AIM To investigate the mechanism by which hepatitis C virus(HCV) core protein-induced mi R-93-5 p up-regulation regulates the interferon(IFN) signaling pathway.METHODS HCV-1 b core protein was exogenously expressed in...AIM To investigate the mechanism by which hepatitis C virus(HCV) core protein-induced mi R-93-5 p up-regulation regulates the interferon(IFN) signaling pathway.METHODS HCV-1 b core protein was exogenously expressed in Huh7 cells using pc DNA3.1(+) vector. The expression of mi R-93-5 p and interferon receptor 1(IFNAR1) was measured using quantitative reverse transcriptionpolymerase chain reaction and Western blot. The protein expression and phosphorylation level of STAT1 were evaluated by Western blot. The overexpression and silencing of mi R-93-5 p and IFNAR1 were performed using mi R-93-5 p agomir and antagomir, and pc DNA3.1-IFNAR1 and IFNAR1 si RNA, respectively. Luciferase assay was used to identify whether IFNAR1 is a target of mi R-93-5 p. Cellular experiments were also conducted.RESULTS Serum mi R-93-5 p level was increased in patients with HCV-1 b infection and decreased to normal level after HCV-1 b clearance, but persistently increased in those with pegylated interferon-α resistance, compared with healthy subjects. Serum mi R-93-5 p expression had an AUC value of 0.8359 in distinguishing patients with pegylated interferon-α resistance from those with pegylated interferon-α sensitivity. HCV-1 b core protein increased mi R-93-5 p expression and induced inactivation of the IFN signaling pathway in Huh7 cells. Furthermore, IFNAR1 was identified as a direct target of mi R-93-5 p, and IFNAR1 restore could rescue mi R-93-5 p-reduced STAT1 phosphorylation, suggesting that the mi R-93-5 p-IFNAR1 axis regulates the IFN signaling pathway.CONCLUSION HCV-1 b core protein-induced mi R-93-5 p up-regulation inhibits the IFN signaling pathway by directly targeting IFNAR1, and the mi R-93-5 p-IFNAR1 axis regulates STAT1 phosphorylation. This axis may be a potential therapeutic target for HCV-1 b infection.展开更多
目的探讨1例疑似孟德尔遗传易感分枝杆菌病（Mendelian susceptibility to mycobacterial disease,MSMD）患儿的临床特征,检测IL-12/23-IFN-γ通路的完整性,并进行IFN-γ受体1（IFNGR1）基因分析。方法根据患儿的临床表现及常规的免疫...目的探讨1例疑似孟德尔遗传易感分枝杆菌病（Mendelian susceptibility to mycobacterial disease,MSMD）患儿的临床特征,检测IL-12/23-IFN-γ通路的完整性,并进行IFN-γ受体1（IFNGR1）基因分析。方法根据患儿的临床表现及常规的免疫学筛查试验排除常见原发性免疫缺陷病,Q-RT-PCR在mRNA水平检测患儿及健康对照者IL-12/23-IFN-γ通路的完整性,通过PCR及RT-PCR分别对患儿及其父母的IFNGR1基因进行扩增并测序。结果患儿有播散性卡介苗（BCG）病及全身多系统损害的表现。患儿全血经BCG和IFN-γ共同刺激48 h后IL-12B的表达水平与单独BCG刺激后比较明显降低（P〈0.05）;基因测序发现患儿IFNGR1第6外显子818-821位杂合缺失4个碱基（c.818-821 del TTAA）,使第276位脯氨酸突变为终止密码（p.Asn274fsX276）,父母此位点正常。结论 MSMD患儿临床特征为BCG病及全身多系统损害。Q-RT-PCR检测IL-12/23-IFN-γ通路完整性是一种有效的筛查MSMD的手段;IFNGR1突变是引起本例患儿发生上述临床特征的根本原因。展开更多
Background Genetic variations in the interferon-gamma （IFN-γ） receptor 1 gene （IFNGR1） may contribute to tuberculosis （TB） risk in different populations.Many studies have investigated the relationship between I...Background Genetic variations in the interferon-gamma （IFN-γ） receptor 1 gene （IFNGR1） may contribute to tuberculosis （TB） risk in different populations.Many studies have investigated the relationship between IFNGR1 56C/T polymorphism and the susceptibility to TB,but have yielded conflicting results.A comprehensive meta-analysis is needed to provide a more accurate estimation of the relationship between them.Methods A literature search based on a combination of manual and computer-based methods was conducted on four English databases （PubMed,Science Direct,SpringerLink,and EBSCO） and three Chinese databases （Wanfang,CQVIP,and Chinese National Knowledge Infrastructure databases）.Pooled odds ratios （ORs） and 95％ confidence intervals （95％ Cls） were calculated using either the fixed-effects model or the random-effects model for different genetic models based on the heterogeneity examination.Results A total of six studies comprising 1 497 confirmed TB cases and 1 802 controls were included in this meta-analysis.Overall,no significant association was observed between IFNGR1-56C/T polymorphism and TB susceptibility （C vs.T,OR=0.90,95％ Cl 0.69-1.17; CC vs.TT,OR=0.87,95％ Cl 0.65-1.18; TC vs.TT,OR=-1.031,95％ Cl 0.872-1.219; CC＋TC vs.TT,OR=0.89,95％ Cl 0.64-1.26; CC vs.TC＋TT,OR=0.92,95％ Cl 0.66-1.29）.In subgroup analysis,a significant association was found in the dominant model （CC＋TC vs.TT,OR=1.24,95％ Cl 1.02-1.51） in Africans,but not in Asians or Caucasians.Conclusions Our meta-analysis did not provide enough powerful evidence to identify a significant association between IFNGR1-56C/T polymorphism and TB susceptibility in the overall population.In subgroup analysis,it indicates that IFNGR1-56C/T is possibly associated with increased TB risk in Africans,but not in Asians or Caucasians.However,larger sample size and better-designed case-control studies are needed to validate these findings.展开更多
基金Supported by National Natural Science Foundation of China,No.81371849the TMMU Key Project for Clinical Research,No.2012XLC05
文摘AIM To investigate the mechanism by which hepatitis C virus(HCV) core protein-induced mi R-93-5 p up-regulation regulates the interferon(IFN) signaling pathway.METHODS HCV-1 b core protein was exogenously expressed in Huh7 cells using pc DNA3.1(+) vector. The expression of mi R-93-5 p and interferon receptor 1(IFNAR1) was measured using quantitative reverse transcriptionpolymerase chain reaction and Western blot. The protein expression and phosphorylation level of STAT1 were evaluated by Western blot. The overexpression and silencing of mi R-93-5 p and IFNAR1 were performed using mi R-93-5 p agomir and antagomir, and pc DNA3.1-IFNAR1 and IFNAR1 si RNA, respectively. Luciferase assay was used to identify whether IFNAR1 is a target of mi R-93-5 p. Cellular experiments were also conducted.RESULTS Serum mi R-93-5 p level was increased in patients with HCV-1 b infection and decreased to normal level after HCV-1 b clearance, but persistently increased in those with pegylated interferon-α resistance, compared with healthy subjects. Serum mi R-93-5 p expression had an AUC value of 0.8359 in distinguishing patients with pegylated interferon-α resistance from those with pegylated interferon-α sensitivity. HCV-1 b core protein increased mi R-93-5 p expression and induced inactivation of the IFN signaling pathway in Huh7 cells. Furthermore, IFNAR1 was identified as a direct target of mi R-93-5 p, and IFNAR1 restore could rescue mi R-93-5 p-reduced STAT1 phosphorylation, suggesting that the mi R-93-5 p-IFNAR1 axis regulates the IFN signaling pathway.CONCLUSION HCV-1 b core protein-induced mi R-93-5 p up-regulation inhibits the IFN signaling pathway by directly targeting IFNAR1, and the mi R-93-5 p-IFNAR1 axis regulates STAT1 phosphorylation. This axis may be a potential therapeutic target for HCV-1 b infection.
文摘目的探讨1例疑似孟德尔遗传易感分枝杆菌病（Mendelian susceptibility to mycobacterial disease,MSMD）患儿的临床特征,检测IL-12/23-IFN-γ通路的完整性,并进行IFN-γ受体1（IFNGR1）基因分析。方法根据患儿的临床表现及常规的免疫学筛查试验排除常见原发性免疫缺陷病,Q-RT-PCR在mRNA水平检测患儿及健康对照者IL-12/23-IFN-γ通路的完整性,通过PCR及RT-PCR分别对患儿及其父母的IFNGR1基因进行扩增并测序。结果患儿有播散性卡介苗（BCG）病及全身多系统损害的表现。患儿全血经BCG和IFN-γ共同刺激48 h后IL-12B的表达水平与单独BCG刺激后比较明显降低（P〈0.05）;基因测序发现患儿IFNGR1第6外显子818-821位杂合缺失4个碱基（c.818-821 del TTAA）,使第276位脯氨酸突变为终止密码（p.Asn274fsX276）,父母此位点正常。结论 MSMD患儿临床特征为BCG病及全身多系统损害。Q-RT-PCR检测IL-12/23-IFN-γ通路完整性是一种有效的筛查MSMD的手段;IFNGR1突变是引起本例患儿发生上述临床特征的根本原因。
文摘目的·扩增干扰素调节因子3(interferon regulator factor 3,IRF3)短发夹RNA(short hairpin RNA,sh RNA)腺病毒,并研究该病毒对脂多糖(lipopolysaccharide,LPS)刺激诱导Raw264.7细胞核内白介素受体相关激酶1结合蛋白1(interleukin-1 receptor associated kinase 1 binding protein 1,Irak1bp1)表达的影响。方法·IRF3 sh RNA腺病毒的扩增在人胚肾293 T(HEK293T)细胞中进行,并采用TCID 50法测定病毒滴度。Raw 264.7细胞随机分为4组,1组为腺病毒(-)LPS(-),2组为腺病毒(-)LPS(+),3组为腺病毒(+)LPS(-),4组为腺病毒(+)LPS(+)。细胞IRF3基因表达采用real-time PCR方法检测;核内IRF3及Irak1bp1的表达采用Western blotting方法检测。结果·经计算扩增腺病毒滴度为2.2×10^(11) PFU/m L,最佳MOI为300。LPS刺激后Raw 264.7细胞内IRF3 m RNA较对照组明显增加,核内IRF3蛋白及Irak1bp1表达也明显增加;IRF3 sh RNA腺病毒应用后,细胞对IRF3 m RNA的组成性表达及LPS刺激诱导的IRF3 m RNA和核内蛋白质表达均明显受抑,但未刺激状态下IRF3蛋白核内组成性表达无明显影响;IRF3 sh RNA腺病毒应用对细胞静息及LPS刺激诱导的核内Irak1bp1表达并无影响。结论·IRF3 sh RNA腺病毒能够有效抑制LPS刺激诱导的核内IRF3的表达,但并不影响核内Irak1bp1的表达。
文摘Background Genetic variations in the interferon-gamma （IFN-γ） receptor 1 gene （IFNGR1） may contribute to tuberculosis （TB） risk in different populations.Many studies have investigated the relationship between IFNGR1 56C/T polymorphism and the susceptibility to TB,but have yielded conflicting results.A comprehensive meta-analysis is needed to provide a more accurate estimation of the relationship between them.Methods A literature search based on a combination of manual and computer-based methods was conducted on four English databases （PubMed,Science Direct,SpringerLink,and EBSCO） and three Chinese databases （Wanfang,CQVIP,and Chinese National Knowledge Infrastructure databases）.Pooled odds ratios （ORs） and 95％ confidence intervals （95％ Cls） were calculated using either the fixed-effects model or the random-effects model for different genetic models based on the heterogeneity examination.Results A total of six studies comprising 1 497 confirmed TB cases and 1 802 controls were included in this meta-analysis.Overall,no significant association was observed between IFNGR1-56C/T polymorphism and TB susceptibility （C vs.T,OR=0.90,95％ Cl 0.69-1.17; CC vs.TT,OR=0.87,95％ Cl 0.65-1.18; TC vs.TT,OR=-1.031,95％ Cl 0.872-1.219; CC＋TC vs.TT,OR=0.89,95％ Cl 0.64-1.26; CC vs.TC＋TT,OR=0.92,95％ Cl 0.66-1.29）.In subgroup analysis,a significant association was found in the dominant model （CC＋TC vs.TT,OR=1.24,95％ Cl 1.02-1.51） in Africans,but not in Asians or Caucasians.Conclusions Our meta-analysis did not provide enough powerful evidence to identify a significant association between IFNGR1-56C/T polymorphism and TB susceptibility in the overall population.In subgroup analysis,it indicates that IFNGR1-56C/T is possibly associated with increased TB risk in Africans,but not in Asians or Caucasians.However,larger sample size and better-designed case-control studies are needed to validate these findings.
