肝细胞癌患者外周血CD3bright T细胞CD27与PD1表达的应用研究
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摘要
目的通过对肝细胞癌(hepatocellular carcinoma, HCC)患者外周血CD3高表达T细胞(CD3bright T细胞)表型特点与治疗前后细胞CD27、PD1表达变化的分析,探讨CD3bright T细胞在肝癌进展中的临床意义。方法选取2018年9月至2019年2月首都医科大学附属北京佑安医院CD3bright T细胞阳性的原发性HCC患者23例为HCC组,收集患者术前与术后1个月时临床当天检测剩余的全血标本,同时收集健康人30例为对照组,在流式细胞平台上进行CD3bright T细胞的T细胞受体类型、CD27分子与PD1表达水平的检测,同时收集患者临床资料用于统计学分析。结果 CD3bright T细胞主要为γδT细胞,HCC组及对照组中T细胞受体(T cell receptor, TCR)表型一致,与γδT细胞呈显著正相关(r=0.938,P<0.01);HCC组外周血CD3bright T细胞主要为CD27阴性,CD3bright CD27~+T细胞比例显著低于CD27阳性的CD3~+T细胞和CD4~+ T细胞,差异有统计学意义(P<0.05);HCC组CD3bright CD27~+T细胞比例显著低于对照组(P<0.05),CD3bright PD1+T细胞显著高于对照组,差异有统计学意义(P<0.05);HCC组治疗前后,常规T细胞CD27与PD1的表达水平无明显变化,CD3bright CD27~+T细胞和CD3bright PD1+T细胞在术后水平均显著升高,差异有统计学意义(P<0.05)。结论 CD3bright T细胞主要为γδT细胞,与γδT细胞呈显著正相关,可在一定程度上反映外周血γδT细胞的水平。HCC患者外周血中,常规T细胞主要为CD27~+T细胞,CD3bright T细胞主要表现为CD27阴性。CD3bright CD27~+T细胞和CD3bright PD1~+T细胞水平在肝癌术后显著上升,细胞活性及功能发生变化,影响HCC的进展与预后。
Objective To investigate the clinical significance of CD3 bright T cells in the progression of liver cancer by analyzing the phenotypic characteristics and the change of activation function of CD3 bright T cells in peripheral blood of patients with hepatocellular carcinoma(HCC) before and after treatment. Methods Twenty-three patients with HCC who were positive for CD3 bright T cells from Beijing Youan Hospital were enrolled as HCC group. The remaining whole blood specimens were collected on the clinical day before and one month after surgery. At the same time, 30 healthy persons were collected as control group. The T cell receptor type, the expression level of CD27 molecule and programmed death molecule 1(PD1)of CD3 bright T cells were detected by flow cytometry, and the clinical data were collected for statistical analysis. Results CD3 bright T cells were mainly γδT cells. The T cell receptor(TCR) phenotypes were consistent in HCC group and control group, CD3 bright T cells had a significant positive correlation with γδT cells(r = 0.938, P<0.01). In peripheral blood of HCC group, CD3 bright T cells were mainly CD27-negative cells, the proportion of CD3 bright CD27+ T cells was significantly lower than that of the CD3~+CD27~+T cells and CD4~+ CD27~+T cells(P<0.05). The proportion of CD3 bright CD27+ T cells in HCC group was significantly lower than that of control group(P<0.05). CD3 bright PD1~+ T cells in HCC group were significantly higher than that of healthy controls, with statistically significant difference(P<0.05). The expression of CD27 and PD1 in conventional T cells did not change significantly before and after treatment in in HCC group. CD3 bright CD27~+T Cells and CD3 bright PD1~+T cells were significantly elevated after surgery(P<0.05). Conclusions CD3 bright T cells were mainlyγδT cells, which have significant positive correlation with γδT cells, and can reflect the level of γδT cells in peripheral blood to some extent. In peripheral blood of patients with HCC, conventional T cells are mainly CD27~+T cells, CD3 bright T cells are mainly CD27 negative, and PD1 expression is higher than that of conventional T cells. The levels of CD3 bright CD27~+T cells and CD3 bright PD1~+T cells increase significantly after treatment, and the capacity of activation has been changed, which probably exert some positive effect to the progression and prognosis of HCC. The dysfunction of CD3 bright T cells should be clarified in HCC patients.
Objective To investigate the clinical significance of CD3 bright T cells in the progression of liver cancer by analyzing the phenotypic characteristics and the change of activation function of CD3 bright T cells in peripheral blood of patients with hepatocellular carcinoma(HCC) before and after treatment. Methods Twenty-three patients with HCC who were positive for CD3 bright T cells from Beijing Youan Hospital were enrolled as HCC group. The remaining whole blood specimens were collected on the clinical day before and one month after surgery. At the same time, 30 healthy persons were collected as control group. The T cell receptor type, the expression level of CD27 molecule and programmed death molecule 1(PD1)of CD3 bright T cells were detected by flow cytometry, and the clinical data were collected for statistical analysis. Results CD3 bright T cells were mainly γδT cells. The T cell receptor(TCR) phenotypes were consistent in HCC group and control group, CD3 bright T cells had a significant positive correlation with γδT cells(r = 0.938, P<0.01). In peripheral blood of HCC group, CD3 bright T cells were mainly CD27-negative cells, the proportion of CD3 bright CD27+ T cells was significantly lower than that of the CD3~+CD27~+T cells and CD4~+ CD27~+T cells(P<0.05). The proportion of CD3 bright CD27+ T cells in HCC group was significantly lower than that of control group(P<0.05). CD3 bright PD1~+ T cells in HCC group were significantly higher than that of healthy controls, with statistically significant difference(P<0.05). The expression of CD27 and PD1 in conventional T cells did not change significantly before and after treatment in in HCC group. CD3 bright CD27~+T Cells and CD3 bright PD1~+T cells were significantly elevated after surgery(P<0.05). Conclusions CD3 bright T cells were mainlyγδT cells, which have significant positive correlation with γδT cells, and can reflect the level of γδT cells in peripheral blood to some extent. In peripheral blood of patients with HCC, conventional T cells are mainly CD27~+T cells, CD3 bright T cells are mainly CD27 negative, and PD1 expression is higher than that of conventional T cells. The levels of CD3 bright CD27~+T cells and CD3 bright PD1~+T cells increase significantly after treatment, and the capacity of activation has been changed, which probably exert some positive effect to the progression and prognosis of HCC. The dysfunction of CD3 bright T cells should be clarified in HCC patients.
引文
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[2]Bruix J,Gores GJ,Mazzaferro V.Hepatocellular carcinoma:clinical frontiers and perspectives[J].Gut,2014,63:844-855.
[3]Heimbach JK,Kulik LM,Finn RS,et al.AASLD guidelines for the treatment of hepatocellular carcinoma[J].Hepatology,2018,67:358-380.
[4]Burchill MA,Tamburini BA,Kedl RM.T cells compete by cleaving cell surface CD27 and blocking access to CD70-bearing APCs[J].Eur J Immunol,2015,45:3140-3149.
[5]Lombes A,Durand A,Charvet C,et al.Adaptive immune-likeγ/δTlymphocytes share many common features with theirα/βT cell counterparts[J].J Immunol,2015,195:1449-1458.
[6]O'Neill RE,Du W,Mohammadpour H,et al.T cell-derived CD70 delivers an immune checkpoint function in inflammatory T cell responses[J].J Immunol(Baltimore),2017,199:3700-3710.
[7]Peperzak V,Veraar EAM,Keller AM,et al.The Pim kinase pathway contributes to survival signaling in primed CD8+T cells upon CD27costimulation[J].J Immunol,2010,185:6670-6678.
[8]Agata Y,Kawasaki A,Nishimura H,et al.Expression of the PD-1 antigen on the surface of stimulated mouse T and B lymphocytes[J].Int Immunol,1996,8:765-772.
[9]Keir ME,Butte MJ,Freeman GJ,et al.PD-1 and its ligands in tolerance and immunity[J].Annu Rev Immunol,2008,26:677-704.
[10]Schietinger A,Greenberg PD.Tolerance and exhaustion:defining mechanisms of T cell dysfunction[J].Trends Immunol,2014,35:51-60.
[11]El-Khoueiry AB,Sangro B,Yau T,et al.Nivolumab in patients with advanced hepatocellular carcinoma(CheckMate 040):an open-label,non-comparative,phase 1/2 dose escalation and expansion trial[J].Lancet,2017,389:2492-2502.
[12]Rizvi NA,Hellmann MD,Snyder A,et al.Mutational landscape determines sensitivity to PD-1 blockade in non-small cell lung cancer[J].Science,2015,348:124-128.
[13]Paget C,Chow MT,Gherardin NA,et al.CD3bright signals onγδTcells identify IL-17A-producing Vγ6Vδ1+T cells[J].Immunol Cell Biol,2015,93:198-212.
[14]Bao Y,Guo L,Mo J.Characterization ofγδT cells in patients with non-small cell lung cancer[J].Oncol Lett,2017,14:1133-1140.