调节性T淋巴细胞在HER2阳性乳腺癌免疫机制中的作用  

Role of regulatory T lymphocytes in the immune mechanism of HER2-positive breast cancer

作　　者：刘济铭 潘桃 全瑶 LIU Jiming;PAN Tao;QUAN Yao(Department of Emergency,Bishan People's Hospital,Chongqing 402760,China)

机构地区：[1]重庆市璧山区人民医院急诊科,402760

出　　处：《重庆医学》2019年第10期1720-1723,共4页Chongqing medicine

基　　金：重庆市璧山区科技计划项目(201625)

摘　　要：目的探讨调节性T淋巴细胞(Treg细胞)在人表皮生长因子受体2(HER2)阳性乳腺癌患中的免疫作用机制。方法梯度离心方法分离健康献血者(对照组)、HER2阳性未转移乳腺癌患者、HER2阳性转移乳腺癌患者外周血单个核细胞(PBMC)。各组均加入刀豆蛋白A(ConA)刺激增殖分化(5μg/mL)。各组作用60h后采用EdU检测各组细胞增殖情况,流式细胞学检测各组CD4^+CD25^+FOXP^(3+)Treg细胞比例和树突状细胞(DC)比例,实时荧光定量聚合酶链反应检测每组细胞中FoxP3 mRNA的表达,酶联免疫吸附试验(ELISA)检测各组上清液中细胞因子血清转化生长因子-β(TGF-β)、白细胞介素-10(IL-10)的水平。结果 ConA可以刺激各组细胞增加,与对照组相比,HER2阳性未转移乳腺癌组、HER2阳性转移乳腺癌组细胞增殖显著(P<0.01),CD4^+CD25^+FOXP^(3+) Treg细胞占CD4+T淋巴细胞的比例和FoxP3mRNA的表达增加(P<0.01)。与对照组相比,HER2阳性未转移乳腺癌组、HER2阳性转移乳腺癌组DC中CD86分子表达下调(P<0.01)。与对照组相比,HER2阳性未转移乳腺癌组、HER2阳性转移乳腺癌组上清液中TGF-β和IL-10的水平明显升高(P<0.01)。结论通过促进Treg细胞及细胞因子的高表达,从而HER2阳性乳腺癌患者发挥免疫抑制作用。Objective To investigate the immune mechanism of regulatory T lymphocytes(Treg)in patients with human epidermal growth factor receptor 2(HER2)-positive breast cancer.Methods Peripheral blood mononuclear cells(PBMC)were isolated by gradient centrifugation from healthy donors(the control group),HER2-positive non-metastatic breast cancer patients,HER2-positive metastatic breast cancer patients.All groups were processed by congestin A(ConA,5μg/mL)which stimulated proliferation and differentiation.After reaction for 60 hin vitro,EdU assay was used to determine the cellular viability.Flow cytometry was used to detect the ratio of CD4^+CD25^+FoxP3^+ Treg and dendritic cells(DC).FoxP3 mRNA was detected by real-time fluorescent quantitation PCR,and the cytokine levels of serum transforming growth factor-β(TGF-β)and interleukin-10(IL-10)in the supernatant were determined by ELISA.Results ConA stimulated cell proliferation in all groups.Comparing with the control group,the HER2-positive non-metastatic breast cancer group and the HER2-positive metastatic breast cancer group had significant cell proliferation(P<0.01),the proportion of CD4^+CD25^+FoxP3^+ Treg cells in CD4^+T lymphocytes and the expression of FoxP3 mRNA increased(P<0.01),the CD86 expression in DC decreased(P<0.01).In addition,the levels of TGF-βand IL-10 in the supernatant of HER2-positive non-metastatic breast cancer group and the HER2-positive metastatic breast cancer group were significantly higher than those in the control group(P <0.01).Conclusion HER2-positive breast cancer patients exert immunosuppressive effects by promoting the high expression of Treg and cytokines.

关 键 词：树突细胞 T淋巴细胞 调节性 细胞因子类 人外周血单个核细胞 

分 类 号：R392-3[医药卫生—免疫学]