PD-1/PD-L1及Treg细胞在JAK2 V617F阳性骨髓增殖性肿瘤中的表达及意义
|
摘要
【目的】探讨程序性死亡受体-1(PD-1)、程序性死亡配体-1(PD-L1)及CD4+CD25+Foxp3+调节性T细胞(Treg)、CD4+/CD8+比值在JAK2 V617F突变阳性骨髓增殖性肿瘤(MPN)患者骨髓中的表达及临床意义。【方法】收集45例JAK2 V617F阳性的MPN患者,其中原发性血小板增多症(ET)17例、真性红细胞增多症(PV)13例、原发性骨髓纤维化(PMF)15例;包括初治组30例、治疗组15例。15例健康志愿者作为对照组。荧光定量聚合酶链反应(FQ-PCR)检测突变型与野生型JAK2比值。免疫组化检测患者及对照骨髓病理切片p-JAK2、PD-1、PD-L1表达情况,流式细胞术检测Treg和淋巴细胞亚群在MPN患者及对照组外周血中的变化。【结果】初治组p-JAK2、PD-1、PD-L1及Treg表达水平明显高于治疗组及对照组(P均<0.05),而CD4+/CD8+比值明显低于治疗组及对照组(P均<0.05)。JAK2 V617F突变量与PD-1及PD-L1正相关,与CD4+/CD8+负相关(r=0.593,P<0.01;r=0.723,P<0.01;r=-0.771,P<0.01)。【结论】p-JAK2、PD-1、PD-L1及Treg、CD4+/CD8+与JAK2 V617F共同参与了骨髓增殖性肿瘤发病。
【Objective】To investigate the expression and clinical significance of programmed death-1(PD-1),programmed death-ligand 1(PD-L1)and CD4+CD25+Foxp3+regulatory T cells(Treg)and CD4+/CD8+ratios in JAK2 V617 F mutation positive myeloroliferative neoplasms patients(MPN).【Methods】45 cases of JAK2 V617 F positive MPN patients were selected including 17 cases of essential thrombocythemia(ET),13 cases of polycythemia vera(PV)and 15 cases of primary myelofibrosis(ET). 30 cases of the newly diagnosed group and 15 cases of treatment group were from them. 15 cases of healthy volunteers were selected as control group. The ratio of mutant and wild type of JAK2 was detected by fluorescence quantitative polymerase chain reaction(FQ-PCR). The expression levels of p-JAK2,PD-1 and PD-L1 in pathological tissues of bone marrow were detected by immunohistochemistry. The changes of treg cells and lymphocyte subsets in peripheral blood of MPN patients and controls were detected by flow cytometry.expression levels of p-JAK2,PD-1,PD-L1,and Treg in the newly diagnosed group were significantly higher than that of treatment group and control group(P<0.05),while the expression levels of CD4+/CD8+ratio were significantly lower than treatment group and control group(P<0.05). JAK2 V617 F mutation burden was positive correlation with PD-1 and PD-L1,and was negative correlation with CD4+/CD8+,the correlation coefficients were r=0.593,P<0.01;r=0.723,P<0.01;r=-0.771,P<0.01,respectively.【Conclusion】p-JAK2,PD-1,PD-L1,Treg,CD4+/CD8+and JAK2 V617 F were involved in the pathogenesis of myeloroliferative neoplasms.
【Objective】To investigate the expression and clinical significance of programmed death-1(PD-1),programmed death-ligand 1(PD-L1)and CD4+CD25+Foxp3+regulatory T cells(Treg)and CD4+/CD8+ratios in JAK2 V617 F mutation positive myeloroliferative neoplasms patients(MPN).【Methods】45 cases of JAK2 V617 F positive MPN patients were selected including 17 cases of essential thrombocythemia(ET),13 cases of polycythemia vera(PV)and 15 cases of primary myelofibrosis(ET). 30 cases of the newly diagnosed group and 15 cases of treatment group were from them. 15 cases of healthy volunteers were selected as control group. The ratio of mutant and wild type of JAK2 was detected by fluorescence quantitative polymerase chain reaction(FQ-PCR). The expression levels of p-JAK2,PD-1 and PD-L1 in pathological tissues of bone marrow were detected by immunohistochemistry. The changes of treg cells and lymphocyte subsets in peripheral blood of MPN patients and controls were detected by flow cytometry.expression levels of p-JAK2,PD-1,PD-L1,and Treg in the newly diagnosed group were significantly higher than that of treatment group and control group(P<0.05),while the expression levels of CD4+/CD8+ratio were significantly lower than treatment group and control group(P<0.05). JAK2 V617 F mutation burden was positive correlation with PD-1 and PD-L1,and was negative correlation with CD4+/CD8+,the correlation coefficients were r=0.593,P<0.01;r=0.723,P<0.01;r=-0.771,P<0.01,respectively.【Conclusion】p-JAK2,PD-1,PD-L1,Treg,CD4+/CD8+and JAK2 V617 F were involved in the pathogenesis of myeloroliferative neoplasms.
引文
[1] Meyer MA,DeNardo DG. Better together:B7S1checkpoint blockade synergizes with anti-PD1[J].Immunity,2018,48(4):621-623.
[2] Hude I,Sasse S,Engert A,et al. The emergingrole of immune checkpoint inhibition in malignantlymphoma[J]. Haematologica,2017,102(1):30-42.
[3] Kamata T,Suzuki A,Mise N,et al. Blockade ofprogrammed death-1/programmed death ligandpathway enhances the antitumor immunity of humaninvariant natural killer T cells[J]. Cancer ImmunolImmunother,2016,65(12):1477-1489.
[4]黄韬,龙兆麟,吴世皓,等. PD-1/PD-L1在膀胱癌患者外周血T细胞和肿瘤细胞表达[J].中山大学学报(医学科学版),2017,38(4):628-631.Huang T,Long ZL,Wu SH,et al. PD-1/PD-L1expressed on T cell and bladder cancer cell[J]. JSun Yat-sen Univ(Med Sci),2017,38(4):628-631.
[5] Downs-Canner S,Berkey S,Delgoffe GM,et al.Suppressive IL-17A+Foxp3+and ex-Th17 IL-17Aneg Foxp3+Treg cells are a source of tumour-associated Treg cells[J]. Nat Commun,2017,8:14649.
[6] Yi T,Li X,Yao S,et al. Host APCs augment invivo expansion of donor natural regulatory T cellsvia B7H1/B7.1 in allogeneic recipients[J]. JImmunol,2011,186(5):2739-2749.
[7] Pianko MJ, Liu Y, Bagchi S, et al. Immunecheckpoint blockade for hematologic malignancies:a review[J]. Stem Cell Investig,2017,4:32-52.
[8] Hao C,Tian J,Liu H,et al. Efficacy and safety ofanti-PD-1 and anti-PD-1 combined with anti-CTLA-4 immunotherapy to advanced melanoma:Asystematic review and meta-analysis of randomizedcontrolled trials[J]. Medicine(Baltimore),2017,96(26):e7325.
[9] Yang H, Bueso-Ramos C, DiNardo C,et al.Expression of PD-L1,PD-L2,PD-1 and CTLA4 inmyelodysplastic syndromes is enhanced by treatmentwith hypomethylating agents[J]. Leukemia,2014,28(6):1280-1288.
[10]付建珠,徐倩,赵亚玲,等.干扰素抑制JAK2V617F阳性骨髓增殖性肿瘤血管新生的机制[J].中华医学杂志,2015,95(46):3727-3732.Fu JZ,Xu Q,Zhao YL,et al. Anti-angiogenic effectof interferon on JAK2V617F positivemyeloproliferative neoplasms and its anti-angiogenicmechanisms[J]. Natl Med J China,2015,95(46):3727-3732.
[11]Chen XF, Fosco D, Douglas E. et al. PD-1regulates extrathymic regulatory T-celldifferentiation[J]. Eur J Immunol,2014,44(9):2603-2616.
[12]Rodríguez JA. HLA-mediated tumor escapemechanisms that may impair immunotherapy clinicaloutcomes via T-cell activation[J]. Oncol Lett,2017,14(4):4415-4427.
[13]Chiu YM,Tsai CL,Kao JT,et al. and PD-L1 up-regulation promotes PD-1 T-cell apoptosis ingastric adenocarcinoma[J]. Anticancer Res,2018,38(4):2069-2078.
[14]朱云霞,陈成,黄建安. PD-1分子在肺癌恶性胸腔积液CD8+T细胞上的表达及其生物学意义[J].肿瘤,2010,30(9):778-781.Zhu YX,Chen C,Hang JA,et al. The expressionof PD-1 on CD8+T cells in malignant pleuraleffusion of lung cancer and its biological significance[J].Tumor,2010,30(9):778-781.
[15]赵亚玲,张丽军,付建珠,等. IFN-α2b对JAK2V617F突变的骨髓增殖性肿瘤COX-2表达及血管新生的影响[J].四川大学学报(医学版),2016,47(4):473-478.Zhao YL,Zhang LJ,Fu JZ,et al. Effect of IFN-α2b on COX-2 and angiogenesis in JAK2V617Fmutation myeloproliferative neoplasms[J]. JSichuan Univ(Med Sci Ed),2016,47(4):473-478.
[2] Hude I,Sasse S,Engert A,et al. The emergingrole of immune checkpoint inhibition in malignantlymphoma[J]. Haematologica,2017,102(1):30-42.
[3] Kamata T,Suzuki A,Mise N,et al. Blockade ofprogrammed death-1/programmed death ligandpathway enhances the antitumor immunity of humaninvariant natural killer T cells[J]. Cancer ImmunolImmunother,2016,65(12):1477-1489.
[4]黄韬,龙兆麟,吴世皓,等. PD-1/PD-L1在膀胱癌患者外周血T细胞和肿瘤细胞表达[J].中山大学学报(医学科学版),2017,38(4):628-631.Huang T,Long ZL,Wu SH,et al. PD-1/PD-L1expressed on T cell and bladder cancer cell[J]. JSun Yat-sen Univ(Med Sci),2017,38(4):628-631.
[5] Downs-Canner S,Berkey S,Delgoffe GM,et al.Suppressive IL-17A+Foxp3+and ex-Th17 IL-17Aneg Foxp3+Treg cells are a source of tumour-associated Treg cells[J]. Nat Commun,2017,8:14649.
[6] Yi T,Li X,Yao S,et al. Host APCs augment invivo expansion of donor natural regulatory T cellsvia B7H1/B7.1 in allogeneic recipients[J]. JImmunol,2011,186(5):2739-2749.
[7] Pianko MJ, Liu Y, Bagchi S, et al. Immunecheckpoint blockade for hematologic malignancies:a review[J]. Stem Cell Investig,2017,4:32-52.
[8] Hao C,Tian J,Liu H,et al. Efficacy and safety ofanti-PD-1 and anti-PD-1 combined with anti-CTLA-4 immunotherapy to advanced melanoma:Asystematic review and meta-analysis of randomizedcontrolled trials[J]. Medicine(Baltimore),2017,96(26):e7325.
[9] Yang H, Bueso-Ramos C, DiNardo C,et al.Expression of PD-L1,PD-L2,PD-1 and CTLA4 inmyelodysplastic syndromes is enhanced by treatmentwith hypomethylating agents[J]. Leukemia,2014,28(6):1280-1288.
[10]付建珠,徐倩,赵亚玲,等.干扰素抑制JAK2V617F阳性骨髓增殖性肿瘤血管新生的机制[J].中华医学杂志,2015,95(46):3727-3732.Fu JZ,Xu Q,Zhao YL,et al. Anti-angiogenic effectof interferon on JAK2V617F positivemyeloproliferative neoplasms and its anti-angiogenicmechanisms[J]. Natl Med J China,2015,95(46):3727-3732.
[11]Chen XF, Fosco D, Douglas E. et al. PD-1regulates extrathymic regulatory T-celldifferentiation[J]. Eur J Immunol,2014,44(9):2603-2616.
[12]Rodríguez JA. HLA-mediated tumor escapemechanisms that may impair immunotherapy clinicaloutcomes via T-cell activation[J]. Oncol Lett,2017,14(4):4415-4427.
[13]Chiu YM,Tsai CL,Kao JT,et al. and PD-L1 up-regulation promotes PD-1 T-cell apoptosis ingastric adenocarcinoma[J]. Anticancer Res,2018,38(4):2069-2078.
[14]朱云霞,陈成,黄建安. PD-1分子在肺癌恶性胸腔积液CD8+T细胞上的表达及其生物学意义[J].肿瘤,2010,30(9):778-781.Zhu YX,Chen C,Hang JA,et al. The expressionof PD-1 on CD8+T cells in malignant pleuraleffusion of lung cancer and its biological significance[J].Tumor,2010,30(9):778-781.
[15]赵亚玲,张丽军,付建珠,等. IFN-α2b对JAK2V617F突变的骨髓增殖性肿瘤COX-2表达及血管新生的影响[J].四川大学学报(医学版),2016,47(4):473-478.Zhao YL,Zhang LJ,Fu JZ,et al. Effect of IFN-α2b on COX-2 and angiogenesis in JAK2V617Fmutation myeloproliferative neoplasms[J]. JSichuan Univ(Med Sci Ed),2016,47(4):473-478.