肝细胞癌组织ICOS~+T细胞的多重免疫组织化学检测及其预后价值研究
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摘要
背景与目的:诱导共刺激分子(inducible costimulator,ICOS)属于B7-CD28免疫球蛋白超家族成员,表达于活化T细胞表面,在T细胞的激活和免疫应答中发挥重要作用;但目前对于ICOS及ICOS~+T细胞在肝细胞癌(hepatocellular carcinoma,HCC)肿瘤组织中的表达及其意义尚不清楚。该研究旨在测定ICOS、ICOS~+T细胞在HCC患者肿瘤组织中的表达情况并探讨其预后价值。方法:应用组织芯片(tissue microarrays,TMAs)和多重免疫组织化学技术(multiplex immunohistochemistry,mIHC)检测2006—2007年在复旦大学附属中山医院接受手术治疗的358例原发性HCC患者肿瘤组织和癌旁组织中ICOS~+细胞、ICOS~+CD4~+及ICOS~+CD8~+T细胞的浸润密度和ICOS~+细胞占CD4~+、CD8~+T细胞的百分比。应用KaplanMeier法和多因素COX回归模型分析上述指标对患者预后的影响。结果:与癌旁组织相比,ICOS~+细胞和ICOS~+CD4~+T细胞在肿瘤组织中浸润数量显著增加(P<0.000 1和P=0.009 1),而ICOS~+CD8~+T细胞则呈相反的降低趋势(P=0.033);在肿瘤中,ICOS~+CD4~+T细胞的浸润程度显著高于ICOS~+CD8~+T细胞。此外,ICOS~+CD4~+和ICOS~+CD8~+T细胞占各自T细胞亚群的百分比在肿瘤组织中均显著增加(P均<0.000 1)。预后分析发现,肿瘤组织中ICOS~+细胞、ICOS~+CD4~+及ICOS~+CD8~+T细胞浸润高的患者总生存期(overall survival,OS)更长,多因素分析证实上述指标是HCC的独立预后良好因素。结论:ICOS在HCC患者的肿瘤组织中高表达,且ICOS~+细胞、ICOS~+CD4~+及ICOS~+CD8~+T细胞是OS的独立预后良好因素,提示上述指标可作为新的HCC预后免疫标志物。
Background and purpose: Inducible costimulator(ICOS) is a member of B7-CD28 immunoglobulin superfamily and expressed on the surface of activated T cells, which plays an important role in T cell activation and effector functions. However, the expression pattern and the significance of ICOS and ICOS~+ T cells in tumor tissue of hepatocellular carcinoma(HCC) are not defined.To this end, the current study was planned to quantitatively detect the expression of ICOS and ICOS~+ T cells in the tumor tissue of HCC patients and evaluate their clinical significance. Methods: Tissue microarrays(TMAs) and multiplex immunohistochemistry(m IHC) were used to detect the expression levels of ICOS~+ cells, ICOS~+CD4~+ and ICOS~+CD8~+ T cells in tumor and paracancerous tissues from HCC patients who received surgical treatment in Zhongshan Hospital, Fudan University from 2006 to 2007(n=358).The clinical prognosis was evaluated by Kaplan-Meier analysis and COX regression analysis. Results: The densities of infiltrating ICOS~+ and ICOS~+CD4~+ T cells were significantly higher in tumor tissue than in paracancerous tissue(P<0.000 1 and P=0.009 1). By contrast, the density of ICOS~+CD8~+ T cells was significantly lower in tumor tissue than in paracancerous tissue(P=0.033). Within the tumor tissue, the density of ICOS~+CD4~+ T cells was significantly higher compared with ICOS~+CD8~+ T cells(P<0.000 1). Moreover,the frequencies of ICOS~+CD4~+ and ICOS~+CD8~+ T cells in tumor tissue were significantly higher compared with their counterparts in paracancerous tissue(P<0.000 1). Multivariate COX regression analysis identified that ICOS~+ cells, ICOS~+CD4~+ and ICOS~+CD8~+ T cells were independent favorable prognostic indices for overall survival(OS). Conclusion: Tumor infiltrating ICOS~+ cells are greatly elevated in the tumor tissue of HCC, and their abundance is associated with prolonged OS. Thus, ICOS~+ cells, ICOS~+CD4~+ and ICOS~+CD8~+ T cells might be used as novel prognostic immune biomarkers for HCC.
Background and purpose: Inducible costimulator(ICOS) is a member of B7-CD28 immunoglobulin superfamily and expressed on the surface of activated T cells, which plays an important role in T cell activation and effector functions. However, the expression pattern and the significance of ICOS and ICOS~+ T cells in tumor tissue of hepatocellular carcinoma(HCC) are not defined.To this end, the current study was planned to quantitatively detect the expression of ICOS and ICOS~+ T cells in the tumor tissue of HCC patients and evaluate their clinical significance. Methods: Tissue microarrays(TMAs) and multiplex immunohistochemistry(m IHC) were used to detect the expression levels of ICOS~+ cells, ICOS~+CD4~+ and ICOS~+CD8~+ T cells in tumor and paracancerous tissues from HCC patients who received surgical treatment in Zhongshan Hospital, Fudan University from 2006 to 2007(n=358).The clinical prognosis was evaluated by Kaplan-Meier analysis and COX regression analysis. Results: The densities of infiltrating ICOS~+ and ICOS~+CD4~+ T cells were significantly higher in tumor tissue than in paracancerous tissue(P<0.000 1 and P=0.009 1). By contrast, the density of ICOS~+CD8~+ T cells was significantly lower in tumor tissue than in paracancerous tissue(P=0.033). Within the tumor tissue, the density of ICOS~+CD4~+ T cells was significantly higher compared with ICOS~+CD8~+ T cells(P<0.000 1). Moreover,the frequencies of ICOS~+CD4~+ and ICOS~+CD8~+ T cells in tumor tissue were significantly higher compared with their counterparts in paracancerous tissue(P<0.000 1). Multivariate COX regression analysis identified that ICOS~+ cells, ICOS~+CD4~+ and ICOS~+CD8~+ T cells were independent favorable prognostic indices for overall survival(OS). Conclusion: Tumor infiltrating ICOS~+ cells are greatly elevated in the tumor tissue of HCC, and their abundance is associated with prolonged OS. Thus, ICOS~+ cells, ICOS~+CD4~+ and ICOS~+CD8~+ T cells might be used as novel prognostic immune biomarkers for HCC.
引文
[1]ZHENG C,ZHENG L,YOO J K,et al.Landscape of infiltrating T cells in liver cancer revealed by single-cell sequencing[J].Cell,2017,169(7):1342-1356.
[2]EL-SERAG H B,MARRERO J A,RUDOLPH L,et al.Diagnosis and treatment of hepatocellular carcinoma[J].Gastroenterology,2008,134(6):1752-1763.
[3]GAO Q,QIU S J,FAN J,et al.Intratumoral balance of regulatory and cytotoxic T cells is associated with prognosis of hepatocellular carcinoma after resection[J].J Clin Oncol,2007,25(18):2586-2593.
[4]BRAHMER J R,TYKODI S S,CHOW L Q,et al.Safety and activity of anti-PD-L1 antibody in patients with advanced cancer[J].N Engl J Med,2012,366(26):2455-2465.
[5]WOLCHOK J D,KLUGER H,CALLAHAN M K,et al.Nivolumab plus ipilimumab in advanced melanoma[J].NEngl J Med,2013,369(2):122-133.
[6]EL-KHOUEIRY A B,SANGRO B,YAU T,et al.Nivolumab in patients with advanced hepatocellular carcinoma(CheckMate040):an open-label,non-comparative,phase 1/2 dose escalation and expansion trial[J].Lancet,2017,389(10088):2492-2502.
[7]WATANABE M,HARA Y,TANABE K,et al.A distinct role for ICOS-mediated co-stimulatory signaling in CD4+and CD8+Tcell subsets[J].Int Immunol,2005,17(3):269-278.
[8]SHARPE A H,FREEMAN G J.The B7-CD28 superfamily[J].Nat Rev Immunol,2002,2(2):116-126.
[9]FAGET J,SISIRAK V,BLAY J Y,et al.ICOS is associated with poor prognosis in breast cancer as it promotes the amplification of immunosuppressive CD4+T cells by plasmacytoid dendritic cells[J].Oncoimmunology,2013,2(3):e23185.
[10]ZHANG Y,LUO Y,QIN S L,et al.The clinical impact of ICOSsignal in colorectal cancer patients[J].Oncoimmunology,2016,5(5):e1141857.
[11]FENG Z,BETHMANN D,KAPPLER M,et al.Multiparametric immune profiling in HPV-oral squamous cell cancer[J].JCIInsight,2017,2(14):93652.
[12]WIKENHEISER D J,STUMHOFER J S.ICOS co-stimulation:friend or foe?[J].Front Immunol,2016,7:304.
[13]ZHANG G,XU Y,ZHOU H.The infiltration of ICOS+cells in nasopharyngeal carcinoma is beneficial for improved prognosis[J].Pathol Oncol Res,2018.[Epub ahead of print]
[14]MCADAM A J,CHANG T T,LUMELSKY A E,et al.Mouse inducible costimulatory molecule(ICOS)expression is enhanced by CD28 costimulation and regulates differentiation of CD4+Tcells[J].J Immunol,2000,165(9):5035-5040.
[15]YAN Z,DANDAN L,HA-JEONG K,et al.A novel role of hematopoietic CCL5 in promoting triple-negative mammary tumor progression by regulating generation of myeloid-derived suppressor cells[J].Cell Res,2013,23(3):394-408.
[16]LEAVENWORTH J W,VERBINNEN B,YIN J,et al.A p85α-osteopontin axis couples the receptor ICOS to sustained Bcl-6expression by follicular helper and regulatory T cells[J].Nat Immunol,2015,16(1):96-106.
[17]BOSSALLER L,BURGER J R,GRIMBACHER B,et al.ICOSdeficiency is associated with a severe reduction of CXCR5+CD4germinal center Th cells[J].J Immunol,2006,177(7):4927-4932.
[18]VALERIE C,CHARLENE T,MARISSA T,et al.Inflammatory tumour microenvironment is associated with superior survival in hepatocellular carcinoma patients[J].J Hepatol,2010,52(3):370-379.
[19]ZHOU G,SPRENGERS D,BOOR P P C,et al.Antibodies against immune checkpoint molecules restore functions of tumor-infiltrating T cells in hepatocellular carcinomas[J].Gastroenterology,2017,153(4):1107-1119.
[20]KIM H D,SONG G W,PARK S,et al.Association between expression level of PD1 by tumor-infiltrating CD8+T cells and features of hepatocellular carcinoma[J].Gastroenterology,2018,155(6):1936-1950.
[21]VONDERHEIDE R H,LORUSSO P M,KHALIL M,et al.Tremelimumab in combination with exemestane in patients with advanced breast cancer and treatment-associated modulation of inducible costimulator expression on patient T cells[J].Clin Cancer Res,2010,16(13):3485-3494.
[22]LIAKOU C I,KAMAT A,TANG D N,et al.CTLA-4 blockade increases IFNgamma-producing CD4+ICOShi cells to shift the ratio of effector to regulatory T cells in cancer patients[J].Proc Natl Acad Sci U S A,2008,105(39):14987-14992.
[23]None.Correction:increased frequency of ICOS+CD4 T cells as a pharmacodynamic biomarker for anti-CTLA-4 therapy[J].Cancer Immunol Res,2017,1(4):229-234.
[24]SLOVIN S F.The need for immune biomarkers for treatment prognosis and response in genitourinary malignancies[J].Biomark Med,2017,11(12):1149-1159.
[2]EL-SERAG H B,MARRERO J A,RUDOLPH L,et al.Diagnosis and treatment of hepatocellular carcinoma[J].Gastroenterology,2008,134(6):1752-1763.
[3]GAO Q,QIU S J,FAN J,et al.Intratumoral balance of regulatory and cytotoxic T cells is associated with prognosis of hepatocellular carcinoma after resection[J].J Clin Oncol,2007,25(18):2586-2593.
[4]BRAHMER J R,TYKODI S S,CHOW L Q,et al.Safety and activity of anti-PD-L1 antibody in patients with advanced cancer[J].N Engl J Med,2012,366(26):2455-2465.
[5]WOLCHOK J D,KLUGER H,CALLAHAN M K,et al.Nivolumab plus ipilimumab in advanced melanoma[J].NEngl J Med,2013,369(2):122-133.
[6]EL-KHOUEIRY A B,SANGRO B,YAU T,et al.Nivolumab in patients with advanced hepatocellular carcinoma(CheckMate040):an open-label,non-comparative,phase 1/2 dose escalation and expansion trial[J].Lancet,2017,389(10088):2492-2502.
[7]WATANABE M,HARA Y,TANABE K,et al.A distinct role for ICOS-mediated co-stimulatory signaling in CD4+and CD8+Tcell subsets[J].Int Immunol,2005,17(3):269-278.
[8]SHARPE A H,FREEMAN G J.The B7-CD28 superfamily[J].Nat Rev Immunol,2002,2(2):116-126.
[9]FAGET J,SISIRAK V,BLAY J Y,et al.ICOS is associated with poor prognosis in breast cancer as it promotes the amplification of immunosuppressive CD4+T cells by plasmacytoid dendritic cells[J].Oncoimmunology,2013,2(3):e23185.
[10]ZHANG Y,LUO Y,QIN S L,et al.The clinical impact of ICOSsignal in colorectal cancer patients[J].Oncoimmunology,2016,5(5):e1141857.
[11]FENG Z,BETHMANN D,KAPPLER M,et al.Multiparametric immune profiling in HPV-oral squamous cell cancer[J].JCIInsight,2017,2(14):93652.
[12]WIKENHEISER D J,STUMHOFER J S.ICOS co-stimulation:friend or foe?[J].Front Immunol,2016,7:304.
[13]ZHANG G,XU Y,ZHOU H.The infiltration of ICOS+cells in nasopharyngeal carcinoma is beneficial for improved prognosis[J].Pathol Oncol Res,2018.[Epub ahead of print]
[14]MCADAM A J,CHANG T T,LUMELSKY A E,et al.Mouse inducible costimulatory molecule(ICOS)expression is enhanced by CD28 costimulation and regulates differentiation of CD4+Tcells[J].J Immunol,2000,165(9):5035-5040.
[15]YAN Z,DANDAN L,HA-JEONG K,et al.A novel role of hematopoietic CCL5 in promoting triple-negative mammary tumor progression by regulating generation of myeloid-derived suppressor cells[J].Cell Res,2013,23(3):394-408.
[16]LEAVENWORTH J W,VERBINNEN B,YIN J,et al.A p85α-osteopontin axis couples the receptor ICOS to sustained Bcl-6expression by follicular helper and regulatory T cells[J].Nat Immunol,2015,16(1):96-106.
[17]BOSSALLER L,BURGER J R,GRIMBACHER B,et al.ICOSdeficiency is associated with a severe reduction of CXCR5+CD4germinal center Th cells[J].J Immunol,2006,177(7):4927-4932.
[18]VALERIE C,CHARLENE T,MARISSA T,et al.Inflammatory tumour microenvironment is associated with superior survival in hepatocellular carcinoma patients[J].J Hepatol,2010,52(3):370-379.
[19]ZHOU G,SPRENGERS D,BOOR P P C,et al.Antibodies against immune checkpoint molecules restore functions of tumor-infiltrating T cells in hepatocellular carcinomas[J].Gastroenterology,2017,153(4):1107-1119.
[20]KIM H D,SONG G W,PARK S,et al.Association between expression level of PD1 by tumor-infiltrating CD8+T cells and features of hepatocellular carcinoma[J].Gastroenterology,2018,155(6):1936-1950.
[21]VONDERHEIDE R H,LORUSSO P M,KHALIL M,et al.Tremelimumab in combination with exemestane in patients with advanced breast cancer and treatment-associated modulation of inducible costimulator expression on patient T cells[J].Clin Cancer Res,2010,16(13):3485-3494.
[22]LIAKOU C I,KAMAT A,TANG D N,et al.CTLA-4 blockade increases IFNgamma-producing CD4+ICOShi cells to shift the ratio of effector to regulatory T cells in cancer patients[J].Proc Natl Acad Sci U S A,2008,105(39):14987-14992.
[23]None.Correction:increased frequency of ICOS+CD4 T cells as a pharmacodynamic biomarker for anti-CTLA-4 therapy[J].Cancer Immunol Res,2017,1(4):229-234.
[24]SLOVIN S F.The need for immune biomarkers for treatment prognosis and response in genitourinary malignancies[J].Biomark Med,2017,11(12):1149-1159.