TIPE2、Foxp3、CTLA-4 mRNA在慢性丙型肝炎患者外周血单个核细胞中的表达及临床意义
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摘要
目的探讨慢性丙型肝炎患者外周血单个核细胞(PBMC)中肿瘤坏死因子α诱导蛋白8样分子2(TIPE2)、调节性T淋巴细胞(Treg)功能分子叉头样转录因子3(Foxp3)和细胞毒性T淋巴细胞相关抗原4(CTLA-4)的表达及临床意义。方法选取2012年5月-2016年12月在河北医科大学第三医院门诊及住院的未经抗病毒药物和免疫调节药物治疗的慢性丙型肝炎患者80例,另选取同期健康志愿者45例作为对照。应用实时荧光定量PCR法检测PBMC中TIPE2、Foxp3、CTLA-4 mRNA表达水平,并观察与肝脏血清生化指标及HCV RNA水平的相关性及TIPE2对Foxp3和CTLA-4的影响。计量资料两组间比较采用t检验;计数资料两组间比较采用χ2检验;相关分析采用Pearson相关性检验。结果慢性丙型肝炎患者PBMC中TIPE2 mRNA表达较健康对照组显著降低(0. 564±0. 232 vs 0. 704±0. 165,t=3. 569,P <0. 001); Foxp3 mRNA和CTLA-4 mRNA表达较健康对照组明显升高(0. 701±0. 405 vs 0. 527±0. 184,t=2. 725,P=0. 007; 0. 638±0. 211vs 0. 448±0. 134,t=5. 449,P <0. 001)。TIPE2 mRNA表达与ALT、AST、TBil水平及HCV RNA载量均呈负相关(r值分别为-0. 368、-0. 417、-0. 416、-0. 327,P值分别为0. 001、<0. 001、<0. 001、0. 003); Foxp3 mRNA表达与TBil水平呈负相关(r=-0. 284,P=0. 011); TIPE2 mRNA表达与Foxp3 mRNA、CTLA-4 mRNA表达均呈正相关(r值分别为0. 461、0. 257,P值分别为<0. 001、0. 021)。结论慢性丙型肝炎患者存在TIPE2基因表达及Treg功能分子Foxp3、CTLA-4 mRNA表达数量和功能的异常,并且与肝细胞损伤程度和病毒复制有一定的关系,TIPE2基因可能通过正性调控Treg介导的免疫反应参与了慢性丙型肝炎的发病过程。
Objective To investigate the mRNA expression of tumor necrosis factor-alpha-induced protein 8-like 2( TIPE2),Foxp3( a functional molecule for regulatory T cells),and cytotoxic T-lymphocyte-associated antigen 4( CTLA-4) in peripheral blood mononuclear cells( PBMCs) in patients with chronic hepatitis C( CHC) and their clinical significance. Methods A total of 80 CHC patients who were hospitalized in The Third Hospital of Hebei Medical University from May 2012 to December 2016 and did not receive antiviral drugs or immunoregulatory drugs were enrolled as CHC group,and 45 healthy volunteers were enrolled as controls. Quantitative real-time PCR was used to measure the mRNA expression of TIPE2,Foxp3,and CTLA-4 in PBMCs; the correlation between liver biochemical parameters and HCV RNA was observed,as well as the influence of TIPE2 on Foxp3 and CTLA-4. The t-test was used for comparison of continuous data between two groups,the chi-square test was used for comparison of categorical data between two groups,and Pearson correlation was used for correlation analysis. Results Compared with the healthy control group,the CHC group had a significant reduction in the mRNA expression of TIPE2( 0. 564 ± 0. 232 vs 0. 704 ± 0. 165,t = 3. 569,P < 0. 001) and significant increases in the mRNA expression of Foxp3( 0. 701 ± 0. 405 vs 0. 527 ± 0. 184,t = 2. 725,P = 0. 007) and CTLA-4( 0. 638 ± 0. 211 vs 0. 448 ± 0. 134,t = 5. 449,P < 0. 001). The mRNA expression of TIPE2 was negatively correlated with serum alanine aminotransferase( r =-0. 368,P = 0. 001),aspartate aminotransferase( r =-0. 417,P < 0. 001),total bilirubin( r =-0. 416,P < 0. 001),and HCV RNA load( r =-0. 327,P = 0. 003) and was positively correlated with the mRNA expression of Foxp3( r = 0. 461,P < 0. 001) and CTLA-4( r = 0. 257,P = 0. 021). The mRNA expression of Foxp3 was negatively correlated with total bilirubin( r =-0. 284,P = 0. 011). Conclusion CHC patients have abnormalities in the mRNA expression of TIPE2 and the mRNA expression and function of Foxp3 and CTLA-4,which is associated with the degree of hepatocellular injury and viral replication. The TIPE2 gene may be involved in the pathogenesis of CHC by positively regulating immune response mediated by regulatory T cells.
Objective To investigate the mRNA expression of tumor necrosis factor-alpha-induced protein 8-like 2( TIPE2),Foxp3( a functional molecule for regulatory T cells),and cytotoxic T-lymphocyte-associated antigen 4( CTLA-4) in peripheral blood mononuclear cells( PBMCs) in patients with chronic hepatitis C( CHC) and their clinical significance. Methods A total of 80 CHC patients who were hospitalized in The Third Hospital of Hebei Medical University from May 2012 to December 2016 and did not receive antiviral drugs or immunoregulatory drugs were enrolled as CHC group,and 45 healthy volunteers were enrolled as controls. Quantitative real-time PCR was used to measure the mRNA expression of TIPE2,Foxp3,and CTLA-4 in PBMCs; the correlation between liver biochemical parameters and HCV RNA was observed,as well as the influence of TIPE2 on Foxp3 and CTLA-4. The t-test was used for comparison of continuous data between two groups,the chi-square test was used for comparison of categorical data between two groups,and Pearson correlation was used for correlation analysis. Results Compared with the healthy control group,the CHC group had a significant reduction in the mRNA expression of TIPE2( 0. 564 ± 0. 232 vs 0. 704 ± 0. 165,t = 3. 569,P < 0. 001) and significant increases in the mRNA expression of Foxp3( 0. 701 ± 0. 405 vs 0. 527 ± 0. 184,t = 2. 725,P = 0. 007) and CTLA-4( 0. 638 ± 0. 211 vs 0. 448 ± 0. 134,t = 5. 449,P < 0. 001). The mRNA expression of TIPE2 was negatively correlated with serum alanine aminotransferase( r =-0. 368,P = 0. 001),aspartate aminotransferase( r =-0. 417,P < 0. 001),total bilirubin( r =-0. 416,P < 0. 001),and HCV RNA load( r =-0. 327,P = 0. 003) and was positively correlated with the mRNA expression of Foxp3( r = 0. 461,P < 0. 001) and CTLA-4( r = 0. 257,P = 0. 021). The mRNA expression of Foxp3 was negatively correlated with total bilirubin( r =-0. 284,P = 0. 011). Conclusion CHC patients have abnormalities in the mRNA expression of TIPE2 and the mRNA expression and function of Foxp3 and CTLA-4,which is associated with the degree of hepatocellular injury and viral replication. The TIPE2 gene may be involved in the pathogenesis of CHC by positively regulating immune response mediated by regulatory T cells.
引文
[1]SUN H,GONG S,CARMODY RJ,et al.TIPE2,a negative regulator of innate and adaptive immunity that maintains immune homeostasis[J].Cell,2008,133(3):415-426.
[2]KONG L,LIU K,ZHANG YZ,et al.Downregulation of TIPE2mRNA expression in peripheral blood mononuclear cells from patients with chronic hepatitis C[J].Hepatol Int,2013,7(3):844-849.
[3]SAKAGUCHI S,SAKAGUCHI N,ASANO M,et al.Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains(CD25).Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases[J].J Immunol,1995,155(3):1151-1164.
[4]NAFADY A,NAFADY-HEGO H,ABDEWAHAB NM,et al.Peripheral lymphocytes analyses in children with chronic hepatitis Cvirus infection[J].Eur J Clin Invest,2018,48(10):e13004.
[5]EBINUMA H,NAKAMOTO N,LI Y,et al.Identification and in vitro expansion of functional antigen-specific CD25+Fox P3+regulatory T cells in hepatitis C virus infection[J].J Virol,2008,82(10):5043-5053.
[6]ZHAI N,LI H,SONG H,et al.Hepatitis C virus induces MD-SCs-like monocytes through TLR2/PI3K/AKT/STAT3 signaling[J].PLo S One,2017,12(1):e0170516.
[7]LUAN YY,YAO YM,ZHANG L,et al.Expression of tumor necrosis factor-αinduced protein 8 like-2 contributes to the immunosuppressive property of CD4+CD25+regulatory T cells in mice[J].Mol Immunol,2011,49(1-2):219-226.
[8]ZHANG L,SHI Y,WANG Y,et al.The unique expression profile of human TIPE2 suggests new functions beyond its role in immune regulation[J].Mol Immunol,2011,48(9-10):1209-1215.
[9]FONTENOT JD,GAVIN MA,RUDENSKY AY.Foxp3 programs the development and function of CD4+CD25+regulatory Tcells[J].Nat Immunol,2003,4(4):330-336.
[10]RUDENSKY AY.Regulatory T cells and Foxp3[J].Immunol Rev,2011,241(1):260-268.
[11]PASSERINI L,BACCHETTA R.Forkhead-box-P3 gene transfer in human CD4+T conventional cells for the generation of stable and efficient regulatory T Cells,suitable for immune modulatory therapy[J].Front Immunol,2017,8:1282.
[12]KITAGAWA Y,SAKAGUCHI S.Molecular control of regulatory T cell development and function[J].Curr Opin Immunol,2017,49:64-70.
[13]SMYK-PEARSON S,GOLDEN-MASON L,KLARQUIST J,et al.Functional suppression by FoxP3+CD4+CD25high regulatory T cells during acute hepatitis C virus infection[J].J Infect Dis,2008,197(1):46-57.
[14]GUO FB,WU JZ,AI LM,et al.Clinical significance of Th17/Treg and associated cytokines in peripheral blood in chronic hepatitis C patients with liver cirrhosis[J].J Clin Hepatol,2017,33(3):479-484.(in Chinese)郭飞波,武军驻,艾黎明,等.慢性丙型肝炎肝硬化患者外周血辅助性T淋巴细胞17/调节性T淋巴细胞及相关因子检测的临床意义[J].临床肝胆病杂志,2017,33(3):479-484.
[15]CLAASSEN MA,de KNEGT RJ,TILANUS HW,et al.Abundant numbers of regulatory T cells localize to the liver of chronic hepatitis C infected patients and limit the extent of fibrosis[J].J Hepatol,2010,52(3):315-321.
[16]ISHIBASHI M,YAMAGUCHI H,HIROTANI Y,et al.Contradictory intrahepatic immune responses activated in high-load hepatitis C virus livers compared with low-load livers[J].Arch Virol,2018,163(4):855-865.
[17]AMORAS EDA S,GOMES ST,FREITAS FB,et al.Intrahepatic mRNA expression of FAS,FASL,and FOXP3 genes is associated with the pathophysiology of chronic HCV infection[J].PLo S One,2016,11(5):e0156604.
[18]JIANG YY,ZHANG XH,ZHENG SJ.Change in immune status after antiviral therapy in patients with hepatitis C virus infection[J].J Clin Hepatol,2018,34(2):403-406.(in Chinese)蒋莹莹,张晓慧,郑素军.HCV感染者抗病毒治疗前后免疫状态的变化[J].临床肝胆病杂志,2018,34(2):403-406.
[19]SHEVACH EM,DIPAOLO RA,ANDERSSON J,et al.The lifestyle of naturally occurring CD4+CD25+Foxp3+regulatory Tcells[J].Immunol Rev,2006,212:60-73.
[20]READ S,GREENWALD R,IZCUE A,et al.Blockade of CTLA-4on CD4+CD25+regulatory T cells abrogates their function in vivo[J].J Immunol,2006,177(7):4376-4383.
[21]LI BT,XIAO L.Research progress of the relationship between CTLA-4,CTLA-4 polymorphism and HCV infection[J/CD].Chin J Clinicians:Electronic Edition,2013,7(23):10905-10908.(in Chinese)李邦涛,肖丽.细胞毒性T淋巴细胞相关抗原4及其基因多态性在丙型病毒性肝炎中的研究进展[J/CD].中华临床医师杂志:电子版,2013,7(23):10905-10908.
[22]DOUMBA PP,SERTI E,BOUTSIKOU M,et al.Phenotypic and functional alterations of primary human PBMCs induced by HCV non-enveloped capsid-like particles uptake[J].Cell Mol Life Sci,2013,70(18):3463-3474.
[23]DING J,SU J,ZHANG L,et al.Crocetin activates Foxp3through TIPE2 in asthma-associated Treg cells[J].Cell Physiol Biochem,2015,37(6):2425-2433.
[24]FAN T,HUANG X,LIU C,et al.Egress of murine regulatory Tcells from the thymus requires TIPE2[J].Biochem Biophys Res Commun,2018,500(2):376-383.
[2]KONG L,LIU K,ZHANG YZ,et al.Downregulation of TIPE2mRNA expression in peripheral blood mononuclear cells from patients with chronic hepatitis C[J].Hepatol Int,2013,7(3):844-849.
[3]SAKAGUCHI S,SAKAGUCHI N,ASANO M,et al.Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains(CD25).Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases[J].J Immunol,1995,155(3):1151-1164.
[4]NAFADY A,NAFADY-HEGO H,ABDEWAHAB NM,et al.Peripheral lymphocytes analyses in children with chronic hepatitis Cvirus infection[J].Eur J Clin Invest,2018,48(10):e13004.
[5]EBINUMA H,NAKAMOTO N,LI Y,et al.Identification and in vitro expansion of functional antigen-specific CD25+Fox P3+regulatory T cells in hepatitis C virus infection[J].J Virol,2008,82(10):5043-5053.
[6]ZHAI N,LI H,SONG H,et al.Hepatitis C virus induces MD-SCs-like monocytes through TLR2/PI3K/AKT/STAT3 signaling[J].PLo S One,2017,12(1):e0170516.
[7]LUAN YY,YAO YM,ZHANG L,et al.Expression of tumor necrosis factor-αinduced protein 8 like-2 contributes to the immunosuppressive property of CD4+CD25+regulatory T cells in mice[J].Mol Immunol,2011,49(1-2):219-226.
[8]ZHANG L,SHI Y,WANG Y,et al.The unique expression profile of human TIPE2 suggests new functions beyond its role in immune regulation[J].Mol Immunol,2011,48(9-10):1209-1215.
[9]FONTENOT JD,GAVIN MA,RUDENSKY AY.Foxp3 programs the development and function of CD4+CD25+regulatory Tcells[J].Nat Immunol,2003,4(4):330-336.
[10]RUDENSKY AY.Regulatory T cells and Foxp3[J].Immunol Rev,2011,241(1):260-268.
[11]PASSERINI L,BACCHETTA R.Forkhead-box-P3 gene transfer in human CD4+T conventional cells for the generation of stable and efficient regulatory T Cells,suitable for immune modulatory therapy[J].Front Immunol,2017,8:1282.
[12]KITAGAWA Y,SAKAGUCHI S.Molecular control of regulatory T cell development and function[J].Curr Opin Immunol,2017,49:64-70.
[13]SMYK-PEARSON S,GOLDEN-MASON L,KLARQUIST J,et al.Functional suppression by FoxP3+CD4+CD25high regulatory T cells during acute hepatitis C virus infection[J].J Infect Dis,2008,197(1):46-57.
[14]GUO FB,WU JZ,AI LM,et al.Clinical significance of Th17/Treg and associated cytokines in peripheral blood in chronic hepatitis C patients with liver cirrhosis[J].J Clin Hepatol,2017,33(3):479-484.(in Chinese)郭飞波,武军驻,艾黎明,等.慢性丙型肝炎肝硬化患者外周血辅助性T淋巴细胞17/调节性T淋巴细胞及相关因子检测的临床意义[J].临床肝胆病杂志,2017,33(3):479-484.
[15]CLAASSEN MA,de KNEGT RJ,TILANUS HW,et al.Abundant numbers of regulatory T cells localize to the liver of chronic hepatitis C infected patients and limit the extent of fibrosis[J].J Hepatol,2010,52(3):315-321.
[16]ISHIBASHI M,YAMAGUCHI H,HIROTANI Y,et al.Contradictory intrahepatic immune responses activated in high-load hepatitis C virus livers compared with low-load livers[J].Arch Virol,2018,163(4):855-865.
[17]AMORAS EDA S,GOMES ST,FREITAS FB,et al.Intrahepatic mRNA expression of FAS,FASL,and FOXP3 genes is associated with the pathophysiology of chronic HCV infection[J].PLo S One,2016,11(5):e0156604.
[18]JIANG YY,ZHANG XH,ZHENG SJ.Change in immune status after antiviral therapy in patients with hepatitis C virus infection[J].J Clin Hepatol,2018,34(2):403-406.(in Chinese)蒋莹莹,张晓慧,郑素军.HCV感染者抗病毒治疗前后免疫状态的变化[J].临床肝胆病杂志,2018,34(2):403-406.
[19]SHEVACH EM,DIPAOLO RA,ANDERSSON J,et al.The lifestyle of naturally occurring CD4+CD25+Foxp3+regulatory Tcells[J].Immunol Rev,2006,212:60-73.
[20]READ S,GREENWALD R,IZCUE A,et al.Blockade of CTLA-4on CD4+CD25+regulatory T cells abrogates their function in vivo[J].J Immunol,2006,177(7):4376-4383.
[21]LI BT,XIAO L.Research progress of the relationship between CTLA-4,CTLA-4 polymorphism and HCV infection[J/CD].Chin J Clinicians:Electronic Edition,2013,7(23):10905-10908.(in Chinese)李邦涛,肖丽.细胞毒性T淋巴细胞相关抗原4及其基因多态性在丙型病毒性肝炎中的研究进展[J/CD].中华临床医师杂志:电子版,2013,7(23):10905-10908.
[22]DOUMBA PP,SERTI E,BOUTSIKOU M,et al.Phenotypic and functional alterations of primary human PBMCs induced by HCV non-enveloped capsid-like particles uptake[J].Cell Mol Life Sci,2013,70(18):3463-3474.
[23]DING J,SU J,ZHANG L,et al.Crocetin activates Foxp3through TIPE2 in asthma-associated Treg cells[J].Cell Physiol Biochem,2015,37(6):2425-2433.
[24]FAN T,HUANG X,LIU C,et al.Egress of murine regulatory Tcells from the thymus requires TIPE2[J].Biochem Biophys Res Commun,2018,500(2):376-383.