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Persistent Infection Promotes Cross-Species Transmissibility of Mouse Hepatitis Virus 总被引:1,自引:4,他引:1
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Ralph S. Baric Eileen Sullivan Lisa Hensley Boyd Yount Wan Chen 《Journal of virology》1999,73(1):638-649
Persistent infection with mouse hepatitis virus (MHV) strain A59 in murine DBT (delayed brain tumor) cells resulted in the emergence of host range variants, designated V51A and V51B, at 210 days postinfection. These host range mutants replicated efficiently in normally nonpermissive Chinese hamster ovary (CHO), in human hepatocarcinoma (HepG2), and to a lesser extent in human breast carcinoma (MCF7) cell lines. Little if any replication was noted in baby hamster kidney (BHK), green African monkey kidney (COS-7), feline kidney (CRFK), and swine testicular (ST) cell lines. By fluorescent antibody (FA) staining, persistent viruses V10B and V30B, isolated at days 38 and 119 days postinfection, also demonstrated very low levels of replication in human HepG2 cells. These data suggest that persistence may rapidly select for host range expansion of animal viruses. Pretreatment of HepG2 cells with a polyclonal antibody directed against human carcinoembryonic antigens (CEA) or with some monoclonal antibodies (Col-1, Col-4, Col-12, and Col-14) that bind human CEA significantly inhibited V51B infection. Under identical conditions, little or no blockade was evident with other monoclonal antibodies (kat4c or Col-6) which also bind the human CEA glycoproteins. In addition, an antibody (EDDA) directed against irrelevant antigens did not block V51B replication. Pretreatment with the Col-4 and Col-14 antibodies did not block Sindbis virus replication in HepG2 cells or MHV infection in DBT cells, suggesting that one or more CEA glycoproteins likely functioned as receptors for V51B entry into human cell lines. To test this hypothesis, the human biliary glycoprotein (Bgp) and CEA genes were cloned and expressed in normally nonpermissive BHK cell lines by using noncytopathic Sindbis virus replicons (pSinRep19). By growth curves and FA staining, human CEA and to a much lesser extent human Bgp functioned as receptors for V51B entry. Furthermore, V51B replication was blocked with polyclonal antiserum directed against human CEA and Bgp. Under identical conditions, the parental MHV strain A59 failed to replicate in BHK cells expressing human Bgp or CEA. These data suggest that MHV persistence may promote virus cross-species transmissibility by selecting for virus variants that recognize phylogenetic homologues of the normal receptor. 相似文献
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新疆地区猪戊型肝炎血清流行病学调查 总被引:5,自引:0,他引:5
The purpose of the present study was to determine the prevalence of swine Hepatitis E virus (HEV) infection in Xinjiang. 813 swine serum samples collected from 1 to 12 months of age at 9 swine farms in Xinjiang region were tested by ELISA for the presence of IgG antibodies against HEV. The recombinant protein pUS 166 containing region 452-617aa of the ORF2 of HEV US strain was used as coating antigen. The result showed that anti -HEV IgG were detected in 265 of 405 pigs (65.43%) in one group and 238 of 408 pigs (58.33%) in another group, and that the seropositivity rate was not related to geographic district and breeds, but differed remarkably by age, being 40% among the 1- to 3-month-old piglets, but 77.33% among ones over 3-month-old. It suggested that swine HEV was widespread in different geographic regions of XinJiang. 相似文献
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新疆地区猪戊型肝炎血清流行病学调查 总被引:11,自引:0,他引:11
戊型肝炎(HE)是一种经粪-口传播的疾病,在发展中国家造成非常严重的健康问题.近年来的研究证实发达国家也存在戊型肝炎问题.该病主要威胁青壮年,孕妇病死率可高达20%.我国自1982年起就有HE的报道,新疆是HE的高流行区.由于HEV的组织培养研究尚不成熟,因此其诊断手段主要是利用RT-PCR检测病毒RNA,或利用酶联免疫吸附试验(ELISA)检测抗体.而用于血清学检测的抗原主要来自HEV ORF2和ORF3的产物,并且用ORF2产物建立的检测法有足够的敏感性和特异性.自Meng[1]1997年从美国猪体内克隆出戊型肝炎病毒(HEV)基因后,我国以及加拿大,西班牙,新西兰,澳大利亚,印度等国家也都克隆出本国猪HEV基因.虽然我国也进行了猪HEV的检测,但在1988年爆发过人源戊型肝炎的新疆地区猪群感染HEV的情况还不清楚.本研究调查了HEV在猪群的感染状况,对新疆不同地区,猪场,年龄段及品种的猪进行HE的血清学检测. 相似文献
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Alicia Solórzano Emanuela Foni Lorena Córdoba Massimiliano Baratelli Elisabetta Razzuoli Dania Bilato María ángeles Martín del Burgo David S. Perlin Jorge Martínez Pamela Martínez-Orellana Lorenzo Fraile Chiara Chiapponi Massimo Amadori Gustavo del Real María Montoya 《Journal of virology》2015,89(22):11190-11202
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Pattaratida Sa-nguanmoo Nawarat Posuwan Preeyaporn Vichaiwattana Norra Wutthiratkowit Somchai Owatanapanich Rujipat Wasitthankasem Thanunrat Thongmee Kittiyod Poovorawan Apiradee Theamboonlers Sompong Vongpunsawad Yong Poovorawan 《PloS one》2015,10(4)
Hepatitis A virus (HAV) and hepatitis E virus (HEV) infection in developing countries are associated with contaminated food or water. Although Thailand is non-endemic for HEV, sporadic infections may occur from zoonotic transmission. Individuals between 7 months to 69 years (mean age = 32.8) from predominantly Islamic Narathiwat (n = 305) and swine farm-dense Lop Buri (n = 416) provinces were screened for anti-HEV and anti-HAV antibodies by commercial enzyme-linked immunosorbent assay and automated chemiluminescent microparticle immunoassay, respectively. Seroprevalence and relative antibody titers were analyzed according to age groups. HAV IgG antibody positive rates in Lop Buri and Narathiwat residents were 39.9% and 58%, respectively (p < 0.001). Greater than 90% of individuals >50 years old in both provinces possessed anti-HAV IgG. In contrast, seroprevalence for anti-HEV IgG was much higher in Lop Buri (37.3%) than in Narathiwat (8.9%) (p < 0.001). Highest anti-HEV IgG prevalence was found among 21-30 year-olds (50%) in Lop Buri and 41-50 year-olds (14.1%) in Narathiwat. In summary, fewer individuals possessed anti-HEV IgG in Narathiwat where most residents abstained from pork and fewer swine farms are present. Therefore, an increased anti-HEV IgG seroprevalence was associated with the density of swine farm and possibly pork consumption. Adults were more likely than children to have antibodies to both HEV and HAV. 相似文献
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戊型肝炎病毒实验感染恒河猴的研究 总被引:6,自引:0,他引:6
报道了用戊型肝炎(Hepatitis E, HE)病人粪便悬液感染恒河猴后的组织病理学、血液生化与免疫学以及病毒学分子生物学检测的结果.三只实验猴在感染后第3~4周均出现ALT异常;粪便以及肝脏与胆囊组织超薄切片中电镜观察到27~34nm大小的病毒样颗粒;病理组织切片观察表明,肝脏组织有典型的急性炎症病灶;粪便与血清经RT-nPCR扩增到戊型肝炎病毒(Hepatitis E Virus, HEV)特异性片段,粪便排毒从感染后第7天持续至第50天左右,病毒血症迟于粪便排毒,出现于感染后两周左右,维持1~2周;ELISA检测发现,实验猴血清中HEV IgG抗体水平在感染后3~4周阳转,4~5个月后转阴.这些实验结果提示,恒河猴作为HEV感染实验动物模型是理想的,建立系统的恒河猴实验模型对探讨HEV感染发病机理、机体免疫应答以及临床诊断与疫苗研制具有重要意义. 相似文献
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戊型肝炎病毒实验感染恒河猴的研究 总被引:1,自引:0,他引:1
报道了用戊型肝炎(HepatitisE,HE)病人粪便悬液感染恒河猴后的组织病理学、血液生化与免疫学以及病毒学分子生物学检测的结果。三只实验猴在感染后第3~4周均出现ALT异常;粪便以及肝脏与胆囊组织超薄切片中电镜观察到27~34nm大小的病毒样颗粒;病理组织切片观察表明,肝脏组织有典型的急性炎症病灶;粪便与血清经RTnPCR扩增到戊型肝炎病毒(HepatitisEVirus,HEV)特异性片段,粪便排毒从感染后第7天持续至第50天左右,病毒血症迟于粪便排毒,出现于感染后两周左右,维持1~2周;ELISA检测发现,实验猴血清中HEVIgG抗体水平在感染后3~4周阳转,4~5个月后转阴。这些实验结果提示,恒河猴作为HEV感染实验动物模型是理想的,建立系统的恒河猴实验模型对探讨HEV感染发病机理、机体免疫应答以及临床诊断与疫苗研制具有重要意义。 相似文献
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Jian-Te Lee Pei-Lan Shao Luan-Yin Chang Ning-Shao Xia Pei-Jer Chen Chun-Yi Lu Li-Min Huang 《PloS one》2013,8(6)
Objectives
Hepatitis E virus (HEV) is an emerging pathogen. We evaluated the seroprevalence of HEV infection among swine farmers and the general population in Taiwan, a nonendemic country.Methods
We conducted a cross-sectional seroepidemiologic study in rural Taiwan in 2012 and 2013. The study included swine farmers, health examination attendees, pregnant women, and students. A commercial enzyme-linked immunosorbent assay was used to detect immunoglobulin G (IgG) and IgM against HEV. Pertinent information was collected using a questionnaire.Results
In total, 660 participants were enrolled in the study, including 156 swine farmers, 314 health examination attendees, 100 pregnant women, and 90 students. IgG anti-HEV was detected in 29.5% of swine farmers, 11.5% of health examination attendees, 2% of pregnant women, and 1.1% of students. Two subjects were positive for IgM anti-HEV. Logistic regression analysis revealed that swine farmers had an approximately 3.5-fold increased risk (odds ratio [OR], 3.46; 95% confidence interval [CI], 1.91–6.27; p<0.0001) for being seropositive for IgG anti-HEV as compared to the general population. Age was positively associated with seropositive rate (OR, 1.07 per year; 95% CI, 1.05–1.09; p<0.0001).Conclusion
HEV infection is prevalent in Taiwan. The seroprevalence of HEV infection is high in swine farmers and in the elderly population. 相似文献10.
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为了探究补体系统与戊型肝炎病毒复制的相关性,分别在HEV感染的A549细胞和BALB/c小鼠中检测C3aR、CD55和CD59蛋白的表达.利用RT-qPCR定量检测细胞和组织中补体的表达,采用免疫组化法检测HEV感染BALB/c小鼠中补体CD59及C5b-9的表达,ELISA检测补体相关炎症因子的变化.HEV感染可以激活补体蛋白C3aR、C5b-9、CD55和CD59的表达,引起补体蛋白相关炎症因子IL-10表达水平下降,IL-12和TNF-α的表达水平的上升,从而导致机体的炎症反应,加剧组织损伤.HEV感染激活补体系统并参与早期的抗病毒反应,HEV感染对补体的持续激活导致炎症因子过度表达,加重机体损伤. 相似文献
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Genotype 1 hepatitis E viruses (HEVs) are restricted to primate hosts, whereas genotype 3 HEVs predominantly infect swine, in addition to primates. In order to identify possible determinants of the host range, infectious recombinant viruses and chimeras of a genotype 1 isolate and a genotype 3 isolate were compared for their ability to infect versus transfect cultured human HepG2/C3A cells and swine LLC-PK cells. The patterns of luciferase expression from virus replicons containing the Gaussia luciferase gene in place of the viral ORF2 or ORF3 genes demonstrated that translation of the ORF2 capsid gene of genotype 1 virus is severely inhibited in swine kidney cells compared to its translation in rhesus macaque kidney or human liver cells. Therefore, this virus may produce insufficient capsid protein for optimal assembly in swine cells. Infectivity assays with a virus containing a chimeric capsid protein confirmed that amino acids 456 to 605 of the virus capsid protein comprised the virus receptor-binding region and suggested that genotype 1 viruses may be prevented from infecting swine because genotype 1 viruses are unable to enter swine cells. Rhesus macaque cells appeared to be better than human cells for growing the genotype 1 virus. These cell and virus combinations may serve as a useful in vitro model with which to study determinants of the natural host range of this virus. 相似文献
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将含有kozak序列及BamH I的上游引物和带有终止密码子及EcoRV酶切位点的下游引物,以猪HEVDQ1 ORF2为模板,进行PCR。将扩增片段和pcDNA3.1质粒以BamH I/EcoRV进行双酶切后进行连接。连接产物转化至大肠杆菌DH5α,经测序证明该序列正确,命名为pcDQ1。进行pcDQ1质粒提取,以Vero细胞为表达细胞进行转染,以间接免疫荧光试验进行验证。以100μg/次/只剂量的pcDQ1对BAL B/C小鼠进行免疫以获取单因子血清。共免疫3次,采集血清,进行ELISA效价测定。结果表明,该核酸疫苗可以免疫使小鼠产生抗体。 相似文献
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Guixia Yu Shigeo Yagi Ricardo Carrion Jr. Eunice C. Chen Maria Liu Kathleen M. Brasky Robert E. Lanford Kristi R. Kelly Karen L. Bales David P. Schnurr Don R. Canfield Jean L. Patterson Charles Y. Chiu 《PloS one》2013,8(7)
Adenoviruses are DNA viruses that infect a number of vertebrate hosts and are associated with both sporadic and epidemic disease in humans. We previously identified a novel adenovirus, titi monkey adenovirus (TMAdV), as the cause of a fulminant pneumonia outbreak in a colony of titi monkeys (Callicebus cupreus) at a national primate center in 2009. Serological evidence of infection by TMAdV was also found in a human researcher at the facility and household family member, raising concerns for potential cross-species transmission of the virus. Here we present experimental evidence of cross-species TMAdV infection in common marmosets (Callithrix jacchus). Nasal inoculation of a cell cultured-adapted TMAdV strain into three marmosets produced an acute, mild respiratory illness characterized by low-grade fever, reduced activity, anorexia, and sneezing. An increase in virus-specific neutralization antibody titers accompanied the development of clinical signs. Although serially collected nasal swabs were positive for TMAdV for at least 8 days, all 3 infected marmosets spontaneously recovered by day 12 post-inoculation, and persistence of the virus in tissues could not be established. Thus, the pathogenesis of experimental inoculation of TMAdV in common marmosets resembled the mild, self-limiting respiratory infection typically seen in immunocompetent human hosts rather than the rapidly progressive, fatal pneumonia observed in 19 of 23 titi monkeys during the prior 2009 outbreak. These findings further establish the potential for adenovirus cross-species transmission and provide the basis for development of a monkey model useful for assessing the zoonotic potential of adenoviruses. 相似文献
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宫颈癌中p53表达和病毒感染的研究 总被引:2,自引:0,他引:2
应用PCR对121例宫颈脱落细胞和组织分别检测HPV-16/18和HSV-2DNA。同时应用免疫组化S-P法检测76例宫颈组织中P53过度表达。结果发现,宫颈癌组织中HPV-16/18和HSV-2阳性率分别为61.3%和32.3%,慢性宫颈炎组HPV-16/18和HSV-2阳性率分别为22.5%和20.0%,与正常宫颈组比较均有显著性差异。宫颈癌中HPV-16/18和HSV-2混合感染率为16.1%。p53过度表达率在慢性宫颈炎、宫颈上皮内瘤变(CIN)、宫颈癌组织中呈梯度递增。另外,宫颈癌组织中P53过度表达与HPV-16/18、HSV-2的感染无相关性。提示:宫颈癌的发生与HPV-16/18关系密切,HSV-2可能与HPV-16/18协同作用导致宫颈癌的发生。宫颈组织中p53过度表达率与宫颈癌的进程有关,这在宫颈癌的防治方面有一定的临床意义。 相似文献
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宫颈癌中p53表达和病毒感染的研究 总被引:5,自引:0,他引:5
应用PCR对121例宫颈脱落细胞和组织分别检测HPV-16/18和HSv-2 DNA.同时应用免疫组化S-P法检测76例宫颈组织中p53过度表达.结果发现,宫颈癌组织中HPV-16/18和HSv-2阳性率分别为61.3%和32.3%,慢性宫颈炎组HPV-16/18和HSV-2阳性率分别为22.5%和20.0%,与正常宫颈组比较均有显著性差异.宫颈癌中HPV-16/18和HSV-2混合感染率为16.1%.p53过度表达率在慢性宫颈炎、宫颈上皮内瘤变(CIN)、宫颈癌组织中呈梯度递增.另外,宫颈癌组织中p53过度表达与HPV-16/18、HSv-2的感染无相关性.提示宫颈癌的发生与HPV-16/18关系密切,HSV-2可能与HPV-16/18协同作用导致宫颈癌的发生.宫颈组织中p53过度表达率与宫颈癌的进程有关,这在宫颈癌的防治方面有一定的临床意义. 相似文献
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从戊型肝炎病毒(Hepatitis Evirus,HEV)IgG检测阳性的新疆某猪场采集70份猪粪便,利用逆转录套式聚合酶链方法(RT-nPCR),检测HEV RNA,其中13份为阳性,阳性率18.57%。将PCR扩增产物克隆到pMD18-T载体上,构建成重组质粒并测序,结果表明,13株猪源HEV分离株在HEV ORF2 348bp核苷酸序列的同源性为97.1%~100%,为同一基因型;与HEVⅠ、Ⅱ、Ⅲ、Ⅳ的同源性分别为74.1%-77.6%,71.6%-74.1%,73.3%~78.2%和82.8%-91.4%,与ⅣA亚型的同源性同源性最高达89.4%-91.4%。以该核苷酸片段绘制的基因进化树显示13株猪源HEV与HEV Ⅳ T1株在同一分支上,属基因Ⅳ型;与国内其他猪源HEV分离株该片段核苷酸序列的同源性为82.6%-91.3%,提示中国猪源HEV的基因型比较一致,同属HEV Ⅳ型。 相似文献
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新疆猪粪便戊型肝炎病毒RNA的检测及序列分析 总被引:9,自引:0,他引:9
从戊型肝炎病毒(Hepatitis E virus,HEV)IgG检测阳性的新疆某猪场采集70份猪粪便,利用逆转录套式聚合酶链方法(RT-nPCR),检测HEV RNA,其中13份为阳性,阳性率18.57%.将PCR扩增产物克隆到pMD18-T载体上,构建成重组质粒并测序,结果表明,13株猪源HEV分离株在HEV ORF2 348bp核苷酸序列的同源性为97.1%~100%,为同一基因型;与HEV Ⅰ、Ⅱ、Ⅲ、Ⅳ的同源性分别为74.1%~77.6%,71.6%~74.1%,73.3%~78.2%和82.8%~91.4%,与ⅣA亚型的同源性同源性最高达89.4%~91.4%.以该核苷酸片段绘制的基因进化树显示13株猪源HEV与HEVⅣT1株在同一分支上,属基因Ⅳ型;与国内其他猪源HEV分离株该片段核苷酸序列的同源性为82.6%~91.3%,提示中国猪源HEV的基因型比较一致,同属HEVⅣ型. 相似文献