Toll样受体在介导危重型手足口病炎性反应中的作用
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摘要
目的研究Toll样受体在介导危重型手足口病(HFMD)炎性反应中的作用。方法选择2016年07月～2017年03月收治的47例危重型手足口病患儿(危重型组)为研究对象,另选取同期感染科收治30例普通型HFMD作为疾病对照组,门诊健康体检30例儿童为健康对照组,采集外周静脉血,分离单个核细胞(PBMCs),测定TLR2、3、4、7、8、9水平及血清TNF-α、IFN-γ、IL-6水平。结果三组受试者PBMCs中TLR3、TLR4、TLR7和TLR8mRNA表达水平差异有统计学意义(P<0.05),TLR2和TLR9mRNA表达水平差异无统计学意义(P>0.05)。其中危重型手足口病组PBMCs中TLR3、TLR4、TLR7和TLR8mRNA表达水平显著高于其他两组,健康对照组最低(P<0.05)。三组受试者血清中TNF-α、IFN-γ、IL-6表达水平有显著性差异;危重型手足口病患儿血清中TNF-α、IFN-γ、IL-6表达水平显著高于其他两组,健康对照组最低(P<0.05)。Pearson相关性分析显示,PBMCs中TLR3、TLR4、TLR7和TLR8mRNA水平与血清中TNF-α、IFN-γ、IL-6水平呈正相关(P<0.05)。结论危重型手足口病TLR3、TLR4、TLR7和TLR8表达明显增高,进而激活相关信号通路,介导机体的炎性反应。
Objective To investigate the inflammatory response of Toll-like receptor in mediating severe hand-footand mouth disease(HFMD).Methods Twenty-seven children with severe HFMD hospitalized in Intensive Care Unit of Xuzhou Children's Hospital were enrolled in the study from July 2016 to January 2017.Thirty children with common HFMD were selected as disease control,and thirty healthy children as healthy control.Peripheral vein blood samples were collected from each individual,and used to isolate peripheral blood mononuclear cells(PBMCs)for mRNA extraction and sera for measuring levels of tumor necrosis factorα,interferonγ,interleukin 6.Results The expression levels of TLR3,TLR4,TLR7 and TLR8mRNA in PBMCs of children with severe HFMD were significantly higher than those in the other children.The lowest values were found in the healthy control group(P<0.05).The expression levels of TLR2 and TLR9mRNA were not significantly different among the groups(P>0.05).Additionally,the expression levels of TNFα,IFNγand IL 6in the serum of the children in the severe group were significantly higher than those in the other two groups,and the lowest values were again found in the healthy control group(P<0.05).Pearson correlation analysis showed that the TLR3,TLR4,TLR7 and TLR8mRNA levels in PBMCs were positively correlated with the TNFα,IFNγ,IL 6levels in the serum(P<0.05).Conclusion The expression levels of TLR3,TLR4,TLR7 and TLR8is higher,which then leads to the activation downstream signaling and affect the inflammatory response of the organism.
Objective To investigate the inflammatory response of Toll-like receptor in mediating severe hand-footand mouth disease(HFMD).Methods Twenty-seven children with severe HFMD hospitalized in Intensive Care Unit of Xuzhou Children's Hospital were enrolled in the study from July 2016 to January 2017.Thirty children with common HFMD were selected as disease control,and thirty healthy children as healthy control.Peripheral vein blood samples were collected from each individual,and used to isolate peripheral blood mononuclear cells(PBMCs)for mRNA extraction and sera for measuring levels of tumor necrosis factorα,interferonγ,interleukin 6.Results The expression levels of TLR3,TLR4,TLR7 and TLR8mRNA in PBMCs of children with severe HFMD were significantly higher than those in the other children.The lowest values were found in the healthy control group(P<0.05).The expression levels of TLR2 and TLR9mRNA were not significantly different among the groups(P>0.05).Additionally,the expression levels of TNFα,IFNγand IL 6in the serum of the children in the severe group were significantly higher than those in the other two groups,and the lowest values were again found in the healthy control group(P<0.05).Pearson correlation analysis showed that the TLR3,TLR4,TLR7 and TLR8mRNA levels in PBMCs were positively correlated with the TNFα,IFNγ,IL 6levels in the serum(P<0.05).Conclusion The expression levels of TLR3,TLR4,TLR7 and TLR8is higher,which then leads to the activation downstream signaling and affect the inflammatory response of the organism.
引文
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[14] Lu XB,Xu Q,Bu XY,et al.Relationship between expression of toll-like receptors 2/4in dendritic cells and chronic hepatitis B virus infection[J].Int J Clin Exp Pathol,2014,7(9):6048-6055.
[15] Dhanasekaran S,Biswas M,Vignesh AR,et al.Toll-like receptor responses to Peste des petits ruminants virus in goats and water buffalo[J].PLoS One,2014,9(11):e111609.
[16] Ohto U,Shibata T,Tanji H,et al.Structural basis of CpG and inhibitory DNA recognition by Toll-like receptor 9[J].Nature,2015,520(7549):702-705.
[17] Zhu K,Yang JH,Luo KM,et al.TLR3signaling in macrophages is indispensable for the protective immunity of invariant natural killer T cells against enterovirus 71infection[J].PLoS Pathog,2015,11(1):1004613.
[18] Lin YL,Hu YC,Liang CC,et al.Enterovirus-71virus-like particles induce the activation and maturation of human monocyte-derived dendritic cells through TLR4signaling[J].PLoS One,2014,9(10):111496.
[19]李斌,马慧敏,王晓雪,等.Toll样受体7和8在EV71感染死亡病例脑及肺组织中的表达及意义[J].中国当代儿科杂志,2015,17(10):1051-1055.
[20] GongX,Zhou J,ZhuW,et al.Excessive proinflammatory cytokine and chemokine responses of human monocyte-derived macrophages to enterovirus 71infection[J].BMC Infect Dis,2012,12:224.
[21] Hsiao HB,Chou AH,Lin SI,et al.Toll-like receptor 9-mediated protection of enterovirus 71infection in mice is due to the release of danger-associated molecular patterns[J].J Virol,2014,88(20):11658-11670.
[22] Lei X,Sun Z,Liu X,et al.Cleavage of the adaptor protein TRIF by enterovirus 71 3Cinhibits antiviral responses mediated by Toll-like receptor 3[J].J Virol,2011,85(17):8811-8818.
[23] Zhang J,Wang J,Pang L,et al.The co-stimulatory effects of MyD88-dependent Toll-like receptor signaling on activation of murineγδT cells[J].PLoS One,2014,9(9):108156.
[2] Yang J,Yang C,Guo N,et al.Type I Interferons Triggered through the Toll-Like Receptor 3-TRIF Pathway Control Coxsackievirus A16Infection in Young Mice[J].J Virol,2015,89(21):10860-10867.
[3]朱磊,齐共健,席晓莹,等.Toll样受体4和核因子-κB在危重型手足口病中的表达及艾司洛尔的干预作用[J].中华微生物学和免疫学杂志,2015,35(11):827-831.
[4]茌静,何雅青,余光清,等.Toll样受体7在肠道病毒71型感染人Jurkat T细胞诱导炎症因子中的作用[J].中华预防医杂志,2016,50(3):266-269.
[5]中华人民共和国卫生部.手足口病诊疗指南(2010年版)[EB/OL].(2010-04-21)[2017-04-02]http://www.nhfpc.gov.cn/yzygj/s3593g/201306/6d935c0f43cd4a1fb46f8f71acf8e245.shtml.
[6]卫生部手足口病临床专家组.肠道病毒7l型(EV71)感染重症病例临床救治专家共识[J].中华儿科杂志,2011,49(9):675-678.
[7] Dang SS,Gao N,Li YP,et al.Dominant CD4-dependent RNAdependent RNA polymerase-specific T-cell responses in children acutely infected with human enterovirus 71and healthy adult controls[J].Immunology,2014,142(1):89-100.
[8] Wei R,Xu L,Zhang N,et al.Elevated antigen-specific Th2type response is associated with the poor prognosis of hand,foot and mouth disease[J].Virus Res,2013,177(1):62-65.
[9] Tan S,Tan X,Sun X,et al.VP2dominated CD4+T cell responses against enterovirus 71and cross-reactivity against coxsackievirus A16and polioviruses in a healthy population[J].J Immunol,2013,191(4):1637-1647.
[10] Liu L,Zhang Y,Wang J,et al.Study of the integrated immune response induced by an inactivated EV71vaccine[J].PLoS One,2013,8(1):54451.
[11] Wei R,Yang C,Zeng M,et al.A dominant EV71-specific CD4+T cell epitope is highly conserved among human enteroviruses[J].PLoS One,2012,7(12):51957.
[12] Liang Y,Zhou XF,Yang E,et al.Analysis of the Th1/Th2Reaction in the Immune Response Induced by EV71Inactivated Vaccine in Neonatal Rhesus Monkeys[J].J Clin Immunol,2012,32(5):1048-1058.
[13] Dong CH,Liu LD,Zhao HL,et al.Immunoprotection elicited by an enterovirus type 71experimental inactivated vaccine in mice and rhesus monkeys[J].Vaccine,2011,29(37):6269-6275.
[14] Lu XB,Xu Q,Bu XY,et al.Relationship between expression of toll-like receptors 2/4in dendritic cells and chronic hepatitis B virus infection[J].Int J Clin Exp Pathol,2014,7(9):6048-6055.
[15] Dhanasekaran S,Biswas M,Vignesh AR,et al.Toll-like receptor responses to Peste des petits ruminants virus in goats and water buffalo[J].PLoS One,2014,9(11):e111609.
[16] Ohto U,Shibata T,Tanji H,et al.Structural basis of CpG and inhibitory DNA recognition by Toll-like receptor 9[J].Nature,2015,520(7549):702-705.
[17] Zhu K,Yang JH,Luo KM,et al.TLR3signaling in macrophages is indispensable for the protective immunity of invariant natural killer T cells against enterovirus 71infection[J].PLoS Pathog,2015,11(1):1004613.
[18] Lin YL,Hu YC,Liang CC,et al.Enterovirus-71virus-like particles induce the activation and maturation of human monocyte-derived dendritic cells through TLR4signaling[J].PLoS One,2014,9(10):111496.
[19]李斌,马慧敏,王晓雪,等.Toll样受体7和8在EV71感染死亡病例脑及肺组织中的表达及意义[J].中国当代儿科杂志,2015,17(10):1051-1055.
[20] GongX,Zhou J,ZhuW,et al.Excessive proinflammatory cytokine and chemokine responses of human monocyte-derived macrophages to enterovirus 71infection[J].BMC Infect Dis,2012,12:224.
[21] Hsiao HB,Chou AH,Lin SI,et al.Toll-like receptor 9-mediated protection of enterovirus 71infection in mice is due to the release of danger-associated molecular patterns[J].J Virol,2014,88(20):11658-11670.
[22] Lei X,Sun Z,Liu X,et al.Cleavage of the adaptor protein TRIF by enterovirus 71 3Cinhibits antiviral responses mediated by Toll-like receptor 3[J].J Virol,2011,85(17):8811-8818.
[23] Zhang J,Wang J,Pang L,et al.The co-stimulatory effects of MyD88-dependent Toll-like receptor signaling on activation of murineγδT cells[J].PLoS One,2014,9(9):108156.