Li Baohua, Chen Haiyan, Shi Mengjiao, Huang Na, Kong Guangyao, Li Zongfang
Core Research Laboratory, 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.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2020 Sep;36(9):802-808.
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-γ in 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-1γ significantly 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-1γ and its receptor CCR1 were obtained by protein chip, and they might be associated with the accumulation of splenic MDSCs in hepatoma H22-bearing mice.
目的 研究荷肝癌H22小鼠脾脏中髓源性抑制细胞（MDSC）相关趋化因子的表达及意义。方法 采用流式细胞术检测脾脏MDSC的百分比。通过流式细胞术分选脾脏MDSC并与活化的脾细胞共培养，然后用ELISA法检测共培养细胞上清中IFN-γ的水平。建立小鼠原位肝癌模型。用蛋白质芯片检测正常小鼠和荷瘤小鼠脾脏中细胞因子的差异表达，并用ELISA法验证脾脏MDSC相关趋化因子。还用流式细胞术检测脾脏MDSC上的趋化因子受体。结果 荷瘤小鼠脾脏中MDSC的百分比明显升高，且脾脏MDSC具有免疫抑制功能。用蛋白质芯片筛选出10种上调的细胞因子和9种下调的细胞因子。上调的细胞因子中，有5种细胞因子为趋化因子，即B淋巴细胞趋化因子（BLC）、趋化因子（C-X-C基序）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MIP-2。荷瘤小鼠脾脏中脾脏MDSC相关趋化因子MIP-1γ的水平显著升高，其受体CCR1也在脾脏MDSC的细胞表面表达。结论 通过蛋白质芯片获得了脾脏MDSC相关趋化因子MIP-1γ及其受体CCR1，它们可能与荷肝癌H22小鼠脾脏中MDSC的积聚有关。
