肝癌患者CD8~+CD28~-Foxp3~+调节性T细胞的变化及意义
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摘要
目的:探讨肝癌患者外周血和癌组织中CD8~+CD28~-Foxp3~+调节性T细胞(CD8~+CD28-Foxp3~+Tregs)的改变及意义。方法:采用流式细胞仪检测72例肝癌患者和22例健康对照人群外周血CD8~+CD28~-Foxp3~+T细与CD8~+T细胞比值(CD8~+CD28-Foxp3~+Tregs/CD8~+T),分析CD8~+CD28~-Foxp3~+Tregs/CD8~+T与肝癌患者临床病理因素的关系;分别用免疫组化和Western blot检测肝癌患者癌组织及癌旁组织中Foxp3阳性细胞与Foxp3蛋白表达水平。结果:与健康对照人群比较,肝癌患者外周血CD8~+CD28~-Foxp3~+Tregs/CD8~+T明显升高(P<0.05);外周血CD8~+CD28~-Foxp3~+Tregs/CD8~+T与患者TNM分期、淋巴结转移、分化程度有关(均P<0.05);肝癌组织中平均Foxp3阳性细胞数与Foxp3蛋白表达量均明显高于癌旁组织(均P<0.05);外周血CD8~+CD28~-Foxp3~+Tregs/CD8~+T,肝癌组织Foxp3阳性细胞数与Foxp3蛋白表达量在高、中、低分化的肝癌中均呈依次升高趋势,但差异均无统计学意义(均P>0.05)。结论:CD8~+CD28~-Foxp3~+Tregs在肝癌患者外周血及癌组织中增多,可能与肿瘤免疫抑制作用有关,其检测对肝癌患者病情有一定评估价值。
Objective: To investigate the changes in CD8~+CD28~–Foxp3~+ regulatory T cells(CD8~+CD28~–Foxp3~+Tregs)in peripheral blood and tumor tissue of patients with hepatocellular carcinoma(HCC) and the significance.Methods: The ratio of CD8~+CD28~–Foxp3~+Tregs to CD8~+T cells(CD8~+CD28~–Foxp3~+Tregs/CD8~+T) in peripheral blood in 72 HCC patients and 22 healthy controls was determined by flow cytometry, and the relations of CD8~+CD28~–Foxp3~+Tregs/CD8~+T with clinicopathologic factors of the HCC patients were analyzed. The Foxp3 positive cells and Foxp3 protein expression in tumor tissues and their adjacent tissues from HCC patientswere determined by immunohistochemical staining and Western blot analysis, respectively.Results: The peripheral blood CD8~+CD28~–Fop3~+Tregs/CD8~+T was significantly increased in HCC patients compared with that in healthy controls(P<0.05), and the peripheral blood CD8~+CD28~–Fop3~+Tregs/CD8~+T was significantly associated with the TNM stage, lymph node metastasis and degree of tumor differentiation of the HCC patients(all P<0.05). The average number of Foxp3 positive cells and Foxp3 protein expression level were significantly higher in tumor tissue than those in adjacent tissue(both P<0.05). The peripheral blood CD8~+CD28~–Fop3~+Tregs/CD8~+T, and number of Foxp3 positive cells and Foxp3 protein expression level in tumor tissue all presented an increasing trend in the order of well, moderately and poorly differentiated HCC, but all differences did not reach a statistical significance(all P>0.05).Conclusion: CD8~+CD28~–Fop3~+Tregs are increased in the peripheral blood and tumor tissue of HCC patients,which may probably be related to tumor immunosuppression, and their detection may have certain value in assessing the disease states of the HCC patients.
Objective: To investigate the changes in CD8~+CD28~–Foxp3~+ regulatory T cells(CD8~+CD28~–Foxp3~+Tregs)in peripheral blood and tumor tissue of patients with hepatocellular carcinoma(HCC) and the significance.Methods: The ratio of CD8~+CD28~–Foxp3~+Tregs to CD8~+T cells(CD8~+CD28~–Foxp3~+Tregs/CD8~+T) in peripheral blood in 72 HCC patients and 22 healthy controls was determined by flow cytometry, and the relations of CD8~+CD28~–Foxp3~+Tregs/CD8~+T with clinicopathologic factors of the HCC patients were analyzed. The Foxp3 positive cells and Foxp3 protein expression in tumor tissues and their adjacent tissues from HCC patientswere determined by immunohistochemical staining and Western blot analysis, respectively.Results: The peripheral blood CD8~+CD28~–Fop3~+Tregs/CD8~+T was significantly increased in HCC patients compared with that in healthy controls(P<0.05), and the peripheral blood CD8~+CD28~–Fop3~+Tregs/CD8~+T was significantly associated with the TNM stage, lymph node metastasis and degree of tumor differentiation of the HCC patients(all P<0.05). The average number of Foxp3 positive cells and Foxp3 protein expression level were significantly higher in tumor tissue than those in adjacent tissue(both P<0.05). The peripheral blood CD8~+CD28~–Fop3~+Tregs/CD8~+T, and number of Foxp3 positive cells and Foxp3 protein expression level in tumor tissue all presented an increasing trend in the order of well, moderately and poorly differentiated HCC, but all differences did not reach a statistical significance(all P>0.05).Conclusion: CD8~+CD28~–Fop3~+Tregs are increased in the peripheral blood and tumor tissue of HCC patients,which may probably be related to tumor immunosuppression, and their detection may have certain value in assessing the disease states of the HCC patients.
引文
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[18]薛旦旦,夏添松,刘晓安,等.乳腺癌肿瘤微环境中CD8+T细胞和调节性T细胞的研究[J].中华实验外科杂志,2014,31(1):149-151.doi:10.3760/cma.j.issn.1001-9030.2014.01.052.Xue DD,Xia TS,Liu XA,et al.Infiltrating CD8+T cells and regulatory T cells in breast cancer microenvironment[J].Chinese Journal of Experimental Surgery,2014,31(1):149-151.doi:10.3760/cma.j.issn.1001-9030.2014.01.052.
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[20]毛丽伟,廖国清,王红梅,等.胃癌患者不同区域淋巴结和外周血CD4+CD25+FoxP3+调节性T细胞的表达和临床意义[J].免疫学杂志,2016,32(11):1001-1004.Mao LW,Liao GQ,Wang HM,et al.The expression of CD4+CD25+FoxP3+regulated T cells in different regional lymph nodes of gastric carcinoma and its clinical significance[J].Immunological Journal,2016,32(11):1001-1004.
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[22]陈中,倪家连,刘鲁岳,等.CD4+CD25+调节性T细胞在肝癌微环境中的分布状况与局部免疫状态的关系[J].中国普通外科杂志,2007,16(7):690-692.doi:10.3969/j.issn.1005-6947.2007.07.022.Chen Z,Ni JL,Liu LY,et al.Relationship of the distribution and local immunology of CD4+CD25+regulartory T cells in tumor microenvironment of hepatocellular carcinoma[J].Chinese Journal of General Surgery,2007,16(7):690-692.doi:10.3969/j.issn.1005-6947.2007.07.022.
[23]Napoletano C,Bellati F,Ruscito I,et al.Immunological and clinical impact of cancer stem cells in vulvar cancer:Role of CD133/CD24/ABCG2-expressing cells[J].Anticancer Res,2016,36(10):5109-5116.doi:10.21873/anticanres.11080.
[24]王文斌,刘三光,刘兵,等.肝外胆管癌组织叉状头/翅膀状螺旋转录因子3阳性调节性T细胞浸润及其临床意义[J].中华实验外科杂志,2015,32(5):1156-1158.doi:10.3760/cma.j.issn.1001-9030.2015.05.075.Wang WB,Liu SG,Liu B,et al.Significance of forkhead/winged helix transcription factor P3+regulatory T cells infiltration in extrahepatic cholangiocarcinoma[J].Chinese Journal of Experimental Surgery,2015,32(5):1156-1158.doi:10.3760/cma.j.issn.1001-9030.2015.05.075.
[25]Wei R,Hu Y,Dong F,Xu X,et al.Hepatoma cell-derived leptin downregulates the immunosuppressive function of regulatory T-cells to enhance the anti-tumor activity of CD8+T-cells[J].Immunol Cell Biol,2016,94(4):388-399.doi:10.1038/icb.2015.110.
[2]Tao Q,Pan Y,Wang Y,et al.Regulatory T cells-derived IL-35promotes the growth of adult acute myeloid leukemia blasts[J].Int JCancer,2015,137(10):2384-2393.doi:10.1002/ijc.29563.
[3]Ouyang Z,Wu H,Li L,et al.Regulatory T cells in the immunotherapy of melanoma[J].TumourBiol,2016,37(1):77-85.doi:10.1007/s13277-015-4315-0.
[4]Yuan CH,Sun XM,Zhu CL,et al.Amphiregulin activates regulatory T lymphocytes and suppresses CD8+T cell-mediated anti-tumor response in hepatocellular carcinoma cells[J].Oncotarget,2015,6(31):32138-32153.doi:10.18632/oncotarget.5171.
[5]Ichikawa A,Miyoshi H,Arakawa F,et al.Detection of Tax-specific CTLs in lymph nodes of adult T-cell leukemia/lymphoma patients and its association with Foxp3 positivity of regulatory T-cell function[J].Oncol Lett,2017,13(6):4611-4618.doi:10.3892/ol.2017.6067.
[6]Jó?wicki W,Bro?yna AA,Siekiera J,et al.Frequency of CD4+CD25+Foxp3+cells in peripheral blood in relation to urinary bladder cancer malignancy indicators before and after surgical removal[J].Oncotarget,2016,7(10):11450-11462.doi:10.18632/oncotarget.7199.
[7]Mishra AK,Kadoishi T,Wang X,et al.Squamous cell carcinomas escape immune surveillance via inducing chronic activation and exhaustion of CD8+T cells co-expressing PD-1 and LAG-3inhibitory receptors[J].Oncotarget,2016,7(49):81341-81356.doi:10.18632/oncotarget.13228.
[8]吉顺荣,姚宛彤,张波,等.胰腺癌患者外周血CD8+CD28+和CD8+CD2 8-T淋巴细胞亚群的分析[J].上海医学,2 0 1 2,35(4):316-319.Ji SR,Yao WT,Zhang B,et al.Expression of CD8+CD28+and CD8+CD28-in the peripheral blood of pancreatic cancer patients[J].Shanghai Medical Journal,2012,35(4):316-319.
[9]Hasanjani Roushan MR,Bayani M,Soleimani Amiri S,et al.Evaluation of CD4+CD25+FoxP3+regulatory T cells during treatment of patients with brucellosis[J].J Biol Regul Homeost Agents,2016,30(3):675-682.
[10]Lexmond WS,Goettel JA,Lyons JJ,et al.FOXP3+Tregs require WASP to restrain Th2-mediated food allergy[J].J Clin Invest,2016,126(10):4030-4044.doi:10.1172/JCI85129.
[11]Chen C,Chen D,Zhang Y,et al.Changes of CD4+CD25+FoxP3+and CD8+CD28-regulatory T cells in non-small cell lung cancer patients undergoing surgery[J].Int Immunopharmacol,2014,18(2):255-261.doi:10.1016/j.intimp.2013.12.004.
[12]Assadiasl S,Ahmadpoor P,Nafar M,et al.Regulatory T cell subtypes and TGF-β1 gene expression in chronic allograft dysfunction[J].Iran J Immunol,2 0 1 4,1 1(3):1 3 9-1 5 2.doi:IJIv11i3A1.
[13]王徐,胡明华,王小明.原发性肝癌患者微环境中CD8+CD28-调节性T细胞表达变化及意义[J].中国普通外科杂志,2014,23(7):976-979.doi:10.7659/j.issn.1005-6947.2014.07.022.Wang X,Hu MH,Wang XM.Change and significance of CD8+CD28-Treg cells expression in the patients suffering from primary hepatocellular carcinoma[J].Chinese Journal of General Surgery,2014,23(7):976-979.doi:10.7659/j.issn.1005-6947.2014.07.022.
[14]Chan IH,Wu V,Bilardello M,et al.PEG-rIL-10 treatment decreases FoxP3(+)Tregs despite upregulation of intratumoral IDO[J].Oncoimmunology,2016,5(7):e1197458.doi:10.1080/2162402X.2016.1197458.
[15]Cheng H,Luo G,Lu Y,et al.The combination of systemic inflammation-based marker NLR and circulating regulatory T cells predicts the prognosis of resectable pancreatic cancer patients[J].Pancreatology,2016,16(6):1080-1084.doi:10.1016/j.pan.2016.09.007.
[16]Beatty PL,van der Geest R,Hashash JG,et al.Immunobiology and immunosurveillance in patients with intraductal papillary mucinous neoplasms(IPMNs),premalignant precursors of pancreatic adenocarcinomas[J].Cancer Immunol Immunother,2016,65(7):771-778.doi:10.1007/s00262-016-1 838-1.
[17]Choi HS,Ha SY,Kim HM,et al.The prognostic effects of tumor infiltrating regulatory T cells and myeloid derived suppressor cells assessed by multicolor flow cytometry in gastric cancer patients[J].Oncotarget,2016,7(7):7940-7951.doi:10.18632/oncotarget.6958.
[18]薛旦旦,夏添松,刘晓安,等.乳腺癌肿瘤微环境中CD8+T细胞和调节性T细胞的研究[J].中华实验外科杂志,2014,31(1):149-151.doi:10.3760/cma.j.issn.1001-9030.2014.01.052.Xue DD,Xia TS,Liu XA,et al.Infiltrating CD8+T cells and regulatory T cells in breast cancer microenvironment[J].Chinese Journal of Experimental Surgery,2014,31(1):149-151.doi:10.3760/cma.j.issn.1001-9030.2014.01.052.
[19]McCoy MJ,Hemmings C,Miller TJ,et al.Low stromal Foxp3+regulatory T-cell density is associated with complete response to neoadjuvant chemoradiotherapy in rectal cancer[J].Br J Cancer,2015,113(12):1677-186.doi:10.1038/bjc.2015.427.
[20]毛丽伟,廖国清,王红梅,等.胃癌患者不同区域淋巴结和外周血CD4+CD25+FoxP3+调节性T细胞的表达和临床意义[J].免疫学杂志,2016,32(11):1001-1004.Mao LW,Liao GQ,Wang HM,et al.The expression of CD4+CD25+FoxP3+regulated T cells in different regional lymph nodes of gastric carcinoma and its clinical significance[J].Immunological Journal,2016,32(11):1001-1004.
[21]Lee HE,Park DJ,Kim WH,et al.High FOXP3+regulatory T-cell density in the sentinel lymph node is associated with downstream non-sentinel lymph-node metastasis in gastric cancer[J].Br JCancer,2011,105(3):413-419.doi:10.1038/bjc.2011.248.
[22]陈中,倪家连,刘鲁岳,等.CD4+CD25+调节性T细胞在肝癌微环境中的分布状况与局部免疫状态的关系[J].中国普通外科杂志,2007,16(7):690-692.doi:10.3969/j.issn.1005-6947.2007.07.022.Chen Z,Ni JL,Liu LY,et al.Relationship of the distribution and local immunology of CD4+CD25+regulartory T cells in tumor microenvironment of hepatocellular carcinoma[J].Chinese Journal of General Surgery,2007,16(7):690-692.doi:10.3969/j.issn.1005-6947.2007.07.022.
[23]Napoletano C,Bellati F,Ruscito I,et al.Immunological and clinical impact of cancer stem cells in vulvar cancer:Role of CD133/CD24/ABCG2-expressing cells[J].Anticancer Res,2016,36(10):5109-5116.doi:10.21873/anticanres.11080.
[24]王文斌,刘三光,刘兵,等.肝外胆管癌组织叉状头/翅膀状螺旋转录因子3阳性调节性T细胞浸润及其临床意义[J].中华实验外科杂志,2015,32(5):1156-1158.doi:10.3760/cma.j.issn.1001-9030.2015.05.075.Wang WB,Liu SG,Liu B,et al.Significance of forkhead/winged helix transcription factor P3+regulatory T cells infiltration in extrahepatic cholangiocarcinoma[J].Chinese Journal of Experimental Surgery,2015,32(5):1156-1158.doi:10.3760/cma.j.issn.1001-9030.2015.05.075.
[25]Wei R,Hu Y,Dong F,Xu X,et al.Hepatoma cell-derived leptin downregulates the immunosuppressive function of regulatory T-cells to enhance the anti-tumor activity of CD8+T-cells[J].Immunol Cell Biol,2016,94(4):388-399.doi:10.1038/icb.2015.110.