贫血对肿瘤免疫反应的影响及机制研究  

Research on impact and mechanism of anemia on tumor immune response

作　　者：张浩 管静芝 ZHANG Hao;GUAN Jingzhi(Hebei North College Graduate School,Zhangjiakou 075000,Hebei Province,China;Department of Medical Oncology,the Fifth Medical Center,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]河北北方学院研究生院,河北张家口075000 [2]解放军总医院第五医学中心肿瘤内科,北京100853

出　　处：《解放军医学院学报》2024年第11期1134-1145,共12页Academic Journal of Chinese PLA Medical School

基　　金：国家自然科学基金项目(82172902)。

摘　　要：背景合并贫血的癌症患者预后较差,且贫血发挥促肿瘤效应的机制尚未明确。目的观察贫血对荷瘤小鼠外周血单个核细胞(peripheral blood mononuclear cell,PBMC)以及脾和肿瘤组织中的抗肿瘤免疫细胞和免疫抑制性细胞的影响。方法小鼠随机分为健康组、对照组和贫血组。通过流式细胞术检测脾和肿瘤组织中的T淋巴细胞和自然杀伤(natural killer,NK)细胞占比以及免疫能力变化。检测外周血、脾和肿瘤组织中红系祖细胞(erythroid progenitor cells,EPCs)和髓源性抑制细胞(myeloid-derived suppressor cells,MDSCs)的细胞丰度。收集各组肿瘤组织,通过转录组测序(RNA sequencing,RNA-seq)检测贫血组与对照组小鼠的基因表达差异。结果流式细胞术结果显示,对照组小鼠脾中T细胞和NK细胞的比例低于健康组小鼠(P<0.05),贫血组脾中T细胞的比例低于对照组(P<0.05)。对照组小鼠脾中CD4+T细胞和NK细胞分泌的肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和干扰素γ(interferon-γ,IFN-γ)以及CD8+T细胞分泌的IFN-γ增加(P<0.01),贫血抑制了脾中T细胞和NK细胞的活化。贫血减少了CD4^(+)T细胞的浸润(P<0.01),增加了CD8+T细胞和NK细胞的浸润(P<0.01)。贫血抑制了肿瘤组织中的CD4^(+)T细胞和CD8^(+)T细胞分泌TNF-α(P<0.01)以及CD4^(+)T细胞和NK细胞分泌IFN-γ(P<0.05)。贫血组小鼠脾、PBMC和肿瘤组织中CD45+EPCs均高于对照组(P<0.01),且贫血组小鼠肿瘤组织中CD45+EPCs可以分泌ARG1和IFN-γ。贫血增加了脾和肿瘤组织中MDSCs的比例(P<0.01)。RNA-seq分析结果显示,贫血减少了M0型巨噬细胞和激活NK细胞的浸润,增加了初始M2型巨噬细胞和TH2细胞的浸润;上调了TNF-α信号通路和NOD样受体信号通路。结论贫血可能通过抑制T淋巴细胞抗肿瘤免疫反应发挥促肿瘤效应。Background Cancer patients with concomitant anemia have a poorer prognosis,and the mechanism by which anemia exerts a pro-tumor effect is not yet clear.Objective The effects of anemia on anti-tumor immune cells and immunosuppressive cells in peripheral blood mononuclear cells(PBMC),spleen and tumor tissues of tumor-bearing mice were observed.Methods The mice in this study were randomly divided into healthy,control and anemic groups.The percentage of T lymphocytes and natural killer cells(NK cells)and changes in immunocompetence in spleen and tumor tissues were detected by flow cytometry.The cell abundance of erythroid progenitor cells(EPCs)and myeloid-derived suppressor cells(MDSCs)in peripheral blood,spleen and tumor tissues were detected.Tumor tissues were collected from each group and gene expression differences between the anemic and control groups were detected by transcriptome sequencing(RNA sequencing,RNA-seq).Results Flow cytometry results showed that the proportion of T cells and NK cells in the spleens of the control group was lower than that of the healthy group(P<0.05),and the proportion of T cells in the spleens of the anemic group was lower than that of the control group(P<0.05).The secretion of TNF-αand IFN-γby CD4^(+)T cells and NK cells as well as the secretion of IFN-γby CD8^(+)T cells was increased in the spleens of the control group(P<0.01),and anemia inhibited the activation of T cells and NK cells in the spleens.Anemia decreased tumor infiltration by CD4^(+)T cells(P<0.01)and increased CD8^(+)T cells and NK cells(P<0.01),while it inhibited TNF-αsecretion by CD4^(+)and CD8^(+)T cells in tumor tissues(P<0.01),and also inhibited IFN-γsecretion by CD4^(+)T cells and NK cells(P<0.05).In addition,CD45+EPCs in spleen,PBMC and tumor tissues of the anemic group were higher than those in the control group(P<0.01),and CD45+EPCs in tumor tissues of mice in the anemic group could secrete ARG1 and IFN-γ.Anemia increased the proportion of MDSCs in spleen and tumor tissues(P<0.01).RNA-seq analyses sho

关 键 词：贫血 肿瘤微环境 红系祖细胞 髓源性抑制细胞 T淋巴细胞 

分 类 号：R730.6[医药卫生—肿瘤]