin;LI Huiyan;HU Huaibin;SONG Zengqing(National Center of Biomedical Analysis,Beijing 100039,China)

机构地区：[1]国家生物医学分析中心,北京100039

出　　处：《军事医学》2024年第6期461-467,共7页Military Medical Sciences

基　　金：国家重点研发计划(2022YFC2505001)。

摘　　要：目的 探究四烯甲萘醌(MK4)对CD8^(+)T细胞激活和抗肿瘤效应的影响。方法 构建小鼠原代CD8^(+)T细胞体外培养与激活体系,通过磁珠法分选脾脏中CD8^(+)T细胞,将细胞置于提前包被CD3/CD28抗体的96孔板中激活培养。流式细胞术检测CD8^(+)T细胞激活受体及效应因子表达情况,并通过酶联免疫吸附实验检测培养上清中效应因子含量,评估MK4对CD8^(+)T细胞激活及产生细胞因子的影响;将CD8^(+)T细胞与小鼠结直肠癌MC38细胞按不同比例共培养进行体外杀伤实验,通过检测肿瘤细胞凋亡情况评估MK4对CD8^(+)T细胞杀伤肿瘤细胞能力的影响。结果通过磁珠分选可得到高纯度的小鼠原代CD8^(+)T细胞(97.5%),CD8^(+)T细胞可被包被的CD3/CD28抗体激活并增殖。与对照组相比,CD8^(+)T细胞经MK4处理后,细胞表面激活受体CD25、CD69、CD44表达升高,效应因子白细胞介素-2、γ干扰素、肿瘤坏死因子α、颗粒酶B产生和分泌水平更高。此外,与对照组相比(29.1%),MK4处理组CD8^(+)T细胞诱导肿瘤细胞凋亡比例更高(36.7%)。结论 MK4可促进CD8^(+)T细胞激活及其杀伤肿瘤细胞的功能。Objective To investigate the effects of menaquinone-4(MK4)on the activation and function of CD8^(+)T cells.Methods An in vitro culture system for primary mouse CD8^(+)T cells was established by isolating these cells from the spleen using CD8a T cell isolation kit.The isolated CD8^(+)T cells were then incubated and activated in a 96-well plate coated with anti-CD3/CD28 antibodies.The impact of MK4 on the activation and cytokine secretion of CD8^(+)T cells was explored by quantifying the expression levels of CD8^(+)T cell activation receptors and cytokines using flow cytometry.Additionally,the concentrations of these cytokines in the culture supernatant were measured by enzyme-linked immunosorbent assay(ELISA).The influence of MK4 on the anti-tumor function of CD8^(+)T cells was evaluated by co-culturing colorectal cancer MC38 cells of mice with CD8^(+)T cells at different ratios,and the effect of MK4 was assessed by detecting tumor cell apoptosis.Results High-purity primary CD8^(+)T cells of mice(97.5%)were isolated using the magnetic bead,which could be activated by pre-coated CD3/CD28 antibodies and showed proliferation.Compared with the control group,the MK4-treated group exhibited increased expressions of CD25,CD69 and CD44 on CD8^(+)T cells,as well as higher production and secretion levels of interleukin-2(IL-2),interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α)and granzyme B.In addition,CD8^(+)T cells in the MK4-treated group induced a higher percentage of tumor cell apoptosis(36.7%)compared with the control group(29.1%).Conclusion MK4 can enhance the activation of CD8^(+)T cells and promote anti-tumor functions.

关 键 词：四烯甲萘醌 CD8^(+)T细胞 抗肿瘤 

分 类 号：R967[医药卫生—药理学] R730.5[医药卫生—药学]