髓源性抑制细胞(MDSC)相关趋化因子MIP-1γ及其受体CCR1在肝癌荷瘤小鼠脾脏的表达及意义  

Expression and significance of myeloid-derived suppressor cells-associated chemokine MIP-1γ and its receptor CCR1 in the spleen of hepatoma H22-bearing mice

作　　者：李宝华 陈海燕[1,2] 石梦娇 黄娜[1,2] 孔光耀 李宗芳[2] LI Baohua;CHEN Haiyan;SHI Mengjiao;HUANG Na;KONG Guangyao;LI Zongfang(Core Research Laboratory,Second Affiliated Hospital,Xi’an Jiaotong University,Xi'an 710004,China;National&Local Joint Engineering Research Center of Biodiagnostics and Biotherapy,Second Affiliated Hospital,Xi’an Jiaotong University,Xi'an 710004,China;Department of Oncology,Second Affiliated Hospital,Xi’an Jiaotong University,Xi'an 710004,China)

机构地区：[1]西安交通大学第二附属医院科研中心实验室,陕西西安710004 [2]西安交通大学第二附属医院生物诊断治疗国家地方联合工程研究中心,陕西西安710004 [3]西安交通大学第二附属医院肿瘤科,陕西西安710004

出　　处：《细胞与分子免疫学杂志》2020年第9期802-808,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81001309);陕西省重点研发计划一般项目-社会发展领域(2018SF-197)。

摘　　要：目的检测髓源性抑制细胞(MDSC)相关趋化因子在肝癌小鼠脾脏中的表达及其意义。方法用流式细胞术检测脾脏MDSC的比例,并采用流式细胞术分选出脾脏MDSC,与活化的脾细胞共培养,ELISA检测共培养上清中IFN-γ的表达水平。制备小鼠原位种植性肝癌模型,用蛋白芯片检测正常小鼠和荷瘤小鼠脾脏的细胞因子差异表达,并用ELISA对脾脏MDSC相关趋化因子进行验证。最后用流式细胞术检测脾脏MDSC表面的趋化因子受体的表达。结果荷瘤小鼠脾脏MDSC的比例显著升高,并且具有免疫抑制功能。蛋白芯片筛选出10个上调的细胞因子和9个下调的细胞因子。在上调的细胞因子中,5个趋化因子是B淋巴细胞趋化因子(BLC)、CXC趋化因子配体16(CXCL16)、巨噬细胞/单核细胞趋化蛋白5(MCP-5)、巨噬细胞炎性蛋白1γ(MIP-1γ)和巨噬细胞炎性蛋白2(MIP-2)。脾脏MDSC相关趋化因子MIP-1γ在荷瘤小鼠脾脏中显著升高,并且脾脏MDSC表面有其受体CC趋化因子受体1(CCR1)的表达。结论用蛋白芯片筛选出脾脏MDSC相关趋化因子MIP-1γ及其受体CCR1,可能与荷瘤小鼠脾脏MDSC聚集有关。Objective To study the expression and significance of myeloid-derived suppressor cell(MDSC)-associated chemokines in the spleen of hepatoma H22-bearing mice.Methods The percentage of splenic MDSCs was examined by flow cytometry.Splenic MDSCs were sorted using flow cytometry and co-cultured with activated splenocytes,and then the level of IFN-γin the supernatant of co-cultured cells was assayed by ELISA.The murine orthotopic hepatoma model was established.The differential cytokine expression in the spleens of normal and tumor-bearing mice was assayed by protein chip,and splenic MDSC-associated chemokines were verified using ELISA.The chemokine receptors on splenic MDSCs were also detected by flow cytometry.Results The percentage of splenic MDSCs was markedly raised in the tumor-bearing mice,and splenic MDSCs exhibited immune-inhibitory function.Ten up-regulated cytokines and nine down-regulated ones were found out using protein chip.Among the up-regulated cytokines,5 cytokines were chemokines,namely B lymphocyte chemoattractant(BLC),chemokine(C-X-C motif)ligand 16(CXCL16),macrophage/monocyte chemotactic protein-5(MCP-5),macrophage inflammatory protein 1γ(MIP-1γ)and MIP-2.The level of splenic MDSC-associated chemokine MIP-1γsignificantly increased in the spleen of tumor-bearing mice,and its receptor CCR1 was also expressed on the cell surface of splenic MDSCs.Conclusion Splenic MDSC-associated chemokine MIP-1γand its receptor CCR1 were obtained by protein chip,and they might be associated with the accumulation of splenic MDSCs in hepatoma H22-bearing mice.

关 键 词：髓源性抑制细胞(MDSC) 趋化因子 肝癌 脾脏 

分 类 号：R735.7[医药卫生—肿瘤] R392.1[医药卫生—临床医学]