目的探讨干扰素诱导基因IFIT1与卵巢癌免疫浸润及预后的相关性。方法通过GEO数据库筛选肿瘤免疫相关基因,Kaplan-Meier与Prognoscan预后数据库筛选与卵巢癌预后显著相关的差异基因。GEPIA、Human Protein Atlas与Timer数据库分析IFIT1...目的探讨干扰素诱导基因IFIT1与卵巢癌免疫浸润及预后的相关性。方法通过GEO数据库筛选肿瘤免疫相关基因,Kaplan-Meier与Prognoscan预后数据库筛选与卵巢癌预后显著相关的差异基因。GEPIA、Human Protein Atlas与Timer数据库分析IFIT1在卵巢癌组织与正常组织中的表达差异。UALCAN数据库分析IFIT1在不同分级和分期的卵巢癌组织中的表达情况。David数据库对String和Genemania数据库筛选的相互作用基因与蛋白进行GO富集分析。Timer和Tisidb数据库相互验证IFIT1与免疫细胞的相关性。IFIT1干扰质粒成功构建后,利用IFIT1干扰稳转株构建裸鼠移植瘤模型,观察IFIT1敲降后肿瘤生长情况,免疫组化检测中性粒细胞浸润。结果GEO数据库、Kaplan-Meier与Prognoscan预后数据库筛选出IFIT1是与卵巢癌预后显著负相关的肿瘤免疫基因。GEPIA、Human Protein Atlas、Timer数据库以及UALCAN数据库表明IFIT1在卵巢癌组织中表达显著高于正常卵巢组织(P<0.05),但与肿瘤的分期、分级并不存在显著相关性。String、Genemania、David数据库分析发现IFIT1的互作基因与蛋白富集于2’-5’寡聚腺苷酸合成酶的活化、病毒防御、先天性免疫等过程。Timer数据库表明IFIT1与卵巢癌中CD8+T细胞、B细胞、树突状细胞、中性粒细胞和巨噬细胞浸润呈正相关,其中与中性粒细胞相关性最为明显(P<0.001)。Tisidb与GSCA均验证了IFIT1与卵巢癌组织的中性粒细胞浸润呈高度正相关(P<0.05)。RT-qPCR和Western blot证明卵巢癌细胞中IFIT1被成功敲降后,IFIT1干扰细胞的移植瘤生长显著减缓(P<0.05),肿瘤中性粒细胞浸润减少。结论IFIT1可能促进中性粒细胞浸润影响卵巢癌的恶性进程。展开更多
目的获取小鼠IFIT1(Interferon-induced protein with tetratricopetide repeats 1)特异性多克隆抗体。方法扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过Amylose Pre-packed column亲和层析柱,将所得纯化的融...目的获取小鼠IFIT1(Interferon-induced protein with tetratricopetide repeats 1)特异性多克隆抗体。方法扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过Amylose Pre-packed column亲和层析柱,将所得纯化的融合蛋白MBP-IFIT1作为抗原,添加福氏佐机剂后免疫兔,收集抗血清,用Western blot和ELISA方法测定抗血清特异性反应和效价。结果纯化的MBP-IFIT1可诱导兔产生特异性免疫应答,所得抗体能够特异性识别融合蛋白中的IFIT1,ELISA结果显示其效价为1∶12800。结论MBP-IFIT1具有良好的抗原性,以该融合蛋白为抗原免疫兔成功制备出IFIT1特异性多克隆抗体。展开更多
目的检测干扰素诱导蛋白IFIT1(interferon-induced protein with tetratricopetide repeats1)在小鼠各器官组织的表达状况,并在此基础上探讨颈交感神经阻滞(SB)对烧伤后早期小鼠肝脏IFIT1表达的影响。方法雄性C57/129小鼠50只,分为对照...目的检测干扰素诱导蛋白IFIT1(interferon-induced protein with tetratricopetide repeats1)在小鼠各器官组织的表达状况,并在此基础上探讨颈交感神经阻滞(SB)对烧伤后早期小鼠肝脏IFIT1表达的影响。方法雄性C57/129小鼠50只,分为对照组和SB治疗组,TBSA15%～20%Ⅲ度小鼠烧伤模型,分别于烧伤前、烧伤后1、6、12h和24h取肝组织,采用免疫印迹法(Western blot)观察IFIT1的蛋白表达情况,同时提取正常小鼠心、肝、脾、肺、肾、小肠、淋巴细胞和骨骼肌组织RNA,以RT-PCR检测IFIT1表达情况。结果RT-PCR结果显示IFIT1在正常小鼠各组织均有表达,但组织间的表达量存在差异。Western blot结果表明对照组烧伤后肝组织IFIT1表达显著增高(P<0.01),烧伤后SB治疗组IFIT1表达量较对照组相应时相点降低非常显著(P<0.01)。结论IFIT1在小鼠体内广泛表达。SB对严重创伤的治疗作用可能是通过抑制肝内IFIT1表达来实现的。展开更多
目的：观察小鼠心、肝、脾、肺、肾等器官干扰素诱导蛋白IFIT1（Interferon—induced protein with tetratfieopetide repeats1）的表达情况，探讨颈交感神经阻滞（cervical sympathetic block，SB）对烧伤后早期小鼠各器官IFIT1表达状...目的：观察小鼠心、肝、脾、肺、肾等器官干扰素诱导蛋白IFIT1（Interferon—induced protein with tetratfieopetide repeats1）的表达情况，探讨颈交感神经阻滞（cervical sympathetic block，SB）对烧伤后早期小鼠各器官IFIT1表达状况的影响．方法：将雄性昆明小鼠18只随机分为正常对照组、烧伤组和颈交感神经阻滞（SB）治疗组、烧伤组和SB治疗组采用体表总面积（Total body surface area，TBSA）15％-20％Ⅲ度小鼠烧伤模型，SB治疗组于烧伤后即刻进行双侧颈交感神经阻滞，各组分别于4h后取心、肝、脾、肺、肾组织，提取RNA，以RT—PCR检测IFIT1表达情况．结果：RT—PCR显示IFIT1在各组的各器官均有表达，并存在差异．烧伤组IFIT1在心、肝、脾表达低于正常组，同时在肺、肾表达高于正常组，差异显著（P〈0．01）．SB治疗组IFIT1的表达均低于正常组（P〈0．01）；同时，SB治疗组IFIT1在心、肝、肺、肾的表达也低于烧伤组（P〈0．01）．另外，在各组内的不同器官的IFIT1表达量存在差异（P〈0．05）．结论：烧伤后早期小鼠各器官IFIT1表达有不同的变化，可能与不同器官在烧伤应激反应过程中损伤程度不同有关．SB可显著降低各器官IFIT1的表达．展开更多
目的：观察小鼠应激相关蛋白IFIT1（Interferon-induced protein with tetratricopetide repeats1）和JAB1（c-Jun平activation共处五项原则domain-binding protein 1）在肝脏的表达的细胞定位及氯胺酮对烧伤后早期肝脏IFIT1和JAB1表达...目的：观察小鼠应激相关蛋白IFIT1（Interferon-induced protein with tetratricopetide repeats1）和JAB1（c-Jun平activation共处五项原则domain-binding protein 1）在肝脏的表达的细胞定位及氯胺酮对烧伤后早期肝脏IFIT1和JAB1表达的影响，初步探讨氯胺酮调节炎症反应的分子机制。方法：将15只健康C57／129小鼠随机分为3组（n=5），正常对照组、烧伤组、烧伤＋氯胺酮组。采用TBSA 15～20％Ⅲ度小鼠烧伤模型；氯胺酮10mg／kg，肌肉注射，烧伤15min给予。分别于烧伤后4h脱臼处死，取肝。采用免疫印迹法（Western blot）察各组肝IFIT1和JAB1的表达情况。取正常对照组肝组织作病理切片，行免疫组织化学方法对IFIT1和JAB1的表达做细胞定位。结果：烧伤组IFIT1表达较正常对照组显著增高（P〈0．01）．烧伤组JAB1表达较正常对照组显著降低（P〈0．01）：氯胺酮治疗组IFIT1表达较正常对照组、烧伤对照组显著降低（P〈0．01）．氯胺酮治疗组JAB1表达较正常对照组显著降低（P〈0．05）、较烧伤组显著升高（P〈0．05）。免疫组织化学方法显示．IFIT1在肝细胞胞浆表达为阳性．JAB1在肝细胞胞浆和胞核均表达为阳性。结论：烧伤早期小鼠肝组织IFIT1表达增高而JJAB1表达降低，氯胺酮可降低烧伤早期小鼠肝组织IFIT1的高表达，增高JAB1表达；氯胺酮可通过调控严重烧伤应激中IFIT1和JAB1的表达而在全身炎性反应综合症（Systemic inflammatory response syndrome SIRS）中发挥作用。展开更多
目的探讨IFIT1(interferon-induced protein with tetratricopetide repeats 1)在放射损伤与感染应激中表达的特点和意义。方法采用5μg/mL的脂多糖(LPS)刺激Raw264.7、3T3和10T1/2细胞株5 h后,收集细胞,提取RNA,行RT-PCR观察IFIT1的表...目的探讨IFIT1(interferon-induced protein with tetratricopetide repeats 1)在放射损伤与感染应激中表达的特点和意义。方法采用5μg/mL的脂多糖(LPS)刺激Raw264.7、3T3和10T1/2细胞株5 h后,收集细胞,提取RNA,行RT-PCR观察IFIT1的表达情况,同时设定空白对照组。另外,将20只C57/BL6小鼠随机分为4组,采用12 Gray60Co一次性全身照射,分别于0 h、1 h、4 h和12 h脱颈处死取肝组织,用Western blot测定肝组织IFIT1表达变化。结果 Raw264.7、3T3和10T1/2细胞株经LPS刺激5 h后,均能诱导IFIT1表达,空白对照组表达为阴性。C57/BL6小鼠放射性损伤后,1 h IFIT1/Actin开始增高,12 h达最高,呈上升趋势,差异有极显著性统计学意义(P<0.01)。结论 LPS可刺激多种细胞株表达IFIT1,提示IFIT1可能参与创伤后毒血症的发生和发展;放射损伤早期能引起小鼠肝组织IFIT1的显著增高。展开更多
AIM To investigate the association between interferoninduced protein with tetratricopeptide repeats 1(IFIT1) polymorphisms and interferon-α(IFNα) treatment efficiency among Chinese hepatitis B virus(HBV) infection p...AIM To investigate the association between interferoninduced protein with tetratricopeptide repeats 1(IFIT1) polymorphisms and interferon-α(IFNα) treatment efficiency among Chinese hepatitis B virus(HBV) infection patients.METHODS Two hundred and twenty five newly diagnosed chronichepatitis B(CHB) patients were enrolled in the study. All of these patients received IFNα treatment for a course of 48 wk, and were followed up for 24 wk after the treatment was end. Clinical information about virological response, hepatitis B e antigen(HBe Ag) seroconversion rate and combined response at the end of the treatment, as well as the sustained response by the time of following up 24 wk after the treatment, was collected. Four tag-single nucleotide polymorphisms(SNPs) of IFIT1 were selected and assessed for their association with these clinical outcomes.RESULTS At the end of the treatment, HBe Ag seroconversion was observed in 27.1% patients. Thirty-six point nine percent patients achieved virological response, and 15.6% patients exhibited combined response. Sustained response was obtained in 26.2% patients. The main HBV genotype of the study was genotype B. Patients who infected with HBV genotype B or C showed better treatment efficiency, no matter which clinical outcome was considered. Among the four SNPs assessed, rs303218(A > G) was found to be significantly associated with the end point virological response when assuming additive model [OR = 0.64(95%CI: 0.42-0.96), P = 0.032]. Patients who carried rs303218 GG genotype had a rather higher rate of achieving virological response(response rate: 52%, OR = 0.40, 95%CI: 0.18-0.91; P = 0.028) when compared to those had AA genotype(response rate: 27%). The most significant interaction was observed in patients who had relative lower baseline aspartate transaminase. No association between SNPs and HBe Ag seroconversion, combined response or sustained response was observed.CONCLUSION IFIT1 involves in the regulation of IFNα treatment for CHB and its polymorphism rs303218 can predict the end point virological response. The finding requires further validation.展开更多
Interferon-induced protein with tetratricopeptide repeats 1(IFIT1), also known as interferon-induced protein 56(IFI56) or Interferon-stimulated protein 56(ISG56), was originally identified as a protein induced upon tr...Interferon-induced protein with tetratricopeptide repeats 1(IFIT1), also known as interferon-induced protein 56(IFI56) or Interferon-stimulated protein 56(ISG56), was originally identified as a protein induced upon treatment with interferon and inhibited by viral replication and translational initiation. In this study, Epinephelus lanceolatus IFIT1(ELIFIT1) gene was cloned for the first time. The complete cDNA of El IFIT1 gene includes 2921 nucleotides, and encodes a 437-amino acid(AA) protein. The putative ELIFIT1 protein has 9 TRP domains and is highly similar with IFIT1 proteins in other teleosts. In healthy fish, ELIFIT1 gene was highly expressed in the blood, which indicate its specific function in the peripheral immune system. Its expression was also observed in various immunity-related tissues including spleen, intestine, and kidney, Inducted with spotted knifejaw iridovirus(SKIV), ELIFIT1 gene expression was upregulated in the spleen, kidney, and liver 24 h after induction and reached its peak at 72 h, indicating that ELIFIT1 may play an important role in antivirus. These findings contribute to the understanding of the antiviral regulation of ELIFIT1 gene in teleost.展开更多
为了研究干扰素刺激基因1(Interferon-induced protein with tetratricopeptide repeats 1,IFIT1)在蓝舌病病毒1型(Bluetongue virus serotype 1,BTV1)感染复制过程中的作用,首先利用实时定量PCR检测到BTV1感染绵羊睾丸细胞后IFIT1基因...为了研究干扰素刺激基因1(Interferon-induced protein with tetratricopeptide repeats 1,IFIT1)在蓝舌病病毒1型(Bluetongue virus serotype 1,BTV1)感染复制过程中的作用,首先利用实时定量PCR检测到BTV1感染绵羊睾丸细胞后IFIT1基因的转录水平明显升高,利用RT-PCR方法扩增获得羊IFIT1基因,测序后进行生物信息学分析,将其克隆到质粒载体pcDNA3.1/(+)上,构建重组表达质粒pcDNA3.1-OV-IFIT1,将其转染BHK-21细胞,观察到IFIT1基因在细胞内的成功表达,然后利用BTV1感染质粒pcDNA3.1-OV-IFIT1转染的细胞,从病毒的mRNA转录、蛋白表达和病毒滴度的变化评价IFIT1对BTV1复制的影响。结果显示,IFIT1在细胞中的过表达可显著抑制BTV1复制,相反,敲低IFIT1的表达可促进BTV1的复制。本研究首次报道了干扰素刺激基因IFIT1在BTV1感染复制过程中的作用,这将有助于揭示BTV1和宿主细胞IFIT1的相互作用机制,同时也为抗病毒药物研发提供了理论指导。展开更多
文摘目的探讨干扰素诱导基因IFIT1与卵巢癌免疫浸润及预后的相关性。方法通过GEO数据库筛选肿瘤免疫相关基因,Kaplan-Meier与Prognoscan预后数据库筛选与卵巢癌预后显著相关的差异基因。GEPIA、Human Protein Atlas与Timer数据库分析IFIT1在卵巢癌组织与正常组织中的表达差异。UALCAN数据库分析IFIT1在不同分级和分期的卵巢癌组织中的表达情况。David数据库对String和Genemania数据库筛选的相互作用基因与蛋白进行GO富集分析。Timer和Tisidb数据库相互验证IFIT1与免疫细胞的相关性。IFIT1干扰质粒成功构建后,利用IFIT1干扰稳转株构建裸鼠移植瘤模型,观察IFIT1敲降后肿瘤生长情况,免疫组化检测中性粒细胞浸润。结果GEO数据库、Kaplan-Meier与Prognoscan预后数据库筛选出IFIT1是与卵巢癌预后显著负相关的肿瘤免疫基因。GEPIA、Human Protein Atlas、Timer数据库以及UALCAN数据库表明IFIT1在卵巢癌组织中表达显著高于正常卵巢组织(P<0.05),但与肿瘤的分期、分级并不存在显著相关性。String、Genemania、David数据库分析发现IFIT1的互作基因与蛋白富集于2’-5’寡聚腺苷酸合成酶的活化、病毒防御、先天性免疫等过程。Timer数据库表明IFIT1与卵巢癌中CD8+T细胞、B细胞、树突状细胞、中性粒细胞和巨噬细胞浸润呈正相关,其中与中性粒细胞相关性最为明显(P<0.001)。Tisidb与GSCA均验证了IFIT1与卵巢癌组织的中性粒细胞浸润呈高度正相关(P<0.05)。RT-qPCR和Western blot证明卵巢癌细胞中IFIT1被成功敲降后,IFIT1干扰细胞的移植瘤生长显著减缓(P<0.05),肿瘤中性粒细胞浸润减少。结论IFIT1可能促进中性粒细胞浸润影响卵巢癌的恶性进程。
文摘目的获取小鼠IFIT1(Interferon-induced protein with tetratricopetide repeats 1)特异性多克隆抗体。方法扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过Amylose Pre-packed column亲和层析柱,将所得纯化的融合蛋白MBP-IFIT1作为抗原,添加福氏佐机剂后免疫兔,收集抗血清,用Western blot和ELISA方法测定抗血清特异性反应和效价。结果纯化的MBP-IFIT1可诱导兔产生特异性免疫应答,所得抗体能够特异性识别融合蛋白中的IFIT1,ELISA结果显示其效价为1∶12800。结论MBP-IFIT1具有良好的抗原性,以该融合蛋白为抗原免疫兔成功制备出IFIT1特异性多克隆抗体。
文摘目的检测干扰素诱导蛋白IFIT1(interferon-induced protein with tetratricopetide repeats1)在小鼠各器官组织的表达状况,并在此基础上探讨颈交感神经阻滞(SB)对烧伤后早期小鼠肝脏IFIT1表达的影响。方法雄性C57/129小鼠50只,分为对照组和SB治疗组,TBSA15%～20%Ⅲ度小鼠烧伤模型,分别于烧伤前、烧伤后1、6、12h和24h取肝组织,采用免疫印迹法(Western blot)观察IFIT1的蛋白表达情况,同时提取正常小鼠心、肝、脾、肺、肾、小肠、淋巴细胞和骨骼肌组织RNA,以RT-PCR检测IFIT1表达情况。结果RT-PCR结果显示IFIT1在正常小鼠各组织均有表达,但组织间的表达量存在差异。Western blot结果表明对照组烧伤后肝组织IFIT1表达显著增高(P<0.01),烧伤后SB治疗组IFIT1表达量较对照组相应时相点降低非常显著(P<0.01)。结论IFIT1在小鼠体内广泛表达。SB对严重创伤的治疗作用可能是通过抑制肝内IFIT1表达来实现的。
文摘目的：观察小鼠心、肝、脾、肺、肾等器官干扰素诱导蛋白IFIT1（Interferon—induced protein with tetratfieopetide repeats1）的表达情况，探讨颈交感神经阻滞（cervical sympathetic block，SB）对烧伤后早期小鼠各器官IFIT1表达状况的影响．方法：将雄性昆明小鼠18只随机分为正常对照组、烧伤组和颈交感神经阻滞（SB）治疗组、烧伤组和SB治疗组采用体表总面积（Total body surface area，TBSA）15％-20％Ⅲ度小鼠烧伤模型，SB治疗组于烧伤后即刻进行双侧颈交感神经阻滞，各组分别于4h后取心、肝、脾、肺、肾组织，提取RNA，以RT—PCR检测IFIT1表达情况．结果：RT—PCR显示IFIT1在各组的各器官均有表达，并存在差异．烧伤组IFIT1在心、肝、脾表达低于正常组，同时在肺、肾表达高于正常组，差异显著（P〈0．01）．SB治疗组IFIT1的表达均低于正常组（P〈0．01）；同时，SB治疗组IFIT1在心、肝、肺、肾的表达也低于烧伤组（P〈0．01）．另外，在各组内的不同器官的IFIT1表达量存在差异（P〈0．05）．结论：烧伤后早期小鼠各器官IFIT1表达有不同的变化，可能与不同器官在烧伤应激反应过程中损伤程度不同有关．SB可显著降低各器官IFIT1的表达．
文摘目的：观察小鼠应激相关蛋白IFIT1（Interferon-induced protein with tetratricopetide repeats1）和JAB1（c-Jun平activation共处五项原则domain-binding protein 1）在肝脏的表达的细胞定位及氯胺酮对烧伤后早期肝脏IFIT1和JAB1表达的影响，初步探讨氯胺酮调节炎症反应的分子机制。方法：将15只健康C57／129小鼠随机分为3组（n=5），正常对照组、烧伤组、烧伤＋氯胺酮组。采用TBSA 15～20％Ⅲ度小鼠烧伤模型；氯胺酮10mg／kg，肌肉注射，烧伤15min给予。分别于烧伤后4h脱臼处死，取肝。采用免疫印迹法（Western blot）察各组肝IFIT1和JAB1的表达情况。取正常对照组肝组织作病理切片，行免疫组织化学方法对IFIT1和JAB1的表达做细胞定位。结果：烧伤组IFIT1表达较正常对照组显著增高（P〈0．01）．烧伤组JAB1表达较正常对照组显著降低（P〈0．01）：氯胺酮治疗组IFIT1表达较正常对照组、烧伤对照组显著降低（P〈0．01）．氯胺酮治疗组JAB1表达较正常对照组显著降低（P〈0．05）、较烧伤组显著升高（P〈0．05）。免疫组织化学方法显示．IFIT1在肝细胞胞浆表达为阳性．JAB1在肝细胞胞浆和胞核均表达为阳性。结论：烧伤早期小鼠肝组织IFIT1表达增高而JJAB1表达降低，氯胺酮可降低烧伤早期小鼠肝组织IFIT1的高表达，增高JAB1表达；氯胺酮可通过调控严重烧伤应激中IFIT1和JAB1的表达而在全身炎性反应综合症（Systemic inflammatory response syndrome SIRS）中发挥作用。
文摘AIM To investigate the association between interferoninduced protein with tetratricopeptide repeats 1(IFIT1) polymorphisms and interferon-α(IFNα) treatment efficiency among Chinese hepatitis B virus(HBV) infection patients.METHODS Two hundred and twenty five newly diagnosed chronichepatitis B(CHB) patients were enrolled in the study. All of these patients received IFNα treatment for a course of 48 wk, and were followed up for 24 wk after the treatment was end. Clinical information about virological response, hepatitis B e antigen(HBe Ag) seroconversion rate and combined response at the end of the treatment, as well as the sustained response by the time of following up 24 wk after the treatment, was collected. Four tag-single nucleotide polymorphisms(SNPs) of IFIT1 were selected and assessed for their association with these clinical outcomes.RESULTS At the end of the treatment, HBe Ag seroconversion was observed in 27.1% patients. Thirty-six point nine percent patients achieved virological response, and 15.6% patients exhibited combined response. Sustained response was obtained in 26.2% patients. The main HBV genotype of the study was genotype B. Patients who infected with HBV genotype B or C showed better treatment efficiency, no matter which clinical outcome was considered. Among the four SNPs assessed, rs303218(A > G) was found to be significantly associated with the end point virological response when assuming additive model [OR = 0.64(95%CI: 0.42-0.96), P = 0.032]. Patients who carried rs303218 GG genotype had a rather higher rate of achieving virological response(response rate: 52%, OR = 0.40, 95%CI: 0.18-0.91; P = 0.028) when compared to those had AA genotype(response rate: 27%). The most significant interaction was observed in patients who had relative lower baseline aspartate transaminase. No association between SNPs and HBe Ag seroconversion, combined response or sustained response was observed.CONCLUSION IFIT1 involves in the regulation of IFNα treatment for CHB and its polymorphism rs303218 can predict the end point virological response. The finding requires further validation.
基金supported by the Shandong Breeding Project (No. 2016LZGC009)the Projects from Laboratory for Marine Fisheries Science and Food Production Processes+2 种基金Pilot National Laboratory for Marine Science and Technology (Qingdao)(Nos. 2018-MFS-T08, 2017A STCP-OS15)the Central Public-interest Scientific Institution Basal Research Fund,CAFS (No. 2020TD20)the Central Public-Interest Scientific Institution Basal Re-search Fund,YSFRI,CAFS (No. 20603022018026)。
文摘Interferon-induced protein with tetratricopeptide repeats 1(IFIT1), also known as interferon-induced protein 56(IFI56) or Interferon-stimulated protein 56(ISG56), was originally identified as a protein induced upon treatment with interferon and inhibited by viral replication and translational initiation. In this study, Epinephelus lanceolatus IFIT1(ELIFIT1) gene was cloned for the first time. The complete cDNA of El IFIT1 gene includes 2921 nucleotides, and encodes a 437-amino acid(AA) protein. The putative ELIFIT1 protein has 9 TRP domains and is highly similar with IFIT1 proteins in other teleosts. In healthy fish, ELIFIT1 gene was highly expressed in the blood, which indicate its specific function in the peripheral immune system. Its expression was also observed in various immunity-related tissues including spleen, intestine, and kidney, Inducted with spotted knifejaw iridovirus(SKIV), ELIFIT1 gene expression was upregulated in the spleen, kidney, and liver 24 h after induction and reached its peak at 72 h, indicating that ELIFIT1 may play an important role in antivirus. These findings contribute to the understanding of the antiviral regulation of ELIFIT1 gene in teleost.
文摘为了研究干扰素刺激基因1(Interferon-induced protein with tetratricopeptide repeats 1,IFIT1)在蓝舌病病毒1型(Bluetongue virus serotype 1,BTV1)感染复制过程中的作用,首先利用实时定量PCR检测到BTV1感染绵羊睾丸细胞后IFIT1基因的转录水平明显升高,利用RT-PCR方法扩增获得羊IFIT1基因,测序后进行生物信息学分析,将其克隆到质粒载体pcDNA3.1/(+)上,构建重组表达质粒pcDNA3.1-OV-IFIT1,将其转染BHK-21细胞,观察到IFIT1基因在细胞内的成功表达,然后利用BTV1感染质粒pcDNA3.1-OV-IFIT1转染的细胞,从病毒的mRNA转录、蛋白表达和病毒滴度的变化评价IFIT1对BTV1复制的影响。结果显示,IFIT1在细胞中的过表达可显著抑制BTV1复制,相反,敲低IFIT1的表达可促进BTV1的复制。本研究首次报道了干扰素刺激基因IFIT1在BTV1感染复制过程中的作用,这将有助于揭示BTV1和宿主细胞IFIT1的相互作用机制,同时也为抗病毒药物研发提供了理论指导。
