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1

Selective Serotonin Reuptake Inhibitor Fluoxetine Inhibits Replication of Human Enteroviruses B and D by Targeting Viral Protein 2C  

PubMed Central

Although the genus Enterovirus contains many important human pathogens, there is no licensed drug for either the treatment or the prophylaxis of enterovirus infections. We report that fluoxetine (Prozac)—a selective serotonin reuptake inhibitor—inhibits the replication of human enterovirus B (HEV-B) and HEV-D but does not affect the replication of HEV-A and HEV-C or human rhinovirus A or B. We show that fluoxetine interferes with viral RNA replication, and we identified viral protein 2C as the target of this compound.

Ulferts, Rachel; van der Linden, Lonneke; Thibaut, Hendrik Jan; Lanke, Kjerstin H. W.; Leyssen, Pieter; Coutard, Bruno; De Palma, Armando M.; Canard, Bruno; Neyts, Johan

2013-01-01

2

Selective serotonin reuptake inhibitor fluoxetine inhibits replication of human enteroviruses B and D by targeting viral protein 2C.  

PubMed

Although the genus Enterovirus contains many important human pathogens, there is no licensed drug for either the treatment or the prophylaxis of enterovirus infections. We report that fluoxetine (Prozac)--a selective serotonin reuptake inhibitor--inhibits the replication of human enterovirus B (HEV-B) and HEV-D but does not affect the replication of HEV-A and HEV-C or human rhinovirus A or B. We show that fluoxetine interferes with viral RNA replication, and we identified viral protein 2C as the target of this compound. PMID:23335743

Ulferts, Rachel; van der Linden, Lonneke; Thibaut, Hendrik Jan; Lanke, Kjerstin H W; Leyssen, Pieter; Coutard, Bruno; De Palma, Armando M; Canard, Bruno; Neyts, Johan; van Kuppeveld, Frank J M

2013-01-18

3

Evidence for Frequent Recombination within Species Human Enterovirus B Based on Complete Genomic Sequences of All Thirty-Seven Serotypes  

Microsoft Academic Search

The species Human enterovirus B (HEV-B) in the family Picornaviridae consists of coxsackievirus A9; coxsackieviruses B1 to B6; echoviruses 1 to 7, 9, 11 to 21, 24 to 27, and 29 to 33; and enteroviruses 69 and 73. We have determined complete genome sequences for the remaining 22 HEV-B serotypes whose sequences were not represented in public databases and analyzed

M. Steven Oberste; Kaija Maher; Mark A. Pallansch

2004-01-01

4

Production of Enterovirus Antigens for Human Enterovirus Types.  

National Technical Information Service (NTIS)

The program has a four-fold purpose: (1) to produce reference seeds of various enteroviruses; (2) to produce seed virus and antiserum to specific simian viruses; (3) to ampoule and test designated reference human picornaviruses; and (4) to test certain si...

S. S. Kalter

1966-01-01

5

Experimental human rhinovirus and enterovirus interspecies recombination.  

PubMed

Human rhinoviruses (HRVs) and enteroviruses (HEVs), two important human pathogens, are non-enveloped, positive-sense RNA viruses of the genus Enterovirus within the family Picornaviridae. Intraspecies recombination is known as a driving force for enterovirus and, to a lesser extent, rhinovirus evolution. Interspecies recombination is much less frequent among circulating strains, and supporting evidence for such recombination is limited to ancestral events, as shown by recent phylogenetic analyses reporting ancient HRV-A/HRV-C, HEV-A/HEV-C and HEV-A/HEV-D recombination mainly at the 5'-untranslated region (5' UTR)-polyprotein junction. In this study, chimeric genomes were artificially generated using the 5' UTR from two different clinical HRV-C strains (HRV-Ca and HRV-Cc), an HRV-B strain (HRV-B37) and an HEV-A strain (HEV-A71), and the remaining part of the genome from an HRV-A strain (HRV-A16). Whilst the chimeric viruses were easily propagated in cell culture, the wild-type HRV-A16 retained a replication advantage, both individually and in competition experiments. Assessment of protein synthesis ability did not show a correlation between translation and replication efficiencies. These results reflect the interchangeability of the 5' UTR, including its functional RNA structural elements implicated in both genome translation and replication among different enterovirus species. The 5' UTR-polyprotein junction therefore represents a theoretic interspecies recombination breakpoint. This recombination potential is probably restricted by the need for co-infection opportunities and the requirement for the progeny chimera to outcompete the parental genomes' fitness, explaining the rare occurrence of such events in vivo. PMID:21940413

Schibler, Manuel; Gerlach, Daniel; Martinez, Yannick; Belle, Sandra Van; Turin, Lara; Kaiser, Laurent; Tapparel, Caroline

2011-09-21

6

Molecular identification of 13 new enterovirus types, EV79–88, EV97, and EV100–101, members of the species Human Enterovirus B  

Microsoft Academic Search

Molecular methods have enabled the rapid identification of new enterovirus (EV) serotypes that are untypeable using existing neutralizing antisera. As a result, sequencing of the VP1 capsid gene has been developed as a surrogate for antigenic typing to distinguish enterovirus types. In this study, 17 enterovirus isolates from four countries were identified as members of 13 new types within the

M. Steven Oberste; Kaija Maher; William A. Nix; Suzanne M. Michele; Moyez Uddin; David Schnurr; Suleiman al-Busaidy; Chantal Akoua-Koffi; Mark A. Pallansch

2007-01-01

7

Enteroviruses  

Microsoft Academic Search

Enteroviruses (EVs) comprise a genus in the Picornaviridae family, so named because they are small, single-stranded ribonucleic\\u000a acid (RNA)-containing viruses. The EVs are divided into the subgroups of polioviruses, coxsackie A viruses, coxsackie B viruses,\\u000a and echoviruses based on replication properties in tissue culture and animal models. Newer EVs are simply designated by number\\u000a (e.g., EV 71), rather than by

Mark J. Abzug

8

Detection of somatic phages, infectious enteroviruses and enterovirus genomes as indicators of human enteric viral pollution in surface water  

Microsoft Academic Search

In the present study, we aimed to determine whether the concentrations of somatic coliphages, infectious enteroviruses or the detection of enterovirus genomes were associated with the detection of human pathogenic viruses in surface water. Four French rivers were sampled monthly or semimonthly for the quantitative detection of somatic coliphages, infectious enteroviruses and the qualitative RT-PCR detection of enterovirus, hepatitis A

D. Hot; O. Legeay; J. Jacques; C. Gantzer; Y. Caudrelier; K. Guyard; M. Lange; L. Andréoletti

2003-01-01

9

Species-specific RT-PCR amplification of human enteroviruses: a tool for rapid species identification of uncharacterized enteroviruses  

Microsoft Academic Search

The 65 serotypes of human enteroviruses are classified into four species, Human enterovirus (HEV) A to D, based largely on phylogenetic relationships in multiple genome regions. The 39-non-translated region of enteroviruses is highly conserved within a species but highly divergent between species. From this information, species-specific RT-PCR primers were developed that can be used to rapidly screen collections of enterovirus

M. Steven Oberste; Kaija Maher; Alford J. Williams; Naomi Dybdahl-Sissoko; Betty A. Brown; Michelle S. Gookin; Silvia Penaranda; Nada Mishrik; Moyez Uddin; Mark A. Pallansch

2006-01-01

10

Accuracy of Diagnostic Methods and Surveillance Sensitivity for Human Enterovirus, South Korea, 1999-2011  

PubMed Central

The epidemiology of enteroviral infection in South Korea during 1999–2011 chronicles nationwide outbreaks and changing detection and subtyping methods used over the 13-year period. Of 14,657 patients whose samples were tested, 4,762 (32.5%) samples were positive for human enterovirus (human EV); as diagnostic methods improved, the rate of positive results increased. A seasonal trend of outbreaks was documented. Genotypes enterovirus 71, echovirus 30, coxsackievirus B5, enterovirus 6, and coxsackievirus B2 were the most common genotypes identified. Accurate test results correlated clinical syndromes to enterovirus genotypes: aseptic meningitis to echovirus 30, enterovirus 6, and coxsackievirus B5; hand, foot and mouth disease to coxsackievirus A16; and hand, foot and mouth disease with neurologic complications to enterovirus 71. There are currently no treatments specific to human EV infections; surveillance of enterovirus infections such as this study provides may assist with evaluating the need to research and develop treatments for infections caused by virulent human EV genotypes.

Hyeon, Ji-Yeon; Hwang, Seoyeon; Kim, Hyejin; Song, Jaehyoung; Ahn, Jeongbae; Kang, Byunghak; Kim, Kisoon; Choi, Wooyoung; Chung, Jae Keun; Kim, Cheon-Hyun; Cho, Kyungsoon; Jee, Youngmee; Kim, Jonghyun; Kim, Kisang; Kim, Sun-Hee; Kim, Min-Ji

2013-01-01

11

Comparative genomics of the coxsackie B viruses and related enteroviruses.  

PubMed

Genomic analysis of the group B coxsackieviruses (CVB) has improved our understanding of CVB evolution, epidemiology, and pathogenesis. Comparison of capsid sequence alignments and virion structures allows correlation of capsid diversity with surface features, such as loops, the receptor canyon, and antigenic sites. Pairwise sequence comparisons and phylogenetic analyses can be used to rapidly identify and classify enteroviruses. Enteroviruses are monophyletic by type only within the capsid region. The CVBs as a group are monophyletic in the capsid region, probably due to their shared use of the coxsackievirus-adenovirus receptor (other members of HEV-B use different receptors). Outside the capsid region, enteroviruses are monophyletic only by species (not by type), reflecting a high frequency of intertypic recombination within a species. Further genomic studies, accompanied by well-characterized clinical outcome/disease data, will facilitate fine-scale mapping of genetic determinants that contribute to virulence. PMID:18357764

Oberste, M S

2008-01-01

12

Cleavage of eukaryotic initiation factor eIF5B by enterovirus 3C proteases  

PubMed Central

The enteroviruses poliovirus (PV), Coxsackie B virus (CVB) and rhinovirus (HRV) are members of Picornaviridae that inhibit host cell translation early in infection. Enterovirus translation soon predominates in infected cells, but eventually also shuts off. This complex pattern of modulation of translation suggests regulation by a multifactorial mechanism. We report here that eIF5B is proteolytically cleaved during PV and CVB infection of cultured cells, beginning at 3 hours post-infection and increasing thereafter. Recombinant PV, CVB and HRV 3Cpro cleaved purified native rabbit eukaryotic initiation factor (eIF) 5B in vitro at a single site (VVEQ?G, equivalent to VMEQ?G479in human eIF5B) that is consistent with the cleavage specificity of enterovirus 3C proteases. Cleavage separates the N-terminal domain of eIF5B from its essential conserved central GTPase and C-terminal domains. 3Cpro-mediated cleavage of eIF5B may thus play an accessory role in the shut-off of translation that occurs in enterovirus-infected cells.

de Breyne, Sylvain; Bonderoff, Jennifer M.; Chumakov, Konstantin M.; Lloyd, Richard E.; Hellen, Christopher U. T.

2008-01-01

13

Human enterovirus 71 epidemics: what's next?  

PubMed

Human enterovirus 71 (EV71) epidemics have affected various countries in the past 40 years. EV71 commonly causes hand, foot and mouth disease (HFMD) in children, but can result in neurological and cardiorespiratory complications in severe cases. Genotypic changes of EV71 have been observed in different places over time, with the emergence of novel genotypes or subgenotypes giving rise to serious outbreaks. Since the late 1990s, intra- and inter-typic recombination events in EV71 have been increasingly reported in the Asia-Pacific region. In particular, 'double-recombinant' EV71 strains belonging to a novel genotype D have been predominant in mainland China and Hong Kong over the last decade, though co-circulating with a minority of other EV71 subgenotypes and coxsackie A viruses. Continuous surveillance and genome studies are important to detect potential novel mutants or recombinants in the near future. Rapid and sensitive molecular detection of EV71 is of paramount importance in anticipating and combating EV71 outbreaks. PMID:24119538

Yip, Cyril C Y; Lau, Susanna K P; Woo, Patrick C Y; Yuen, Kwok-Yung

2013-09-10

14

Cytokine and chemokine production by human pancreatic islets upon enterovirus infection.  

PubMed

Enteroviruses of the human enterovirus B species (HEV-Bs) (e.g., coxsackie B viruses [CVBs] and echoviruses) have been implicated as environmental factors that trigger/accelerate type 1 diabetes, but the underlying mechanism remains elusive. The aim of this study was to gain insight into the cytokines and chemokines that are produced by human pancreatic islets upon infection with CVBs. To this end, we studied the response of human islets of Langerhans upon mock or CVB3 infection. Using quantitative PCR, we showed that upon CVB3 infection, transcription of interferon (IFN), IFN-stimulated genes, and inflammatory genes was induced. Analysis of secreted cytokines and chemokines by Luminex technology confirmed production and secretion of proinflammatory cytokines (e.g., interleukin [IL]-6 and tumor necrosis factor-?) as well as various chemotactic proteins, such as IFN-?-induced protein 10, macrophage inflammatory protein (MIP)-1?, MIP-1?, and IL-8. Infection with other HEV-Bs induced similar responses, yet their extent depended on replication efficiency. Ultra violet-inactivated CVB3 did not induce any response, suggesting that virus replication is a prerequisite for antiviral responses. Our data represent the first comprehensive overview of inflammatory mediators that are secreted by human islets of Langerhans upon CVB infection and may shed light on the role of enteroviruses in type 1 diabetes pathogenesis. PMID:22596052

Schulte, Barbara M; Lanke, Kjerstin H W; Piganelli, Jon D; Kers-Rebel, Esther D; Bottino, Rita; Trucco, Massimo; Huijbens, Richard J F; Radstake, Timothy R D J; Engelse, Marten A; de Koning, Eelco J P; Galama, Jochem M; Adema, Gosse J; van Kuppeveld, Frank J M

2012-05-17

15

Cellular tropism of human enterovirus D species serotypes EV-94, EV-70, and EV-68 in vitro: implications for pathogenesis.  

PubMed

Enterovirus 94 (EV-94) is an enterovirus serotype described recently which, together with EV-68 and EV-70, forms human enterovirus D species. This study investigates the seroprevalences of these three serotypes and their abilities to infect, replicate, and damage cell types considered to be essential for enterovirus-induced diseases. The cell types studied included human leukocyte cell lines, primary endothelial cells, and pancreatic islets. High prevalence of neutralizing antibodies against EV-68 and EV-94 was found in the Finnish population. The virus strains studied had wide leukocyte tropism. EV-94 and EV-68 were able to produce infectious progeny in leukocyte cell lines with monocytic, granulocytic, T-cell, or B-cell characteristics. EV-94 and EV-70 were capable of infecting primary human umbilical vein endothelial cells, whereas EV-68 had only marginal progeny production and did not induce cytopathic effects in these cells. Intriguingly, EV-94 was able to damage pancreatic islet ?-cells, to infect, replicate, and cause necrosis in human pancreatic islets, and to induce proinflammatory and chemoattractive cytokine expression in endothelial cells. These results suggest that HEV-D viruses may be more prevalent than has been thought previously, and they provide in vitro evidence that EV-94 may be a potent pathogen and should be considered a potentially diabetogenic enterovirus type. PMID:20872722

Smura, Teemu; Ylipaasto, Petri; Klemola, Päivi; Kaijalainen, Svetlana; Kyllönen, Lauri; Sordi, Valeria; Piemonti, Lorenzo; Roivainen, Merja

2010-11-01

16

Complete Genomic Sequencing Shows that Polioviruses and Members of Human Enterovirus Species C Are Closely Related in the Noncapsid Coding Region  

Microsoft Academic Search

The 65 human enterovirus serotypes are currently classified into five species: Poliovirus (3 serotypes), Human enterovirus A (HEV-A) (12 serotypes), HEV-B (37 serotypes), HEV-C (11 serotypes), and HEV-D (2 serotypes). Coxsackie A virus (CAV) serotypes 1, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 24 constitute HEV-C. We have determined the complete genome sequences for the remaining nine

Betty Brown; M. Steven Oberste; Kaija Maher; Mark A. Pallansch

2003-01-01

17

Detection of human enterovirus 71 and coxsackievirus A16 in children with hand, foot and mouth disease in China.  

PubMed

The aims of the present study were to investigate the genetic characteristics of enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) strains in China and to evaluate the relationship between the genotypes of CVA16 and EV71 and their geographical distribution. A total of 399 stool specimens were collected from children with symptoms of hand, foot and mouth disease (HFMD) in Zhejiang Province. The presence of enteroviruses was determined using reverse transcription-semi-nested PCR targeted to the VP1 gene of all human enteroviruses and DNA sequencing. EV71 and CVA16, the major etiological agents of HFMD, were detected in 38.4% (38/99) and 35.4% (35/99) of HEV-A species-positive cases, respectively. Based on the phylogenetic analysis of the VP1 gene, EV71 strains identified in this study belong to subgenotype C4, and CVA16 strains herein were classified into clusters B2a and B2b within the genotype B2. Taking into consideration other published data, we conclude that the genetic characteristics of enteroviruses in China reflect the pattern of the endemic circulation of the subgenotype C4 to EV71 and clusters B2a and B2b within genotype B2 to CVA16, which have been continuously circulating in China since 1997. This observation indicates that the genetic characteristics of enteroviruses in China seem to depend on their special geographical and climatical features allowing them to be sustained with little external effect. PMID:22218731

Chen, Ling; Mou, Xiaozhou; Zhang, Qiong; Li, Yifei; Lin, Jian; Liu, Fanlong; Yuan, Li; Tang, Yiming; Xiang, Charlie

2012-01-03

18

Detection of human enterovirus 71 and coxsackievirus A16 in children with hand, foot and mouth disease in China  

PubMed Central

The aims of the present study were to investigate the genetic characteristics of enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) strains in China and to evaluate the relationship between the genotypes of CVA16 and EV71 and their geographical distribution. A total of 399 stool specimens were collected from children with symptoms of hand, foot and mouth disease (HFMD) in Zhejiang Province. The presence of enteroviruses was determined using reverse transcription-semi-nested PCR targeted to the VP1 gene of all human enteroviruses and DNA sequencing. EV71 and CVA16, the major etiological agents of HFMD, were detected in 38.4% (38/99) and 35.4% (35/99) of HEV-A species-positive cases, respectively. Based on the phylogenetic analysis of the VP1 gene, EV71 strains identified in this study belong to subgenotype C4, and CVA16 strains herein were classified into clusters B2a and B2b within the genotype B2. Taking into consideration other published data, we conclude that the genetic characteristics of enteroviruses in China reflect the pattern of the endemic circulation of the subgenotype C4 to EV71 and clusters B2a and B2b within genotype B2 to CVA16, which have been continuously circulating in China since 1997. This observation indicates that the genetic characteristics of enteroviruses in China seem to depend on their special geographical and climatical features allowing them to be sustained with little external effect.

CHEN, LING; MOU, XIAOZHOU; ZHANG, QIONG; LI, YIFEI; LIN, JIAN; LIU, FANLONG; YUAN, LI; TANG, YIMING; XIANG, CHARLIE

2012-01-01

19

Antiviral effect of geraniin on human enterovirus 71 in vitro and in vivo.  

PubMed

Human enterovirus 71 infection causes hand, foot and mouth disease in children under 6 years of age and has caused mortalities in large-scale outbreaks in the Asia-Pacific region. No effective vaccine or antiviral drugs currently exist against enterovirus 71 in the clinic. In this study, we investigated the antiviral effect of geraniin on enterovirus 71 both in vitro and in vivo. The results showed that geraniin effectively inhibited virus replication in rhabdomyosarcoma cells with an IC(50) of 10 ?g/ml. Moreover, geraniin treatment of mice that were challenged with a lethal dose of enterovirus 71 resulted in a reduction of mortality, relieved clinical symptoms, and inhibited virus replication in muscle tissues. The results suggest that geraniin may be used as a potential drug for anti-enterovirus 71. PMID:22342145

Yang, Yajun; Zhang, Lianfeng; Fan, Xiaoxu; Qin, Chuan; Liu, Jiangning

2012-02-02

20

Structure of human enterovirus 71 in complex with a capsid-binding inhibitor  

PubMed Central

Human enterovirus 71 is a picornavirus causing hand, foot, and mouth disease that may progress to fatal encephalitis in infants and small children. As of now, no cure is available for enterovirus 71 infections. Small molecule inhibitors binding into a hydrophobic pocket within capsid viral protein 1 were previously shown to effectively limit infectivity of many picornaviruses. Here we report a 3.2-Å-resolution X-ray structure of the enterovirus 71 virion complexed with the capsid-binding inhibitor WIN 51711. The inhibitor replaced the natural pocket factor within the viral protein 1 pocket without inducing any detectable rearrangements in the structure of the capsid. Furthermore, we show that the compound stabilizes enterovirus 71 virions and limits its infectivity, probably through restricting dynamics of the capsid necessary for genome release. Thus, our results provide a structural basis for development of antienterovirus 71 capsid-binding drugs.

Plevka, Pavel; Perera, Rushika; Yap, Moh Lan; Cardosa, Jane; Kuhn, Richard J.; Rossmann, Michael G.

2013-01-01

21

Identification and Typing of Human Enterovirus: A Genomic Barcode Approach  

PubMed Central

Identification and typing of human enterovirus (HEVs) are important to pathogen detection and therapy. Previous phylogeny-based typing methods are mainly based on multiple sequence alignments of specific genes in the HEVs, but the results are not stable with respect to different choices of genes. Here we report a novel method for identification and typing of HEVs based on information derived from their whole genomes. Specifically, we calculate the k-mer based barcode image for each genome, HEV or other human viruses, for a fixed k, 1human viruses and 395 HEV sequences. The tree shows a clear separation of the HEV viruses from all the non-HEV viruses with 100% accuracy and a separation of the HEVs into four distinct clads with 93.4% consistency with a multiple sequence alignment-based phylogeny. Our detailed analyses of the HEVs having different typing results by the two methods indicate that our results are in better agreement with known information about the HEVs.

Chen, Xin; Huang, Honglan; Liu, Bin; Xu, Ying; Li, Fan

2011-01-01

22

Genetic diversity of enterovirus subgroups  

Microsoft Academic Search

Summary Enterovirus serotypes were studied using nucleic acid hybridization and nucleotide sequence analysis. A great majority of enteroviruses could be roughly divided into two larger subgroups the first consisting of poliovirus and certain coxsackievirus A serotypes. The second subgroup included coxsackie B viruses, most ECHO viruses, enterovirus 71 and representatives of coxsackie A viruses. Enterovirus 70 showed low homology to

P. Auvinen; G. Stanway; T. Hyypiä

1989-01-01

23

Structural basis for antiviral inhibition of the main protease, 3C, from human enterovirus 93.  

PubMed

Members of the Enterovirus genus of the Picornaviridae family are abundant, with common human pathogens that belong to the rhinovirus (HRV) and enterovirus (EV) species, including diverse echo-, coxsackie- and polioviruses. They cause a wide spectrum of clinical manifestations ranging from asymptomatic to severe diseases with neurological and/or cardiac manifestations. Pandemic outbreaks of EVs may be accompanied by meningitis and/or paralysis and can be fatal. However, no effective prophylaxis or antiviral treatment against most EVs is available. The EV RNA genome directs the synthesis of a single polyprotein that is autocatalytically processed into mature proteins at Gln?Gly cleavage sites by the 3C protease (3C(pro)), which has narrow, conserved substrate specificity. These cleavages are essential for virus replication, making 3C(pro) an excellent target for antivirus drug development. In this study, we report the first determination of the crystal structure of 3C(pro) from an enterovirus B, EV-93, a recently identified pathogen, alone and in complex with the anti-HRV molecules compound 1 (AG7404) and rupintrivir (AG7088) at resolutions of 1.9, 1.3, and 1.5 Å, respectively. The EV-93 3C(pro) adopts a chymotrypsin-like fold with a canonically configured oxyanion hole and a substrate binding pocket similar to that of rhino-, coxsackie- and poliovirus 3C proteases. We show that compound 1 and rupintrivir are both active against EV-93 in infected cells and inhibit the proteolytic activity of EV-93 3C(pro) in vitro. These results provide a framework for further structure-guided optimization of the tested compounds to produce antiviral drugs against a broad range of EV species. PMID:21835784

Costenaro, Lionel; Kaczmarska, Zuzanna; Arnan, Carme; Janowski, Robert; Coutard, Bruno; Solà, Maria; Gorbalenya, Alexander E; Norder, Heléne; Canard, Bruno; Coll, Miquel

2011-08-10

24

Impaired glutamate uptake and EAAT2 downregulation in an enterovirus chronically infected human glial cell line.  

PubMed

Rapid and efficient uptake of glutamate via the high-affinity glutamate transporter EAAT2 is important for limiting glutamate-mediated excitotoxicity involved in neuronal death. Furthermore, there is evidence of altered glutamate uptake and catabolism in motor neuron diseases. Such a defect has been reported in amyotrophic lateral sclerosis, the major motor neuron disease, and was associated with impairment in EAAT2 processing. We recently reported the presence of enterovirus genome specifically in the anterior horn of amyotrophic lateral sclerosis cases, suggesting the involvement of a chronic/persistent enterovirus infection in amyotrophic lateral sclerosis. To investigate a putative link between enterovirus infection and the glutamate-mediated excitotoxicity observed in amyotrophic lateral sclerosis, we developed an in vitro model consisting of a human glial cell line infected with ECHOvirus 6, one of the enteroviruses with sequences closely related to those detected in patients with amyotrophic lateral sclerosis. In these glial cells, an ECHOvirus 6 chronic infection was established, resulting in altered extracellular glutamate uptake. This correlated with an aberrant splicing of the EAAT2 pre-messenger ribonucleic acid and a significant loss of EAAT2 protein expression, similar to that observed in patients with amyotrophic lateral sclerosis. These results provide convincing evidence that an enterovirus chronic/persistent infection may alter glial glutamate uptake and catabolism. As enteroviruses are extremely common human pathogens, they may act as a trigger in the development of certain motor neuron diseases, such as amyotrophic lateral sclerosis. PMID:12752781

Legay, Vincent; Deleage, Christelle; Beaulieux, Frederik; Giraudon, Pascale; Aymard, Michelle; Lina, Bruno

2003-05-01

25

Molecular phylogeny of all human enterovirus serotypes based on comparison of sequences at the 5? end of the region encoding VP2  

Microsoft Academic Search

Sixty-six human enterovirus serotypes have been described using antibody neutralization, with antigenic variants defined within several serotypes. Despite the availability of sequence data for numerous enteroviruses, the molecular basis of serotype is unknown. Previous studies by others have identified four major phylogenetic groups within the human enteroviruses, but there has been no complete database of homologous sequences for all human

M. Steven Oberste; Kaija Maher; Mark A. Pallansch

1998-01-01

26

Human bocavirus and rhino-enteroviruses in childhood otitis media with effusion  

Microsoft Academic Search

BackgroundViral respiratory infections play an important role in the pathogenesis of otitis media with effusion (OME) in children. The most common human rhinoviruses (HRVs) have been detected in middle ear effusions (MEE), but there is only limited data available about the closely related human enteroviruses (HEVs). The newly discovered human bocavirus (HBoV) has not, however, been identified in MEE of

Szilárd Rezes; Maria Söderlund-Venermo; Merja Roivainen; Kaisa Kemppainen; Zsolt Szabó; István Sziklai; Anne Pitkäranta

2009-01-01

27

Enteroviruses and the pathogenesis of type 1 diabetes revisited: cross-reactivity of enterovirus capsid protein (VP1) antibodies with human mitochondrial proteins.  

PubMed

Current or recent enteroviral infections show an association with type 1 diabetes. However, evidence for this has mainly been generated using a particular mouse monoclonal antibody (clone 5-D8/1) which binds the viral capsid protein VP1. Difficulty in confirming these findings using other independent methods has led to the concern that this might be artefactual. To address this, we examined the potential cross-reactivity of clone 5-D8/1 with normal islet proteins. Western blotting, two-dimensional gel electrophoresis, and mass spectrometry were used to identify human islet proteins bound by the clone 5-D8/1. We found a distinct reactivity with two mitochondrial proteins, creatine kinase B-type and ATP synthase beta subunit. Immunohistochemistry using the clone 5-D8/1 revealed a granular cytoplasmic staining pattern in mitochondria-rich cells, ie hepatocytes, ductal epithelial cells, vascular endothelial cells, skeletal muscle cells, and the neoplastic salivary gland oncocytoma cells, whereas connective tissue and infiltrating immune cells were negative. Staining on islets of Langerhans from subjects with recent-onset type 1 diabetes, but not on isolated human islets infected in vitro with enteroviruses, could be blocked after mixing the clone 5-D8/1 with the mitochondrial proteins. Collectively, our data show that the clone 5-D8/1 detects two human mitochondrial enzymes in addition to enteroviral VP1. The notion that the previously reported VP1 positivity in islets of recent-onset type 1 diabetes patients could reflect cross-reactivity to native islet proteins and not the presence of EV is supported by difficulties in demonstrating EV infection by independent techniques such as PCR or in situ hybridization. These findings call for revisiting the presence of enteroviruses in pancreatic islets of patients with type 1 diabetes. PMID:23335350

Hansson, Sara F; Korsgren, Stella; Pontén, Fredrik; Korsgren, Olle

2013-04-01

28

Early human enterovirus infections in healthy Swedish children participating in the PRODIA pilot study.  

PubMed

Human enteroviruses (HEV) are common, especially in childhood and during the enterovirus season, causing mainly asymptomatic infections but also mild and severe illnesses. Numerous studies have shown the association between HEV infections and type 1 diabetes. Here, the prevalence of HEV infections was studied in healthy Swedish children with increased HLA-associated risk for type 1 diabetes participating in the PRODIA pilot study in which children were randomized to receive probiotics or placebo during the first 6 months of life. Stool specimens collected from 197 children in every 3 months from the age of 3 to 24 months were screened for HEV using traditional viral culturing method and identified with reverse transcriptase polymerase chain reaction (RT-PCR) and sequencing of the partial VP1 coding part of the viral genome. Altogether 4.8% (52/1,094) of the specimens were HEV-positive and 22.3% (44/197) of the children excreted HEV during the follow-up. HEV-A and HEV-B were present in 2.1 and 2.7% of the specimens, respectively. HEV-C and HEV-D viruses were not detected. In total, 17 different HEV serotypes were detected and the most common findings were CV-A9 (13.5%), CV-A16 (11.5%), and CV-A2 (9.6%). The majority of the infections (92.3%) were during the enterovirus season extending from July to December. Probiotic treatment did not affect significantly the risk of HEV infections during the 2-year follow-up although a trend for transient decrease for HEV positivity (HEV-A and/or HEV-B) by the age of 12 months was observed in children who received probiotics [OR 0.40; 95% confidence interval 0.15 to 1.08; P-value 0.071, generalized estimating (GEE) analysis]. According to the results, HEV-A findings were nearly as common as HEV-B findings among the healthy children participating in this study. Also it was shown that serotypes belonging to HEV-A species can be detected by means of viral culturing. PMID:22499016

Simonen-Tikka, Marja-Leena; Hiekka, Anna-Kaisa; Klemola, Päivi; Poussa, Tuija; Ludvigsson, Johnny; Korpela, Riitta; Vaarala, Outi; Roivainen, Merja

2012-06-01

29

Meta-analysis of the association of enteroviruses with human heart disease  

PubMed Central

The role of viruses in the genesis of both dilated cardiomyopathy (DCM) and acute myocarditis remains uncertain. Modern molecular techniques such as polymerase chain reaction (PCR) and in situ hybridisation are sensitive means of detecting viral genomic material in human myocardial tissue and may help to resolve the quest. Meta-analysis of the papers in the literature records studies of both acute myocarditis and DCM where molecular techniques were used to demonstrate enteroviruses. This review studies information from the published literature as well as statistical analysis of the cumulative molecular data relating enteroviruses to DCM, and to compare these findings with the information available on the role of enteroviruses in acute myocarditis. Twelve papers reported studies in acute myocarditis, of which 11 found higher percentages of enteroviral RNA positivity in the diseased population, giving an overall odds ratio of 4.4. Seventeen papers reported studies in DCM, with 11 recording higher positivity rates in these patients. Cumulative analysis of these data suggests an overall odds ratio of 3.8. The causative role of enteroviruses in acute myocarditis, particularly in children, is supported by meta-analysis of the available literature. The data on DCM is suggestive of an association but a proportion of the studies are negative.?? Keywords: enteroviruses;  dilated cardiomyopathy;  myocarditis;  meta-analyses

Baboonian, C.; Treasure, T.

1997-01-01

30

Enterovirus 71 Infection of Human Immune Cells Induces the Production of Proinflammatory Cytokines  

Microsoft Academic Search

A proinflammatory cytokine storm has been proposed to explain the pathogenesis of enterovirus 71 (EV71)-induced fatalities; however, the mechanism to induce these cytokines during EV71 infection remains unclear. Since most of the proinflammatory cytokines are produced by immune cells, we tested whether EV71 infects human immune cells and induces cytokine production. EV71 infection of a human T cell line (Jurkat),

Lien-Cheng Chen; Trai-Ming Yeh

2009-01-01

31

Serological and molecular evidence of enterovirus infection in patients with end-stage dilated cardiomyopathy.  

PubMed Central

OBJECTIVE: To study the relative diagnostic value of enterovirus-specific molecular biological and serological assays in patients with end-stage dilated cardiomyopathy, and to investigate the possible role of other cardiotropic viruses in dilated cardiomyopathy. DESIGN: Analysis of recipient myocardial tissue and serum from patients with dilated cardiomyopathy and controls undergoing cardiac transplantation for end-stage cardiac disease. SETTING: University virology department and transplantation unit. METHODS: Reverse transcriptase-polymerase chain reaction and nucleotide sequence analysis of myocardial RNA and DNA; enterovirus-specific in situ hybridization; enterovirus-specific immunoglobulin M detection. RESULTS: Enterovirus RNA was detected in myocardial tissue from only a small proportion of (five of 75) hearts. However, although enterovirus-specific immunoglobulin M responses were detected in 22 (28%) of 39 controls patients, a significantly higher prevalence was observed among patients with dilated cardiomyopathy (22 (56%) of 39 patients; P < 0.005). All enteroviruses detected in myocardium showed greatest nucleotide sequence homology with coxsackievirus type B3. Detection of enterovirus RNA in myocardium by the polymerase chain reaction and by in situ hybridisation gave comparable results. Other potentially cardiotropic virus genomes, including human cytomegalovirus, influenzaviruses, and coronaviruses were not detected in myocardium. CONCLUSION: This study found that enterovirus-specific immunoglobulin M responses provided the strongest evidence of enterovirus involvement in patients with end-stage dilated cardiomyopathy. However, the high background prevalence of these responses limits their diagnostic value. The finding that enteroviruses detected in myocardium were coxsackievirus type B3 accords with recent findings in patients with acute myocarditis, and indicates that this serotype is the major cardiotropic human enterovirus. Images

Muir, P.; Nicholson, F.; Illavia, S. J.; McNeil, T. S.; Ajetunmobi, J. F.; Dunn, H.; Starkey, W. G.; Reetoo, K. N.; Cary, N. R.; Parameshwar, J.; Banatvala, J. E.

1996-01-01

32

Immunology in the clinic review series; focus on type 1 diabetes and viruses: the enterovirus link to type 1 diabetes: critical review of human studies.  

PubMed

The hypothesis that under some circumstances enteroviral infections can lead to type 1 diabetes (T1D) was proposed several decades ago, based initially on evidence from animal studies and sero-epidemiology. Subsequently, enterovirus RNA has been detected more frequently in serum of patients than in control subjects, but such studies are susceptible to selection bias and reverse causality. Here, we review critically recent evidence from human studies, focusing on longitudinal studies with potential to demonstrate temporal association. Among seven longitudinal birth cohort studies, the evidence that enterovirus infections predict islet autoimmunity is quite inconsistent in our interpretation, due partially, perhaps, to heterogeneity in study design and a limited number of subjects studied. An association between enterovirus and rapid progression from autoimmunity to T1D was reported by one longitudinal study, but although consistent with evidence from animal models, this novel observation awaits replication. It is possible that a potential association with initiation and/or progression of islet autoimmunity can be ascribed to a subgroup of the many enterovirus serotypes, but this has still not been investigated properly. There is a need for larger studies with frequent sample intervals and collection of specimens of sufficient quality and quantity for detailed characterization of enterovirus. More research into the molecular epidemiology of enteroviruses and enterovirus immunity in human populations is also warranted. Ultimately, this knowledge may be used to devise strategies to reduce the risk of T1D in humans. PMID:22385232

Stene, L C; Rewers, M

2012-04-01

33

Global transcriptomic analysis of human neuroblastoma cells in response to enterovirus type 71 infection.  

PubMed

Human enterovirus type 71 (EV71) is the major pathogen of hand-foot-and-mouth disease (HFMD) and has been associated with severe neurological disease and even death in infants and young children. The pathogenesis of EV71 infection in the human central nervous system remains unclear. In this study, human whole genome microarray was employed to perform transcriptome profiling in SH-SY5Y human neuroblastoma cells infected with EV71. The results indicated that EV71 infection lead to altered expression of 161 human mRNAs, including 74 up-regulated genes and 87 down-regulated genes. Bioinformatics analysis indicated the possible roles of the differentially regulated mRNAs in selected pathways, including cell cycle/proliferation, apoptosis, and cytokine/chemokine responses. Finally, the microarray results were validated using real-time RT-PCR with high identity. Overall, our results provided fundamental information regarding the host response to EV71 infection in human neuroblastoma cells, and this finding will help explain the pathogenesis of EV71 infection and virus-host interaction. PMID:23861741

Xu, Li-Juan; Jiang, Tao; Zhang, Fu-Jun; Han, Jian-Feng; Liu, Juan; Zhao, Hui; Li, Xiao-Feng; Liu, Rui-Ju; Deng, Yong-Qiang; Wu, Xiao-Yan; Zhu, Shun-Ya; Qin, E-De; Qin, Cheng-Feng

2013-07-05

34

Diabetogenic effects of the most prevalent enteroviruses in Finnish sewage.  

PubMed

Common enterovirus infections appear to initiate or facilitate the pathogenetic processes leading to type 1 diabetes (T1D) and also sometimes precipitate the clinical disease. We have recently demonstrated that (1) enterovirus-positive islet cells were seen on postmortem pancreatic specimens of several T1D patients but not in the corresponding samples of nondiabetic controls, and (2) several different enteroviruses can be associated with T1D. Enterovirus infections are transmitted from person to person by fecal-oral or respiratory routes, which means that infections usually start from the respiratory or gastrointestinal mucosa. Regardless of the clinical symptoms of the disease, viral replication continues in the submucosal lymphatic tissue for several weeks, up to a couple of months, and during that time the virus is excreted into the feces and translocated to the environment. Monitoring of sewage samples for enteroviruses can be used as a tool in epidemiologic studies of enterovirus. Finland has successfully used environmental control data in poliovirus surveillance for decades. About 24 samples have been collected annually from the Helsinki region, which covers about 20% of the population. In the present study, we have reanalyzed the sewage samples of the years 1993-2004 for nonpolio enteroviruses by inoculating them into five different continuous cell lines known to cover a wide range of serotypes. Isolated strains were identified by RT-PCR and VP1 sequencing. The most commonly detected serotypes were coxsackie B viruses (CBV1-5) and echoviruses (E6, 7, 11, 25, 30). Diabetogenic effects of the most prevalent enterovirus serotypes were studied in primary human beta cells. PMID:19120297

Klemola, Paivi; Kaijalainen, Svetlana; Ylipaasto, Petri; Roivainen, Merja

2008-12-01

35

Full Genome Sequence of a Novel Human Enterovirus C (EV-C118) Isolated from Two Children with Acute Otitis Media and Community-Acquired Pneumonia in Israel  

PubMed Central

The new enterovirus C strain EV-C118 belongs to the human enterovirus C species of the Picornaviridae family. We report the complete genome sequence of this strain, which was identified in respiratory specimens of two children hospitalized in Israel because of acute otitis media and community-acquired pneumonia who were enrolled in the Community-Acquired Pneumonia Pediatric Research Initiative (CAP-PRI) study.

Daleno, Cristina; Piralla, Antonio; Scala, Alessia; Baldanti, Fausto; Greenberg, David; Principi, Nicola

2013-01-01

36

Display of VP1 on the Surface of Baculovirus and Its Immunogenicity against Heterologous Human Enterovirus 71 Strains in Mice  

Microsoft Academic Search

BackgroundHuman Enterovirus 71 (EV71) is a common cause of hand, foot and mouth disease (HFMD) in young children. It is often associated with severe neurological diseases and has caused high mortalities in recent outbreaks across the Asia Pacific region. Currently, there is no effective vaccine and antiviral agents available against EV71 infections. VP1 is one of the major immunogenic capsid

Tao Meng; Annasaheb B. Kolpe; Tanja K. Kiener; Vincent T. K. Chow; Jimmy Kwang; Ravi Jhaveri

2011-01-01

37

Adoptive transfer of macrophages from adult mice reduces mortality in mice infected with human enterovirus 71.  

PubMed

Human enterovirus 71 (EV71) causes hand, foot and mouth disease in children under 6 years of age, and the neurological complications of this virus can lead to death. Until now, no vaccines or drugs have been available for the clinical control of this epidemic. Macrophages can engulf pathogens and mediate a series of host immune responses that play a role in the defence against infectious diseases. Using immunohistochemistry, we observed the localizations of virus in muscle tissues of EV71-infected mice. The macrophages isolated from the adult mice could kill the virus gradually in vitro, as shown using quantitative real-time PCR (qRT-PCR) and virus titration. Co-localisation of lysosomes and virus within macrophages suggested that the lysosomes were possibly responsible for the phagocytosis of EV71. Activation of the macrophages in the peritoneal cavity of mice four days pre-infection reduced the mortality of mice upon lethal EV71 infection. The adoptive transfer of macrophages from adult mice inhibited virus replication in the muscle tissues of infected mice, and this was followed by a relief of symptoms and a significant reduction of mortality, which suggested that the adoptive transfer of macrophages from adult humans represents a potential strategy to treat EV71-infected patients. PMID:23065110

Liu, Jiangning; Li, Xiaoying; Fan, Xiaoxu; Ma, Chunmei; Qin, Chuan; Zhang, Lianfeng

2012-10-11

38

Resequencing microarray probe design for typing genetically diverse viruses: human rhinoviruses and enteroviruses  

PubMed Central

Background Febrile respiratory illness (FRI) has a high impact on public health and global economics and poses a difficult challenge for differential diagnosis. A particular issue is the detection of genetically diverse pathogens, i.e. human rhinoviruses (HRV) and enteroviruses (HEV) which are frequent causes of FRI. Resequencing Pathogen Microarray technology has demonstrated potential for differential diagnosis of several respiratory pathogens simultaneously, but a high confidence design method to select probes for genetically diverse viruses is lacking. Results Using HRV and HEV as test cases, we assess a general design strategy for detecting and serotyping genetically diverse viruses. A minimal number of probe sequences (26 for HRV and 13 for HEV), which were potentially capable of detecting all serotypes of HRV and HEV, were determined and implemented on the Resequencing Pathogen Microarray RPM-Flu v.30/31 (Tessarae RPM-Flu). The specificities of designed probes were validated using 34 HRV and 28 HEV strains. All strains were successfully detected and identified at least to species level. 33 HRV strains and 16 HEV strains could be further differentiated to serotype level. Conclusion This study provides a fundamental evaluation of simultaneous detection and differential identification of genetically diverse RNA viruses with a minimal number of prototype sequences. The results demonstrated that the newly designed RPM-Flu v.30/31 can provide comprehensive and specific analysis of HRV and HEV samples which implicates that this design strategy will be applicable for other genetically diverse viruses.

Wang, Zheng; Malanoski, Anthony P; Lin, Baochuan; Kidd, Carolyn; Long, Nina C; Blaney, Kate M; Thach, Dzung C; Tibbetts, Clark; Stenger, David A

2008-01-01

39

21 CFR 866.3225 - Enterovirus nucleic acid assay.  

Code of Federal Regulations, 2013 CFR

...from individuals who have signs and symptoms consistent with meningitis or meningoencephalitis. The detection of enterovirus RNA...tests, aids in the clinical laboratory diagnosis of viral meningitis caused by enterovirus. (b) Classification . Class...

2013-04-01

40

The essential role of clathrin-mediated endocytosis in the infectious entry of human enterovirus 71.  

PubMed

Little is currently known about the infectious entry process of human enterovirus 71 (HEV71) into host cells, which may represent potential anti-viral targeting sites. In this study a targeted small-interfering RNA (siRNA) screening platform assay was established and validated to identify and profile key cellular genes involved in processes of endocytosis, cytoskeletal dynamics, and endosomal trafficking essential for HEV71 infection. Screen evaluation was conducted via the expression of well characterized dominant-negative mutants, bioimaging studies (double-labeled immunofluorescence assays, transmission electron microscopy analysis), secondary siRNA-based dosage dependence studies, and drug inhibition assays. The infectious entry of HEV71 into rhabdomyosarcoma cells was shown to be significantly inhibited by siRNAs targeting genes associated with clathrin-mediated endocytosis (CME) that include AP2A1, ARRB1, CLTC, CLTCL1, SYNJ1, ARPC5, PAK1, ROCK1, and WASF1. The functional role of CME was verified by the observation of strong co-localization between HEV71 particles and clathrin as well as dose-dependent inhibition of HEV71 infection upon siRNA knockdown of CME-associated genes. HEV71 entry by CME was further confirmed via inhibition by dominant-negative EPS15 mutants and treatment of CME drug inhibitors, with more than 80% inhibition observed at 20 ?m chlorpromazine. Furthermore, HEV71 infection was shown to be sensitive to the disruption of human genes in regulating early to late endosomal trafficking as well as endosomal acidic pH. The identification of clathrin-mediated endocytosis as the entry pathway for HEV71 infection of susceptible host cells contributes to a better understanding of HEV71 pathogenesis and enables future development of anti-viral strategies against HEV71 infection. PMID:20956521

Hussain, Khairunnisa' Mohamed; Leong, Kim Lian Janet; Ng, Mary Mah-Lee; Chu, Justin Jang Hann

2010-10-18

41

The Essential Role of Clathrin-mediated Endocytosis in the Infectious Entry of Human Enterovirus 71*  

PubMed Central

Little is currently known about the infectious entry process of human enterovirus 71 (HEV71) into host cells, which may represent potential anti-viral targeting sites. In this study a targeted small-interfering RNA (siRNA) screening platform assay was established and validated to identify and profile key cellular genes involved in processes of endocytosis, cytoskeletal dynamics, and endosomal trafficking essential for HEV71 infection. Screen evaluation was conducted via the expression of well characterized dominant-negative mutants, bioimaging studies (double-labeled immunofluorescence assays, transmission electron microscopy analysis), secondary siRNA-based dosage dependence studies, and drug inhibition assays. The infectious entry of HEV71 into rhabdomyosarcoma cells was shown to be significantly inhibited by siRNAs targeting genes associated with clathrin-mediated endocytosis (CME) that include AP2A1, ARRB1, CLTC, CLTCL1, SYNJ1, ARPC5, PAK1, ROCK1, and WASF1. The functional role of CME was verified by the observation of strong co-localization between HEV71 particles and clathrin as well as dose-dependent inhibition of HEV71 infection upon siRNA knockdown of CME-associated genes. HEV71 entry by CME was further confirmed via inhibition by dominant-negative EPS15 mutants and treatment of CME drug inhibitors, with more than 80% inhibition observed at 20 ?m chlorpromazine. Furthermore, HEV71 infection was shown to be sensitive to the disruption of human genes in regulating early to late endosomal trafficking as well as endosomal acidic pH. The identification of clathrin-mediated endocytosis as the entry pathway for HEV71 infection of susceptible host cells contributes to a better understanding of HEV71 pathogenesis and enables future development of anti-viral strategies against HEV71 infection.

Hussain, Khairunnisa' Mohamed; Leong, Kim Lian Janet; Ng, Mary Mah-Lee; Chu, Justin Jang Hann

2011-01-01

42

A study of the virulence in mice of high copying fidelity variants of human enterovirus 71.  

PubMed

Polioviruses with a G64S mutation in the 3D polymerase have enhanced replication fidelity and are attenuated in animal models. Here we describe the mouse virulence properties of high replication fidelity 3D polymerase variants of human enterovirus 71 (HEV71), with mutations at positions 3D-S264L, 3D-G64R or at 3D-S264L plus 3D-G64R. Mouse-adapted strains (MP-G64R, MP-S264L and MP-S264L-G64R) were constructed in order to compare the virulence of the 3D polymerase variants with that of mouse-adapted parental virus (MP-26M). MP-S264L and MP-S264L-G64R were attenuated in mice (mean survival time 7.0 and 7.5 days p.i., respectively) compared to MP-G64R and MP-26M (mean survival time 6.5 and 6.0 days p.i., respectively). MP-26M and MP-G64R infection induced early onset, severe generalised necrotising myositis, whereas MP-S264L and MP-S264L-G64R infection induced a later onset, mild and focal skeletal muscle myositis. Our findings demonstrate that only the 3D-S264L mutation attenuates HEV71 in mice, suggesting that the high replication fidelity phenotype is not essential for virulence attenuation in this model. PMID:23856384

Sadeghipour, Sara; McMinn, Peter C

2013-07-12

43

A Dominant EV71-Specific CD4+ T Cell Epitope Is Highly Conserved among Human Enteroviruses  

PubMed Central

CD4+ T cell-mediated immunity plays a central role in determining the immunopathogenesis of viral infections. However, the role of CD4+ T cells in EV71 infection, which causes hand, foot and mouth disease (HFMD), has yet to be elucidated. We applied a sophisticated method to identify promiscuous CD4+ T cell epitopes contained within the sequence of the EV71 polyprotein. Fifteen epitopes were identified, and three of them are dominant ones. The most dominant epitope is highly conserved among enterovirus species, including HFMD-related coxsackieviruses, HFMD-unrelated echoviruses and polioviruses. Furthermore, the CD4+ T cells specific to the epitope indeed cross-reacted with the homolog of poliovirus 3 Sabin. Our findings imply that CD4+ T cell responses to poliovirus following vaccination, or to other enteroviruses to which individuals may be exposed in early childhood, may have a modulating effect on subsequent CD4+ T cell response to EV71 infection or vaccine.

Wei, Ruicheng; Yang, Chunfu; Zeng, Mei; Terry, Frances; Zhu, Kai; Yang, Chunhui; Altmeyer, Ralf; Martin, William; De Groot, Anne S.; Leng, Qibin

2012-01-01

44

Selective human enterovirus and rhinovirus inhibitors: An overview of capsid-binding and protease-inhibiting molecules.  

PubMed

The absence of effective vaccines for most viral infections highlights an urgent necessity for the design and development of effective antiviral drugs. Due to the advancement in virology since the late 1980s, several key events in the viral life cycle have been well delineated and a number of molecular targets have been validated, culminating in the emergence of many new antiviral drugs in recent years. Inhibitors against enteroviruses and rhinoviruses, responsible for about half of the human common colds, are currently under active investigation. Agents targeted at either viral protein 1 (VP1), a relatively conserved capsid structure mediating viral adsorption/uncoating process, or 3C protease, which is highly conserved among different serotypes and essential for viral replication, are of great potential to become antipicornavirus drugs. PMID:15170592

Shih, Shin-Ru; Chen, Shu-Jen; Hakimelahi, Gholam Hossein; Liu, Hsing-Jang; Tseng, Chen-Tso; Shia, Kak-Shan

2004-07-01

45

Molecular detection and identification of enteroviruses in children admitted to a university hospital in Greece.  

PubMed

Although enteroviral infections occur frequently during childhood, the circulation of particular serotypes has never been studied in Greece. The objectives of the present report were molecular detection and identification of human enteroviruses in children admitted with nonspecific febrile illness or meningitis to a university hospital during a 22-month period. A one-step Real-Time RT-PCR protocol was used for rapid enterovirus detection in genetic material extracted directly from clinical samples, and a sensitive reverse transcription-semi-nested PCR targeting part of the VP1-coding region was used for genotypic identification of the different serotypes. Twenty-one enterovirus strains were detected and identified in 20 stool samples, one cerebrospinal fluid (CSF) sample, one whole blood sample and one throat swab from 21 out of 134 febrile patients (15.7%). Ten strains belonged to Human Enterovirus Species B (HEV-B) (six serotypes) and eleven to HEV-A (four serotypes). Most of the strains were closely associated with virulent strains circulating in Europe and elsewhere. Detection of the emerging pathogen enterovirus 71 for a first time in Greece was particularly important. PMID:21803150

Siafakas, Nikolaos; Attilakos, Ahilleas; Vourli, Sofia; Stefos, Efstathios; Meletiadis, Joseph; Nikolaidou, Polyxeni; Zerva, Loukia

2011-07-23

46

Animal "orphan" enteroviruses  

PubMed Central

Since the discovery, some ten years ago, of the pathogenic effect of polioviruses on non-nervous-tissue cells, tissue-culture methods have come to be widely used in virological research. Through these improved techniques for studying viruses, a large number of new cytopathogenic agents have been isolated from the intestinal tract of man. Many of these agents have been obtained from persons suffering from polio-like disease, but others have been isolated from apparently normal persons. The term ”orphans” is used to designate those viruses which cannot definitely be associated with any recognized disease syndrome. The existence of these enteric pathogenic human orphan (ECHO) viruses, and their association with clinical disease in certain cases, stimulated interest in their animal counter-parts, which might constitute a serious threat to both human and animal health. In this paper, the author reviews the information at present available on the occurrence of the so-called ”orphan” enteroviruses in monkeys, cattle, swine, and other animals in various parts of the world, and discusses the possible interrelationships of these animal viruses with each other and with the human enteroviruses. ImagesFIG. 1FIG. 2FIG. 3

Kalter, Seymour S.

1960-01-01

47

Diverse apoptotic pathways in enterovirus 71-infected cells  

Microsoft Academic Search

Mechanisms related to the neuropathogenesis of enterovirus 71 infection remain unclear. This investigation conducts a comprehensive\\u000a study of the apoptotic pathways in neural and non-neural cells following enterovirus 71 infection. Infections with enterovirus\\u000a 71 not only induce classical cytopathic effects in SF268 (human glioblastoma), SK-N-MC (human neuroblastoma), RD, and Vero\\u000a cells, but also induce classic signs of apoptosis in all

Shih-Cheng Chang; Jing-Yi Lin; Lily Yen-Cheng Lo; Mei-Ling Li; Shin-Ru Shih

2004-01-01

48

Genetic Characteristics of Human Enterovirus 71 and Coxsackievirus A16 Circulating from 1999 to 2004 in Shenzhen, People's Republic of China  

Microsoft Academic Search

The genetic and phylogenetic characteristics of human enterovirus 71 (EV71) and coxsackievirus A16 (CA16) sampled from children with hand, foot, and mouth disease in Shenzhen, People's Republic of China, over a 6-year period (1999 to 2004) were examined with reverse transcription-PCR and DNA sequencing. Out of 147 stool specimens, 60 showed positive signals when screened with EV71- and CA16-specific primers.

Linlin Li; Yaqing He; Hong Yang; Junping Zhu; Xingye Xu; Jie Dong; Yafang Zhu; Qi Jin

2005-01-01

49

Detection of herpes simplex virus (1 and 2), varicella-zoster virus, cytomegalovirus, human herpesvirus 6 and enterovirus in immunocompetent Tunisian patients with acute neuromeningeal disorder.  

PubMed

Enteroviruses (EVs) and human herpesviruses (HHVs) are involved frequently in acute neurological disorders of viral etiology. This study aimed to investigate the incidence of herpes simplex virus types-1 (HSV-1) and 2 (HSV-2), varicella-zoster virus (VZV), cytomegalovirus (CMV), human herpesvirus 6 (HHV-6) and human enteroviruses (EVs) in cerebrospinal fluid (CSF) samples of Tunisian immunocompetent patients with neuromeningeal disorders. The patients had been hospitalized at the Fattouma Bourguiba University Hospital (Monastir, Tunisia) between September 2007 and June 2009. At least one viral genome was detected in 58 (46%) out of 126 CSF samples collected. Enterovirus was detected in 31 of the positive samples (53.4%), CMV in 20 (34.5%), HSV-1 in 3 (5.2%), HSV-2 in 6 (10.3%), VZV in 4 (6.9%), HHV-6 in 2 (3.4%). More than one viral genome was detected in seven CSF samples, including CMV DNA in six of the samples. The high frequency of enteroviral infections in aseptic meningitis was confirmed. The detection of CMV DNA only suggests a direct role of this virus in the etiology of acute neuromeningeal disorder. PMID:22170549

Nahdi, Imen; Boukoum, Hanen; Nabil Ben Salem, Abid; Ben Romdane, Fouad; Hammami, Saber; Chebel, Saber; Mahbouba, Frih-Ayed; Guediche, Mohamed Neji; Chakroun, Mohamed; Aouni, Mahjoub; Imbert-Marcille, Berthe-Marie; Bressollette-Bodin, Celine

2012-02-01

50

Molecular Determinants of Enterovirus 71 Viral Entry  

PubMed Central

Enterovirus 71 (EV71) is one of the major pathogens that cause hand, foot, and mouth disease outbreaks in young children in the Asia-Pacific region in recent years. Human scavenger receptor class B 2 (SCARB2) is the main cellular receptor for EV71 on target cells. The requirements of the EV71-SCARB2 interaction have not been fully characterized, and it has not been determined whether SCARB2 serves as an uncoating receptor for EV71. Here we compared the efficiency of the receptor from different species including human, horseshoe bat, mouse, and hamster and demonstrated that the residues between 144 and 151 are critical for SCARB2 binding to viral capsid protein VP1 of EV71 and seven residues from the human receptor could convert murine SCARB2, an otherwise inefficient receptor, to an efficient receptor for EV71 viral infection. We also identified that EV71 binds to SCARB2 via a canyon of VP1 around residue Gln-172. Soluble SCARB2 could convert the EV71 virions from 160 S to 135 S particles, indicating that SCARB2 is an uncoating receptor of the virus. The uncoating efficiency of SCARB2 significantly increased in an acidic environment (pH 5.6). These studies elucidated the viral capsid and receptor determinants of enterovirus 71 infection and revealed a possible target for antiviral interventions.

Chen, Pan; Song, Zilin; Qi, Yonghe; Feng, Xiaofeng; Xu, Naiqing; Sun, Yinyan; Wu, Xing; Yao, Xin; Mao, Qunyin; Li, Xiuling; Dong, Wenjuan; Wan, Xiaobo; Huang, Niu; Shen, Xinliang; Liang, Zhenglun; Li, Wenhui

2012-01-01

51

Detection of enterovirus in human skeletal muscle from patients with chronic inflammatory muscle disease or fibromyalgia and healthy subjects  

Microsoft Academic Search

Enterovirus RNA has been found previously in specimens of muscle biopsy from patients with idiopathic dilated cardiomyopathy, chronic in- flammatory muscle diseases, and fibromyalgia or chronic fatigue syndrome (fibromyalgia\\/chronic fatigue syndrome). These results suggest that skeletal muscle may host enteroviral persistent infection.Totestthishypothesis,weinvestigated by reverse transcription-polymerase chain reac- tion (RT-PCR) assay the presence of enterovirus in skeletal muscle of patients with

Fatima Douche-Aourik; Willy Berlier; Thomas Bourlet; Rafik Harrath; Shabir Omar; Florence Grattard; Christian Denis; Bruno Pozzetto

2003-01-01

52

The survey of porcine teschoviruses, sapeloviruses and enteroviruses B infecting domestic pigs and wild boars in the Czech Republic between 2005 and 2011.  

PubMed

This study presents results of epidemiological survey and genetic characterisation of porcine enteric picornaviruses belonging to the genera Teschovirus, Sapelovirus, and Porcine enterovirus B. Faecal or gut content samples from domestic pigs (Sus scrofa f. domestica) and the cecal content of wild boars (Sus scrofa) of different ages (collected between 2005 and 2011) were analysed by molecular methods. Porcine enterovirus B was the most prevalent virus detected in both domestic pigs and wild boars (50.2% and 69.4%, respectively), followed by Porcine teschovirus and Porcine sapelovirus. The majority of positive domestic pigs (69.4%) and wild boars (64.3%) were infected with two or three tested viruses. There was no significant difference in prevalences of teschoviruses, sapeloviruses, and enteroviruses among healthy and diarrhoeic pigs. Results of epidemiological survey demonstrated that all target viral genera are common in Czech farms producing pigs and wild boars. Amplified nucleotide fragments of VP2 region obtained from randomly selected both historical and recent Teschovirus isolates were sequenced. Based on sequence data, historical Porcine teschovirus isolate CAPM V-180, previously determined as serotype 1 was reclassified into serotype 11. Moreover, another recent Porcine teschovirus isolate OH264/2010 was described and classified into serotype 11. Four nontypeable PTV strains (historical isolate CAPM V-182/1976 and recent isolates JA247/2010, NI429/2010, and BR1576/2007) identified in this study might represent novel serotypes. To the best of our knowledge, our study represents the first description of this serotype in the Czech Republic. PMID:22579481

Prod?lalová, Jana

2012-05-03

53

A water extract of Pueraria lobata inhibited cytotoxicity of enterovirus 71 in a human foreskin fibroblast cell line.  

PubMed

Enterovirus 71 (EV71) can cause brain encephalitis and mortality. However, effective vaccines or chemotherapeutic agents are not yet available. We tested the hypothesis that Pueraria lobata could inhibit the cytotoxic effect of EV71 in a human foreskin fibroblast cell line by the XTT method. Our results showed that the water extract of P. lobata could inhibit cytopathy induced by EV71 when given before (p < 0.0001), simultaneously with (p < 0.0001), or after viral infection (p < 0.0001). Water extract of P. lobata was effective and its minimal concentration that inhibited 50% of the cytopathic effect (IC50) was 0.028 microg/mL. P. lobata was also safe with a selectivity index greater than 107,000. Water extract of P. lobata appeared to inhibit viral attachment (p < 0.0001) and penetration (p < 0.0001). The anti-EV71 activity of the water extract of P. lobata was not mediated by interferons. In conclusion, the water extract of P. lobata was effective in the management of the disease induced by EV71 infection. PMID:19181583

Su, Fu-Min; Chang, Jung-San; Wang, Kuo-Chih; Tsai, Jih-Jin; Chiang, Lien-Chai

2008-10-01

54

Human astrocytic cells support persistent coxsackievirus b3 infection.  

PubMed

Enteroviruses can frequently target the human central nervous system to induce a variety of neurological diseases. Although enteroviruses are highly cytolytic, emerging evidence has shown that these viruses can establish persistent infections both in vivo and in vitro. Here, we investigated the susceptibility of three human brain cell lines, CCF-STTG1, T98G, and SK-N-SH, to infection with three enterovirus serotypes: coxsackievirus B3 (CVB3), enterovirus 71, and coxsackievirus A9. Persistent infection was observed in CVB3-infected CCF-STTG1 cells, as evidenced by prolonged detection of infectious virions, viral RNA, and viral antigens. Of note, infected CCF-STTG1 cells expressed the nonfunctional canonical viral receptors coxsackievirus-adenovirus receptor and decay-accelerating factor, while removal of cell surface chondroitin sulfate from CCF-STTG1 cells inhibited the replication of CVB3, suggesting that receptor usage was one of the major limiting factors in CVB3 persistence. In addition, CVB3 curtailed the induction of beta interferon in infected CCF-STTG1 cells, which likely contributed to the initiation of persistence. Furthermore, proinflammatory chemokines and cytokines, such as vascular cell adhesion molecule 1, interleukin-8 (IL-8), and IL-6, were upregulated in CVB3-infected CCF-STTG1 cells and human progenitor-derived astrocytes. Our data together demonstrate the potential of CCF-STTG1 cells to be a novel cell model for studying CVB3-central nervous system interactions, providing the basis toward a better understanding of CVB3-induced chronic neuropathogenesis. PMID:24027313

Zhang, Xiaowei; Zheng, Zhenhua; Shu, Bo; Liu, Xijuan; Zhang, Zhenfeng; Liu, Yan; Bai, Bingke; Hu, Qinxue; Mao, Panyong; Wang, Hanzhong

2013-09-11

55

Flow cytometric detection and serotyping of enterovirus for the clinical laboratory.  

PubMed

Culture and serotyping of human enteroviruses by fluorescence microscopy are time-consuming and labor-intensive. Flow cytometry has the potential of being more rapid, sensitive, and objective but has not been used for these purposes in a clinical laboratory. Primary rhesus monkey kidney (PMK) cells were inoculated with several enterovirus serotypes and stained with enterovirus-specific antibodies for flow cytometry and indirect fluorescence antibody testing (IFA). Kinetic studies of coxsackievirus B1 and echovirus 30 infection of PMK cells were performed on days 1-4 after inoculation. Flow cytometry results for echovirus 6, 9, 11, and 30 and coxsackievirus B1 correlated with IFA in all cases. Coxsackievirus B1 and echovirus 30 infections were detected 1 day earlier by flow cytometry than IFA. Flow cytometry can be effectively used for detecting enterovirus-infected cells in a clinical laboratory with the advantages of better quantitation of low levels of infection and earlier detection of virally infected cells in culture systems. PMID:19733594

She, Rosemary C; Preobrazhensky, Sergey N; Taggart, Edward W; Petti, Cathy A; Bahler, David W

2009-09-04

56

Detection and whole genome sequence analysis of an enterovirus 68 cluster  

PubMed Central

Background Enteroviruses are a common cause of human disease and are associated with a wide range of clinical manifestations. Enterovirus 68 is rarely detected yet was reported in many countries in 2010. Here enterovirus 68 was identified for the first time in New Zealand in 2010 and was detected in a further fourteen specimens over a six month period. Objectives To genetically characterise enterovirus 68 specimens identified in New Zealand in 2010. Study design The genome sequence of a New Zealand representative enterovirus 68 isolate was obtained. Ten clinical specimens were analysed by sequencing the VP1 region of the enterovirus 68 genome. Results Based on sequence analysis of the VP1 region and the full genome of one representative isolate, the New Zealand enterovirus 68 isolates clustered with contemporary enterovirus 68 viruses and do not show any clear distinguishing genetic diversity when compared to other strains. All fifteen specimens showed high similarity with enterovirus 68 by VP1 sequencing. The majority of New Zealand patients suffered from bronchiolitis, were less than two years of age and were of Pacific Island or Maori descent. Conclusions We document the rare occurrence of an enterovirus 68 cluster in New Zealand in 2010. These viruses shared similarity with other clusters of enterovirus 68 that occurred globally in 2010. A greater awareness in enterovirus 68 infection may help detect this virus with increased frequency and enable us to better understand the role this strain plays in disease and the reasons behind this global emergence in 2010.

2013-01-01

57

Production of Enterovirus Antisera.  

National Technical Information Service (NTIS)

The production of enterovirus antisera in horses and of rhinovirus antisera in goats is reported. Eight other coxsackie virus, one echo virus, and eight rhinovirus antigens are in progress. Data on inoculations, bleedings, and stored and transferred sera ...

R. W. Brown

1967-01-01

58

Enteroviruses in the pathogenesis of type 1 diabetes.  

PubMed

The question if enteroviruses could cause beta-cell damage and type 1 diabetes has become more and more relevant when recent studies have provided new evidence supporting this scenario. One important observation is the recent discovery of IFIH1 as a risk gene for type 1 diabetes. This gene is an innate immune system receptor for enteroviruses offering one possible mechanism for the diabetogenic effect of enteroviruses. This is further emphasized by the observations suggesting that the innate immune system is activated in the pancreatic islets of type 1 diabetic patients and that the innate immune system is important for the defense against the virus and for the regulation of adaptive immune system. Important progress has also been gained in studies analyzing pancreas tissue for possible presence of enteroviruses. Several studies have found enteroviruses in the pancreatic islets of type 1 diabetic patients using various methods. The virus seems to be located in the islets while exocrine pancreas is mostly uninfected. One recent study found the virus in the intestinal mucosa in the majority of diabetic patients. Enteroviruses can also infect cultured human pancreatic islets causing either rapid cell destruction or a persistent-like noncytolytic infection. Combined with all previous, epidemiological findings indicating the risk effect of enteroviruses in cross-sectional and prospective studies, these observations fit to a scenario where certain diabetogenic enterovirus variants establish persistent infection in gut mucosa and in the pancreatic islets. This in turn could lead to a local inflammation and the breakdown of tolerance in genetically susceptible individuals. This is also supported by mouse experiments showing that enteroviruses can establish prolonged infection in the pancreas and intestine, and some virus strains cause beta-cell damage and diabetes. In conclusion, recent studies have strengthened the hypothesis that enteroviruses play a role in the pathogenesis of type 1 diabetes. These findings open also new opportunities to explore the underlying mechanism and get closer to causal relationship. PMID:20424841

Tauriainen, Sisko; Oikarinen, Sami; Oikarinen, Maarit; Hyöty, Heikki

2010-04-28

59

The detection of enteroviruses in sewage using Caco-2 cells.  

PubMed

The work presented here demonstrates the utility of Caco-2 cells to detect enteroviruses in sewage. Viruses were concentrated by beef extract elution and organic flocculation prior to analysis by cell culture assays and RT-PCR. Enteroviruses were detected in all sewage samples, but only one sample was positive solely in RT-PCR assay. We proved that Caco-2 cells were more effective than RD and L20B cells in enterovirus isolation, depending on procedures used in the inoculation process. PMID:23829085

Wieczorek, Magdalena; Kuryk, ?ukasz; Witek, Agnieszka; Diuwe, Anna; Litwi?ska, Bogumi?a

2013-01-01

60

Molecular Typing of Enteroviruses: Current Status and Future Requirements  

PubMed Central

Human enteroviruses have traditionally been typed according to neutralization serotype. This procedure is limited by the difficulty in culturing some enteroviruses, the availability of antisera for serotyping, and the cost and technical complexity of serotyping procedures. Furthermore, the impact of information derived from enterovirus serotyping is generally perceived to be low. Enteroviruses are now increasingly being detected by PCR rather than by culture. Classical typing methods will therefore no longer be possible in most instances. An alternative means of enterovirus typing, employing PCR in conjunction with molecular genetic techniques such as nucleotide sequencing or nucleic acid hybridization, would complement molecular diagnosis, may overcome some of the problems associated with serotyping, and would provide additional information regarding the epidemiology and biological properties of enteroviruses. We argue the case for developing a molecular typing system, discuss the genetic basis of such a system, review the literature describing attempts to identify or classify enteroviruses by molecular methods, and suggest ways in which the goal of molecular typing may be realized.

Muir, Peter; Kammerer, Ulrike; Korn, Klaus; Mulders, Mick N.; Poyry, Tuija; Weissbrich, Benedikt; Kandolf, Reinhard; Cleator, Graham M.; van Loon, Anton M.

1998-01-01

61

Echovirus 22 is an atypical enterovirus.  

PubMed Central

Although echovirus 22 (EV22) is classified as an enterovirus in the family Picornaviridae, it is atypical of the enterovirus paradigm, typified by the polioviruses and the coxsackie B viruses. cDNA reverse transcribed from coxsackievirus B3 (CVB3) RNA does not hybridize to genomic RNA of EV22, and conversely, cDNA made to EV22 does not hybridize to CVB3 genomic RNA or to molecular clones of CVB3 or poliovirus type 1. EV22 cDNA does not hybridize to viral RNA of encephalomyocarditis virus or to a molecular clone of Theiler's murine encephalomyelitis virus, members of the cardiovirus genus. The genomic RNA of EV22 cannot be detected by the polymerase chain reaction using generic enteroviral primers. EV22 does not shut off host cell protein synthesis, and the RNA of EV22 is efficiently translated in vitro in rabbit reticulocyte lysates. Murine enterovirus-immune T cells recognize and proliferate against EV22 as an antigen in vitro, demonstrating that EV22 shares an epitope(s) common to enteroviruses but not found among other picornaviruses. Images

Coller, B A; Chapman, N M; Beck, M A; Pallansch, M A; Gauntt, C J; Tracy, S M

1990-01-01

62

Expression of the Coxsackievirus and Adenovirus Receptor in Cultured Human Umbilical Vein Endothelial Cells: Regulation in Response to Cell Density  

Microsoft Academic Search

The six serotypes of the group B coxsackieviruses (CVB1 to -6) are human enteroviruses that cause a variety of acute dis- eases and are strongly implicated as causative agents in several chronic diseases (1, 10). These enteroviruses presumably gain access to the heart, as well as to other tissues distant from the gut, via the blood during viremia. They must

STEVEN D. CARSON; JUSTIN T. HOBBS; STEVEN M. TRACY; NORA M. CHAPMAN

1999-01-01

63

Quantitative analysis of viral RNA kinetics in coxsackievirus B3-induced murine myocarditis: biphasic pattern of clearance following acute infection, with persistence of residual viral RNA throughout and beyond the inflammatory phase of disease  

Microsoft Academic Search

Although the association remains controversial, enteroviruses have been implicated in the aetiology of several chronic diseases of humans. To further understand the mechanism of enterovirus persistence and its relationship to organ pathology, virus infectivity and viral RNA kinetics in the heart and other target organs during acute and persistent phases of murine coxsackievirus B3 infection were investigated. These studies revealed

K. Nundita Reetoo; Shabina A. Osman; Shirin J. Illavia; Charlotte L. Cameron-Wilson; Jangu E. Banatvala; Peter Muir

64

Enterovirus 71 VP1 Activates Calmodulin-Dependent Protein Kinase II and Results in the Rearrangement of Vimentin in Human Astrocyte Cells.  

PubMed

Enterovirus 71 (EV71) is one of the main causative agents of foot, hand and mouth disease. Its infection usually causes severe central nervous system diseases and complications in infected infants and young children. In the present study, we demonstrated that EV71 infection caused the rearrangement of vimentin in human astrocytoma cells. The rearranged vimentin, together with various EV71 components, formed aggresomes-like structures in the perinuclear region. Electron microscopy and viral RNA labeling indicated that the aggresomes were virus replication sites since most of the EV71 particles and the newly synthesized viral RNA were concentrated here. Further analysis revealed that the vimentin in the virus factories was serine-82 phosphorylated. More importantly, EV71 VP1 protein is responsible for the activation of calmodulin-dependent protein kinase II (CaMK-II) which phosphorylated the N-terminal domain of vimentin on serine 82. Phosphorylation of vimentin and the formation of aggresomes were required for the replication of EV71 since the latter was decreased markedly after phosphorylation was blocked by KN93, a CaMK-II inhibitor. Thus, as one of the consequences of CaMK-II activation, vimentin phosphorylation and rearrangement may support virus replication by playing a structural role for the formation of the replication factories. Collectively, this study identified the replication centers of EV71 in human astrocyte cells. This may help us understand the replication mechanism and pathogenesis of EV71 in human. PMID:24073199

Haolong, Cong; Du, Ning; Hongchao, Tian; Yang, Yang; Wei, Zhang; Hua, Zhang; Wenliang, Zhang; Lei, Song; Po, Tien

2013-09-20

65

Enterovirus 71 VP1 Activates Calmodulin-Dependent Protein Kinase II and Results in the Rearrangement of Vimentin in Human Astrocyte Cells  

PubMed Central

Enterovirus 71 (EV71) is one of the main causative agents of foot, hand and mouth disease. Its infection usually causes severe central nervous system diseases and complications in infected infants and young children. In the present study, we demonstrated that EV71 infection caused the rearrangement of vimentin in human astrocytoma cells. The rearranged vimentin, together with various EV71 components, formed aggresomes-like structures in the perinuclear region. Electron microscopy and viral RNA labeling indicated that the aggresomes were virus replication sites since most of the EV71 particles and the newly synthesized viral RNA were concentrated here. Further analysis revealed that the vimentin in the virus factories was serine-82 phosphorylated. More importantly, EV71 VP1 protein is responsible for the activation of calmodulin-dependent protein kinase II (CaMK-II) which phosphorylated the N-terminal domain of vimentin on serine 82. Phosphorylation of vimentin and the formation of aggresomes were required for the replication of EV71 since the latter was decreased markedly after phosphorylation was blocked by KN93, a CaMK-II inhibitor. Thus, as one of the consequences of CaMK-II activation, vimentin phosphorylation and rearrangement may support virus replication by playing a structural role for the formation of the replication factories. Collectively, this study identified the replication centers of EV71 in human astrocyte cells. This may help us understand the replication mechanism and pathogenesis of EV71 in human.

Haolong, Cong; Du, Ning; Hongchao, Tian; Yang, Yang; Wei, Zhang; Hua, Zhang; Wenliang, Zhang; Lei, Song; Po, Tien

2013-01-01

66

Detection of Infectious Enteroviruses, Enterovirus Genomes, Somatic Coliphages, and Bacteroides fragilis Phages in Treated Wastewater  

Microsoft Academic Search

In this study, three types of treated wastewater were tested for infectious enteroviruses, the enterovirus genome, somatic coliphages, and Bacteroides fragilis phages. The aim of this work was to determine whether the presence of the two types of bacteriophages or of the enterovirus genome was a good indicator of infectious enterovirus contamination. The enterovirus genome was detected by reverse transcription-polymerase

C. GANTZER; A. MAUL; J. M. AUDIC; L. SCHWARTZBROD; Facultede Pharmacie

1998-01-01

67

Fatal enterovirus 71 encephalomyelitis  

Microsoft Academic Search

During an outbreak of hand-foot-mouth disease caused by enterovirus 71 (EV-71) in 1997, 4 children presented with sudden cardiopulmonary collapse and minimal neurologic features. All children received cardiopulmonary resuscitation but died within a few hours of admission. Postmortem studies showed infection by EV-71 with extensive damage to the medulla and pons. We postulate an etiologic link between EV-71 and brainstem

Lucy C. S. Lum; K. T. Wong; S. K. Lam; K. B. Chua; A. Y. T. Goh; W. L. Lim; B. B. Ong; G. Paul; S. AbuBakar; M. Lambert

1998-01-01

68

Genetic clustering of all 102 human rhinovirus prototype strains: serotype 87 is close to human enterovirus 70  

Microsoft Academic Search

Human rhinoviruses (HRV), common agents of respiratory infections, comprise 102 designated serotypes. The genetic relationships of HRV prototype strains and the possibility of using genetic identification of a given HRV field strain were studied. Genomic sequences in the VP4\\/VP2 region were obtained from all 102 prototype strains. Phylogenetic analysis included 61 recently isolated Finnish field strains. Seventy-six out of the

Carita Savolainen; Soile Blomqvist; Mick N. Muldersã; Tapani Hovi

2002-01-01

69

Detection of enteroviruses in the intestine of type 1 diabetic patients.  

PubMed

Enterovirus infections have been diagnosed more frequently in type 1 diabetic patients than in the healthy population, and enteroviruses have also been found in the pancreas of diabetic patients. Primary replication of the virus occurs in the gut, but there are no previous studies evaluating possible presence of virus in the intestine of diabetic patients. The purpose of this study was to investigate if enteroviruses can be found in small intestinal tissue of type 1 diabetic patients. Formalin-fixed, paraffin-embedded upper intestinal biopsy samples were analysed for the presence of enterovirus using in situ hybridization and immunohistochemistry. Enterovirus was detected by in situ hybridization in six (50%) of the type 1 diabetic patients (n = 12) but in none of the control subjects (n = 10, P = 0.015). Immunohistochemistry identified enterovirus in nine (75%) of the patients and one (10%) control subject (P = 0.004). The presence of the virus was confirmed by reverse transcription-polymerase chain reaction in one of the four patients from whom a frozen and unfixed sample was available. Intestinal morphology was normal in all study subjects. The results suggest that a substantial proportion of type 1 diabetic patients have an ongoing enterovirus infection in gut mucosa, possibly reflecting persistent enterovirus infection. This observation opens new avenues for further studies on the possible role of enteroviruses in human type 1 diabetes. PMID:17991291

Oikarinen, M; Tauriainen, S; Honkanen, T; Oikarinen, S; Vuori, K; Kaukinen, K; Rantala, I; Mäki, M; Hyöty, H

2007-11-07

70

Relationship between numbers of enteroviruses and bacteriophages infecting bacterowes fragilis in different environmental samples  

Microsoft Academic Search

Bacteriophages infecting Bacteroides fragilis HSP40 have been detected in waters and sediments with varying levels of domestic sewage pollution. They were not found in environmental samples without human pollution. Usually they outnumbered enteroviruses by at least a factor of ten. Although there was not any correlation among the values of phages and enteroviruses, there was a significant concurrence of the

C. Tartera; J. Jofre; F. Lucena

1988-01-01

71

Evaluation of human enterovirus 71 and coxsackievirus A16 specific immunoglobulin M antibodies for diagnosis of hand-foot-and-mouth disease  

PubMed Central

Background Hand-foot-and-mouth disease (HFMD) is caused mainly by the human enterovirus type 71 (HEV71) and the Coxsackievirus A group type 16 (CVA16). Large outbreaks of disease have occurred frequently in the Asia-Pacific region. Reliable methods are needed for diagnosis of HFMD in childen. IgM-capture ELISA, with its notable advantages of convenience and low cost, provides a potentially frontline assay. We aimed to evaluate the newly developed IgM-capture ELISAs for HEV71 and CVA16 in the diagnosis of HFMD, and to measure the kinetics of IgM over the course of HEV71 or CVA16 infections. Results We mapped, for the first time, the kinetics of IgM in HEV71 and CVA16 infection. HEV71- and CVA16-IgM were both detectable in some patients on day 1 of illness, and in 100% of patients by day 5 (HEV71) and day 8 (CVA16) respectively; both IgMs persisted for several weeks. The IgM detection rates were 90.2% (138 of 153 sera) and 68.0% (66 of 97 sera) for HEV71 and CVA16 infections, respectively, during the first 7 days of diseases. During the first 90 days after onset these values were 93.6% (233 of 249 sera) and 72.8% (91 of 125 sera) for HEV71 and CVA16 infections, respectively. Some cross-reactivity was observed between HEV71- and CVA16-IgM ELISAs. HEV71-IgM was positive in 38 of 122 (31.1%) CVA16 infections, 14 of 49 (28.6%) other enteroviral infections and 2 of 105 (1.9%) for other respiratory virus infected sera. Similarly, CVA16-IgM was apparently positive in 58 of 211 (27.5%) HEV71 infections, 16 of 48 (33.3%) other enterovirus infections and 3 of 105 (2.9%) other respiratory virus infected sera. Nevertheless, the ELISA yielded the higher OD450 value of main antibody than that of cross-reaction antibody, successfully identifying the enteroviral infection in 96.6% (HEV71) and 91.7% (CVA16) cases. When blood and rectal swabs were collected on the same day, the data showed that the agreement between IgM-capture ELISA and real-time RT-PCR in HEV71 was high (Kappa value = 0.729) while CVA16 somewhat lower (Kappa value = 0.300). Conclusions HEV71- and CVA16-IgM ELISAs can be deployed successfully as a convenient and cost-effective diagnostic tool for HFMD in clinical laboratories.

2012-01-01

72

Human B Lymphotropic Virus.  

National Technical Information Service (NTIS)

The invention is related generally to the isolation and characterization of a new virus. More particularly, it is related to providing a biologically pure, isolated human B lymphotropic virus, molecular clones, nucleic acid, distinctive antigenic proteins...

S. Salahuddin

1988-01-01

73

Detection of human enterovirus 71 and Coxsackievirus A16 in an outbreak of hand, foot, and mouth disease in Henan Province, China in 2009.  

PubMed

During 2009, an outbreak of hand, foot, and mouth disease (HFMD) enrolled 490 people in Henan Province, causing the death of two children. In order to investigate the pathogens responsible for this outbreak and characterize their genetic characteristics, a total of 508 clinical specimens (stool, throat swab, and vesicle fluid) were collected from the Center for Disease Control and Prevention of Henan Province. Virological investigations (virus isolation, conventional reverse transcription PCR, and real-time reverse transcription PCR) and phylogenetic analysis were performed. It was found that human enterovirus 71 (EV71) was the main pathogen causing this outbreak, while Coxsackievirus A16 (CoxA16) played only a subsidiary role. Phylogenetic analysis of 24 EV71 isolates collected during the period from March 11 to July 24, 2009 showed that they belonged to subgenotypes C4 and C5. Our study for the first time characterizes the epidemiology of HFMD and EV71 infection in Henan Province in 2009 and provides the first direct evidence of the genotype of EV71 circulating in Henan Province at that time. Our study should facilitate the development of public health measures for the control and prevention of HFMD and EV71 infection in at-risk individuals in China. PMID:23080402

Fan, Xingliang; Jiang, Jun; Liu, Yanjing; Huang, Xueyong; Wang, Pengzhi; Liu, Licheng; Wang, Junzhi; Chen, Weijun; Wu, Weili; Xu, Bianli

2012-10-19

74

Development of a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) system for a highly sensitive detection of enterovirus in the stool samples of acute flaccid paralysis cases  

PubMed Central

Background In the global eradication program for poliomyelitis, the laboratory diagnosis plays a critical role by isolating poliovirus (PV) from the stool samples of acute flaccid paralysis (AFP) cases. In this study, we developed a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) system for a rapid and highly sensitive detection of enterovirus including PV to identify stool samples positive for enterovirus including PV. Methods A primer set was designed for RT-LAMP to detect enterovirus preferably those with PV-like 5'NTRs of the viral genome. The sensitivity of RT-LAMP system was evaluated with prototype strains of enterovirus. Detection of enterovirus from stool extracts was examined by using RT-LAMP system. Results We detected at least 400 copies of the viral genomes of PV(Sabin) strains within 90 min by RT-LAMP with the primer set. This RT-LAMP system showed a preference for Human enterovirus species C (HEV-C) strains including PV, but exhibited less sensitivity to the prototype strains of HEV-A and HEV-B (detection limits of 7,400 to 28,000 copies). Stool extracts, from which PV, HEV-C, or HEV-A was isolated in the cell culture system, were mostly positive by RT-LAMP method (positive rates of 15/16 (= 94%), 13/14 (= 93%), and 4/4 (= 100%), respectively). The positive rate of this RT-LAMP system for stool extracts from which HEV-B was isolated was lower than that of HEV-C (positive rate of 11/21 (= 52%)). In the stool samples, which were negative for enterovirus isolation by the cell culture system, we found that two samples were positive for RT-LAMP (positive rates of 2/38 (= 5.3%)). In these samples, enterovirus 96 was identified by sequence analysis utilizing a seminested PCR system. Conclusions RT-LAMP system developed in this study showed a high sensitivity comparable to that of the cell culture system for the detection of PV, HEV-A, and HEV-C, but less sensitivity to HEV-B. This RT-LAMP system would be useful for the direct detection of enterovirus from the stool extracts.

2009-01-01

75

Differential display RT-PCR analysis of enterovirus-71-infected rhabdomyosarcoma cells reveals mRNA expression responses of multiple human genes with known and novel functions.  

PubMed

In order to better understand cellular responses to viral infection at the transcriptional level, we employed differential display RT-PCR to analyze mRNAs from RD rhabdomyosarcoma cells following infection with a neurovirulent enterovirus 71 (EV71) strain, compared with mRNAs from uninfected cells. Of 250 expressed sequence tags (ESTs) isolated, sequenced, and identified, all were of cellular origin except 1 that was of viral origin. Of these, 156 were individual distinctive clones, comprising 45 mRNAs showing unaltered expression and 111 mRNAs exhibiting upregulation or downregulation. Of the 45 uniformly expressed mRNAs, 14 represented unknown genes. Of the 111 differentially expressed mRNAs, 63 did not match any known genes. Forty-eight of the 111 mRNAs modified by EV71 infection matched known genes, including those encoding components of cell cycle, cytoskeleton, and cell death mediators; protein degradation mediators; mitochondrial-related proteins; components of protein translation and modification; and cellular transport proteins. The altered expression profiles of representative genes were authenticated by semiquantitative RT-PCR and real-time RT-PCR. We also identified a novel alternatively spliced transcript of TRIP7 thyroid receptor interactor protein; the putative human homolog of murine mc7 mRNA predominantly expressed in the brain; and a novel mRNA similar to that encoding vacuolar protein 8 involved in protein targeting. These results underscore the applicability of the mRNA differential display technique for elucidating the expression profiles of known and even novel genes in response to cellular infection with pathogenic viruses. PMID:12033773

Leong, Peter W F; Liew, Kingsley; Lim, William; Chow, Vincent T K

2002-03-30

76

Evidence for an enterovirus as the cause of encephalitis lethargica  

PubMed Central

Background The epidemic of encephalitis lethargica (EL), called classical EL, was rampant throughout the world during 1917–1926, affecting half a million persons. The acute phase was lethal for many victims. Post-encephalitic parkinsonism (PEP) affected patients for decades. Our purpose was to investigate the cause of classical EL by studying the few available brain specimens. Cases of PEP and modern EL were also studied. Transmission electron microscopy (TEM) and immunohistochemistry were employed to examine brain from four classical EL cases, two modern EL cases and one PEP case. Methods Standard methods for TEM, immunohistochemistry and RTPCR were applied. Results 27?nm virus-like particles (VLP) were observed in the cytoplasm and nuclei of midbrain neurons in all classical EL cases studied. Large (50?nm) VLP and 27?nm intranuclear VLP were observed in the modern EL cases and the PEP case. Influenza virus particles were not found. VLP were not observed in the control cases. TEM of cell cultures inoculated with coxsackievirus B4 and poliovirus revealed both small and large intranuclear virus particles and small cytoplasmic particles, similar to the VLP in EL neurons. In the EL brains, nascent VLP were embedded in putative virus factories and on endoplasmic reticulum (ER). The VLP in the cases of classical EL survived, whereas ribosomes underwent autolysis due to the lack of refrigeration and slow formaldehyde fixation of whole brain. The VLP were larger than ribosomes from well preserved brain. Immunohistochemistry of classical EL cases using anti-poliovirus and anti-coxsackievirus B polyclonal antibodies showed significant staining of cytoplasm and nuclei of neurons as well as microglia and neuropil. Purkinje cells were strongly stained. A 97-bp RNA fragment of a unique virus was isolated from brain tissue from acute EL case #91558. Sequence analysis revealed up to 95% identity to multiple human Enteroviruses. Additional cases had Enterovirus positive reactions by real time PCR. Conclusions The data presented here support the hypothesis that the VLP observed in EL tissue is an Enterovirus.

2012-01-01

77

Coxsackievirus B1-based antibody-capture enzyme-linked immunosorbent assay for detection of immunoglobulin G (IgG), IgM, and IgA with broad specificity for enteroviruses.  

PubMed Central

An antibody-capture enzyme-linked immunosorbent assay (ELISA) with coxsackievirus B1 as the antigen was evaluated for detection of immunoglobulin G (IgG), IgM, and IgA antibodies and showed broad specificity for enteroviruses. In total, 116 serum or cerebrospinal fluid samples from 62 patients were tested by ELISA and the complement fixation test (CFT). Additionally, 15 serum samples that contained poliovirus-specific IgM antibody were tested. Serum samples from 200 healthy blood donors were used for standardization of the assays. The sensitivity of the ELISA varied with time of serum sampling, with a relatively low sensitivity when serum was collected within 3 days after the onset of symptoms (23%; 5 of 22) but good sensitivity when serum was collected later (83%; 20 of 24). The sensitivity was better than that of the CFT. The ELISAs were broadly reactive as concluded from typing of virus isolates that were simultaneously obtained. The assay did, furthermore, detect antibody against poliovirus type 3. Sera that contained rheumatoid factor, antinuclear antibody, or cardiolipin antibody (by the Venereal Disease Research Laboratory test) did not react in this ELISA. Nonspecific reactivity did occur, however, in cases of infectious mononucleosis and in Mycoplasma pneumoniae infection. The enterovirus-specific ELISA is found to be simple to perform, more sensitive than the CFT, and far less laborious than the neutralization test.

Swanink, C M; Veenstra, L; Poort, Y A; Kaan, J A; Galama, J M

1993-01-01

78

Enterovirus 75 Encephalitis in Children, Southern India  

PubMed Central

Recent outbreaks of enterovirus in Southeast Asia emphasize difficulties in diagnosis of this infection. To address this issue, we report 5 (4.7%) children infected with enterovirus 75 among 106 children with acute encephalitis syndrome during 2005–2007 in southern India. Throat swab specimens may be useful for diagnosis of enterovirus 75 infection.

Perera, David; Ooi, Mong How; Last, Anna; Kumar, Ravi; Desai, Anita; Begum, Ashia; Ravi, Vasanthapuram; Shankar, M. Veera; Tio, Phaik Hooi; Cardosa, Mary Jane; Solomon, Tom

2010-01-01

79

Formaldehyde-inactivated human enterovirus 71 vaccine is compatible for co-immunization with a commercial pentavalent vaccine.  

PubMed

In this study we tested the effectiveness of a formaldehyde-inactivated EV71 vaccine and its compatibility for co-immunization with a pentavalent vaccine that contained inactivated poliovirus (PV) vaccine. The inactivated EV71 vaccine (C2 genogroup) elicited an antibody response which broadly neutralized homologous and heterologous genogroups, including B4, C4, and B5. Pups from vaccinated dams were resistant to the EV71 challenge and had a high survival rate and a low tissue viral burden when compared to those from non-vaccinated counterparts. Co-immunization with pentavalent and inactivated EV71 vaccines elicited antibodies against the major components of the pentavalent vaccine including the PV, Bordetella pertussis, Haemophilus influenzae type b, diphtheria toxoid, and tetanus toxoid at the same levels as in mice immunized with pentavalent vaccine alone. Likewise, EV71 neutralizing antibody titers were comparable between EV71-vaccinated mice and mice co-immunized with the two vaccines. These results indicate that formaldehyde-inactivated whole virus EV71 vaccine is feasible for designing multivalent vaccines. PMID:21315698

Chen, Chun-Wei; Lee, Yi-Ping; Wang, Ya-Fang; Yu, Chun-Keung

2011-02-11

80

Detection of Precytopathic Effect of Enteroviruses in Clinical Specimens by Centrifugation-Enhanced Antigen Detection  

Microsoft Academic Search

Rapid enterovirus detection is important for decisions about antibiotic administration and length of hospital stay. The efficacy of rapid antigen detection-cell culture amplification (Ag-CCA) was evaluated with monoclonal antibodies (MAbs) 5-D8\\/1 (DAKO) and Pan-Enterovirus clone 2E11 (Chemicon) with 10 poliovirus, echovirus, and coxsackievirus type A and B stock isolates and College of American Pathologists check samples. By using Ag-CCA technology,

STEVEN M. LIPSON; KATHRYN DAVID; FATIMA SHAIKH; LIAN QIAN

2001-01-01

81

Molecular epidemiology of enterovirus 71 in peninsular Malaysia, 1997–2000  

Microsoft Academic Search

Summary. Human enterovirus 71 (EV71) (genus Enterovirus, family Picornaviridae) has been responsible for sporadic cases and outbreaks of hand-foot-and-mouth disease (HFMD), aseptic meningitis, encephalitis and poliomyelitis-like disease in Europe, the U.S.A., Australia and Asia. Recently, there has been an increase in EV71 activity in the Asia-Pacific region, with many outbreaks of HFMD associated with brainstem encephalitis manifesting as neurogenic pulmonary

L. J. Herrero; C. S. M. Lee; R. J. Hurrelbrink; B. H. Chua; K. B. Chua; P. C. McMinn

2003-01-01

82

World Health Organization International Reference Centre for Enteroviruses.  

National Technical Information Service (NTIS)

Contents: Coordination of and participation in studies on reference reagents; Testing program on enterovirus horse sera; Other enterovirus horse sera for further cooperative testing; Identification and characterization of candidates for new enterovirus ty...

J. L. Melnick

1966-01-01

83

21 CFR 866.3225 - Enterovirus nucleic acid assay.  

Code of Federal Regulations, 2010 CFR

...2009-04-01 2009-04-01 false Enterovirus nucleic acid assay. 866.3225...Serological Reagents § 866.3225 Enterovirus nucleic acid assay. (a) Identification . An enterovirus nucleic acid assay is a device...

2009-04-01

84

21 CFR 866.3225 - Enterovirus nucleic acid assay.  

Code of Federal Regulations, 2010 CFR

...2010-04-01 2010-04-01 false Enterovirus nucleic acid assay. 866.3225...Serological Reagents § 866.3225 Enterovirus nucleic acid assay. (a) Identification . An enterovirus nucleic acid assay is a device...

2010-04-01

85

Enterovirus infections: Diagnosis and treatment  

Microsoft Academic Search

Enteroviruses cause infections that present in diverse ways and affect people of all ages. Infections peak during summer and fall epidemics and cause 10 to 15 million symptomatic infections annually in the United States. The 70 enteroviral serotypes cause illness that ranges from nonspecific fevers and rashes to life-threatening myocarditis or central nervous system disease. These common infections create a

Mark H Sawyer

2002-01-01

86

Coxsackievirus B4 can infect human pancreas ductal cells and persist in ductal-like cell cultures which results in inhibition of Pdx1 expression and disturbed formation of islet-like cell aggregates.  

PubMed

The role of enteroviruses, especially Coxsackievirus B (CVB), in type 1 diabetes is suspected, but the mechanisms of the virus-induced or aggravated pathogenesis of the disease are unknown. The hypothesis of an enterovirus-induced disturbance of pancreatic ?-cells regeneration has been investigated in the human system. The infection of human pancreas ductal cells and pancreatic duct cell line, PANC-1, with CVB4E2 has been studied. Primary ductal cells and PANC-1 cells were infectable with CVB4E2 and a RT-PCR assay without extraction displayed that a larger proportion of cells harbored viral RNA than predicted by the detection of the viral capsid protein VP1 by indirect immunofluorescence. The detection of intracellular positive- and negative-strands of enterovirus genomes in cellular extracts by RT-PCR and the presence of infectious particles in supernatant fluids during the 37 weeks of monitoring demonstrated that CVB4E2 could persist in the pancreatic duct cell line. A persistent infection of these cells resulted in an impaired expression of Pdx1, a transcription factor required for the formation of endocrine pancreas, and a disturbed formation of islet-like cell aggregates of which the viability was decreased. These data support the hypothesis of an impact of enteroviruses onto pancreatic ductal cells which are involved in the renewal of pancreatic ?-cells. PMID:23775130

Sane, Famara; Caloone, Delphine; Gmyr, Valéry; Engelmann, Ilka; Belaich, Sandrine; Kerr-Conte, Julie; Pattou, François; Desailloud, Rachel; Hober, Didier

2013-06-18

87

Development of an enterovirus specific PCR method for the quantification of enterovirus genomes in blood of diabetes patients  

Microsoft Academic Search

Background: Insulin-dependent diabetes mellitus or type 1 diabetes is a disease with a diverse aetiology. Epidemiological studies examining newly diagnosed, recent onset IDDM patients have suggested a role for viruses in the aetiology of IDDM (Yoon, 1995, Diabetes\\/Metabolism Reviews 11, 83–107). Important candidates are the enteroviruses, in particular coxsackieviruses B3 and B4. The latter can cause diabetes in animals (Clements

S Lauwers; V Bissay; B Rombaut

1998-01-01

88

Pathogenesis of type 1 diabetes mellitus: interplay between enterovirus and host.  

PubMed

Enteroviruses are believed to contribute to the pathogenesis of type 1 diabetes mellitus (T1DM). In this Review, the interplay between infection with enteroviruses, the immune system and host genes is discussed. Data from retrospective and prospective epidemiological studies strongly suggest the involvement of enteroviruses, such as coxsackievirus B, in the development of T1DM. Enteroviral RNA and/or proteins can be detected in tissues of patients with T1DM. Isolation of coxsackievirus B4 from the pancreas of patients with T1DM or the presence of enteroviral components in their islets strengthens the hypothesis of a relationship between the virus and the disease. Enteroviruses can play a part in the early phase of T1DM through the infection of beta cells and the activation of innate immunity and inflammation. In contrast with its antiviral role, virus-induced interferon alpha can be deleterious, acting as an initiator of the autoimmunity directed against beta cells. Enteroviruses, through persistent and/or successive infections, can interact with the adaptive immune system. Host genes, such as IFIH1, that influence susceptibility to T1DM are associated with antiviral activities. An increased activity of the IFIH1 protein may promote the development of T1DM. An improved knowledge of the pathogenic mechanisms of enterovirus infections should help to uncover preventive strategies for T1DM. PMID:20351698

Hober, Didier; Sauter, Pierre

2010-03-30

89

Enterovirus 74 Infection in Children  

PubMed Central

Enterovirus 74 (EV74) is a rarely detected viral infection of children. In 2010, EV74 was identified in New Zealand in a 2 year old child with acute flaccid paralysis (AFP) through routine polio AFP surveillance. A further three cases of EV74 were identified in children within six months. These cases are the first report of EV74 in New Zealand. In this study we describe the near complete genome sequence of four EV74 isolates from New Zealand, which shows only limited sequence identity in the non-structural proteins when compared to the other two known EV74 sequences. As is typical of enteroviruses multiple recombination events were evident, particularly in the P2 region and P3 regions. This is the first complete EV74 genome sequenced from a patient with acute flaccid paralysis.

Peacey, Matthew; Hall, Richard J.; Wang, Jing; Todd, Angela K.; Yen, Seiha; Chan-Hyams, Jasmine; Rand, Christy J.; Stanton, Jo-Ann; Huang, Q. Sue

2013-01-01

90

Enterovirus 74 infection in children.  

PubMed

Enterovirus 74 (EV74) is a rarely detected viral infection of children. In 2010, EV74 was identified in New Zealand in a 2 year old child with acute flaccid paralysis (AFP) through routine polio AFP surveillance. A further three cases of EV74 were identified in children within six months. These cases are the first report of EV74 in New Zealand. In this study we describe the near complete genome sequence of four EV74 isolates from New Zealand, which shows only limited sequence identity in the non-structural proteins when compared to the other two known EV74 sequences. As is typical of enteroviruses multiple recombination events were evident, particularly in the P2 region and P3 regions. This is the first complete EV74 genome sequenced from a patient with acute flaccid paralysis. PMID:24098514

Peacey, Matthew; Hall, Richard J; Wang, Jing; Todd, Angela K; Yen, Seiha; Chan-Hyams, Jasmine; Rand, Christy J; Stanton, Jo-Ann; Huang, Q Sue

2013-10-02

91

[Fruit of the emergence of an enterovirus: acute haemorrhagic conjunctivitis].  

PubMed

First seen in Ghana and Indonesia in the early 70's, acute haemorrhagic conjunctivitis or "Apollo 11" disease is an eye infection caused by Enterovirus type 70 (EV70). The disease appeared to be a highly contagious conjunctivitis which spread rapidly all over the world. EV70 has been considered as an emerging virus and was classified as a new Enterovirus. No human or animal virus genetically similar to EV70 was known before the sudden outcome of the disease in Ghana, West Africa. EV70 appeared as a pretty demonstrative example of virus emergence and virus spreading. Studies of virus genetic mutations emphasized the variations of RNA virus within a short time period. The current review presents the EV70 infection and the genetic profile of the virus from its emergence to nowadays. PMID:18957336

Sane, F; Sauter, P; Fronval, S; Goffard, A; Dewilde, A; Hober, D

92

Symmetry-Related Clustering of Positive Charges Is a Common Mechanism for Heparan Sulfate Binding in Enteroviruses  

PubMed Central

Coxsackievirus A9 (CAV9), a member of the Picornaviridae family, uses an RGD motif in the VP1 capsid protein to bind to integrin ?v?6 during cell entry. Here we report that two CAV9 isolates can bind to the heparan sulfate/heparin class of proteoglycans (HSPG). Sequence analysis identified an arginine (R) at position 132 in VP1 in these two isolates, rather than a threonine (T) as seen in the nonbinding strains tested. We introduced a T132R substitution into the HSPG-nonbinding strain Griggs and recovered infectious virus capable of binding to immobilized heparin, unlike the parental Griggs strain. The known CAV9 structure was used to identify the location of VP1 position 132, 5 copies of which were found to cluster around the 5-fold axis of symmetry, presumably producing a region of positive charge which can interact with the negatively charged HSPG. Analysis of several enteroviruses of the same species as CAV9, Human enterovirus B (HEV-B), identified examples from 5 types in which blocking of infection by heparin was coincident with an arginine (or another basic amino acid, lysine) at a position corresponding to 132 in VP1 in CAV9. Together, these data show that membrane-associated HSPG can serve as a (co)receptor for some CAV9 and other HEV-B strains and identify symmetry-related clustering of positive charges as one mechanism by which HSPG binding can be achieved. This is a potentially powerful mechanism by which a single amino acid change could generate novel receptor binding capabilities, underscoring the plasticity of host-cell interactions in enteroviruses.

McLeish, Nigel J.; Williams, Cigdem H.; Kaloudas, Dimitrios; Roivainen, Merja M.

2012-01-01

93

Immunology in the clinic review series; focus on type 1 diabetes and viruses: enterovirus, thymus and type 1 diabetes pathogenesis  

PubMed Central

OTHER THEMES PUBLISHED IN THIS IMMUNOLOGY IN THE CLINIC REVIEW SERIES Metabolic diseases, host responses, cancer, autoinflammatory diseases, allergy. Thymus dysfunction, especially immune suppression, is frequently associated with various virus infections. Whether viruses may disturb the thymus function and play a role in the pathogenesis of autoimmune diseases is an open issue. Enteroviruses, especially Coxsackievirus B4 (CV-B4), have been largely suggested as potential inducers or aggravating factors of type 1 diabetes (T1D) pathogenesis in genetically predisposed individuals. Several pathogenic mechanisms of enterovirus-induced T1D have been suggested. One of these mechanisms is the impairment of central self-tolerance due to viral infections. Coxsackievirus-B4 is able to infect murine thymus in vitro and in vivo and to infect human thymus in vitro. Thymic epithelial cells and thymocytes are targets of infection with this virus, and several abnormalities, especially disturbance of maturation/differentiation processes, were observed. Altogether, these data suggest that CV-B infection of thymus may be involved in the pathogenesis of T1D. Further investigations are needed to explore this hypothesis.

Jaidane, H; Sane, F; Hiar, R; Goffard, A; Gharbi, J; Geenen, V; Hober, D

2012-01-01

94

Immunology in the clinic review series; focus on type 1 diabetes and viruses: enterovirus, thymus and type 1 diabetes pathogenesis.  

PubMed

Thymus dysfunction, especially immune suppression, is frequently associated with various virus infections. Whether viruses may disturb the thymus function and play a role in the pathogenesis of autoimmune diseases is an open issue. Enteroviruses, especially Coxsackievirus B4 (CV-B4), have been largely suggested as potential inducers or aggravating factors of type 1 diabetes (T1D) pathogenesis in genetically predisposed individuals. Several pathogenic mechanisms of enterovirus-induced T1D have been suggested. One of these mechanisms is the impairment of central self-tolerance due to viral infections. Coxsackievirus-B4 is able to infect murine thymus in vitro and in vivo and to infect human thymus in vitro. Thymic epithelial cells and thymocytes are targets of infection with this virus, and several abnormalities, especially disturbance of maturation/differentiation processes, were observed. Altogether, these data suggest that CV-B infection of thymus may be involved in the pathogenesis of T1D. Further investigations are needed to explore this hypothesis. PMID:22385235

Jaïdane, H; Sané, F; Hiar, R; Goffard, A; Gharbi, J; Geenen, V; Hober, D

2012-04-01

95

A novel enterovirus and parechovirus multiplex one-step real-time PCR-validation and clinical experience.  

PubMed

As the number of new enteroviruses and human parechoviruses seems ever growing, the necessity for updated diagnostics is relevant. We have updated an enterovirus assay and combined it with a previously published assay for human parechovirus resulting in a multiplex one-step RT-PCR assay. The multiplex assay was validated by analysing the sensitivity and specificity of the assay compared to the respective monoplex assays, and a good concordance was found. Furthermore, the enterovirus assay was able to detect 42 reference strains from all 4 species, and an additional 9 genotypes during panel testing and routine usage. During 15 months of routine use, from October 2008 to December 2009, we received and analysed 2187 samples (stool samples, cerebrospinal fluids, blood samples, respiratory samples and autopsy samples) were tested, from 1546 patients and detected enteroviruses and parechoviruses in 171 (8%) and 66 (3%) of the samples, respectively. 180 of the positive samples could be genotyped by PCR and sequencing and the most common genotypes found were human parechovirus type 3, echovirus 9, enterovirus 71, Coxsackievirus A16, and echovirus 25. During 2009 in Denmark, both enterovirus and human parechovirus type 3 had a similar seasonal pattern with a peak during the summer and autumn. Human parechovirus type 3 was almost invariably found in children less than 4 months of age. In conclusion, a multiplex assay was developed allowing simultaneous detection of 2 viruses, which can cause similar clinical symptoms. PMID:23845901

Nielsen, Alex Christian Yde; Böttiger, Blenda; Midgley, Sofie Elisabeth; Nielsen, Lars Peter

2013-07-08

96

High Susceptibility for Enterovirus Infection and Virus Excretion Features in Tunisian Patients with Primary Immunodeficiencies  

PubMed Central

To estimate the susceptibility to enterovirus infection and the frequency of long-term poliovirus excreters in Tunisian patients with primary immunodeficiencies (PIDs), enteroviruses were assessed in stool specimens of 82 patients with humoral, combined, and other PIDs. Isolated viruses were typed and intratyped by standard molecular techniques, and the whole VP1 region of poliovirus isolates was sequenced. Polioviruses were detected in 6 patients; all isolates were vaccine related. Five patients rapidly stopped excretion; one excreted a poliovirus type 1 isolate for several months, and the isolate accumulated up to 14 mutations in the VP1 region. Nonpolio enteroviruses were identified in 6 patients; 4 of them kept excreting the same strain for more than 6 months. The rate of enterovirus infection was 13.4% of the PID patients and 20.7% of those with an IgG defect; it greatly exceeded the rates generally found in Tunisian supposed-immunocompetent individuals (4.1% during the study period; P = 0.001 and P < 0.0001, respectively). Interestingly, patients with combined immunodeficiencies were at a higher risk for enterovirus infection than those with an exclusively B cell defect. A major histocompatibility complex (MHC) class II antigen expression defect was found in 54% of enterovirus-positive patients and in the unique long-term poliovirus excreter. The study results also suggest that substitutive immunoglobulin therapy may help clearance of a poliovirus infection and that most PID patients have the ability to stop poliovirus excretion within a limited period. However, the high susceptibility of these patients to enterovirus infection reinforces the need for enhanced surveillance of these patients until the use of oral poliovirus vaccine (OPV) is stopped.

Driss, Nadia; Ben-Mustapha, Imen; Mellouli, Fethi; Ben Yahia, Ahlem; Touzi, Henda; Bejaoui, Mohamed; Ben Ghorbel, Mohamed; Barbouche, Mohamed-Ridha

2012-01-01

97

Transgenic mouse model for the study of enterovirus 71 neuropathogenesis.  

PubMed

Enterovirus 71 (EV71) typically causes mild hand-foot-and-mouth disease in children, but it can also cause severe neurological disease. Recently, epidemic outbreaks of EV71 with significant mortality have been reported in the Asia-Pacific region, and EV71 infection has become a serious public health concern worldwide. However, there is little information available concerning EV71 neuropathogenesis, and no vaccines or anti-EV71 drugs have been developed. Previous studies of this disease have used monkeys and neonatal mice that are susceptible to some EV71 strains as models. The monkey model is problematic for ethical and economical reasons, and mice that are more than a few weeks old lose their susceptibility to EV71. Thus, the development of an appropriate small animal model would greatly contribute to the study of this disease. Mice lack EV71 susceptibility due to the absence of a receptor for this virus. Previously, we identified the human scavenger receptor class B, member 2 (hSCARB2) as a cellular receptor for EV71. In the current study, we generated a transgenic (Tg) mouse expressing hSCARB2 with an expression profile similar to that in humans. Tg mice infected with EV71 exhibited ataxia, paralysis, and death. The most severely affected cells were neurons in the spinal cord, brainstem, cerebellum, hypothalamus, thalamus, and cerebrum. The pathological features in these Tg mice were generally similar to those of EV71 encephalomyelitis in humans and experimentally infected monkeys. These results suggest that this Tg mouse could represent a useful animal model for the study of EV71 infection. PMID:23959904

Fujii, Ken; Nagata, Noriyo; Sato, Yuko; Ong, Kien Chai; Wong, Kum Thong; Yamayoshi, Seiya; Shimanuki, Midori; Shitara, Hiroshi; Taya, Choji; Koike, Satoshi

2013-08-19

98

Production of Enterovirus Antigens for Human Enterovirus Types.  

National Technical Information Service (NTIS)

The program has a three-fold purpose: (1) to produce reference seeds for Echo viruses 9, 13, 15, 21, 24, 25, 26, 27, 29, 30 and 31 and Coxsackieviruses A-20A, 2, 3, 8, 10, 11, 12, 13 and 16; (2) to produce seed virus and antiserum in rabbits to specific s...

S. S. Kalter

1965-01-01

99

Environmental surveillance of enterovirus in Northern India using an integrated shell vial culture with a semi-nested RT PCR and partial sequencing of the VP1 gene.  

PubMed

Enteroviruses have been reported in epidemic form during last 10 years in northern India. Environmental surveillance of sewage is the method of choice in limited resources countries for detection of enterovirus serotypes circulating in the community. Twenty-four sewage samples collected between January, 2009 and December, 2010 were tested for enterovirus by using a new modified integrated shell vial culture (ISVC) with a semi-nested RT-PCR of a partial VP1 gene and virus isolation integrated with semi-nested RT-PCR of a partial VP1 gene. Twenty-one (87.5%) out of 24 samples were positive for enterovirus by the conventional method and all samples (100%) by the ISVC-RT-PCR. The additional positive samples detected by ISVC-RT-PCR was typed as six different enterovirus serotypes (Sabin poliovirus 3, Coxsackievirus B3, Coxsackievirus A13, Coxsackievirus A17, Echovirus 33, and Enterovirus 75). Phylogenetic analysis of a partial VP1 gene of Echovirus 19 showed that one genetic lineage clustered with isolates from Georgia suggesting their importation into northern India. Detection of wild poliovirus in the absence of clinical cases with 16 different co-circulating enterovirus serotypes supports the need of increased molecular surveillance of sewage. Rapid identification and characterization of enterovirus serotypes is necessary to study their transmission and evolution in different geographical regions to prevent future outbreak. PMID:23341372

Shukla, Deepti; Kumar, Arvind; Srivastava, Shalini; Idris, Mohammad Z; Dhole, Tapan N

2013-03-01

100

Molecular epidemiology of enterovirus 71 in Taiwan  

Microsoft Academic Search

Summary.  ?Taiwan suffered a severe and widespread outbreak of enterovirus infection in 1998. More than 400 children were hospitalized,\\u000a with seventy-eight fatalities due to central nerve system (CNS) involvement and cardiopulmonary collapse. Enterovirus 71 (EV71)\\u000a was incriminated as the causative agent for the fatal cases. To understand the viral molecular epidemiology in this outbreak,\\u000a fragments of 207-bp length of the VP4

P.-Y. Chu; K.-H. Lin; K.-P. Hwang; L.-C. Chou; C.-F. Wang; S.-R. Shih; J.-R. Wang; Y. Shimada; H. Ishiko

2001-01-01

101

Fatal Case of Enterovirus 71 Infection, France, 2007  

PubMed Central

A fatal case of enterovirus 71 infection with pulmonary edema and rhombencephalitis occurred in Brest, France, in April 2007. The virus was identified as subgenogroup C2. This highly neurotropic enterovirus merits specific surveillance outside the Asia-Pacific region.

Legrand-Quillien, Marie-Christine; Dailland, Thomas; Podeur, Gaetan; Gouriou, Stephanie; Schuffenecker, Isabelle; Payan, Christopher; Marcorelles, Pascale

2009-01-01

102

Serotype-specific identification of enterovirus 71 by PCR  

Microsoft Academic Search

Background: Enterovirus 71 and coxsackievirus A16 are closely related genetically and are causative agents of hand foot and mouth disease. Because enterovirus 71 is more often associated with severe neurological disease, there is a need to rapidly discriminate between enterovirus 71 and coxsackievirus A16 during hand, foot, and mouth disease outbreaks. Objectives: Our goal was to develop and evaluate a

Betty A Brown; David R Kilpatrick; M. Steven Oberste; Mark A Pallansch

2000-01-01

103

Multicenter Quality Assessment of PCR Methods for Detection of Enteroviruses  

Microsoft Academic Search

We conducted a multicenter evaluation of commercial and in-house PCR methods for the detection of enteroviruses. Three coded panels of test and control RNA samples, artificial clinical specimens, and repre- sentative enterovirus serotypes were used to assess amplification methods, RNA extraction methods, and reactivities with different enterovirus serotypes. Despite several differences between PCR methods, there was good agreement, although some

PETER MUIR; ALBERT RAS; PAUL E. KLAPPER; GRAHAM M. CLEATOR; KLAUS KORN; CHRISTIAN AEPINUS; ANDERS FOMSGAARD; PIERRE PALMER; AGNETA SAMUELSSON; ANTONIO TENORIO; BENEDIKT WEISSBRICH; A. M. VAN LOON

1999-01-01

104

Evaluation of methods using celite to concentrate norovirus, adenovirus and enterovirus from wastewater  

EPA Science Inventory

Enteroviruses, noroviruses and adenoviruses are among the most common viruses infecting humans worldwide. These viruses are shed in the feces of infected individuals and can accumulate in wastewater. Therefore, wastewater is a source of a potentially diverse group of enteric viru...

105

Development and assay of RNA transcripts of enterovirus species A to D, rhinovirus species a to C, and human parechovirus: assessment of assay sensitivity and specificity of real-time screening and typing methods.  

PubMed

Nucleic acid amplification methods such as the PCR have had a major impact on the diagnosis of viral infections, often achieving greater sensitivities and shorter turnaround times than conventional assays and an ability to detect viruses refractory to conventional isolation methods. Their effectiveness is, however, significantly influenced by assay target sequence variability due to natural diversity and rapid sequence changes in viruses that prevent effective binding of primers and probes. This was investigated for a diverse range of enteroviruses (EVs; species A to D), human rhinoviruses (HRVs; species A to C), and human parechovirus (HPeV) in a multicenter assay evaluation using a series of full-length prequantified RNA transcripts. RNA concentrations were quantified by absorption (NanoDrop) and fluorescence methods (RiboGreen) prior to dilution in buffer supplemented with RNase inhibitors and carrier RNA. RNA transcripts were extremely stable, showing minimal degradation after prolonged storage at temperatures between ambient and -20°C and after multiple freeze-thaw cycles. Transcript dilutions distributed to six referral laboratories were screened by real-time reverse transcriptase PCR assays using different primers and probes. All of the laboratories reported high assay sensitivities for EV and HPeV transcripts approaching single copies and similar amplification kinetics for all four EV species. HRV detection sensitivities were more variable, often with substantially impaired detection of HRV species C. This could be accounted for in part by the placement of primers and probes to genetically variable target regions. Transcripts developed in this study provide reagents for the ongoing development of effective diagnostics that accommodate increasing knowledge of genetic heterogeneity of diagnostic targets. PMID:22740708

McLeish, Nigel J; Witteveldt, Jeroen; Clasper, Lucy; McIntyre, Chloe; McWilliam Leitch, E Carol; Hardie, Alison; Bennett, Susan; Gunson, Rory; Carman, William F; Feeney, Susan A; Coyle, Peter V; Vipond, Barry; Muir, Peter; Benschop, Kimberley; Wolthers, Katja; Waris, Matti; Osterback, Riikka; Johannessen, Ingo; Templeton, Kate; Harvala, Heli; Simmonds, Peter

2012-06-27

106

Enteroviruses and type 1 diabetes: towards a better understanding of the relationship.  

PubMed

Environmental factors, especially viruses, are involved in the initiation or the acceleration of type 1 diabetes (T1D) pathogenesis. Epidemiological data strongly suggest that enteroviruses, such as coxsackievirus B4 (CV-B4), can be associated with T1D. It has been demonstrated that enterovirus infections were significantly more prevalent in at risk individuals, such as siblings of diabetic patients, when they developed anti-beta-cell autoantibodies or T1D, and in recently diagnosed diabetic patients, compared with control subjects. The isolation of CV-B4 from the pancreas of diabetic patients strengthened the hypothesis of a relationship between the virus and the disease. Studies performed in vitro and in vivo in animal models helped to discover mechanisms of the infection of pancreas and other tissues, potentially able to play a role in the pathogenesis of T1D. Interestingly, it cannot be excluded that enteroviruses behave as half-devil half-angel since experimental studies suggest that, in certain conditions, these agents would be able to protect individuals against the disease. All of the plausible mechanisms by which enterovirus may be related to T1D will be reviewed here. PMID:20629044

Jaïdane, Hela; Sauter, Pierre; Sane, Famara; Goffard, Anne; Gharbi, Jawhar; Hober, Didier

2010-09-01

107

Management of Atrial Tachycardia in the Newborn With Enterovirus Myocarditis  

PubMed Central

Neonatal enterovirus myocarditis is a rare but serious infection that is often an underrecognized cause of cardiovascular collapse. Enterovirus myocarditis in patients with such collapse should be suspected when signs of congestive heart failure and tachyarrhythmia are present. The majority of reported electrical disturbances associated with enterovirus myocarditis are ventricular in origin, but the infection can present as atrial tachyarrhythmia. Atrial tachyarrhythmias associated with enterovirus myocarditis are difficult to manage because of their resistance to conventional antiarrhythmic therapy. We present 2 cases of neonates with atrial tachycardia associated with enterovirus myocarditis who responded to a combination of amiodarone and flecainide.

Petroni, Daniel H.; Yang, Song G.; Kattash, Mudar M.; Snyder, Christopher S.

2012-01-01

108

The structure of coxsackievirus B3 at 3.5 å resolution  

Microsoft Academic Search

Background: Group B coxsackieviruses (CVBs) are etiologic agents of a number of human diseases that range in severity from asymptomatic to lethal infections. They are small, single-stranded RNA icosahedral viruses that belong to the enterovirus genus of the picornavirus family. Structural studies were initiated in light of the information available on the cellular receptors for this virus and to assist

Jodi K Muckelbauer; Marcia Kremer; Iwona Minor; Guy Diana; Frank J Dutko; James Groarke; Daniel C Pevear; Michael G Rossmann

1995-01-01

109

B-1B Human Factors Baseline Study Report.  

National Technical Information Service (NTIS)

A Human Factors study was conducted on B-1B Blocks D, E, and F. The 1st study objective was to evaluate B-1B Blocks D, E, and F crew workload and situational awareness. The 2nd objective was to establish a measurable crew performance baseline for future B...

W. G. Kalman J. M. Kline S. Provost B. A. Gable C. R. Taylor

1999-01-01

110

Optimization of a Combined Human Parechovirus-Enterovirus Real-Time Reverse Transcription-PCR Assay and Evaluation of a New Parechovirus 3-Specific Assay for Cerebrospinal Fluid Specimen Testing  

PubMed Central

Human parechoviruses (HPeVs), particularly type 3 (HPeV3), are known central nervous system (CNS) pathogens, causing serious infections in infants similar to those caused by enteroviruses (EVs). The primary aim of this study was to combine and validate HPeV and EV real-time reverse transcription-PCR (RT-PCR) detection assays with the best available RT-PCR reagents and conditions for parallel detection of HPeV and EV on a single platform. The secondary aim was to develop and validate a newly developed HPeV3-specific real-time RT-PCR assay. Five commercially available RT-PCR kits were evaluated with the pan-HPeV and EV assays in one-step and two-step RT-PCRs. Two-step RT-PCR with the AgPath ID RT-PCR (AGP) kit performed best for both pan-HPeV and EV assays. The pan-HPeV-specific assay performed best with the AGP kit in a one-step RT-PCR. Frozen aliquots of 145 (for HPeV, n = 70; for EV, n = 75) previously characterized cerebrospinal fluid (CSF) specimens were tested by EV-, pan-HPeV-, and HPeV3-specific (HPeV specimens only) assays. The pan-HPeV and EV assays demonstrated 100% analytical sensitivity and specificity compared to historic results, while the HPeV3-specific assay demonstrated 97% sensitivity and 100% specificity. We propose a real-time pan-HPeV, EV two-step RT-PCR algorithm for simultaneous detection of HPeV and EV from CSF specimens on a single platform. The HPeV3-specific one-step RT-PCR assay can be used as a rapid and cost-effective assay to detect and identify HPeV3 in pan-HPeV RT-PCR assay-positive CSF specimens.

Selvaraju, Suresh B.; Nix, W. Allan; Oberste, M. Steven

2013-01-01

111

Enterovirus markers and serum CXCL10 in children with type 1 diabetes.  

PubMed

Most patients with type 1 diabetes are considered to have a T-cell mediated autoimmune disease. The chemokine CXCL10 promotes the migration of activated T-cells. Virus infections might contribute to the pathogenesis of type 1 diabetes and enterovirus protein and/or genome have been detected in beta-cells from a majority of tested newly diagnosed children with type 1 diabetes. The chemokine CXCL10 is induced in human islet cells by enterovirus infections in vivo and in vitro, but is not expressed in islets from normal organ donors. Since CXCL10 is a chemokine known to be induced by virus infections and/or cellular damage, our aim was to study if levels of CXCL10 are elevated in serum from children with type 1 diabetes and whether it correlates to the presence of enterovirus markers. CXCL10, neutralizing antibody titer rises against certain enterovirus, and antibodies against GAD65 were measured in serum, and enterovirus PCR was performed on whole blood from 83 type 1 diabetes patients at onset, 48 siblings and 69 controls. CXCL10 was also measured in serum from 46 patients with proven enterovirus infection and in serum from 46 patients with other proven virus infections. The CXCL10 serum levels were not elevated in children at onset of type 1 diabetes and there was a considerable overlap between the groups with 99 (8-498) pg/ml in serum from children with type 1 diabetes, 120 (17-538) pg/ml in serum from controls, and 117 (7-448) pg/ml in siblings of the children with type 1 diabetes. The CXCL10 serum levels in patients with proven enterovirus infection were slightly increased compared to the levels in the other groups, 172 (0-585) pg/ml but there was no statistically significant difference. In contrast, CXCL10 serum levels in patients with other proven virus infections were clearly elevated 418 (34-611) pg/ml. Despite that elevated CXCL10 levels have been demonstrated in some groups of patients with type 1 diabetes, in this study the mean CXCL10 serum levels were not elevated in patients with type 1 diabetes neither in patients with proven enterovirus infection. In contrast, in patients with other virus infections the CXCL10 levels were elevated, presumably reflecting the severity or the site of infection. This suggests that local production of CXCL10 in the affected organ cannot be measured reproducible in serum and that its potential use in clinical practice is limited. PMID:20648615

Berg, Anna-Karin; Tuvemo, Torsten; Frisk, Gun

2010-09-01

112

Enteroviruses and Bacteriophages in Bathing Waters  

Microsoft Academic Search

A new procedure for detecting and counting enteroviruses based on the VIRADEN method applied to 10 liters of seawater was examined. It improved the efficiency of detection by taking into account both the number of positive isolations and numbers found with traditional methods. It was then used to quantify viruses in bathing waters. A number of bacterial indicators and bacteriophages

L. Moce-Llivina; Francisco Lucena; Juan Jofre

2005-01-01

113

Enterovirus 71 Outbreaks, Taiwan: Occurrence and Recognition  

Microsoft Academic Search

Enterovirus 71 (EV71) caused a large outbreak in Taiwan in 1998 with 78 deaths, and smaller outbreaks recurred in 2000 and 2001. The outbreak was recognized because of a large number of hand, foot, and mouth disease cases and the rapid deaths of children with the disease. Virologic and pathologic studies indicated that EV71 was the most important agent relat-

Tzou-Yien Lin; Shiing-Jer Twu; Mei-Shang Ho; Luan-Yin Chang; Chin-Yun Lee

2003-01-01

114

Detection of adenoviruses and enteroviruses in polluted waters by nested PCR amplification.  

PubMed Central

A procedure has been developed for the rapid detection of enteroviruses and adenoviruses in environmental samples. Several systems for virus concentration and extraction of nucleic acid were tested by adding adenovirus type 2 and poliovirus type 1 to different sewage samples. The most promising method for virus recovery involved the concentration of viruses by centrifugation and elution of the virus pellets by treatment with 0.25 N glycine buffer, pH 9.5. Nucleic acid extraction by adsorption of RNA and DNA to silica particles was the most efficient. One aliquot of the extracted nucleic acids was used for a nested two-step PCR, with specific primers for all adenoviruses; and another aliquot was used to synthesize cDNA for a nested two-step PCR with specific primers for further detection of seeded polioviruses or all enteroviruses in the river water and sewage samples. The specificity and sensitivity were evaluated, and 24 different enterovirus strains and the 47 human adenovirus serotypes were recognized by the primers used. The sensitivity was estimated to be between 1 and 10 virus particles for each of the species tested. Twenty-five samples of sewage and polluted river water were analyzed and showed a much higher number of positive isolates by nested PCR than by tissue culture analysis. The PCR-based detection of enteroviruses and adenoviruses shows good results as an indicator of possible viral contamination in environmental wastewater. Images

Puig, M; Jofre, J; Lucena, F; Allard, A; Wadell, G; Girones, R

1994-01-01

115

Enteroviruses in water environment--a potential threat to public health.  

PubMed

Enteroviruses belong to the Picornaviridae family and are the smallest, nonenveloped viruses known to infect both humans and animals. The spread of enteroviral infections is mainly by the faecal-oral and oral-oral route, but also through direct contact with secretions from ophthalmic and dermal lesions. Water, food and soil contaminated by infected faeces are an exogenous infection source which creates many opportunities for the transfer of the infection, and cause an epidemic outbreak in a short period of time. Enteroviruses are being isolated from all types of water: ground, sea, sewage and fresh water environments but also--and what is the most important from the epidemiological point of view--drinking water. They are resilient organisms, able to withstand high concentrations of sodium chloride (NaCl) and large changes in temperature. These abilities allow the viruses to flourish in a water environment, their natural reservoir. The number of infections in temperate climate regions peak in summer months and early autumn. Detection of enteroviruses in the water environment is performed by virus isolation in cell cultures and the use of molecular techniques. Many researches conducted in different countries with the use of methods mentioned above, reveal widespread environmental contamination by enteroviruses. PMID:19061255

Rajtar, Barbara; Majek, Magdalena; Pola?ski, ?ukasz; Polz-Dacewicz, Ma?gorzata

2008-12-01

116

Presence of enteroviruses in recreational water in Wuhan, China.  

PubMed

Contaminated recreational waters pose a public health concern, as the potential for waterborne diseases exists in water contaminated with human fecal waste. Worldwide, bacterial indicators such as Escherichia coli, enterococci, and total and fecal coliform are used as indicators of water quality. However, enteric viruses also present a public health concern and their presence cannot always be determined based on bacterial indicators. This study explores the use of molecular detection methods of enteric viruses as indicators of fecal contamination. Four viruses, enterovirus, norovirus genogroups I and II, and male-specific FRNA coliphage, were tested in this study. Highly sensitive RT-PCR methods developed at the University of Hawaii at Manoa were utilized to evaluate environmental samples collected from three lakes in Wuhan, Hubei Province, China. Sixteen of twenty-five sites tested positive for at least one virus. Enterovirus was the most commonly detected virus, followed by norovirus genogroup I. These findings support the use of molecular detection methods to test for enteric virus presence in recreational freshwater sources in China as alternative water quality indicators, and utilize recently developed, highly sensitive methods of detection of these viruses. In addition, these findings suggest that there is substantial fecal contamination of the three lakes tested in this study. PMID:23827950

Allmann, Erin; Pan, Lei; Li, Lu; Li, Dejia; Wang, Suqing; Lu, Yuanan

2013-07-01

117

Characterization of Full-Length Enterovirus 71 Strains from Severe and Mild Disease Patients in Northeastern China  

PubMed Central

Human enterovirus 71 (EV71)-associated hand, foot, and mouth disease (HFMD) has been a leading cause of childhood infection in China since 2008. Epidemic and molecular characteristics of HFMD have been examined in many areas of China, including the central and southern regions. However, clinical and genetic characterization of EV71 in the northeastern region of China is scarce. In this study, a series of analyses were performed on seven full-length EV71 sequences from HFMD patients who had either severe or mild disease. We have determined that these seven circulating EV71 viruses from Changchun, China are actually complex recombinant viruses involving multiple type A human enterovirus (HEV). Classified as EV71 subtype C4 (EV71 C4), these Changchun EV71 viruses contain genetic recombination events between the CA4, CA5, EV71B4 and EV71C1 strains. Most of the structural protein region (P1) of these viruses resembled that of the prototype EV71 C1 strains. The non-structural protein domains (P2 and P3) showed a high degree of similarity with CA4, CA5 and EV71 B4 in different regions. The 5?UTR had unclassified recombination,while partial 3D region of these viruses showed a high degree of similarity to CA16. Phylogenetic analysis of full-length or partial sequences of isolates from severe or mild disease patients in Changchun always formed a single cluster in various phylogenetic analyses of different genomic regions, suggesting that all seven strains originated from one single common ancestor. There was no correlation between viral genomic sequence and virulence. Thus, we found that circulating recombinant forms of EV71 are prevalent among HFMD patients in Northeastern China. The existence of a unique cluster of EV71 related viruses in Northeast China has important implications for vaccine development that would address the increasing prevalence of HFMD.

Bao, Wanguo; Zhao, Ke; Niu, Junqi; Yu, Xiao-Fang; Zhang, Wenyan

2012-01-01

118

Serologic Evidence of an Association between Enteroviruses and the Onset of Type 1 Diabetes Mellitus  

Microsoft Academic Search

Serum was collected from 128 patients ?18 years of age admitted to the Children's Hospital of Pittsburgh with new-onset insulin-dependent diabetes mellitus (IDDM) and from 120 control-patients who were frequency-matched to case-patients for age, sex, and date of bleed. Serum was tested for IgM against 14 enterovirus serotypes: coxsackieviruses B1-B6 and A9, echoviruses 4, 6, 9, 11, 30, and 34,

Rita F. Helfand; Howard E. Gary Jr.; Charlotte Y. Freeman; Larry J. Anderson; Mark A. Pallansch

1995-01-01

119

Passive protection effect of chicken egg yolk immunoglobulins on enterovirus 71 infected mice  

Microsoft Academic Search

The objective of this study is to evaluate the passive protective efficiency of immunoglobulin in yolk (IgY) specific against human enterovirus type 71 (EV71). The antibody was raised by intramuscular immunization to 10 White Leghorn hens, with inactivated human EV71 serving as the antigen. The titer and specificity of the antibody were analyzed from purified IgY in the egg yolks

Jenn-Fa Liou; Chih-Wei Chang; Jui-jane Tailiu; Chun-Keung Yu; Huan-Yao Lei; Lih-Ren Chen; Chein Tai

2010-01-01

120

[Outbreak of acute enterovirus intestinal infection in Sakhalin region in August 2010].  

PubMed

The investigation of cases of acute intestinal infections in the Sakhalin region of Russia in August, 2010 is described. Epidemiological and molecular biological studies were conducted. After initial PCR screening and determining the nucleotide sequences of the positive samples the following enteroviruses were found: Coxsackie A2 - 42 samples (45%), Coxsackie A4--31 sample (34%), Enterovirus 71--6 samples (6,5%), Coxsackievirus B5--6 samples (6,5%), Coxsackie B3--4 samples (4%) and Coxsackie B1--4 samples (4%). The phylogenetic analysis of sequences showed that the closest analogues for the nucleotide sequences of these genotypes were previously identified in Japan, Korea and China in 2000-2010. PMID:22642180

Demina, A V; Ternovo?, V A; Darizhapov, B B; Iakubich, T V; Sementsova, A O; Demina, O K; Protopopova, E V; Loktev, V B; Agafonov, A P; Netesov, S V

2012-01-01

121

Quantitative real-time RT-PCR assay for research studies on enterovirus infections in the central nervous system.  

PubMed

Human enteroviruses are the most frequent cause of aseptic meningitis and are involved in other neurological infections. Qualitative detection of enterovirus genomes in cerebrospinal fluid is a prerequisite in diagnosing neurological diseases. The pathogenesis of these infections is not well understood and research in this domain would benefit from the availability of a quantitative technique to determine viral load in clinical specimens. This study describes the development of a real-time RT-qPCR assay using hydrolysis TaqMan probe and a competitive RNA internal control. The assay has high specificity and can be used for a large sample of distinct enterovirus strains and serotypes. The reproducible limit of detection was estimated at 1875 copies/ml of quantitative standards composed of RNA transcripts obtained from a cloned echovirus 30 genome. Technical performance was unaffected by the introduction of a competitive RNA internal control before RNA extraction. The mean enterovirus RNA concentration in an evaluation series of 15 archived cerebrospinal fluid specimens was determined at 4.78 log(10)copies/ml for the overall sample. The sensitivity and reproducibility of the real time RT-qPCR assay used in combination with the internal control to monitor the overall specimen process make it a valuable tool with applied research into enterovirus infections. PMID:22766179

Volle, Romain; Nourrisson, Céline; Mirand, Audrey; Regagnon, Christel; Chambon, Martine; Henquell, Cécile; Bailly, Jean-Luc; Peigue-Lafeuille, Hélène; Archimbaud, Christine

2012-07-02

122

Enterovirus infection and type 1 diabetes mellitus: systematic review and meta-analysis of observational molecular studies  

PubMed Central

Objective To review the association between current enterovirus infection diagnosed with molecular testing and development of autoimmunity or type 1 diabetes. Design Systematic review and meta-analysis of observational studies, analysed with random effects models. Data sources PubMed (until May 2010) and Embase (until May 2010), no language restrictions, studies in humans only; reference lists of identified articles; and contact with authors. Study eligibility criteria Cohort or case-control studies measuring enterovirus RNA or viral protein in blood, stool, or tissue of patients with pre-diabetes and diabetes, with adequate data to calculate an odds ratio and 95% confidence intervals. Results The 24 papers and two abstracts (all case-control studies) that met the eligibility criteria included 4448 participants. Study design varied greatly, with a high level of statistical heterogeneity. The two separate outcomes were diabetes related autoimmunity or type 1 diabetes. Meta-analysis showed a significant association between enterovirus infection and type 1 diabetes related autoimmunity (odds ratio 3.7, 95% confidence interval 2.1 to 6.8; heterogeneity ?2/df=1.3) and clinical type 1 diabetes (9.8, 5.5 to 17.4; ?2/df=3.2). Conclusions There is a clinically significant association between enterovirus infection, detected with molecular methods, and autoimmunity/type 1 diabetes. Larger prospective studies would be needed to establish a clear temporal relation between enterovirus infection and the development of autoimmunity and type 1 diabetes.

2011-01-01

123

Les Enterovirus responsables de conjonctivite aiguë hémorragique  

Microsoft Academic Search

Acute hemorrhagic conjunctivitis (AHC) is an epidemic form of highly contagious conjunctivitis, characterized by conjunctival hemorrhages. The first AHC outbreak was described in 1969 in Ghana, West Africa, and was called Apollo disease, from the Apollo landing on the moon. This outbreak was caused by Enterovirus 70 (EV70) together with a Coxsackievirus A24 (CVA24v) variant, which are the major etiological agents

N. Lévêque; P. Huguet; H. Norder; J.-J. Chomel

2010-01-01

124

Identification and characterization of a cross-neutralization epitope of Enterovirus 71  

Microsoft Academic Search

Enterovirus 71 (EV71) infections in children manifest as exanthema and are most commonly known as hand-foot-and-mouth disease (HFMD). Because it can cause severe neurological complications like poliomyelitis, EV71 has now emerged as an important neurotropic virus in Asia. EV71 virus has been shown to consist of 3 (A, B and C) genotypes and many subgenotypes. Although EV71 vaccine development has

Chia-Chyi Liu; Ai-Hsiang Chou; Shu-Pei Lien; Hsiao-Yu Lin; Shih-Jen Liu; Jui-Yuan Chang; Meng-Shin Guo; Yen-Hung Chow; Wun-Syue Yang; Kate Hsuen-Wen Chang; Charles Sia; Pele Chong

2011-01-01

125

Enterovirus infections and insulin dependent diabetes mellitus—evidence for causality  

Microsoft Academic Search

Background: Insulin-dependent diabetes mellitus (IDDM) has a long subclinical period characterised by gradually progressing autoimmune damage of insulin producing beta-cells. Clinical IDDM is manifested when 90% of beta-cells have been destroyed. Several studies have indicated that enterovirus infections, coxsackievirus B (CVB) infections especially, are frequent at the manifestation of clinical IDDM suggesting that they can precipitate the symptoms of IDDM

Heikki Hyöty; Merja Hiltunen; Maria Lönnrot

1998-01-01

126

Worldwide emergence of multiple clades of enterovirus 68  

PubMed Central

Human enterovirus 68 (EV-D68) is a historically rarely reported virus linked with respiratory disease. In the past 3 years, a large increase in respiratory disease associated with EV-D68 has been reported, with documented outbreaks in North America, Europe and Asia. In several outbreaks, genetic differences were identified among the circulating strains, indicating the presence of multiple clades. In this report, we analyse archived and novel EV-D68 strains from Africa and the USA, obtained from patients with respiratory illness. Phylogenetic analysis of all EV-D68 sequences indicates that, over the past two decades, multiple clades of the virus have emerged and spread rapidly worldwide. All clades appear to be currently circulating and contributing to respiratory disease.

Firth, Cadhla; Madhi, Shabir A.; Howie, Stephen R. C.; Wu, Winfred; Sall, Amadou Alpha; Haq, Saddef; Briese, Thomas; Lipkin, W. Ian

2012-01-01

127

Fatal case of enterovirus 71 infection, France, 2007.  

PubMed

A fatal case of enterovirus 71 infection with pulmonary edema and rhombencephalitis occurred in Brest, France, in April 2007. The virus was identified as subgenogroup C2. This highly neurotropic enterovirus merits specific surveillance outside the Asia-Pacific region. PMID:19891879

Vallet, Sophie; Legrand Quillien, Marie Christine; Dailland, Thomas; Podeur, Gaëtan; Gouriou, Stéphanie; Schuffenecker, Isabelle; Payan, Christopher; Marcorelles, Pascale

2009-11-01

128

Environmental surveillance of non-polio enteroviruses in Iran  

Microsoft Academic Search

BACKGROUND: Enteroviruses can shed in feces for several weeks, so many excrete viruses can remain infectious for a long time in environment. Therefore, by detecting enteroviruses in environmental specimens and sewage, we can understand this virus circulation, the approximate ratio of contaminated persons in society and they are suitable indicators for environmental surveillance. METHODS: Since March 2006 to February 2007,

Mohammad Kargar; Sara Sadeghipour; Rakhshandeh Nategh

2009-01-01

129

Outbreak of severe neurologic involvement associated with enterovirus 71 infection  

Microsoft Academic Search

Enterovirus 71 has been associated with several outbreaks, as well as sporadic cases, of central nervous system infection and has a worldwide distribution. Seven children with encephalitis and five with aseptic meningitis caused by Enterovirus 71 were seen at Otsu Municipal Hospital during the summer of 1997. The infections were confirmed serologically, although detection of the viral genome in cerebrospinal

Hiroshi Komatsu; Yoshitaka Shimizu; Yoshihiro Takeuchi; Hiroaki Ishiko; Hiroshi Takada

1999-01-01

130

Norovirus, hepatitis A virus and enterovirus presence in shellfish from high quality harvesting areas in Portugal.  

PubMed

This is the first report on the screening of shellfish from Portugal for the presence of human enteropathogenic viruses. Approximately 2000 shellfish (Curbicula fluminea, Ruditapes decussatus, Tellina crassa, Spisula solida, Dosinia exoleta, Ensis spp., Mytilus spp., Ostrea edulis and Cerastoderma edule), organized in 49 batches, were collected between March 2008 and February 2009. They were tested for norovirus (NoV), hepatitis A virus (HAV) and enterovirus (EV) by RT-PCR followed by nucleotide sequencing. Bacterial contamination was also evaluated by Escherichia coli counts. Viral contamination was detected throughout the year in all shellfish species and in all collection areas, independently of their harvesting areas classification. Overall, 67% of all analyzed batches were contaminated by at least one of the studied viruses while the simultaneous presence of two and three viruses was detected in 22% and 6% batches, respectively. Of the three viruses, NoV was detected in 37% of the batches, followed by EV in 35%, and HAV in 33%. Nucleotide sequencing of the NoV and HAV RT-PCR products demonstrated that all strains belonged to NoV genotype GII.4 and HAV subgenotype 1B. The presence of NoV and HAV in shellfish from "A class" harvesting areas of Portugal can represent a potential health risk. PMID:21569936

Mesquita, João R; Vaz, Lisa; Cerqueira, Sónia; Castilho, Fernanda; Santos, Ricardo; Monteiro, Sílvia; Manso, Carmen F; Romalde, Jesus L; Nascimento, Maria São José

2011-01-25

131

Primary Infection by Human Parvovirus B19  

Microsoft Academic Search

We describe a case of primary infection by human parvovirus B19 in a 20-year-old woman; it manifested as erythemato-maculo-papular lesions, pharyngotonsillitis, lymphadenopathy, fever, arthralgia and myalgia, asthenia and anorexia. Laboratory tests revealed anaemia, leucopenia, thrombocytopaenia and a rise in some inflammatory indices. Elisa test was positive for anti-human parvovirus B19 IgM. Clinical symptoms spontaneously regressed in 2 weeks. Thirty days

S. Veraldi; G. Rizzitelli; G. Lunghi; R. Cardone

1993-01-01

132

Pathogenic parasites and enteroviruses in wastewater: support for a regulation on water reuse.  

PubMed

Brazilian regulations for nonpotable reuse are being established using World Health Organization guidelines, however, they should be developed based on local monitoring studies. This study intended to analyze enteroviruses, protozoa and viable Ascaris sp. eggs in raw (24) and treated (24) effluents from four Wastewater Treatment Plants of São Paulo State, Brazil. The protozoa were detected with the US Environmental Protection Agency (USEPA) Method 1623 in the treated effluents and by centrifugation/Immunomagnetic Separation in the raw influent samples. Viable Ascaris sp. eggs were analyzed according to a modified USEPA method. Enteroviruses were quantified by using human rhabdomyosarcoma cells after adequate concentration procedures. All wastewater influents were positive for Giardia sp. whereas Cryptosporidium sp. was detected in 58.3% of the samples. Giardia sp. and Cryptosporidium sp. were present in 79.2 and 25.0% respectively, of the treated wastewater samples. Viable Ascaris sp. eggs were detected in 50.0 and 12.5% of influent and treated wastewater samples. Enteroviruses were isolated in the 24 raw influent samples and in 46% of the treated samples. Taking into account the densities of Giardia sp. in some treated wastewaters intended to be used as reclaimed water, Quantitative Microbial Risk Assessment studies should be conducted to establish pathogen quantitative criteria for a future Brazilian regulation for water reuse. PMID:23552239

Hachich, Elayse M; Galvani, Ana T; Padula, Jose A; Stoppe, Nancy C; Garcia, Suzi C; Bonanno, Vilma M S; Barbosa, Mikaela R F; Sato, Maria Inês Z

2013-01-01

133

HLA-DRB1-DQA1-DQB1 genotype and frequency of enterovirus in longitudinal monthly fecal samples from healthy infants.  

PubMed

Enterovirus infections may be involved in the etiology of type 1 diabetes (T1D), which is strongly associated with certain human leukocyte antigen (HLA) class II haplotypes. Our aim was to assess whether HLA genotypes conferring varying degrees of risk for T1D were associated with enterovirus gut infections. From the general Norwegian population, 190 healthy infants at high-risk for T1D (DR4-DQ8/DR3-DQ2), and 383 infants without this genotype were identified. Non-DR4-DQ8/DR3-DQ2 genotypes were further categorized as conferring either an increased-to-moderate risk (DR4-DQ8 or DR3-DQ2), were protective (DQB1*06:02), or were neutral (all other genotypes). A total of 4626 monthly fecal samples taken between age 3 and 12?mo were tested for enterovirus RNA using real-time PCR. Enterovirus prevalence was 11.5% among high-risk children, and 12.2% in other children (adjusted odds ratio: 1.23, p=0.12). The prevalence was 11.3% in those with increased-to-moderate risk, 13.0% in the protective group, and 12.6% in the neutral group (likelihood ratio test, 3 d.f.: p=0.37). In conclusion, there was no statistically significant association between HLA genotype and the occurrence of human enterovirus gut infections. PMID:22691100

Witsø, Elisabet; Cinek, Ondrej; Tapia, German; Rasmussen, Trond; Stene, Lars C; Rønningen, Kjersti S

2012-06-01

134

[Enteroviruses responsible for acute hemorrhagic conjunctivitis].  

PubMed

Acute hemorrhagic conjunctivitis (AHC) is an epidemic form of highly contagious conjunctivitis, characterized by conjunctival hemorrhages. The first AHC outbreak was described in 1969 in Ghana, West Africa, and was called Apollo disease, from the Apollo landing on the moon. This outbreak was caused by Enterovirus 70 (EV70) together with a Coxsackievirus A24 (CVA24v) variant, which are the major etiological agents involved in AHC outbreaks worldwide. AHC is known to be directly transmitted by close person-to-person contact or indirectly through soiled ophthalmological materials or unsafe recreational water. Recently, a possible airborne virus spread was suggested which could explain the high transmission rate of the disease. In the absence of a specific antiviral therapy, a rapid diagnosis of the causative agent is required to distinguish AHC due to enteroviruses from other ocular infectious diseases, for there are active drugs, or to quickly implement proper public health measures to limit the extension of the outbreak. However, virus identification remains difficult and time-consuming. Moreover, virological diagnosis is difficult to implement in developing countries where AHC has recently become a major problem for public health. PMID:19836177

Lévêque, N; Huguet, P; Norder, H; Chomel, J-J

2010-04-01

135

Advances in Human B Cell Phenotypic Profiling  

PubMed Central

To advance our understanding and treatment of disease, research immunologists have been called-upon to place more centralized emphasis on impactful human studies. Such endeavors will inevitably require large-scale study execution and data management regulation (“Big Biology”), necessitating standardized and reliable metrics of immune status and function. A well-known example setting this large-scale effort in-motion is identifying correlations between eventual disease outcome and T lymphocyte phenotype in large HIV-patient cohorts using multiparameter flow cytometry. However, infection, immunodeficiency, and autoimmunity are also characterized by correlative and functional contributions of B lymphocytes, which to-date have received much less attention in the human Big Biology enterprise. Here, we review progress in human B cell phenotyping, analysis, and bioinformatics tools that constitute valuable resources for the B cell research community to effectively join in this effort.

Kaminski, Denise A.; Wei, Chungwen; Qian, Yu; Rosenberg, Alexander F.; Sanz, Ignacio

2012-01-01

136

Viral Pollution in the Environment and in Shellfish: Human Adenovirus Detection by PCR as an Index of Human Viruses  

PubMed Central

A study of the presence of human viruses (adenoviruses, enteroviruses, and hepatitis A viruses [HAVs]) in environmental and shellfish samples was carried out by applying DNA and cDNA amplification techniques by PCR. The detection of human adenoviruses by PCR was also examined as a potential molecular test to monitor viral pollution. The samples studied were urban and slaughterhouse sewage, river water, seawater, and shellfish. Enteroviruses were quantified by PFU in Buffalo green monkey kidney cells and fecal coliforms and phages of Bacteroides fragilis HSP40 were also evaluated in some of the samples. The amplification of viral DNA and cDNA has shown a high prevalence of human viruses that would not be detected by the use of classical techniques, such as the quantification of PFU in cell lines. The results of the analysis of slaughterhouse sewage samples together with the test of farm animal feces indicate that the adenoviruses and the HAVs detected in the environment are mostly of human origin. A significative correlation between the detection of human viruses by PCR and the values of bacteriophages of B. fragilis HSP40 in urban raw sewage was observed. Human adenoviruses were the viruses most frequently detected throughout the year, and all the samples that were positive for enteroviruses or HAVs were also positive for human adenoviruses. The results suggest that the detection of adenoviruses by PCR could be used as an index of the presence of human viruses in the environment where a molecular index is acceptable.

Pina, Sonia; Puig, Montserrat; Lucena, Francisco; Jofre, Joan; Girones, Rosina

1998-01-01

137

Prognostic implications of myoclonic jerk in children with enterovirus infection  

Microsoft Academic Search

To determine the prognostic value of myoclonic jerk in children with enterovirus 71 (EV71) infection, a retrospective study was conducted on 665 enterovirus culture-confirmed patients admitted to Chang Gung Children's Hospital from January 2000 to September 2001. The mean age was 35.0 months ± 32.2 months, ranging from 1 day to 15 years and 416 (62.6%) of them were male.

Hsiao-Kuo Lu; Tzou-Yien Lin; Shao-Hsuan Hsia; Cheng-Hsun Chiu; Yhu-Chering Huang; Kuo-Chien Tsao; Luan-Yin Chang

138

Complete nucleotide sequence of enterovirus 71 is distinct from poliovirus  

Microsoft Academic Search

Enterovirus 71 (EV71) is capable of causing paralytic disease indistinguishable from poliomyelitis due to poliovirus. To determine the relationship of EV71 to poliovirus and other enteroviruses, two strains of EV71 have been cloned and sequenced. The EV71 strains had only 46% amino acid identity with the polioviral P1 capsid region and 55% with the entire polyprotein. There were no regions

Betty A. Brown; Mark A. Pallansch

1995-01-01

139

Heat Shock protein 90: Role in Enterovirus 71 Entry and Assembly and Potential Target for Therapy.  

PubMed

Although several factors participating in enterovirus 71 (EV71) entry and replication had been reported, the precise mechanisms associated with these events are far from clear. In the present study, we showed that heat shock protein 90 (HSP90) is a key element associated with EV71 entry and replication in a human rhabdomyosarcoma of RD cells. Inhibition of HSP90 by pretreating host cells with HSP90? siRNA or blocking HSP90 with a HSP90-specific antibody or geldanamycin (GA), a specific inhibitor of HSP90, as well as recombinant HSP90? resulted in inhibiting viral entry and subsequent viral replication. Co-immunprecipitation of EV71 with recombinant HSP90? and colocalization of EV71-HSP90 in the cells demonstrated that HSP90 was physically associated with EV71 particles. HSP90 seems to mediate EV71 replication by preventing proteosomal degradation of the newly synthesized capsid proteins, but does not facilitate viral gene expression at transcriptional level. This was evident by post-treatment of host cells with GA, which did not affect the expression of viral transcripts but accelerated the degradation of viral capsid proteins and interfered with the formation of assembled virions. In vivo studies were carried out using human SCARB2-transgenic mice to evaluate the protection conferred by HSP90 inhibitor, 17-allyamino-17-demethoxygeldanamycin (17-AAG), an analog of geldanamycin, that elicited similar activity but with less toxicity. The results showed that the administration of 17-AAG twice conferred the resistance to hSCARB2 mice challenged with C2, C4, and B4 genotypes of EV71. Our data supports HSP90 plays an important role in EV71 infection. Targeting of HSP90 with clinically available drugs might provide a feasible therapeutic approach to treat EV71 infection. PMID:24098578

Tsou, Yueh-Liang; Lin, Yi-Wen; Chang, Hsuen-Wen; Lin, Hsiang-Yin; Shao, Hsiao-Yun; Yu, Shu-Ling; Liu, Chia-Chyi; Chitra, Ebenezer; Sia, Charles; Chow, Yen-Hung

2013-10-02

140

Heat Shock protein 90: Role in Enterovirus 71 Entry and Assembly and Potential Target for Therapy  

PubMed Central

Although several factors participating in enterovirus 71 (EV71) entry and replication had been reported, the precise mechanisms associated with these events are far from clear. In the present study, we showed that heat shock protein 90 (HSP90) is a key element associated with EV71 entry and replication in a human rhabdomyosarcoma of RD cells. Inhibition of HSP90 by pretreating host cells with HSP90? siRNA or blocking HSP90 with a HSP90-specific antibody or geldanamycin (GA), a specific inhibitor of HSP90, as well as recombinant HSP90? resulted in inhibiting viral entry and subsequent viral replication. Co-immunprecipitation of EV71 with recombinant HSP90? and colocalization of EV71-HSP90 in the cells demonstrated that HSP90 was physically associated with EV71 particles. HSP90 seems to mediate EV71 replication by preventing proteosomal degradation of the newly synthesized capsid proteins, but does not facilitate viral gene expression at transcriptional level. This was evident by post-treatment of host cells with GA, which did not affect the expression of viral transcripts but accelerated the degradation of viral capsid proteins and interfered with the formation of assembled virions. In vivo studies were carried out using human SCARB2-transgenic mice to evaluate the protection conferred by HSP90 inhibitor, 17-allyamino-17-demethoxygeldanamycin (17-AAG), an analog of geldanamycin, that elicited similar activity but with less toxicity. The results showed that the administration of 17-AAG twice conferred the resistance to hSCARB2 mice challenged with C2, C4, and B4 genotypes of EV71. Our data supports HSP90 plays an important role in EV71 infection. Targeting of HSP90 with clinically available drugs might provide a feasible therapeutic approach to treat EV71 infection.

Tsou, Yueh-Liang; Lin, Yi-Wen; Chang, Hsuen-Wen; Lin, Hsiang-Yin; Shao, Hsiao-Yun; Yu, Shu-Ling; Liu, Chia-Chyi; Chitra, Ebenezer; Sia, Charles; Chow, Yen-Hung

2013-01-01

141

Folate, vitamin B12 and human health  

Technology Transfer Automated Retrieval System (TEKTRAN)

During the past decade the role of folate and vitamin B12 in human nutrition have been under constant re-examination. Basic knowledge on the metabolism and interactions between these essential nutrients has expanded and multiple complexities have been unraveled. These micronutrients have shared func...

142

Taxonomic Classification of Human Hepatitis B Virus  

Microsoft Academic Search

Summary Sufficient data have accumulated to permit the ICTV Study Group on the Nomenclature of Hepatitis Viruses to recognize human hepatitis B virus as a member of a unique group of viruses and to classify it, together with a number of related animal viruses, into a new family called the Hepadnaviridae. Over the past decade, the International Committee on Taxonomy

Ian D. Gust; Christopher J. Burrell; Anthony G. Coulepis; William S. Robinson; Arie J. Zuckerman

1986-01-01

143

Persistence of human parvovirus B19 in human tissues  

Microsoft Academic Search

Human parvovirus B19 infection causes various clinical symptoms, such as rash, arthropathy, anemias and fetal death, but it can also remain asymptomatic. The arthropathies and anemias can become chronic for several years, not infrequently resembling autoimmune syndromes. B19 replicates only in red blood cell precursors of bone marrow or fetal liver, resulting in high-titred short-lived viremia, but viral DNA is

Maria Söderlund-Venermo; Kati Hokynar; Janne Nieminen; Henna Rautakorpi; Klaus Hedman

2002-01-01

144

Enteroviruses, type 1 diabetes and hygiene: a complex relationship.  

PubMed

Type 1 diabetes (T1D) is an autoimmune disease in which the immune system mounts an attack on the host's insulin-producing beta cells. Because most cases of T1D cannot be attributed only to individual genetics, it is strongly inferred that there is a significant environmental contribution, such as infection, impacting disease development. The human enteroviruses (HEV) are common picornaviruses often implicated as triggers of human T1D, although precisely which of the numerous HEV may be involved in human T1D development is unknown. Experiments using non-obese diabetic (NOD) mice, commonly used to model T1D, show that induction of T1D by HEV infection in NOD mice is a multifactorial process involving both the virus and the host. Interestingly, results demonstrate that HEV infection of NOD mice can also induce long-term protection from T1D under certain conditions, suggesting that a similar mechanism may occur in humans. Based upon both experimental animal and observational human studies, we postulate that HEV have a dual role in T1D development and can either cause or prevent autoimmune disease. Whichever outcome occurs depends upon multiple variables in the host-virus equation, many of which can be deduced from results obtained from NOD mouse studies. We propose that the background to the sharply rising T1D incidences observed in the 20th century correlates with increased levels of hygiene in human societies. Viewing T1D in this perspective suggests that potential preventative options could be developed. PMID:20049905

Tracy, S; Drescher, K M; Jackson, J D; Kim, K; Kono, K

2010-03-01

145

Persistence of enteroviruses in sewage sludge*  

PubMed Central

Sewage from residential areas often contains viruses pathogenic for man and significant amounts are probably associated with solids in sewage sludge. Information on the survival of viruses in sewage sludge is necessary in order to develop guidelines for recycling programmes that involve spreading the sludge on land. In the present study, a number of enteroviruses were added to sewage sludge and the artificially contaminated sludges were tested for viruses at intervals over a 12-week period. Most of the viruses survived for many weeks at room temperature. It is clear that sewage sludge destined for land application should be adequately treated for virus inactivation. In interpreting these results, it should be borne in mind that the survival of hepatitis A virus might be similar. Recent reports about the reappearance of poliomyelitis in regions with immunization programmes should also be taken into consideration.

Subrahmanyan, T. P.

1977-01-01

146

Enterovirus infection in Korean children and anti-enteroviral potential candidate agents.  

PubMed

Although most enterovirus infections are not serious enough to be life threatening, several enteroviruses such as enterovirus 71 are responsible for severe, potentially life-threatening disease. The epidemic patterns of enteroviruses occur regularly during the year, but they may change due to environmental shifts induced by climate change due to global warming. Therefore, enterovirus epidemiological studies should be performed continuously as a basis for anti-viral studies. A great number of synthesized antiviral compounds that work against enteroviruses have been developed but only a few have demonstrated effectiveness in vivo. No proven effective antiviral agents are available for enterovirus disease therapy. The development of a new antiviral drug is a difficult task due to poor selective toxicity and cost. To overcome these limitations, one approach is to accelerate the availability of other existing antiviral drugs approved for antiviral effect against enteroviruses, and the other way is to screen traditional medicinal plants. PMID:23133481

Park, Kwi Sung; Choi, Young Jin; Park, Joon Soo

2012-10-29

147

Adaptation of enterovirus 71 to adult interferon deficient mice.  

PubMed

Non-polio enteroviruses, including enterovirus 71 (EV71), have caused severe and fatal cases of hand, foot and mouth disease (HFMD) in the Asia-Pacific region. The development of a vaccine or antiviral against these pathogens has been hampered by the lack of a reliable small animal model. In this study, a mouse adapted EV71 strain was produced by conducting serial passages through A129 (?/? interferon (IFN) receptor deficient) and AG129 (?/?, ? IFN receptor deficient) mice. A B2 sub genotype of EV71 was inoculated intraperitoneally (i.p.) into neonatal AG129 mice and brain-harvested virus was subsequently passaged through 12 and 15 day-old A129 mice. When tested in 10 week-old AG129 mice, this adapted strain produced 100% lethality with clinical signs including limb paralysis, eye irritation, loss of balance, and death. This virus caused only 17% mortality in same age A129 mice, confirming that in the absence of a functional IFN response, adult AG129 mice are susceptible to infection by adapted EV71 isolates. Subsequent studies in adult AG129 and young A129 mice with the adapted EV71 virus examined the efficacy of an inactivated EV71 candidate vaccine and determined the role of humoral immunity in protection. Passive transfer of rabbit immune sera raised against the EV71 vaccine provided protection in a dose dependent manner in 15 day-old A129 mice. Intramuscular injections (i.m.) in five week-old AG129 mice with the alum adjuvanted vaccine also provided protection against the mouse adapted homologous strain. No clinical signs of disease or mortality were observed in vaccinated animals, which received a prime-and-boost, whereas 71% of control animals were euthanized after exhibiting systemic clinical signs (P<0.05). The development of this animal model will facilitate studies on EV71 pathogenesis, antiviral testing, the evaluation of immunogenicity and efficacy of vaccine candidates, and has the potential to establish correlates of protection studies. PMID:23527208

Caine, Elizabeth A; Partidos, Charalambos D; Santangelo, Joseph D; Osorio, Jorge E

2013-03-19

148

Adaptation of Enterovirus 71 to Adult Interferon Deficient Mice  

PubMed Central

Non-polio enteroviruses, including enterovirus 71 (EV71), have caused severe and fatal cases of hand, foot and mouth disease (HFMD) in the Asia-Pacific region. The development of a vaccine or antiviral against these pathogens has been hampered by the lack of a reliable small animal model. In this study, a mouse adapted EV71 strain was produced by conducting serial passages through A129 (?/? interferon (IFN) receptor deficient) and AG129 (?/?, ? IFN receptor deficient) mice. A B2 sub genotype of EV71 was inoculated intraperitoneally (i.p.) into neonatal AG129 mice and brain-harvested virus was subsequently passaged through 12 and 15 day-old A129 mice. When tested in 10 week-old AG129 mice, this adapted strain produced 100% lethality with clinical signs including limb paralysis, eye irritation, loss of balance, and death. This virus caused only 17% mortality in same age A129 mice, confirming that in the absence of a functional IFN response, adult AG129 mice are susceptible to infection by adapted EV71 isolates. Subsequent studies in adult AG129 and young A129 mice with the adapted EV71 virus examined the efficacy of an inactivated EV71 candidate vaccine and determined the role of humoral immunity in protection. Passive transfer of rabbit immune sera raised against the EV71 vaccine provided protection in a dose dependent manner in 15 day-old A129 mice. Intramuscular injections (i.m.) in five week-old AG129 mice with the alum adjuvanted vaccine also provided protection against the mouse adapted homologous strain. No clinical signs of disease or mortality were observed in vaccinated animals, which received a prime-and-boost, whereas 71% of control animals were euthanized after exhibiting systemic clinical signs (P<0.05). The development of this animal model will facilitate studies on EV71 pathogenesis, antiviral testing, the evaluation of immunogenicity and efficacy of vaccine candidates, and has the potential to establish correlates of protection studies.

Caine, Elizabeth A.; Partidos, Charalambos D.; Santangelo, Joseph D.; Osorio, Jorge E.

2013-01-01

149

Effect of enteroviruses on adherence to and invasion of HEp-2 cells by Campylobacter isolates.  

PubMed Central

Coinfection of HEp-2 epithelial cells with coxsackievirus B3, echovirus 7, poliovirus (LSc type 1), porcine enterovirus, and Campylobacter isolates was performed to determine if a synergistic effect could be obtained. The invasiveness of Campylobacter jejuni ATCC 33560 was significantly increased for HEp-2 cells preinfected with echovirus 7, coxsackievirus B3, and UV-inactivated (noninfectious) coxsackievirus B3 particles. Additionally, the invasiveness of C. jejuni M96, a clinical isolate, was significantly increased for HEp-2 cells preinfected with coxsackievirus B3. Poliovirus and porcine enterovirus had no effect on C. jejuni ATCC 33560 adherence and invasiveness. Furthermore, poliovirus had no effect on the ability of C. jejuni M96 to adhere to and invade HEp-2 cells. Campylobacter hyointestinalis and Campylobacter mucosalis, two noninvasive isolates, did not invade virus-infected HEp-2 cells. The increase in the invasiveness of C. jejuni appeared to be the result of specific interactions between the virus and the HEp-2 cell membrane. The data suggest that the invasiveness of Campylobacter spp. is dependent upon the inherent properties of the organism. Virus-induced cell alterations can potentiate the invasiveness of virulent Campylobacter spp. but are not sufficient to allow internalization of noninvasive bacteria.

Konkel, M E; Joens, L A

1990-01-01

150

Survival of enteroviruses and coliform bacteria in a sludge lagoon.  

PubMed Central

Enteroviruses associate with aerobically and anaerobically digested sludge were determined before the addition of the sludge to a sludge lagoon. The fate of sludge-associated viruses was followed during detention of sludge in the lagoon and after application of sludge to land for disposal. While digested sludge was being added to the lagoon, enteroviruses were readily detected in grab samples of sludge from the lagoon. Sludge-associated viruses dropped to low or undetectable levels after disposal of sludge on land and during periods when addition of digested sludge to the lagoon was suspended. Changes in the levels of fecal coliforms in the lagooned sludge paralleled changes in the numbers of enteroviruses. Enteroviruses were not detected in water from deep wells located on the sludge disposal site or near the lagoon. During the initial part of the study, poliovirus serotypes accounted for greater than 90% of the viruses identified. Later, poliovirus serotypes comprised less than 40% of the virus isolates, and echoviruses and Coxsackieviruses were the most common enteroviruses identified.

Farrah, S R; Bitton, G; Hoffmann, E M; Lanni, O; Pancorbo, O C; Lutrick, M C; Bertrand, J E

1981-01-01

151

Enterovirus 71 blocks selectively type I interferon production through the 3C viral protein in mice.  

PubMed

Type I interferons (IFNs) represent an essential innate defense mechanism for controlling enterovirus 71 (EV 71) infection. Mice inoculated with EV 71 produced a significantly lower amount of type I IFNs than those inoculated with poly (I:C), adenovirus type V, or coxsackievirus B3 (CB3). EV 71 infection, however, mounted a proinflammatory response with a significant increase in the levels of serum and brain interleukin (IL)-6, monocyte chemoattractant protein-1, tumor necrosis factor, and IFN-?. EV 71 infection abolished both poly (I:C)- and CB3-induced type I IFN production of mice. Such effect was not extended to other enteroviruses including coxsackievirus A24, B2, B3, and echovirus 9, as mice infected with these viruses retained type I IFN responsiveness upon poly (I:C) challenge. In addition, EV 71-infected RAW264.7 cells produced significantly lower amount of type I IFNs than non-infected cells upon poly (I:C) stimulation. The inhibitory effect of EV 71 on type I IFN production was attributed to the viral protein 3C, which was confirmed using over-expression systems in both mice and RAW264.7 cells. The 3C over-expression, however, did not interfere with poly (I:C)-induced proinflammatory cytokine production. These findings indicate that EV 71 can hamper the host innate defense by blocking selectively type I IFN synthesis through the 3C viral protein. PMID:22997081

Lee, Yi-Ping; Wang, Ya-Fang; Wang, Jen-Ren; Huang, Szu-Wei; Yu, Chun-Keung

2012-11-01

152

Autoprocessing: an essential step for expression and purification of enterovirus 71 3C(pro) in Escherichia coli.  

PubMed

A gene encoding the 3BC of human enterovirus 71 (EV71) was cloned and inserted into a derivative of plasmid pET-32a(+) driven by T7 promoter. The expressed 3C protease (3C(pro)) autocatalytically cleaved itself from the recombinant protein Trx-3BC and the mature 3C(pro) partitioned in the soluble fraction of bacterial lysate. The 13-amino-acid peptide substrates with the junction of 3B/3C were used to verify the proteolysis activity of the purified 3C(pro). The EV71 3C(pro) had a Km value of 63 ?M (measured by a continuous fluorescence assay). The other solid-phase activity assay of the EV71 3C(pro) was developed using HPLC to analyze the proteolytic products. The combination of two activity assays contributes to promote the identification of the specific inhibitors targeted to the EV71 3C(pro). PMID:23881322

Huang, Shuqiong; Lyu, Yanning; Qing, Xianyun; Wang, Weiwei; Tang, Liang; Cheng, Kedi; Wang, Wei

2013-07-24

153

[Folate, vitamin B12 and human health].  

PubMed

During the past decade the role of folate and vitamin B12 in human nutrition have been under constant re-examination. Basic knowledge on the metabolism and interactions between these essential nutrients has expanded and multiple complexities have been unraveled. These micronutrients have shared functions and intertwined metabolic pathways that define the size of the "methyl donor" pool utilized in multiple metabolic pathways; these include DNA methylation and synthesis of nucleic acids. In Chile, folate deficiency is virtually nonexistent, while vitamin B12 deficiency affects approximately 8.5-51% depending on the cut-off value used to define deficiency. Folate is found naturally mainly in vegetables or added as folic acid to staple foods. Vitamin B12 in its natural form is present only in foods of animal origin, which is why deficit is more common among strict vegetarians and populations with a low intake of animal foods. Poor folate status in vulnerable women of childbearing age increases the risk of neural tube birth defects, so the critical time for the contribution of folic acid is several months before conception since neural tube closure occurs during the first weeks of life. The absorption of vitamin B12 from food is lower in older adults, who are considered to have higher risk of gastric mucosa atrophy, altered production of intrinsic factor and acid secretion. Deficiency of these vitamins is associated with hematological disorders. Vitamin B12 deficiency can also induce clinical and sub-clinical neurological and of other disorders. The purpose of this review is to provide an update on recent advances in the basic and applied knowledge of these vitamins relative to human health. PMID:23677195

Brito, Alex; Hertrampf, Eva; Olivares, Manuel; Gaitán, Diego; Sánchez, Hugo; Allen, Lindsay H; Uauy, Ricardo

2012-11-01

154

RAG Mutations in Human B Cell-Negative SCID  

Microsoft Academic Search

Patients with human severe combined immunodeficiency (SCID) can be divided into those with B lymphocytes (B^+ SCID) and those without (B^- SCID). Although several genetic causes are known for B^+ SCID, the etiology of B^- SCID has not been defined. Six of 14 B^- SCID patients tested were found to carry a mutation of the recombinase activating gene 1 (RAG-1),

Klaus Schwarz; George H. Gauss; Leopold Ludwig; Ulrich Pannicke; Zhong Li; Doris Lindner; Wilhelm Friedrich; Reinhard A. Seger; Thomas E. Hansen-Hagge; Stephen Desiderio; Michael R. Lieber; Claus R. Bartram

1996-01-01

155

Co-circulation and evolution of polioviruses and species C enteroviruses in a district of Madagascar.  

PubMed

Between October 2001 and April 2002, five cases of acute flaccid paralysis (AFP) associated with type 2 vaccine-derived polioviruses (VDPVs) were reported in the southern province of the Republic of Madagascar. To determine viral factors that favor the emergence of these pathogenic VDPVs, we analyzed in detail their genomic and phenotypic characteristics and compared them with co-circulating enteroviruses. These VDPVs appeared to belong to two independent recombinant lineages with sequences from the type 2 strain of the oral poliovaccine (OPV) in the 5'-half of the genome and sequences derived from unidentified species C enteroviruses (HEV-C) in the 3'-half. VDPV strains showed characteristics similar to those of wild neurovirulent viruses including neurovirulence in poliovirus-receptor transgenic mice. We looked for other VDPVs and for circulating enteroviruses in 316 stools collected from healthy children living in the small area where most of the AFP cases occurred. We found vaccine PVs, two VDPVs similar to those found in AFP cases, some echoviruses, and above all, many serotypes of coxsackie A viruses belonging to HEV-C, with substantial genetic diversity. Several coxsackie viruses A17 and A13 carried nucleotide sequences closely related to the 2C and the 3D(pol) coding regions of the VDPVs, respectively. There was also evidence of multiple genetic recombination events among the HEV-C resulting in numerous recombinant genotypes. This indicates that co-circulation of HEV-C and OPV strains is associated with evolution by recombination, resulting in unexpectedly extensive viral diversity in small human populations in some tropical regions. This probably contributed to the emergence of recombinant VDPVs. These findings give further insight into viral ecosystems and the evolutionary processes that shape viral biodiversity. PMID:18085822

Rakoto-Andrianarivelo, Mala; Guillot, Sophie; Iber, Jane; Balanant, Jean; Blondel, Bruno; Riquet, Franck; Martin, Javier; Kew, Olen; Randriamanalina, Bakolalao; Razafinimpiasa, Lalatiana; Rousset, Dominique; Delpeyroux, Francis

2007-12-01

156

The Interplays between Autophagy and Apoptosis Induced by Enterovirus 71  

PubMed Central

Background Enterovirus 71 (EV71) is the causative agent of human diseases with distinct severity, from mild hand, foot and mouth disease to severe neurological syndromes, such as encephalitis and meningitis. The lack of understanding of viral pathogenesis as well as lack of efficient vaccine and drugs against this virus impedes the control of EV71 infection. EV71 virus induces autophagy and apoptosis; however, the relationship between EV71-induced autophagy and apoptosis as well as the influence of autophagy and apoptosis on virus virulence remains unclear. Methodology/Principal Findings In this study, it was observed that the Anhui strain of EV71 induced autophagy and apoptosis in human rhabdomyosarcoma (RD-A) cells. Additionally, by either applying chemical inhibitors or knocking down single essential autophagic or apoptotic genes, inhibition of EV71 induced autophagy inhibited the apoptosis both at the autophagosome formation stage and autophagy execution stage. However, inhibition of autophagy at the stage of autophagosome and lysosome fusion promoted apoptosis. In reverse, the inhibition of EV71-induced apoptosis contributed to the conversion of microtubule-associated protein 1 light chain 3-I (LC3-I) to LC3-II and degradation of sequestosome 1 (SQSTM1/P62). Furthermore, the inhibition of autophagy in the autophagsome formation stage or apoptosis decreased the release of EV71 viral particles. Conclusions/Significance In conclusion, the results of this study not only revealed novel aspect of the interplay between autophagy and apoptosis in EV71 infection, but also provided a new insight to control EV71 infection.

Wang, Bei; Wang, Tao; Wang, Ji; Huang, He; Wang, Jianwei; Jin, Qi; Zhao, Zhendong

2013-01-01

157

Replication strategy of human hepatitis B virus  

SciTech Connect

To study the replication strategy of the human hepatitis B virus, the 5' end of the RNA pregenome and the initiation sites of DNA plus and minus strands have been mapped. The RNA pregenome was found to be terminally redundant by 120 nucleotides; it is initiated within the pre-C region and may also function as mRNA for synthesis of the major core protein and the hepatitis B virus reverse transcriptase. The hepatitis B virus DNA minus strand is initiated within the direct repeat sequence DR1, it contains a terminal redundancy of up to eight nucleotides, and its synthesis does not require any template switch. The DNA plus strand is primed by a short oligoribonucleotide probably derived from the 5' end of the RNA pregenome, and its synthesis is initiated close to the direct repeat sequence DR2. For its elongation to pass the discontinuity in the DNA minus strand an intramolecular template switch occurs using the terminal redundancy of this template. Thus, the route of reverse transcription and DNA replication of hepatitis B viruses is fundamentally different from that of retroviruses.

Will, H.; Reiser, W.; Weimer, T.; Pfaff, E.; Buescher, M.; Sprengel, R.; Cattaneo, R.; Schaller, H.

1987-03-01

158

Human B cell defects in perspective  

PubMed Central

While primary immune defects are generally considered to lead to severe and easily recognized disease in infants and children, a number of genetic defects impairing B cell function may not be clinically apparent or diagnosed until adult life. The commonest of these is common variable immune deficiency, the genetic origins of which are beginning to be at least partially understood. CVID affects ? 1/25,000 Caucasians and is characterized by a marked reduction in serum IgG, almost always in serum IgA, and reduced serum IgM in about half of all cases; these defects continue to provide an opportunity to investigate the genes necessary for B cell function in humans. Recently, a small number of genes necessary for normal B cell function have been identified in consanguineous families leading to varying degrees of hypogammaglobulinemia and loss of antibody production. In other studies, whole-exome sequencing and copy number variation, applied to large cohorts, have extended research into understanding both the genetic basis of this syndrome and the clinical phenotypes of CVID.

2012-01-01

159

Identification of Bulgarian Strain 258 of Enterovirus 71  

Microsoft Academic Search

Summary In 1975 in Bulgaria a severe epidemic of central nervous system (CNS) disease occurred. Clinically, histopathologically, and epidemiologically the cases resembled poliomyelitis, aseptic meningitis, meningoencephalitis, and, in some cases, encephalomyocarditis. About 21% of the 700 reported cases developed paralysis, 44 with fatal outcome [ref. 1]. In 65 cases, 92 strains of enterovirus of the same serologic type were isolated:

J. L. Melnick; N. J. Schmidt; R. R. Mirkovic; M. P. Chumakov; I. K. Lavrova; M. K. Voroshilova

1979-01-01

160

The Complete Nucleotide Sequence of a Bovine Enterovirus  

Microsoft Academic Search

SUMMARY The complete nucleotide sequence of the genome of a bovine enterovirus (strain VG- 5-27) has been determined using molecular cloning and DNA sequencing techniques. Excluding the poly(A) tract the genome was 7414 nucleotides long and contained a 5\\

J. A. P. Earle; R. A. Skuce; C. S. Fleming; E. M. Hoey; S. J. Martin

1988-01-01

161

ENTEROVIRUSES IN SLUDGE: MULTIYEAR EXPERIENCE WITH FOUR WASTEWATER TREATMENT PLANTS  

EPA Science Inventory

The authors describe their experience with the isolation of viruses from four treatment plants located in different geographic areas. Over a period of 3 years, 297 enteroviruses were isolated from 307 sludge samples. The highest frequency of viral isolation (92%), including multi...

162

Natural genetic recombination between co-circulating heterotypic enteroviruses  

Microsoft Academic Search

Natural recombination in poliovirus is a frequent phenomenon. In practice, whenever different genotypes have the opportunity to infect the same individual, a high proportion of viruses with recombinant genomes are excreted. To determine whether enteroviruses other than poliovirus can naturally produce viable virions with recombinant genomes, we studied the molecular features of two distant regions of the viral genomes -

Gabriela Oprisan; Mariana Combiescu; Sophie Guillot; Valerie Caro; Andrei Combiescu; Francis Delpeyroux; Radu Crainic

2002-01-01

163

RT-PCR and cell culture infectivity assay to detect enteroviruses during drinking water treatment processes.  

PubMed

In this study, 62 water samples were collected from two water treatment plants (WTPs) in Suez Canal cities (Port Said and Ismaillia) and one plant in Cairo (Giza WTP) in addition to the beginning of the two Nile river branches (Rosetta and Damietta). Viruses were concentrated by adsorption-elution ethod sing 142 mm-diameter nitrocellulose membrane of 0.45 microm pore size and eluted with 3% beef extract at pH 9.5. The concentrated samples were inoculated for 3 successive passages in three cell culture types (Vero, BGM and RD). Enterovirus RNAs in CPE-induced samples were extracted by guanidinium thiocyanate/ phenol/chloroform and heat shock methods and detected by RT-PCR and neutralization test. The results showed that eight samples [14.5% (8/62)] contained enteroviruses most of them were polioviruses [87.5% (7/8)] and coxsackievirus type B2 [12.5% (1/8)]. The three cell cultures were of the same sensitivity to detect the isolated viruses. Also, RT-PCR followed by neutralization assay facilitates and accelerate the results. The guanidinium thiocyanate extraction method was more sensitive than heat shock method. The results turned our attention to review our technology of water treatment and disinfection step in addition to the selection of suitable intake for the drinking water treatment plants. PMID:17219867

Ali, M A; El-Esnawy, N A; Shoaeb, A R; Ibraheim, M; El-Hawaary, S E

1999-01-01

164

Isolation of enterovirus and reovirus from sewage and treated effluents in selected Puerto Rican communities.  

PubMed

Sewage treatment plant effluents were surveyed for viral contributions to gastroenteritis outbreaks in Puerto Rico. Of the 15 sewage treatment plants studied, all discharged their effluents upstream from water treatment plant intakes. No base-line data on the degree of viral challenge to these sewage treatment plants or the subsequent reduction of viruses before discharge existed. Enterovirus counts were generally much higher than those found in the continental United States. At four plants, viruses in the incoming sewage exceeded 100,000 PFU/liter, and one of these, a trickling filter plant, was discharging 24,000 PFU/liter to receiving waters. Virus identification showed that more than 80% of the enterovirus isolates were coxsackievirus B5. These overwhelming viral numbers pointed to defects in the sewage treatment processes. Without reasonable barriers to protect receiving waters, several of the downstream communities were using raw waters that posed extraordinary demands on the ability of their water treatment plants to supply virologically safe drinking water. PMID:2541664

Dahling, D R; Safferman, R S; Wright, B A

1989-02-01

165

RapidDiagnosis ofEnterovirus Infection byMagneticBead Extraction andPolymerase ChainReaction Detection of Enterovirus RNA inClinical Specimens  

Microsoft Academic Search

We describe a rapid methodforextraction anddetection ofenterovirus RNA inclinical samples. Byusing magnetic beadtechnology, enterovirus RNA was efficiently andrapidly extracted fromcerebrospinal fluid, stool, saliva, blood, pericardial fluid, urine, andcryopreserved orformalin-fixed solid tissue. Enterovirus RNA was thendetected byreversetranscription followed bypolymerase chainreaction amplification withprimers designed toallow detection ofmostenterovirus serotypes. Fordetection ofenteroviruses inspecimens from patients withacuteenteroviral disease, theoverallsensitivity ofenzymatic RNA amplification was greater than

PETER MUIR; FELICITY NICHOLSON; MAHMOOD JHETAM; SHEILA NEOGI; JANGU E. BANATVALA

1993-01-01

166

Differential apoptosis gene expressions of rhabdomyosarcoma cells in response to enterovirus 71 infection  

PubMed Central

Background Enterovirus 71 (EV71) infection can induce the apoptosis of infected cells. The aim of this study is to explore the effect of EV71 infection on apoptosis mechanisms in virus-infected human rhabdomyosarcoma (RD) cells. Methods The apoptosis of RD cells was examined using annexin V-FITC/PI by flow cytometry and cytokines were detected by ELISA. Cellular RNA was extracted and transcribed to cDNA. PCR array was employed to analyze the expressions of 84 apoptotic genes from EV71-infected RD cells at 8 and 20 h postinfection, respectively. In addition, the expressions of FasL, caspase, AKT2, JNK1/2, c-Jun and NF-?B proteins were detected by western blotting. Results Flow cytometry demonstrated that the apoptosis or death of EV71-infected RD cells was increased by 37.1% with a multiplicity of infection (MOI) of 5 at 20 h postinfection. The production of IL-4, IL-10 and TNF-? was enhanced by the subsequent EV71 infection. PCR array revealed significant changes in the expressions of apoptotic genes. Among 84 genes, 42 genes were down-regulated after EV71 infection at 8 h, whereas 32 genes were up-regulated at 20 h postinfection. Moreover, the ligands of TNF superfamily such as FasL, CD40L and TNF-? were significantly up-regulated and enhanced the expressions of apoptosis-related cysteine peptidases, including caspase-10, -8, -7 and -3. In addition, EV71 infection induces the phosphorylation of AKT2, JNK1/2, c-Jun and NF-?B at 20 h postinfection. Conclusion PCR array for the determination of apoptosis gene expressions is an informative assay in elucidating biological pathways. During the early stage of EV71 infection, the apoptotic process of RD cells is significantly delayed. EV71 infection can also induce the expressions of FasL, TNF-? and CD40L, which contribute to the apoptosis of RD cells.

2012-01-01

167

REPSIM-B: A Computer Simulation Model of Human Reproduction.  

National Technical Information Service (NTIS)

The monograph presents illustrative applications and technical details of the computer simulation model REPSIM-B, which was developed for studying a number of important social and biological factors affecting the human reproductive process. REPSIM-B simul...

J. C. Ridley A. S. Clague J. C. Ott S. S. Pastroff C. M. Suchindran

1979-01-01

168

The Human Fetal Omentum: A Site of B Generation  

Microsoft Academic Search

Summary The fetal mouse omentum has been shown to be a source of precursors that exclusively reconstitutes Lyl + B cells and the closely related Lyl- sister population, but not conventional B cells or T cells. We have extended these studies to compare B cell development in the human fetal omentum, liver, and spleen, and to demonstrate that the pro\\/pre-B

Nanette Solvason; John F. Kearney

1992-01-01

169

Degradation of laminin by human tumor cathepsin B  

Microsoft Academic Search

Cathepsin B activity, including that of a plasma membrane-associated cathepsin B, has been linked to tumor malignancy. As cathepsin B at the tumor cell surface has been hypothesized to play a role in the focal degradation of basement membrane during the metastatic cascade, we have examined the ability of human tumor cathepsin B to degrade laminin, an adhesive glycoprotein found

Tamara T. Lah; Michael R. Buck; Kenneth V. Honn; John D. Crissman; Nagesawa C. Rao; Lance A. Liotta; Bonnie F. Sloane

1989-01-01

170

Human cytomegalovirus glycoprotein B genotyping from bronchoalveolar lavage specimens.  

PubMed

The genes encoding glycoprotein complexes of human cytomegalovirus are often polymorphic; in particular, glycoprotein B (gB), which is essential for both in vivo and in vitro replication, is encoded by the highly polymorphic gene UL55. In this study, the distribution of gB genotypes was investigated in 44 bronchoalveolar lavage specimens from adult patients positive for human cytomegalovirus DNA by a multiplex nested fast PCR able to amplify 5 gB genotypes (gB1-gB5). The distribution of gB genotypes was as follows: 12 (27.3%) gB1, 11 (25%) gB2, 9 (20.4%) gB3, 4 (9.1%) gB4, 0 gB5, and 8 (18.2%) mixed genotypes. No difference in prevalence was found in relation to clinical features, including immunological status, non-transplant or transplant condition, and type of transplanted organ, or in follow-up specimens; while gB4 and gB3 were shown to be significantly more prevalent in patients with respiratory insufficiency, and gB4 and gB2 in those with pneumonia. The prevalence of gB genotypes in the lower respiratory tract was similar to that previously reported using other specimen types and patients, with gB1 found to be the most prevalent. The association of gB genotypes with specific clinical features should be further investigated. PMID:21491978

Bergallo, Massimiliano; Costa, Cristina; Gambarino, Stefano; Tornicelli, Alessandra; Astegiano, Sara; Terlizzi, Maria Elena; Solidoro, Paolo; Cavallo, Rossana

2011-04-01

171

Functional Comparison of SCARB2 and PSGL1 as Receptors for Enterovirus 71  

PubMed Central

Human scavenger receptor class B, member 2 (SCARB2), and P-selectin glycoprotein ligand-1 (PSGL1) have been identified to be the cellular receptors for enterovirus 71 (EV71). We compared the EV71 infection efficiencies of mouse L cells that expressed SCARB2 (L-SCARB2) and PSGL1 (L-PSGL1) and the abilities of SCARB2 and PSGL1 to bind to the virus. L-SCARB2 cells bound a reduced amount of EV71 compared to L-PSGL1 cells. However, EV71 could infect L-SCARB2 cells more efficiently than L-PSGL1 cells. The results suggested that the difference in the binding capacities of the two receptors was not the sole determinant of the infection efficiency and that SCARB2 plays an essential role after attaching to virions. Therefore, we examined the viral entry into L-SCARB2 cells and L-PSGL1 cells by immunofluorescence microscopy. In both cells, we detected internalized EV71 virions that colocalized with an early endosome marker. We then performed a sucrose density gradient centrifugation analysis to evaluate viral uncoating. After incubating the EV71 virion with L-SCARB2 cells or soluble SCARB2 under acidic conditions below pH 6.0, we observed that part of the native virion was converted into an empty capsid that lacked both genomic RNA and VP4 capsid proteins. The results suggested that the uncoating of EV71 requires both SCARB2 and an acidic environment and occurs after the internalization of the virus-receptor complex into endosomes. However, the empty capsid formation was not observed after incubation with L-PSGL1 cells or soluble PSGL1 under any of the tested pH conditions. These results indicated that SCARB2 is capable of viral binding, viral internalization, and viral uncoating and that the low infection efficiency of L-PSGL1 cells is due to the inability of PSGL1 to induce viral uncoating. The characterization of SCARB2 as an uncoating receptor greatly contributes to the understanding of the early steps of EV71 infection.

Yamayoshi, Seiya; Ohka, Seii; Fujii, Ken

2013-01-01

172

Expression, refolding, and purification of recombinant human granzyme B  

Microsoft Academic Search

Granzyme B (GrB) is a member of a family of serine proteases involved in cytotoxic T-lymphocyte-mediated killing of potentially harmful cells, where GrB induces apoptosis by cleavage of a limited number of substrates. To investigate the suitability of GrB as an enzyme for specific fusion protein cleavage, two derivatives of human GrB, one dependent on blood coagulation factor Xa (FXa)

Rikke H. Lorentsen; Charlotte H. Fynbo; Hans C. Thøgersen; Michael Etzerodt; Thor L. Holtet

2005-01-01

173

Diagnosis of neonatal enterovirus infection by polymerase chain reaction  

Microsoft Academic Search

A 5-hour colorimetric polymerase chain reaction (PCR) assay was more sensitive than viral culture in identifying viral infection in initial serum (13\\/16 vs 5\\/16; p = 0.008) and urine (10\\/16 vs 5\\/16; p = 0.2) specimens from 16 enterovirus- infected newborn infants, and remained more sensitive throughout their illnesses. Combined sensitivity of serum and urine PCR was 14 of 16

Mark J. Abzug; Michael Loeffelholz; Harley A. Rotbart

1995-01-01

174

Reverse transcription PCR to detect enteroviruses in surface water.  

PubMed Central

We have developed a simple, fast, and efficient procedure to detect enteroviruses in water samples. Aliquots of water are subjected to two-step filtration, with the second filter containing a positively charged nylon membrane that holds back virus particles. Viruses thus adsorbed are directly lysed, and RNA is isolated by hybridization to specific oligonucleotides bound to magnetic beads. The solution used contains guanidine thiocyanate, which lyses virus particles, inactivates enzymes, e.g., RNases, allows mild hybridization conditions, and does not influence biotin-streptavidin interaction on magnetic beads. Detection and specific identification are accomplished by reverse transcription PCR of the highly conserved noncoding region at the 5' end of virus RNA combined with Southern hybridization. The system was tested with tap water artificially spiked with poliovirus vaccine and yielded a detection limit of 20 50% tissue culture infective doses per liter. We used the same procedure to investigate the water quality of surface water at public beaches by rivers and lakes. Of 40 samples tested, 7 were positive for enteroviruses. A comparison with enterobacterial contamination determined by PCR and classical microbiological methods in parallel showed that enteroviruses were found only in samples also positive for Escherichia coli. In conclusion, this procedure can easily be adapted to test large water samples and is simple enough to be used for routine determinations of water quality in terms of virus contamination.

Gilgen, M; Wegmuller, B; Burkhalter, P; Buhler, H P; Muller, U; Luthy, J; Candrian, U

1995-01-01

175

T cell Responses to Enterovirus Antigens and to ?-cell Autoantigens in Unaffected Children Positive for IDDM-Associated Autoantibodies  

Microsoft Academic Search

Enterovirus infections have been implicated in the pathogenesis of IDDM in a number of studies. The aim of the present study was to evaluate whether the cellular immune response to enterovirus antigens is abnormal in children who test positive for IDDM-associated autoantibodies. Lymphocyte proliferation responses were analysed to enterovirus antigens and to a panel of ?-cell autoantigen preparations in 31

S Juhela; H Hyöty; A Hinkkanen; J Elliott; M Roivainen; P Kulmala; J Rahko; M Knip; J Ilonen

1999-01-01

176

Enterovirus Co-infections and Onychomadesis after Hand, Foot, and Mouth Disease, Spain, 2008  

PubMed Central

Hand, foot, and mouth disease (HFMD), a common disease caused by enteroviruses (EVs), usually affects children. Clustered and sporadic HFMD cases, followed by onychomadesis (nail shedding), occurred during summer and fall 2008 in Valencia, Spain. Fecal samples from onychomadesis patients, who did or did not have previous HFMD, and from healthy children exposed to onychomadesis patients tested positive for EV. The complete viral protein 1 capsid gene sequence was obtained for typing and phylogenetic analysis. Two EV serotypes, coxsackievirus A10 and coxsackievirus B1 (CVB1), were mainly detected as a monoinfection or co-infection in a childcare center where an onychomadesis outbreak occurred. On the basis of our results, and detection of CVB1 in 2 other contemporary onychomadesis outbreaks in childcare centers in Spain, we propose that mixed infection of an EV serotype that causes HFMD, plus the serotype CVB1, could explain the emergence after HFMD of onychomadesis, a rare and late complication.

Gonzalez-Candelas, Fernando; Valero, Ana; Cordoba, Juan; Salazar, Antonio

2011-01-01

177

High frequency and diversity of species C enteroviruses in Cameroon and neighboring countries.  

PubMed

Human enteroviruses (HEVs) are endemic worldwide and among the most common viruses infecting humans. Nevertheless, there are very limited data on the circulation and genetic diversity of HEVs in developing countries and sub-Saharan Africa in particular. We investigated the circulation and genetic diversity of HEVs among 436 healthy children in a limited area of the far north region of Cameroon in 2008 and 2009. We also characterized the genetic biodiversity of 146 nonpolio enterovirus (NPEV) isolates obtained throughout the year 2008 from stool specimens of patients with acute flaccid paralysis (AFP) in Cameroon, Chad, and Gabon. We found a high rate of NPEV infections (36.9%) among healthy children in the far north region of Cameroon. Overall, 45 different HEV types were found among healthy children and AFP patients. Interestingly, this study uncovered a high rate of HEVs of species C (HEV-C) among all typed NPEVs: 63.1% (94/149) and 39.5% (49/124) in healthy children and AFP cases, respectively. Besides extensive circulation, the most prevalent HEV-C type, coxsackievirus A-13, featured a tremendous intratypic diversity. Africa-specific HEV lineages were discovered, including HEV-C lineages and the recently reported EV-A71 "genogroup E." Virtually all pathogenic circulating vaccine-derived polioviruses (cVDPVs) that have been fully characterized were recombinants between oral poliovaccine (OPV) strains and cocirculating HEV-C strains. The extensive circulation of diverse HEV-C types and lineages in countries where OPV is massively used constitutes a major viral factor that could promote the emergence of recombinant cVDPVs in the Central African subregion. PMID:23254123

Sadeuh-Mba, Serge Alain; Bessaud, Maël; Massenet, Denis; Joffret, Marie-Line; Endegue, Marie-Claire; Njouom, Richard; Reynes, Jean-Marc; Rousset, Dominique; Delpeyroux, Francis

2012-12-19

178

EBNA3B-deficient EBV promotes B cell lymphomagenesis in humanized mice and is found in human tumors  

PubMed Central

Epstein-Barr virus (EBV) persistently infects more than 90% of the human population and is etiologically linked to several B cell malignancies, including Burkitt lymphoma (BL), Hodgkin lymphoma (HL), and diffuse large B cell lymphoma (DLBCL). Despite its growth transforming properties, most immune-competent individuals control EBV infection throughout their lives. EBV encodes various oncogenes, and of the 6 latency-associated EBV-encoded nuclear antigens, only EBNA3B is completely dispensable for B cell transformation in vitro. Here, we report that infection with EBV lacking EBNA3B leads to aggressive, immune-evading monomorphic DLBCL-like tumors in NOD/SCID/?c–/– mice with reconstituted human immune system components. Infection with EBNA3B-knockout EBV (EBNA3BKO) induced expansion of EBV-specific T cells that failed to infiltrate the tumors. EBNA3BKO-infected B cells expanded more rapidly and secreted less T cell–chemoattractant CXCL10, reducing T cell recruitment in vitro and T cell–mediated killing in vivo. B cell lines from 2 EBV-positive human lymphomas encoding truncated EBNA3B exhibited gene expression profiles and phenotypic characteristics similar to those of tumor-derived lines from the humanized mice, including reduced CXCL10 secretion. Screening EBV-positive DLBCL, HL, and BL human samples identified additional EBNA3B mutations. Thus, EBNA3B is a virus-encoded tumor suppressor whose inactivation promotes immune evasion and virus-driven lymphomagenesis.

White, Robert E.; Ramer, Patrick C.; Naresh, Kikkeri N.; Meixlsperger, Sonja; Pinaud, Laurie; Rooney, Cliona; Savoldo, Barbara; Coutinho, Rita; Bodor, Csaba; Gribben, John; Ibrahim, Hazem A.; Bower, Mark; Nourse, Jamie P.; Gandhi, Maher K.; Middeldorp, Jaap; Cader, Fathima Z.; Murray, Paul; Munz, Christian; Allday, Martin J.

2012-01-01

179

Left ventricular dysfunction in children with fulminant enterovirus 71 infection: an evaluation of the clinical course.  

PubMed

We describe 2 children with typical hand, foot, and mouth disease due to enterovirus 71 infection, 1 of whom died. Both cases were complicated by acute fulminant shock syndrome; the patients had remarkable acute left ventricular dysfunction. The clinical experience indicates that the rapid death associated with fulminant enterovirus rhombencephalitis is the result of rapid cardiogenic shock rather than neurogenic pulmonary edema. PMID:11880970

Huang, Fang-Liang; Jan, Sheng-Ling; Chen, Po-Yen; Chi, Ching-Shiang; Wang, Teh-Ming; Fu, Yun-Ching; Tsai, Chi-Ren; Chang, Yan

2002-03-04

180

A Study of Porcine Enteroviruses Isolated from Swine in the Toronto Area  

PubMed Central

A number of enteric viruses isolated from swine in the Connaught Medical Research Laboratories and the Mimico Reformatory herds were grouped serologically and compared with previously described porcine enteroviruses. Four of five antigenic types defined were related to other porcine enteroviruses. The fifth was unrelated to any viruses with which it was compared in this study.

Thorsen, J.; Macpherson, L. W.

1966-01-01

181

Improvement of Enterovirus Neutralization by Treatment with Sodium Deoxycholate or Chloroform  

Microsoft Academic Search

Summary Enterovirus strains which could be neutralized only partially or not at all by specific antiserum were readily accessible to the antibodies after treatment with sodium deoxycholate or chloroform. The latter treatment is simple and can be performed with crude virus suspensions of low titer. The method was particularly successful for routine typing of enterovirus 71 and coxsackievirus types A7

Jacoba G. Kapsenberg; Albert Ras; Jantje Korte

1979-01-01

182

Rapid Diagnosis of Enterovirus Infection by a New One-Step Reverse Transcription-PCR Assay  

Microsoft Academic Search

The AMPLICOR Enterovirus Test was evaluated with 103 cerebrospinal fluid (CSF) specimens. Twenty- seven CSF specimens were culture positive. With the AMPLICOR test, enterovirus RNA was detected in 34 specimens. Compared with culture, the AMPLICOR test gave a sensitivity of 96.3% and a specificity of 100%. The sensitivity of culture was 79.4% in comparison with the AMPLICOR test. Clinical presentation

HARALD H. KESSLER; BRIGITTE SANTNER; HOLGER RABENAU; ANNEMARIE BERGER; ADRIANA VINCE; CAROL LEWINSKI; BERNARD WEBER; KAREN PIERER; DORIS STUENZNER; EGON MARTH; ANDHANS W. DOERR

183

Enterovirus 71 isolated from cases of epidemic poliomyelitis-like disease in Bulgaria  

Microsoft Academic Search

Summary Virological and serological studies of an epidemic disease in Bulgaria, 1975, were carried out. Epidemiologically, clinically and pathomorphologically, the disease simulated almost all known forms of poliomyelitis, acute stem encephalitis, encephalomyocarditis and aseptic meningitis. The studies completely ruled out the participation of polioviruses and provided comprehensive evidence for the etiological role of a peculiar enterovirus subsequently identified as enterovirus

M. Chumakov; M. VOROSttILOVA; L. Shindarov; I. Lavrova; L. Gracheva; G. Koroleva; S. Vasilenko; I. Brodvarova; M. Nikolova; S. Gyurova; M. Gacheva; G. Mitov; N. Ninov; E. Tsylka; I. Robinson; M. Frolova; V. Bashkirtsev; L. Martiyanova; V. Rodin

1979-01-01

184

Epidemic of Hand, Foot and Mouth Disease Associated with Enterovirus 71 Infection  

Microsoft Academic Search

Summary Viruses isolated from patients with hand, foot and mouth disease in widespread outbreaks in Japan in 1973 were identified as enterovirus 71. Although cases with aseptic meningitis were observed concurrently, the main clinical symptom associated with enterovirus 71 infection was hand, foot and mouth disease.Copyright © 1978 S. Karger AG, Basel

Akio Hagiwara; Isamu Tagaya; Tetsuo Yoneyama

1978-01-01

185

Detection of enterovirus specific RNA sequences in muscle biopsy specimens from patients with adult onset myositis  

Microsoft Academic Search

A subgenomic cDNA probe with broad specificity for a range of enteroviruses was used to test by an in situ hybridisation technique for the presence of enterovirus specific genomic sequences in muscle biopsy samples obtained from patients with chronic adult myositis. Virus specific RNA sequences were detected in 6\\/13 (46%) patients with idiopathic polymyositis or dermatomyositis. Control samples obtained from

G E Yousef; D A Isenberg; J F Mowbray

1990-01-01

186

Reverse Transcription Multiplex PCR for Differentiation between Polio and Enteroviruses from Clinical and Environmental Samples  

Microsoft Academic Search

For the rapid detection of polioviruses and their differentiation from nonpoliovirus enteroviruses, we developed a protocol in which clinical or environmental specimens are first inoculated onto cell cultures in tubes. After overnight incubation, the cultures are subjected to reverse transcription multiplex PCR with a primer pair which detects all enteroviruses (T. Hyypia¨, P. Auvinen, and M. Maaronen, J. Gen. Virol.

DENISE EGGER; LUIS PASAMONTES; MARIANNE OSTERMAYER; ANDKURT BIENZ

1995-01-01

187

Rapid and sensitive detection of enteroviruses in specimens from patients with aseptic meningitis.  

PubMed Central

A 5-h PCR assay (Amplicor enterovirus test) was compared with viral culture for the detection of enteroviruses in cerebrospinal fluid. Of the cerebrospinal fluid specimens collected during a summer outbreak of aseptic meningitis, 34% were positive by viral culture whereas 66% were positive by the Amplicor PCR, suggesting that this technique improves the diagnosis of enteroviral meningitis.

Yerly, S; Gervaix, A; Simonet, V; Caflisch, M; Perrin, L; Wunderli, W

1996-01-01

188

Detection of human parvovirus B19 in papillary thyroid carcinoma  

Microsoft Academic Search

To evaluate whether parvovirus B19, a common human pathogen, was also involved in papillary thyroid carcinoma (PTC), 112 paraffin-embedded thyroid specimens of benign nodules, papillary, medullary and follicular carcinomas, and normal controls were examined for B19 DNA and capsid protein by nested PCR, in situ hybridisation (ISH) and immunohistochemistry (IHC). The expression of the nuclear factor-?B (NF-?B) was investigated by

J H Wang; W P Zhang; H X Liu; D Wang; Y F Li; W Q Wang; L Wang; F R He; Z Wang; Q G Yan; L W Chen; G S Huang; GS Huang

2008-01-01

189

New insights in the regulation of human B cell differentiation  

PubMed Central

B lymphocytes provide the cellular basis of the humoral immune response. All stages of this process, from B cell activation to formation of germinal centers and differentiation into memory B cells or plasma cells, are influenced by extrinsic signals and controlled by transcriptional regulation. Compared to naïve B cells, memory B cells display a distinct expression profile, which allows for their rapid secondary responses. Indisputably, many B cell malignancies result from aberrations in the circuitry controlling B cell function, particularly during the GC reaction. Here we review new insights into memory B cell subtypes, recent literature on transcription factors regulating human B cell differentiation, and further evidence for B cell lymphomagenesis emanating from errors during the GC cell reactions.

Schmidlin, Heike; Diehl, Sean A.; Blom, Bianca

2009-01-01

190

Metabolic activation of aflatoxin B1 by human placental microsomes.  

PubMed

The metabolic activation of aflatoxin B1 by human placental microsomes was studied. Aflatoxin B1 showed relatively high mutagenic activity in Ames test when incubated with human placental microsomes. Addition of alpha-naphthoflavone or aminoglutethimide, known inhibitors of cytochrome P450 1A and P450 19, respectively, into the test system partially inhibited the mutagen-producing activity. It was suggested that the activation of aflatoxin B1 in human placental microsomes is mediated by at least these two forms of cytochrome P450. PMID:8331693

Sawada, M; Kitamura, R; Norose, T; Kitada, M; Itahashi, K; Kamataki, T

1993-05-01

191

Human B Lymphocytes and B Lymphomas Express PPAR-? and Are Killed by PPAR-? Agonists  

Microsoft Academic Search

This paper evaluates the expression and functional significance of PPAR-? on human B cells. Recent interest in PPAR-? has focused on its adipogenic effects on non-bone marrow-derived cells. PPAR-? agonists also have been proposed as anti-inflammatory agents owing to inhibition of NF-?B activation. We report herein the first study evaluating PPAR-? expression and functional significance in human B lineage cells.

Josué Padilla; Edmund Leung; Richard P. Phipps

2002-01-01

192

Bovine lactoferrin digested with human gastrointestinal enzymes inhibits replication of human echovirus 5 in cell culture.  

PubMed

Many infant formulas are enriched with lactoferrin (Lf) because of its claimed beneficial effects on health. Native bovine Lf (bLf) is known to inhibit in vitro replication of human enteroviruses, a group of pathogenic viruses that replicate in the gut as their primary infection site. On the basis of a model digestion and human gastrointestinal enzymes, we hypothesized that bLf could retain its antiviral properties against enterovirus in the gastrointestinal tract, either as an intact protein or through bioactive peptide fragments released by digestive enzymes. To test our hypothesis, bLf was digested with human gastric juice and duodenal juice in a 2-step in vitro digestion model. Two gastric pH levels and reduction conditions were used to simulate physiological conditions in adults and infants. The antiviral activity of native bLf and of the digested fractions was studied on echovirus 5 in vitro, using various assay conditions, addressing several mechanisms for replication inhibition. Both native and digested bLf fractions revealed a significant inhibitory effect, when added before or simultaneously with the virus onto the cells. Furthermore, a significant stronger sustained antiviral effect was observed when bLf was fully digested in the gastric phase with fast pH reduction to 2.5, compared with native bLf, suggesting the release of antiviral peptides from bLf during the human digestion process. In conclusion, this study demonstrates that bLf may have a role in the prevention of human gastrointestinal virus infection under physiological conditions and that food containing bLf may protect against infection in vivo. PMID:22901558

Furlund, Camilla B; Kristoffersen, Anja B; Devold, Tove G; Vegarud, Gerd E; Jonassen, Christine M

2012-07-18

193

Amphotericin B induced abnormalities in human platelets  

PubMed Central

Aims—To investigate in vitro the effect of amphotericin B on platelets in order to understand poor platelet recovery in patients receiving platelet transfusions and amphotericin B simultaneously. Methods—Washed platelets were isolated from platelet concentrates and exposed to amphotericin B (4 ?g/ml) for one hour. Platelet function was assessed by aggregation response to thrombin (0-0.6 U/ml), serotonin release, response to hypotonic stress, and mean platelet volume. The expression of surface membrane glycoprotein (GP) Ib-IX complex, GPIIb-IIIa complex and CD62P (P-selectin) was examined by flow cytometry using fluorescence labelled monoclonal antibodies. Heterotypic cell adhesion was measured in amphotericin B treated platelets coincubated with isolated, autologous polymorphonuclear leucocytes (PMN) by flow cytometric analysis. Results—Amphotericin B induced platelet dysfunction. The rate of aggregation by thrombin, serotonin uptake and thrombin induced release of serotonin, and the response of platelets to hypotonic stress were inhibited. There was up to a two-fold increase in the mean platelet volume. The expression of platelet surface GPIb-IX and GPIIb-IIIa was not affected. P-selectin, normally expressed only on the surface of activated platelets, was also expressed on unactivated platelets. Amphotericin B increased platelet adherence to PMN and the number of platelets bound per PMN. Conclusions—In vitro, amphotericin B induces P-selectin expression on the surface of unactivated platelets and increases platelet adhesion to PMN, which is exacerbated by storage. Platelet dysfunction resulting from exposure to amphotericin B may contribute to poor platelet recovery in vivo when amphotericin B is administered concomitantly with platelet transfusion.

Pastakia, K B; Brownson, N E; Terle, D A; Poindexter, B J

1996-01-01

194

Aging Affects Human B Cell Responses  

Microsoft Academic Search

Aging represents a complex remodeling in which both innate and adaptive immunities deteriorate. Age-related changes in humoral\\u000a immunity are responsible for the reduced vaccine responses observed in elderly individuals. Although T cell alterations play\\u000a a significant role in age-related humoral immune changes, alterations in B cells also occur. We here provide an overview of\\u000a age-related changes in B cell markers

Daniela Frasca; Bonnie B. Blomberg

195

Therapeutic Human Hyperimmune Polyclonal Antibodies Against Staphylococcal Enterotoxin B.  

National Technical Information Service (NTIS)

This SBIR project aims to develop hyperimmune human polyclonal antibody that neutralizes the toxic activity of staphylococcal enterotoxin B (SEB) as treatment for toxic shock induced by SEB disseminated as aerosol in a biowarfare scenario. The primary goa...

M. J. Aman

2008-01-01

196

bZIPDB : A database of regulatory information for human bZIP transcription factors  

PubMed Central

Background Basic region-leucine zipper (bZIP) proteins are a class of transcription factors (TFs) that play diverse roles in eukaryotes. Malfunctions in these proteins lead to cancer and various other diseases. For detailed characterization of these TFs, further public resources are required. Description We constructed a database, designated bZIPDB, containing information on 49 human bZIP TFs, by means of automated literature collection and manual curation. bZIPDB aims to provide public data required for deciphering the gene regulatory network of the human bZIP family, e.g., evaluation or reference information for the identification of regulatory modules. The resources provided by bZIPDB include (1) protein interaction data including direct binding, phosphorylation and functional associations between bZIP TFs and other cellular proteins, along with other types of interactions, (2) bZIP TF-target gene relationships, (3) the cellular network of bZIP TFs in particular cell lines, and (4) gene information and ontology. In the current version of the database, 721 protein interactions and 560 TF-target gene relationships are recorded. bZIPDB is annually updated for the newly discovered information. Conclusion bZIPDB is a repository of detailed regulatory information for human bZIP TFs that is collected and processed from the literature, designed to facilitate analysis of this protein family. bZIPDB is available for public use at .

Ryu, Taewoo; Jung, Juhyun; Lee, Sunjae; Nam, Ho Jung; Hong, Sun Woo; Yoo, Jae Wook; Lee, Dong-ki; Lee, Doheon

2007-01-01

197

Rapid determination of vitamin B2 and B12 in human urine by isocratic liquid chromatography.  

PubMed

A simple and rapid method for the identification and quantification of vitamin B(2) and B(12) in human urine has been developed using reverse phase high performance liquid chromatography (HPLC) and the peaks identity were confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). HPLC separation was performed in single wavelength detector (lambda(365)) mode and separated isocratically using mobile phase methanol: 1mM aqueous TFA (1:4) in C18 column. The calibration graphs plotted with different concentrations of vitamin B(2) and B(12) was linear with a correlation coefficients (r(2))=0.9975 and 0.9985, respectively. The recoveries of vitamin B(2) and B(12) were above 87% and 90%, respectively. The results of this present study suggest that the proposed method may be simple and convenient way of identifying and quantifying vitamin B(2) and B(12) from human urine. PMID:19362629

Mandal, Santi M; Mandal, Mahitosh; Ghosh, Ananta K; Dey, Satyahari

2009-03-17

198

Inhibition of human neutrophil chemotaxis and chemiluminescence by amphotericin B.  

PubMed Central

The effect of the antifungal drugs amphotericin B, fluocytosine, miconazole, griseofulvin, and nystatin on the chemotactic responsiveness of human neutrophils was studied. Amphotericin B in a concentration of 2 mug/ml inhibited chemotatic responsiveness, and in a concentration of 5 mug/ml it also inhibited chemiluminescence. The inhibition of chemotaxis could be reversed by washing the cells. The other antifungal drugs did not inhibit chemotaxis even in concentrations much higher than those obtained in human serum during treatment.

Bjorksten, B; Ray, C; Quie, P G

1976-01-01

199

Acute cerebellar ataxia with human parvovirus B19 infection  

Microsoft Academic Search

A 2 year old boy developed acute cerebellar ataxia in association with erythema infectiosum. During the disease, genomic DNA and antibodies against human parvovirus B19 were detected in serum but not in cerebrospinal fluid. Parvovirus B19 associated acute cerebellar ataxia might occur due to transient vascular reaction in the cerebellum during infection.

Yoshitaka Shimizu; Tamaki Ueno; Hiroshi Komatsu; Hiroshi Takada; Tadasu Nunoue

1999-01-01

200

Streptozotocin is not toxic to the human fetal B cell  

Microsoft Academic Search

Summary  It has been generally assumed that because streptozotocin is toxic to the adult B cell of most species, it should also damage B cells obtained at earlier stages of development. This paper examines whether this is true for human fetal pancreata obtained from the therapeutic termination of pregnancies during the first half of the second trimester. Experiments were carried out

B. E. Tuch; J. R. Turtle; C. J. Simeonovic

1989-01-01

201

Mutant human cells with constitutive activation of NF-B  

Microsoft Academic Search

We have used a genetic approach to generate eight different mutant human cell lines in which NF-B is constitutively activated. These independent clones have different phenotypes and belong to several different genetic complementation groups. In one clone inhibitor of B(IB) kinase is constitutively active, but in the seven others it is not, despite the fact that IB is degraded in

Swati S. Sathe; Nywana Sizemore; Xiaoxia Li; Kalpa Vithalani; Mairead Commane; Shannon M. Swiatkowski; George R. Stark

2004-01-01

202

Predominant Autoantibody Production by Early Human B Cell Precursors  

Microsoft Academic Search

During B lymphocyte development, antibodies are assembled by random gene segment reassortment to produce a vast number of specificities. A potential disadvantage of this process is that some of the antibodies produced are self-reactive. We determined the prevalence of self-reactive antibody formation and its regulation in human B cells. A majority (55 to 75%) of all antibodies expressed by early

Hedda Wardemann; Sergey Yurasov; Anne Schaefer; James W. Young; Eric Meffre; Michel C. Nussenzweig

2003-01-01

203

Human BLyS Facilitates Engraftment of Human PBL Derived B Cells in Immunodeficient Mice  

PubMed Central

The production of fully immunologically competent humanized mice engrafted with peripheral lymphocyte populations provides a model for in vivo testing of new vaccines, the durability of immunological memory and cancer therapies. This approach is limited, however, by the failure to efficiently engraft human B lymphocytes in immunodeficient mice. We hypothesized that this deficiency was due to the failure of the murine microenvironment to support human B cell survival. We report that while the human B lymphocyte survival factor, B lymphocyte stimulator (BLyS/BAFF) enhances the survival of human B cells ex vivo, murine BLyS has no such protective effect. Although human B cells bound both human and murine BLyS, nuclear accumulation of NF-?B p52, an indication of the induction of a protective anti-apoptotic response, following stimulation with human BLyS was more robust than that induced with murine BLyS suggesting a fundamental disparity in BLyS receptor signaling. Efficient engraftment of both human B and T lymphocytes in NOD rag1?/? Prf1?/? immunodeficient mice treated with recombinant human BLyS is observed after adoptive transfer of human PBL relative to PBS treated controls. Human BLyS treated recipients had on average 40-fold higher levels of serum Ig than controls and mounted a de novo antibody response to the thymus-independent antigens in pneumovax vaccine. The data indicate that production of fully immunologically competent humanized mice from PBL can be markedly facilitated by providing human BLyS.

Schmidt, Madelyn R.; Appel, Michael C.; Giassi, Lisa J.; Greiner, Dale L.; Shultz, Leonard D.; Woodland, Robert T.

2008-01-01

204

Stimulation of human B lymphocytes by Listeria cell wall fraction.  

PubMed Central

Cell wall fraction of Listeria monocytogenes (LCWF), a B cell mitogen for mouse spleen cells, is also mitogenic for human adult and cord peripheral blood lymphocytes. Purified B-cell suspensions responded to LCWF in vitro proliferation, to a similar extent as the unfractionated suspensions. Furthermore, LCWF-induced B cell differentiation into IgM-containing cells and their percentage correlated significantly with the extent of lymphocyte proliferation.

Ivanyi, L

1978-01-01

205

The Evolutionary Dynamics of Human Influenza B Virus  

Microsoft Academic Search

Despite their close phylogenetic relationship, type A and B influenza viruses exhibit major epidemiological differences in\\u000a humans, with the latter both less common and less often associated with severe disease. However, it is unclear what processes\\u000a determine the evolutionary dynamics of influenza B virus, and how influenza viruses A and B interact at the evolutionary scale.\\u000a To address these questions

Rubing Chen; Edward C. Holmes

2008-01-01

206

Genomic Analysis of the Human B-Lymphotropic Virus (HBLV)  

Microsoft Academic Search

The human B-lymphotropic virus (HBLV) has a double-stranded DNA genome of greater than 110 kilobase pairs, which is consistent with its morphological classification as a herpesvirus. A 9000--base pair cloned probe of HBLV detected specific sequences in DNA and RNA of infected cells but did not hybridize to the genomic DNA of other human herpesviruses including the Epstein-Barr virus, human

Steven F. Josephs; S. Zaki Salahuddin; Dharam V. Ablashi; Francois Schachter; Flossie Wong-Staal; Robert C. Gallo

1986-01-01

207

Detection of human parvovirus B19 in papillary thyroid carcinoma  

PubMed Central

To evaluate whether parvovirus B19, a common human pathogen, was also involved in papillary thyroid carcinoma (PTC), 112 paraffin-embedded thyroid specimens of benign nodules, papillary, medullary and follicular carcinomas, and normal controls were examined for B19 DNA and capsid protein by nested PCR, in situ hybridisation (ISH) and immunohistochemistry (IHC). The expression of the nuclear factor-?B (NF-?B) was investigated by IHC. The results showed B19 DNA commonly exists in human thyroid tissues; however, there were significant differences between PTC group and normal controls, and between PTC and nonneoplastic adjacent tissues (P<0.001). The presence of viral DNA in PTC neoplastic epithelium was confirmed by laser-capture microdissection and sequencing of nested PCR products. B19 capsid protein in PTC group was significantly higher than that of all the control groups and nonneoplastic adjacent tissues (P?0.001). Compared with control groups, the activation of NF-?B in PTC group was significantly increased (P?0.02), except for medullary carcinomas, and the activation of NF-?B was correlated with the viral protein presence (P=0.002). Moreover, NF-?B was colocalised with B19 DNA in the neoplastic epithelium of PTC by double staining of IHC and ISH. These results indicate for the first time a possible role of B19 in pathogenesis of PTC.

Wang, J H; Zhang, W P; Liu, H X; Wang, D; Li, Y F; Wang, W Q; Wang, L; He, F R; Wang, Z; Yan, Q G; Chen, L W; Huang, G S

2008-01-01

208

Preparation of Immune Serum to Enteroviruses.  

National Technical Information Service (NTIS)

Certification tests for infectivity, identity and purity were completed on 11 reference seeds. These were Mg-stabilized seeds of coxsackievirus B5, echoviruses 13, 15, 24, 25, 27, 31, and original fluid seeds of coxsackievirus A16, echoviruses 9 (Vispo), ...

J. L. Melnick B. Hampil

1966-01-01

209

Diagnosis of Enterovirus Infection by Genus-Specific PCR and Enzyme-Linked Immunosorbent Assays  

PubMed Central

PCR for the diagnosis of enterovirus infections is resource intensive but is increasingly used due to wide availability. Enzyme-linked immunosorbent assays (ELISAs) that detect heterotypical antibodies against enterovirus immunoglobulin M (IgM), IgA, and IgG were compared with reverse transcription-PCR by using primers specific to the 5? untranslated regions of 60 enterovirus species. The ELISAs were less sensitive than the PCR, and only the ELISA for IgM was highly specific. When retrospective diagnosis is important or when specimens are unsuitable for PCR, the ELISA has a limited role if PCR is not available.

Craig, Maria E.; Robertson, Peter; Howard, Neville J.; Silink, Martin; Rawlinson, William D.

2003-01-01

210

Evidence of association between type 1 diabetes and exposure to enterovirus in Cuban children and adolescents.  

PubMed

A possible etiologic role of enteroviruses for type 1 diabetes has been researched for 40 years, but evidence to date is inconclusive. This article summarizes new evidence from Cuban research supporting a role for enteroviruses, both in preclinical stages of autoimmune reactions against pancreatic ? cells and at clinical onset, in a population with low type 1 diabetes incidence. Possible pathogenetic mechanisms are also discussed, such as acute cytolytic damage and molecular mimicry. Although direct causal effects of enteroviruses in type 1 diabetes are difficult to demonstrate, arguments supporting their role in type 1 diabetes pathogenesis should not be ignored; and confirmation could contribute to development of more effective preventive strategies. PMID:23396239

Sarmiento, Luis; Cubas-Dueñas, Ileana; Cabrera-Rode, Eduardo

2013-01-01

211

Age effects on B cells and humoral immunity in humans  

PubMed Central

Both humoral and cellular immune responses are impaired in aged individuals, leading to decreased vaccine responses. Although T cell defects occur, defects in B cells play a significant role in age-related humoral immune changes. The ability to undergo class switch recombination (CSR), the enzyme for CSR, AID (activation-induced cytidine deaminase) and the transcription factor E47 are all decreased in aged stimulated B cells. We here present an overview of age-related changes in human B cell markers and functions, and also discuss some controversies in the field of B cell aging.

Frasca, Daniela; Diaz, Alain; Romero, Maria; Landin, Ana Marie; Blomberg, Bonnie B

2010-01-01

212

Development of a method for detection of enteroviruses in shellfish by PCR with poliovirus as a model.  

PubMed Central

The application of the PCR to complex samples is hindered by amplification inhibitors. We describe a reverse transcription-PCR-based method capable of inhibitor removal for the detection of enteroviruses in shellfish. Initial virus extraction stages based on a modified polyethylene glycol precipitation technique (G.D. Lewis and T.G. Metcalf, Appl. Environ. Microbiol. 54:1983-1988, 1988) were followed by virus purification with 1,1,2-trichloro,2,2,1-trifluoroethane and concentration by ultrafiltration. A guanidine isothiocyanate-glass powder extraction system was utilized for sample lysis, RNase protection, and nucleic acid purification. Removal of PCR inhibitors and method sensitivity were quantified in shellfish (oysters and mussels) seeded with poliovirus. PCR sample tolerance exceeded 4 g for depurated shellfish; however, polluted field samples were more inhibitory. Virus recoveries of 31% for oyster extracts and 17% for mussel extracts and nucleic acid extraction reverse transcription-PCR detection limits down to 1 PFU yielded an overall sensitivity limit of < 10 PFU of poliovirus in up to 5 g of shellfish. PCR-positive results were obtained from a variety of polluted field samples naturally contaminated with human enteroviruses. The methods developed for virus recovery and PCR inhibitor removal should be equally applicable to detection of other RNA viruses such as hepatitis A virus, Norwalk virus, and other small round-structured viruses in shellfish. Images

Lees, D N; Henshilwood, K; Dore, W J

1994-01-01

213

B lymphocytes in human subcutaneous adipose crown-like structures  

PubMed Central

Accumulation of macrophages and T cells within crown-like structures (CLS) in subcutaneous adipose tissue predicts disease severity in obesity related insulin resisance (OIR). While rodent data suggest the B cell is an important feature of these lesions, B cells have not been described within the human CLS. In order to identify B cells in the human subcutaneous CLS (sCLS) in obese subjects and determine if the presence of B cells predict insulin resistance, we examined archived samples of subcutaneous and omental fat from 32 obese men and women and related findings to clinical parameters. Using immunohistochemistry we identified B (CD19+) and T cells (CD3+) within the sCLS and perivascular space. B cell presence and density (B cells pHPF, T cells pHPF and B:T cell ratio) were compared with measures of insulin resistance (HOMA) and other variables. In sixteen of thirty-two subjects (50%) CD19+ B cells were localized within sCLS and were relatively more numerous than T cells. HOMA was not different between subjects with CD19+ vs. CD19? sCLS (5.5 vs. 5.3, p= 0.88). After controlling for diabetes and glycemia (HA1c), the B:T cell ratio correlated with current metformin treatment (r=0.89, p = 0.001). These results indicate that in human OIR, B cells are an integral component of organized inflammation in subcutaneous fat, and defining their role will lead to a better understanding of OIR pathogenesis and potentially impact treatment.

McDonnell, Marie E.; Ganley-Leal, Lisa M.; Mehta, Ankeeta; Bigornia, Sherman; Mott, Melanie; Rehman, Qasim; Farb, Melissa G; Hess, Donald; Joseph, Lija; Gokce, Noyan; Apovian, Caroline M.

2013-01-01

214

Expression and Initial Characterization of Human ALDH3B1  

PubMed Central

Aldehyde dehydrogenases (ALDHs) are critical enzymes in the metabolism of endogenous and exogenous aldehydes. The human genome contains nineteen putatively functional ALDH genes; ALDH3B1 belongs to the ALDH3 family. While recent studies have linked the ALDH3B1 locus to schizophrenia, nothing was known, until now, about the properties and significance of the ALDH3B1 protein. The aim of this study was to characterize the ALDH3B1 protein. Human ALDH3B1 was baculovirus-expressed and found to be catalytically active towards medium- and long-chain aliphatic aldehydes and the aromatic aldehyde benzaldehyde. Western blot analyses indicate that ALDH3B1 is highly expressed in kidney and liver and moderately expressed in various brain regions. ALDH3B1-transfected HEK293 cells were significantly protected against cytotoxicity induced by the lipid-peroxidation product octanal when compared to vector-transfected cells. This study shows for the first time the functionality, expression and protective role of ALDH3B1 and indicates a potential physiological role of ALDH3B1 against oxidative stress.

Marchitti, Satori A.; Orlicky, David J.; Vasiliou, Vasilis

2007-01-01

215

Oral immunization of mice using Bifidobacterium longum expressing VP1 protein from enterovirus 71.  

PubMed

Bifidobacterium longum is an attractive candidate for delivering biologically active proteins by the mucosal route due to its non-pathogenic and colonizing properties. Enterovirus 71 (EV71) has aroused widespread attention recently due to several epidemics, and great attention should be paid to the fact that there are currently no effective antiviral drugs or vaccines against EV71 infection. In this report, we described a recombinant B. longum that could be used to develop an oral vaccine against EV71 infection. A VP1 expression vector (pBBADs-VP1) was constructed by amplifying the EV71 VP1 gene and inserting it into the E. coli-Bifidobacterium shuttle expression vector pBBAD/Xs. Then, the expression of VP1 protein in pBBADs-VP1-transformed bacteria was demonstrated by western blot. In vivo studies indicated that oral immunization of BALB/c mice with pBBADs-VP1-transformed bacteria induced potent immune responses against EV71 infection, including virus-neutralising titers, anti-EV71-VP1 antibody and the induction of Th1 immune responses in the spleen and Peyer's patches. Importantly, immunization of mother mice with this recombinant VP1-expressing B. longum conferred protection to neonatal mice. These results demonstrate that the novel oral vaccine utilizing B. longum expressing the VP1 protein might successfully elicit a specific immune response against EV71 infection. PMID:23275129

Yu, Zhijian; Huang, Zhen; Sao, Chongwen; Huang, Yuanjian; Zhang, Fan; Ma, Guihong; Chen, Zhong; Zeng, Zhongming; Qiwen, Deng; Zeng, Weiseng

2013-01-01

216

AN INTEGRATED CELL CULTURE/RT-PCR METHOD FOR DETECTING ENTEROVIRUS IN WATER  

EPA Science Inventory

Echovirus and coxsackievirus can cause mild to severe disease following consumption of contaminated drinking water. However, comprehensive occurrence studies of enteroviruses in drinking water matrices are limited, in part because of the lack of available methods that are rapid, ...

217

Combined 5' UTR RFLP analysis and VP1 sequencing for epidemic investigation of enteroviruses.  

PubMed

Enteroviruses, the main cause of aseptic meningitis, consist of 100 serotypes, and many of them have been associated with large outbreaks. In the present study, a comparison of RFLP analysis of the 5' untranslated region (5'UTR) and sequencing of both the 5'UTR and VP1 regions was conducted for epidemiological linkage of 27 clinical enterovirus strains. The clinical enterovirus strains were clustered into five restriction profile groups. Even though the restriction profile clusters of clinical isolates were not related to those of the respective prototype strains, epidemiological relationships between the members of each cluster were observed. The restriction profile clusters in the 5'UTR corresponded to the phylogenetic clusters in the VP1 genomic region. The incongruence between the topology of Gior strain in 5'UTR and VP1 phylogenetic trees indicates a recombination event. The proposed RFLP assay in combination with VP1 sequencing can offer crucial epidemiological information about the circulating enteroviruses. PMID:22983155

Kyriakopoulou, Zaharoula; Tsolis, Kostas; Pliaka, Vaia; Tsakogiannis, Dimitris; Ruether, Irina Georgia Anna; Gartzonika, Constantina; Levidiotou-Stefanou, Stamatina; Markoulatos, Panayotis

2012-09-16

218

A sensor-adaptor mechanism for enterovirus uncoating from structures of EV71  

PubMed Central

Enterovirus 71 (EV71), a major agent of hand-foot-and-mouth disease in children, can cause severe central nervous system disease and mortality. At present no vaccine or antiviral therapy is available. We have determined high-resolution structures for the mature virus and natural empty particles. The structure of the mature virus is similar to that of other enteroviruses, whilst the empty particles are dramatically expanded, with notable fissures, resembling elusive enterovirus uncoating intermediates not previously characterized in atomic detail. Hydrophobic capsid pockets within the EV71 capsid are collapsed in this expanded particle, providing a detailed explanation of the mechanism for receptor-binding triggered virus uncoating. The results provide a paradigm for enterovirus uncoating, in which the VP1 GH loop acts as an adaptor-sensor for the attachment of cellular receptors, converting heterologous inputs to a generic uncoating mechanism, spotlighting novel points for therapeutic intervention.

Wang, Xiangxi; Peng, Wei; Ren, Jingshan; Hu, Zhongyu; Xu, Jiwei; Lou, Zhiyong; Li, Xumei; Yin, Weidong; Shen, Xinliang; Porta, Claudine; Walter, Thomas S.; Evans, Gwyndaf; Axford, Danny; Owen, Robin; Rowlands, David J.; Wang, Junzhi; Stuart, David I.; Fry, Elizabeth E.; Rao, Zihe

2012-01-01

219

Comparison of Heart Failure in Children with Enterovirus 71 Rhombencephalitis and Cats with Norepinephrine Cardiotoxicity  

Microsoft Academic Search

The mechanism of heart failure in patients with enterovirus 71 rhombencephalitis (brain stem encephalitis) remains unknown.\\u000a Our previous reports hypothesized that a catecholamine storm induced by rhombencephalitis may account for the heart failure.\\u000a The aim of this study was to develop a novel feline model of norepinephrine cardiotoxicity and compare the resulting heart\\u000a failure to that in children with enterovirus

Y.-C. Fu; C. Cheng; N.-N. Lin; S.-L. Jan; B. Hwang; S.-L. Hsu; C.-L. Gong; Y.-T. Chen; Y.-T. Chiu

2006-01-01

220

Enterovirus RNA in Blood Is Linked to the Development of Type 1 Diabetes  

PubMed Central

OBJECTIVE To assess whether the detection of enterovirus RNA in blood predicts the development of clinical type 1 diabetes in a prospective birth cohort study. Further, to study the role of enteroviruses in both the initiation of the process and the progression to type 1 diabetes. RESEARCH DESIGN AND METHODS This was a nested case-control study where all case children (N = 38) have progressed to clinical type 1 diabetes. Nondiabetic control children (N = 140) were pairwise matched for sex, date of birth, hospital district, and HLA-DQ–conferred genetic susceptibility to type 1 diabetes. Serum samples, drawn at 3- to 12-month intervals, were screened for enterovirus RNA using RT-PCR. RESULTS Enterovirus RNA–positive samples were more frequent among the case subjects than among the control subjects. A total of 5.1% of the samples (17 of 333) in the case group were enterovirus RNA–positive compared with 1.9% of the samples (19 of 993) in the control group (P < 0.01). The strongest risk for type 1 diabetes was related to enterovirus RNA positivity during the 6-month period preceding the first autoantibody-positive sample (odds ratio 7.7 [95% CI 1.9–31.5]). This risk effect was stronger in boys than in girls. CONCLUSIONS The present study supports the hypothesis that enteroviruses play a role in the pathogenesis of type 1 diabetes, especially in the initiation of the ?-cell damaging process. The enterovirus-associated risk for type 1 diabetes may be stronger in boys than in girls.

Oikarinen, Sami; Martiskainen, Mika; Tauriainen, Sisko; Huhtala, Heini; Ilonen, Jorma; Veijola, Riitta; Simell, Olli; Knip, Mikael; Hyoty, Heikki

2011-01-01

221

Characterization of a novel porcine enterovirus in domestic pig in Hungary  

Microsoft Academic Search

Porcine enteroviruses (PEVs) of genus Enterovirus are small, non-enveloped viruses with single-stranded, positive sense genomic RNA, belonging to the family Picornaviridae. The discovery of two distinct serotypes (PEV9 and 10) was first reported in 1979. Despite the sporadic detection and partial genome sequences of these viruses our knowledge about the prevalence and molecular epidemiology of PEV types in domestic pigs

Ákos Boros; Péter Pankovics; Gábor Reuter

2011-01-01

222

Genetic Analysis with Man-Mouse Somatic Cell Hybrids: Linkage between Human Lactate Dehydrogenase B and Peptidase B Genes  

Microsoft Academic Search

Evidence of an active linkage between the human genes that control lactate dehydrogenase B and peptidase B is presented. It is also concluded that there is no link between the genes for lactate dehydrogenase A and lactate dehydrogenase B.

A. Silvana Santachiara; M. Nabholz; V. Miggiano; A. J. Darlington; W. Bodmer

1970-01-01

223

Telomere lengthening and telomerase activation during human B cell differentiation  

PubMed Central

The function of the immune system is highly dependent on cellular differentiation and clonal expansion of antigen-specific lymphocytes. However, little is known about mechanisms that may have evolved to protect replicative potential in actively dividing lymphocytes during immune differentiation and response. Here we report an analysis of telomere length and telomerase expression, factors implicated in the regulation of cellular replicative lifespan, in human B cell subsets. In contrast to previous observations, in which telomere shortening and concomitant loss of replicative potential occur in the process of somatic cell differentiation and cell division, it was found that germinal center (GC) B cells, a compartment characterized by extensive clonal expansion and selection, had significantly longer telomeric restriction fragments than those of precursor naive B cells. Furthermore, it was found that telomerase, a telomere-synthesizing enzyme, is expressed at high levels in GC B cells (at least 128-fold higher than those of naive and memory B cells), correlating with the long telomeres in this subset of B cells. Finally, both naive and memory B cells were capable of up-regulating telomerase activity in vitro in response to activation signals through the B cell antigen receptor in the presence of CD40 engagement and/or interleukin 4. These observations suggest that a novel process of telomere lengthening, possibly mediated by telomerase, functions in actively dividing GC B lymphocytes and may play a critical role in humoral immune response by maintaining the replicative potential of GC and descendant memory B cells.

Weng, Nan-ping; Granger, Lawrence; Hodes, Richard J.

1997-01-01

224

Type 1 diabetes is associated with enterovirus infection in gut mucosa.  

PubMed

Enterovirus infections have been linked to type 1 diabetes in several studies. Enteroviruses also have tropism to pancreatic islets and can cause ?-cell damage in experimental models. Viral persistence has been suspected to be an important pathogenetic factor. This study evaluates whether gut mucosa is a reservoir for enterovirus persistence in type 1 diabetic patients. Small-bowel mucosal biopsy samples from 39 type 1 diabetic patients, 41 control subjects, and 40 celiac disease patients were analyzed for the presence of enterovirus using in situ hybridization (ISH), RT-PCR, and immunohistochemistry. The presence of virus was compared with inflammatory markers such as infiltrating T cells, HLA-DR expression, and transglutaminase 2-targeted IgA deposits. Enterovirus RNA was found in diabetic patients more frequently than in control subjects and was associated with a clear inflammation response in the gut mucosa. Viral RNA was often detected in the absence of viral protein, suggesting defective replication of the virus. Patients remained virus positive in follow-up samples taken after 12 months' observation. The results suggest that a large proportion of type 1 diabetic patients have prolonged/persistent enterovirus infection associated with an inflammation process in gut mucosa. This finding opens new opportunities for studying the viral etiology of type 1 diabetes. PMID:22315304

Oikarinen, Maarit; Tauriainen, Sisko; Oikarinen, Sami; Honkanen, Teemu; Collin, Pekka; Rantala, Immo; Mäki, Markku; Kaukinen, Katri; Hyöty, Heikki

2012-02-07

225

Type 1 Diabetes Is Associated With Enterovirus Infection in Gut Mucosa  

PubMed Central

Enterovirus infections have been linked to type 1 diabetes in several studies. Enteroviruses also have tropism to pancreatic islets and can cause ?-cell damage in experimental models. Viral persistence has been suspected to be an important pathogenetic factor. This study evaluates whether gut mucosa is a reservoir for enterovirus persistence in type 1 diabetic patients. Small-bowel mucosal biopsy samples from 39 type 1 diabetic patients, 41 control subjects, and 40 celiac disease patients were analyzed for the presence of enterovirus using in situ hybridization (ISH), RT-PCR, and immunohistochemistry. The presence of virus was compared with inflammatory markers such as infiltrating T cells, HLA-DR expression, and transglutaminase 2–targeted IgA deposits. Enterovirus RNA was found in diabetic patients more frequently than in control subjects and was associated with a clear inflammation response in the gut mucosa. Viral RNA was often detected in the absence of viral protein, suggesting defective replication of the virus. Patients remained virus positive in follow-up samples taken after 12 months’ observation. The results suggest that a large proportion of type 1 diabetic patients have prolonged/persistent enterovirus infection associated with an inflammation process in gut mucosa. This finding opens new opportunities for studying the viral etiology of type 1 diabetes.

Oikarinen, Maarit; Tauriainen, Sisko; Oikarinen, Sami; Honkanen, Teemu; Collin, Pekka; Rantala, Immo; Maki, Markku; Kaukinen, Katri; Hyoty, Heikki

2012-01-01

226

Maternal Enterovirus Infection as a Risk Factor for Type 1 Diabetes in the Exposed Offspring  

PubMed Central

OBJECTIVE Maternal enterovirus infections during pregnancy have been linked to an increased risk of type 1 diabetes in the offspring. The aim of this study was to evaluate this association in a unique series of pregnant mothers whose child progressed to clinical type 1 diabetes. RESEARCH DESIGN AND METHODS Maternal and in utero enterovirus infections were studied in 171 offspring who presented with type 1 diabetes before the age of 11 years and in 316 control subjects matched for date and place of birth, sex, and HLA-DQ risk alleles for diabetes. Acute enterovirus infections were diagnosed by increases in enterovirus IgG and IgM in samples taken from the mother at the end of the first trimester of pregnancy and cord blood samples taken at delivery. RESULTS Signs of maternal enterovirus infection were observed in altogether 19.3% of the mothers of affected children and in 12.0% of the mothers of control children (P = 0.038). This difference was seen in different HLA risk groups and in both sexes of the offspring, and it was unrelated to the age of the child at the diagnosis of diabetes or the age of the mother at delivery. CONCLUSIONS These results suggest that an enterovirus infection during pregnancy is not a major risk factor for type 1 diabetes in childhood but may play a role in some susceptible subjects.

Viskari, Hanna; Knip, Mikael; Tauriainen, Sisko; Huhtala, Heini; Veijola, Riitta; Ilonen, Jorma; Simell, Olli; Surcel, Helja-Marja; Hyoty, Heikki

2012-01-01

227

An enterovirus strain isolated from diabetic child belongs to a genetic subcluster of echovirus 11, but is also neutralised with monotypic antisera to coxsackievirus A9.  

PubMed

An enterovirus strain (designated D207) isolated from a Slovakian diabetic child and originally serotyped as coxsackievirus A9 (CAV-9) was found to cause rapid cytolysis coinciding with severe functional damage of the surviving cells in primary cultures of human pancreatic islets. This finding prompted us to clone the isolate for full-length genome sequencing and molecular characterization as the prototype strain of CAV-9 is known to cause only minimal damage to insulin-producing beta-cells. Based on capsid-coding sequence comparisons, the isolate turned out to be echovirus 11 (E-11). Phylogenetic analyses demonstrated that E-11/D207 was closely related to a specific subgroup B of E-11 strains known to cause uveitis. To study further antigenic properties of isolate E-11/D207 and uveitis-causing E-11 strains, neutralization experiments were carried out with CAV-9- and E-11-specific antisera. Unlike the prototype strains, the isolate E-11/D207 and uveitis-causing E-11 strains were well neutralized with both CAV-9- and E-11-specific antisera. Attempts to identify recombination of the capsid coding sequences as a reason for double-reactivity using the Simplot analysis failed to reveal major transferred motifs. However, peptide scanning technique was able to identify antigenic regions of capsid proteins of E-11/D207 as well as regions cross-reacting with an antiserum raised to CAV-9. Thus, double specificity of E-11/D207 seems to be a real characteristic shared by the phylogenetically closely related virus strains in the genetic subgroup B of E-11. PMID:18632967

Al-Hello, Haider; Paananen, Anja; Eskelinen, Mervi; Ylipaasto, Petri; Hovi, Tapani; Salmela, K; Lukashev, Alexander N; Bobegamage, Shubhada; Roivainen, Merja

2008-08-01

228

Passive protection effect of chicken egg yolk immunoglobulins on enterovirus 71 infected mice.  

PubMed

The objective of this study is to evaluate the passive protective efficiency of immunoglobulin in yolk (IgY) specific against human enterovirus type 71 (EV71). The antibody was raised by intramuscular immunization to 10 White Leghorn hens, with inactivated human EV71 serving as the antigen. The titer and specificity of the antibody were analyzed from purified IgY in the egg yolks of immunized hens. Results indicate that the titer of IgY specific against EV71 increased from the third week after the first immunization. The content of total IgY was 190 ± 26 mg/yolk, with an average concentration of specific IgY of 6.34 ± 3.38 mg/yolk in the eggs from 3 to 18 wk after immunization. The results of the neutralization effect of specific IgY in EV71-challenged mice demonstrate that the EV71-specific IgY, either by intraperitoneal injection or oral administration, was able to significantly reduce the morbidity and mortality in EV71 infected mice pups. PMID:20937321

Liou, Jenn-Fa; Chang, Chih-Wei; Tailiu, Jui-jane; Yu, Chun-Keung; Lei, Huan-Yao; Chen, Lih-Ren; Tai, Chein

2010-10-29

229

The relationship between arthritis and human parvovirus B19 infection  

Microsoft Academic Search

In order to evaluate the role of human parvovirus B19 in the etiopathogenesis of autoimmune diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), synovial fluid and blood specimens were collected at 1-month intervals from 20 patients with early synovitis (ES) and 31 with RA. Blood specimens were also collected from 25 patients with SLE, 25 with osteoarthritis

R. Caliskan; S. Masatlioglu; M. Aslan; S. Altun; S. Saribas; S. Ergin; E. Uckan; V. Koksal; V. Oz; K. Altas; I. Fresko; B. Kocazeybek

2005-01-01

230

Manipulation of human B cells to confer immortality.  

PubMed

Electroporation was used to deliver genomic DNA from a lymphoid tumor to activated/stimulated human peripheral blood lymphocytes to create immortalized lymphoid cell lines. Activation of the recipient lymphocytes was essential for efficient immortalization. A panel of human B cell transfectant clones, each phenotypically representing specific stages of differentiation, resulted from the transfection. Monoclonal antibody production was measured, and the level produced depended on the phenotype of the cells, with the more mature B cell transfectants secreting up to 10 micrograms/mL of immunoglobulin. The transfectants were stable with respect to their morphological appearance, growth rate, and antibody production. Chromosome analysis indicated that the transfectants displayed a normal karyotype, devoid of abnormalities. We have shown that electroporation is an effective method of immortalizing human lymphocytes at different stages of differentiation. The transfectants provide a panel of cells that can readily be studied with respect to their phenotypic/karyotypic stability, regulation, and production of immunoglobulin, lymphokines, and growth factors. These data demonstrate the feasibility of generating immortalized human B cells to provide an important resource for the study of B cell differentiation and immortalization. PMID:1335787

Jonak, Z L; Henri, E; Trulli, S; Jensen, C P; Muirhead, K A

1992-10-01

231

B iochemical mechanisms of hibernation and stunning in the human  

Microsoft Academic Search

a ¨¨ ¨ Institute of Pharmacology and Toxicology , Hospital of the Westf alische Wilhelms -Universitat, Domagkstraße 12, D-48149 Munster, Germany b Abstract Background: Myocardial hibernation and stunning are characterized by depressed cardiac function in the presence of reduced or normal coronary blood flow. The underlying biochemical mechanisms are widely unknown and only limited data are available in human hearts.

Hartmut Luss; Michael Schafers; Joachim Neumann; Dieter Hammel; Christian Vahlhaus; Hideo A. Baba; Frauke Janssen; Hans H. Scheld; Otmar Schober; Gunter Breithardt; Wilhelm Schmitz; Thomas Wichter

232

Neurological manifestations of human parvovirus B19 infection.  

PubMed

Since its discovery, human parvovirus B19 has been linked with a broad spectrum of clinical syndromes. An aetiological role for the virus has been confirmed in erythema infectiosum, transient aplastic crisis, persistent infection manifesting as pure red cell aplasia in immunocompromised persons, non-immune hydrops fetalis and arthritis. Less commonly recognised, but receiving increasing attention recently, are the neurological manifestations, a variety of which have been described in patients with either clinically diagnosed or laboratory confirmed B19 infection. The purpose of this review is to summarise present knowledge of B19, its known and potential pathogenic mechanisms and its association with human diseases, particularly those with neurological manifestations. The outcome of the review supports an aetiological role of the virus in neurological disease. However, the pathogenesis remains unknown and elucidating this is a priority. PMID:12740833

Barah, Faraj; Vallely, Pamela J; Cleator, Graham M; Kerr, Jonathan R

233

CD23b Isoform Expression in Human Schistosomiasis Identifies a Novel Subset of Activated B Cells ?  

PubMed Central

Resistance to schistosomiasis is associated with increased levels of serum parasite-specific IgE. IgE exerts its functions through its cellular receptors, Fc?RI and Fc?RII/CD23; however, its functional significance in humans requires further characterization. We previously reported that increased levels of CD23+ B cells correlate with resistance to schistosomiasis in hyperexposed populations and sought to define their potential function and relationship with IgE. We found that CD23+ B cells are a heterogeneous cell population with functional and phenotypic differences. Circulating CD23+ B cells are uniquely activated in schistosomiasis and express the CD23b isoform and CXCR5, the homing receptor for lymphoid follicles. High CXCR5 expression by CD23+ B cells was associated with the capacity to home to the cognate ligand CXCL13. CD23-bound IgE cross-linking increased surface expression of CXCR5, suggesting that CD23+ B cells home directly into the lymphoid follicles upon antigen capture. As human schistosomiasis is an intravascular parasitic infection associated with a high antigenic burden in the blood, circulating CD23+ B cells may play a role in the capture and shuttling of antigens directly to splenic follicles, highlighting a new role for circulating B cells. This function likely plays an important role in the development of protective immunity to infection with schistosomes.

Onguru, Daniel; Liang, YanMei; Elliot, Jennifer; Mwinzi, Pauline; Ganley-Leal, Lisa

2011-01-01

234

CD23b isoform expression in human schistosomiasis identifies a novel subset of activated B cells.  

PubMed

Resistance to schistosomiasis is associated with increased levels of serum parasite-specific IgE. IgE exerts its functions through its cellular receptors, Fc?RI and Fc?RII/CD23; however, its functional significance in humans requires further characterization. We previously reported that increased levels of CD23(+) B cells correlate with resistance to schistosomiasis in hyperexposed populations and sought to define their potential function and relationship with IgE. We found that CD23(+) B cells are a heterogeneous cell population with functional and phenotypic differences. Circulating CD23(+) B cells are uniquely activated in schistosomiasis and express the CD23b isoform and CXCR5, the homing receptor for lymphoid follicles. High CXCR5 expression by CD23(+) B cells was associated with the capacity to home to the cognate ligand CXCL13. CD23-bound IgE cross-linking increased surface expression of CXCR5, suggesting that CD23(+) B cells home directly into the lymphoid follicles upon antigen capture. As human schistosomiasis is an intravascular parasitic infection associated with a high antigenic burden in the blood, circulating CD23(+) B cells may play a role in the capture and shuttling of antigens directly to splenic follicles, highlighting a new role for circulating B cells. This function likely plays an important role in the development of protective immunity to infection with schistosomes. PMID:21708991

Onguru, Daniel; Liang, YanMei; Elliot, Jennifer; Mwinzi, Pauline; Ganley-Leal, Lisa

2011-06-27

235

Alterations in Peripheral Blood B Cell Subsets and Dynamics of B Cell Responses during Human Schistosomiasis  

PubMed Central

Antibody responses are thought to play an important role in control of Schistosoma infections, yet little is known about the phenotype and function of B cells in human schistosomiasis. We set out to characterize B cell subsets and B cell responses to B cell receptor and Toll-like receptor 9 stimulation in Gabonese schoolchildren with Schistosoma haematobium infection. Frequencies of memory B cell (MBC) subsets were increased, whereas naive B cell frequencies were reduced in the schistosome-infected group. At the functional level, isolated B cells from schistosome-infected children showed higher expression of the activation marker CD23 upon stimulation, but lower proliferation and TNF-? production. Importantly, 6-months after 3 rounds of praziquantel treatment, frequencies of naive B cells were increased, MBC frequencies were decreased and with the exception of TNF-? production, B cell responsiveness was restored to what was seen in uninfected children. These data show that S. haematobium infection leads to significant changes in the B cell compartment, both at the phenotypic and functional level.

Labuda, Lucja A.; Ateba-Ngoa, Ulysse; Feugap, Eliane Ngoune; Heeringa, Jorn J.; van der Vlugt, Lucien E. P. M.; Pires, Regina B. A.; Mewono, Ludovic; Kremsner, Peter G.; van Zelm, Menno C.; Adegnika, Ayola A.; Yazdanbakhsh, Maria; Smits, Hermelijn H.

2013-01-01

236

Alterations in peripheral blood B cell subsets and dynamics of B cell responses during human schistosomiasis.  

PubMed

Antibody responses are thought to play an important role in control of Schistosoma infections, yet little is known about the phenotype and function of B cells in human schistosomiasis. We set out to characterize B cell subsets and B cell responses to B cell receptor and Toll-like receptor 9 stimulation in Gabonese schoolchildren with Schistosoma haematobium infection. Frequencies of memory B cell (MBC) subsets were increased, whereas naive B cell frequencies were reduced in the schistosome-infected group. At the functional level, isolated B cells from schistosome-infected children showed higher expression of the activation marker CD23 upon stimulation, but lower proliferation and TNF-? production. Importantly, 6-months after 3 rounds of praziquantel treatment, frequencies of naive B cells were increased, MBC frequencies were decreased and with the exception of TNF-? production, B cell responsiveness was restored to what was seen in uninfected children. These data show that S. haematobium infection leads to significant changes in the B cell compartment, both at the phenotypic and functional level. PMID:23505586

Labuda, Lucja A; Ateba-Ngoa, Ulysse; Feugap, Eliane Ngoune; Heeringa, Jorn J; van der Vlugt, Luciën E P M; Pires, Regina B A; Mewono, Ludovic; Kremsner, Peter G; van Zelm, Menno C; Adegnika, Ayola A; Yazdanbakhsh, Maria; Smits, Hermelijn H

2013-03-07

237

Association between human parvovirus B19 infection and arthritis.  

PubMed Central

OBJECTIVE--To gain information concerning the association between parvovirus B19 infection and arthritis. METHODS--Blood or synovial fluid, or both, from a total of 77 adult patients with various arthropathies (rheumatoid arthritis 13; mechanical arthropathies 11; crystal induced arthritis 13; idiopathic mono/oligoarthritis 25; suspicion of viral arthritis 15) were tested for the presence of the viral genome and anti-B19 antibodies. B19 DNA in blood and synovial fluid was investigated by nested polymerase chain reaction, and anti-B19 IgM and IgG antibodies were detected in blood by enzyme immunoassay. RESULTS--A recent parvovirus infection was documented by the presence of anti-B19 IgM antibodies in the blood of 13 patients. B19 DNA, together with anti-B19 IgM and IgG antibodies, were detected in the blood of seven patients who had an acute transient arthritis, putatively of viral origin. Viral DNA was detected in a synovial fluid sample and in the blood of one patient with monoarthritis who had an anti-B19 IgG response only. CONCLUSIONS--The prevalence of anti-B19 IgG antibody in these patients with various forms of arthritis (63%) was within the same range as that in the general population (blood donors). However, for the patients with clinical suspicion of viral arthritis, the increased seroprevalence of anti-B19 IgM and the presence of the B19 genome point to an association between human parvovirus infections and acute forms of arthritis.

Cassinotti, P; Bas, S; Siegl, G; Vischer, T L

1995-01-01

238

Effect of B-activin on human T suppressor cells  

SciTech Connect

The authors studied the influence of B-activin on the effect of human concanavalin A (con A)-induced T suppressor cells and also on the process of induction of T suppressor cells by con A and stimulation of proliferative activity of lymphocytes by phytohemagglutinin (PHA). Con A-induced suppression and the effect of B-activin on it were studied in a system in which the test cell culture and the culture for induction of suppressors were prepared simultaneously. Peripheral blood was obtained from blood donors for the experiments and during the preparation of the experiments, /sup 3/H-thymidine was added. The results of investigation of the influence of B-activin on the effect of con A-induced suppressors and also on the process of their induction are given. It is concluded that B-activin blocks the effect of con A-induced human suppressor cells but does not affect their induction, and B-activin does not affect proliferative activity of lymphocytes induced by PHA.

Gambarov, S.S.; Khzardzhyan, A.M.; Adamyan, N.V.; Shakhsuvarov, A.V.; Suzdal'tseva, A.A.; Rakhmanova, G.A.

1986-10-01

239

Human parvovirus B19-associated disease in bone marrow transplantation.  

PubMed

Human parvovirus B19 can persist in immunocompromised patients and may produce severe clinical illness. In this retrospective study the incidence of B19-associated infections in bone marrow transplant patients was investigated. During 1 year 60 patients received bone marrow grafts (eight autografts and 52 allogeneic transplantations). In case of early onset, atypical or steroid-resistant erythrodermia the patients' blood and/or tissue specimens were screened for B19 infection by polymerase chain reaction (PCR). Additionally, specimens of patients with severe organ failure were tested. A total of 64 PCRs was performed in 27 patients. Seven patients with erythrodermia and one with vulvovaginitis proved to be PCR positive. In patients with organ failure B19 DNA was detected in the myocardium and liver. The incidence of B19 infections in this cohort was 15% and the B19-associated mortality rate 7%. In conclusion, parvovirus B19-associated infections may be more common in immunocompromised patients than previously anticipated. PMID:10219641

Schleuning, M; Jäger, G; Holler, E; Hill, W; Thomssen, C; Denzlinger, C; Lorenz, T; Ledderose, G; Wilmanns, W; Kolb, H J

240

Molecular epidemiology of enteroviruses with special reference to their potential role in the etiology of insulin-dependent diabetes mellitus (IDDM). A review  

Microsoft Academic Search

Background: Several lines of evidence suggest that enterovirus infections may be involved in the etiology of the insulin-dependent diabetes mellitus (IDDM). Often in the literature, a reference is given to specifically diabetogenic strains of enterovirus but there is no systematic assessment about the generation of such strains in the course of evolution or about their abundance among the 64 enterovirus

Tapani Hovi

1998-01-01

241

Enterovirus 71 Disrupts Interferon Signaling by Reducing the Level of Interferon Receptor 1  

PubMed Central

The recent outbreak of enterovirus 71 (EV71) infected millions of children and caused over 1,000 deaths. To date, neither an effective vaccine nor antiviral treatment is available for EV71 infection. Interferons (IFNs) have been successfully applied to treat patients with hepatitis B and C viral infections for decades but have failed to treat EV71 infections. Here, we provide the evidence that EV71 antagonizes type I IFN signaling by reducing the level of interferon receptor 1 (IFNAR1). We show that the host cells could sense EV71 infection and stimulate IFN-? production. However, the induction of downstream IFN-stimulated genes is inhibited by EV71. Also, only a slight interferon response and antiviral effects could be detected in cells treated with recombinant type I IFNs after EV71 infection. Further studies reveal that EV71 blocks the IFN-mediated phosphorylation of STAT1, STAT2, Jak1, and Tyk2 by reducing IFNAR1. Finally, we identified the 2A protease encoded by EV71 as an antagonist of IFNs and show that the protease activity is required for reducing IFNAR1 levels. Taken together, our study for the first time uncovers a mechanism used by EV71 to antagonize type I IFN signaling and provides new targets for future antiviral strategies.

Lu, Jing; Yi, Lina; Zhao, Jin; Yu, Jun; Chen, Ying; Lin, Marie C.; Kung, Hsiang-Fu

2012-01-01

242

Enterovirus 71 disrupts interferon signaling by reducing the level of interferon receptor 1.  

PubMed

The recent outbreak of enterovirus 71 (EV71) infected millions of children and caused over 1,000 deaths. To date, neither an effective vaccine nor antiviral treatment is available for EV71 infection. Interferons (IFNs) have been successfully applied to treat patients with hepatitis B and C viral infections for decades but have failed to treat EV71 infections. Here, we provide the evidence that EV71 antagonizes type I IFN signaling by reducing the level of interferon receptor 1 (IFNAR1). We show that the host cells could sense EV71 infection and stimulate IFN-? production. However, the induction of downstream IFN-stimulated genes is inhibited by EV71. Also, only a slight interferon response and antiviral effects could be detected in cells treated with recombinant type I IFNs after EV71 infection. Further studies reveal that EV71 blocks the IFN-mediated phosphorylation of STAT1, STAT2, Jak1, and Tyk2 by reducing IFNAR1. Finally, we identified the 2A protease encoded by EV71 as an antagonist of IFNs and show that the protease activity is required for reducing IFNAR1 levels. Taken together, our study for the first time uncovers a mechanism used by EV71 to antagonize type I IFN signaling and provides new targets for future antiviral strategies. PMID:22258259

Lu, Jing; Yi, Lina; Zhao, Jin; Yu, Jun; Chen, Ying; Lin, Marie C; Kung, Hsiang-Fu; He, Ming-Liang

2012-01-18

243

Enhancement of enterovirus infectivity in vitro by pretreating host cell monolayers with the cationic polymer polyethyleneimine.  

PubMed Central

Laboratory strains of enteroviruses, as well as viruses isolated from raw wastewater, were found to exhibit enhanced infectivity in vitro when BGM cell monolayers were pretreated with the cationic polymer polyethyleneimine (PEI). Viruses were assayed by the cytopathic effect technique and as PFU under methylcellulose and agar overlays with monolayers treated with 0 to 5.0 x 10(-3)% (wt/vol) PEI in phosphate-buffered saline supplemented with 2% fetal bovine serum. Poliovirus type 1 cytopathic effect occurred at an enhanced rate in cells treated with 5.0 x 10(-3)% PEI compared with untreated cells. PEI-treated cells were found to adsorb viruses much more effectively than untreated cells did. When the methylcellulose overlay procedure was used, rates of infectivity were enhanced as follows: poliovirus type 1, 5.5-fold; echovirus type 1, 1.2-fold; echovirus type 5, 5.2-fold; and coxsackievirus type B5, 4.9-fold. Viruses concentrated from raw wastewater showed a 3.8-fold increase in titer when quantitated by the most-probable-number method and a 3.3-fold increase when quantitated as PFU under an agar overlay.

Preston, D R; Bitton, G; Farrah, S R

1990-01-01

244

Rupintrivir is a promising candidate for treating severe cases of Enterovirus-71 infection  

PubMed Central

AIM: To evaluate the suitability of rupintrivir against Enterovirus 71 (EV71) induced severe clinical symptoms using computational methods. METHODS: The structure of EV71 3C protease was predicted by homology modeling. The binding free energies between rupintrivir and EV71 3C and human rhinovirus 3C protease were computed by molecular dynamics and molecular mechanics Poisson-Boltzmann/surface area and molecular mechanics generalized-born/surface area methods. EV71 3C fragments obtained from clinical samples collected during May to July 2008 in Shanghai were amplified by reverse-transcription and polymerase chain reaction and sequenced. RESULTS: We observed that rupintrivir had favorable binding affinity with EV71 3C protease (-10.76 kcal/mol). The variability of the 3C protein sequence in isolates of various outbreaks, including those obtained in our hospital from May to July 2008, were also analyzed to validate the conservation of the drug binding pocket. CONCLUSION: Rupintrivir, whose safety profiles had been proved, is an attractive candidate and can be quickly utilized for treating severe EV71 infection.

Zhang, Xiao-Nan; Song, Zhi-Gang; Jiang, Ting; Shi, Bi-Sheng; Hu, Yun-Wen; Yuan, Zheng-Hong

2010-01-01

245

Chlorogenic Acid inhibits the replication and viability of enterovirus 71 in vitro.  

PubMed

Enterovirus 71 (EV71) is an etiology for a number of diseases in humans. Traditional Chinese herbs have been reported to be effective for treating EV71 infection. However, there is no report about the antiviral effects of CHA against EV71. In this study, plaque reduction assay demonstrated that the inhibitory concentration 50% (IC50) of CHA on EV71 replication is 6.3 µg/ml. When both CHA (20 µg/ml) and EV71 were added, or added post-infection at different time points, CHA was able to effectively inhibit EV71 replication between 0 and 10 h. In addition, CHA inhibited EV71 2A transcription and translation in EV71-infected RD cells, but did not affect VP1, 3C, and 3D expression. Furthermore, CHA inhibited secretions of IL-6, TNF-?, IFN-? and MCP-1 in EV71-infected RD cells. Altogether, these results revealed that CHA may have antiviral properties for treating EV71 infection. PMID:24098754

Li, Xiang; Liu, Yuanyuan; Hou, Xueling; Peng, Hongjun; Zhang, Li; Jiang, Qingbo; Shi, Mei; Ji, Yun; Wang, Yuyue; Shi, Weifeng

2013-09-30

246

Chlorogenic Acid Inhibits the Replication and Viability of Enterovirus 71 In Vitro  

PubMed Central

Enterovirus 71 (EV71) is an etiology for a number of diseases in humans. Traditional Chinese herbs have been reported to be effective for treating EV71 infection. However, there is no report about the antiviral effects of CHA against EV71. In this study, plaque reduction assay demonstrated that the inhibitory concentration 50% (IC50) of CHA on EV71 replication is 6.3 µg/ml. When both CHA (20 µg/ml) and EV71 were added, or added post-infection at different time points, CHA was able to effectively inhibit EV71 replication between 0 and 10 h. In addition, CHA inhibited EV71 2A transcription and translation in EV71-infected RD cells, but did not affect VP1, 3C, and 3D expression. Furthermore, CHA inhibited secretions of IL-6, TNF-?, IFN-? and MCP-1 in EV71-infected RD cells. Altogether, these results revealed that CHA may have antiviral properties for treating EV71 infection.

Hou, Xueling; Peng, Hongjun; Zhang, Li; Shi, Mei; Ji, Yun; Wang, Yuyue

2013-01-01

247

The thiazolobenzimidazole TBZE-029 inhibits enterovirus replication by targeting a short region immediately downstream from motif C in the nonstructural protein 2C  

Microsoft Academic Search

TBZE-029 {1-(2,6-difluorophenyl)-6-trifluoromethyl-1H,3H-thiazolo[3,4-a]benzimidazole} is a novel selective inhibitor of the replication of several enteroviruses. We show that TBZE-029 exerts its antiviral activity through inhibition of viral RNA replication, without affecting polyprotein processing. To identify the viral target of TBZE-029, drug-resistant coxsackievirus B3 (CVB3) was selected. Genotyping of resistant clones led to the identification of three amino acid mutations in nonstructural protein

Armando M. De Palma; Ward Heggermont; Kjerstin Lanke; Bruno Coutard; Mirko Bergmann; Anna-Maria Monforte; Bruno Canard; Erik De Clercq; Alba Chimirri; Gerhard Purstinger; Jacques Rohayem; Frank van Kuppeveld; Johan Neyts

2008-01-01

248

CRH inhibits NF-?B signaling in human melanocytes  

PubMed Central

Corticotropin releasing hormone (CRH), a messenger of stress at the central level, is expressed in the epidermis where it operates within local equivalent of hypothalamo-pituitary axis. CRH inhibits NF-?B activity in human immortalized epidermal (PIG1) melanocytes. In melanocytes CRH stimulates pro-opiomelanocortin (POMC) mRNA and adrenocorticotropin (ACTH) peptide production. Knockdown of POMC levels by transfecting cells with antisense oligonucleotides blocks the effect of CRH on NF-?B signaling indicating that the above inhibition is indirect, e.g. through activation of POMC. We suggest that induction of POMC by CRH serves as a feedback mechanism to self-restrict inflammatory response in the skin.

Zbytek, Blazej; Pfeffer, Lawrence M.; Slominski, Andrzej T.

2007-01-01

249

Coxsackievirus B4 and type 1 diabetes pathogenesis: contribution of animal models.  

PubMed

The role of enteroviruses, in particular type B coxsackieviruses (CV-B), in type 1 diabetes (T1D) pathogenesis is supported by epidemiological, clinical and experimental observations.The investigation of T1D pathogenesis benefits from the contribution of animal models called spontaneously diabetic. Among these animals the non-obese diabetic (NOD) mouse and the bio-breeding diabetes-prone (BBDP) rat present a genetic susceptibility manifested by the expression of an autoimmune diabetes similar to the pathology observed in human beings. Other models whose genetic predisposition is less known are of considerable contribution as well. Numerous major observations relative to several aspects of T1D pathogenesis in the context of CV-B infections, such as susceptibility, diabetogenicity, pancreatotropism, mechanisms of beta cells destruction and others, have been deduced thanks to investigations with animal models. Despite their limits, these models are necessary in improving our knowledge of the role of enteroviruses, like CV-B4, in the pathogenesis of T1D, and the recent advances ensuing from their contribution may have important therapeutic and preventive spin-offs. PMID:19621354

Jaïdane, H; Sané, F; Gharbi, J; Aouni, M; Romond, M B; Hober, D

2009-10-01

250

Comparative Study of the Cytokine/Chemokine Response in Children with Differing Disease Severity in Enterovirus 71-Induced Hand, Foot, and Mouth Disease  

PubMed Central

Background Enterovirus 71 (EV71) infection can lead to a rapidly progressing, life-threatening, and severe neurological disease in young children, including the development of human hand, foot, and mouth disease (HFMD). This study aims to further characterize the specific immunological features in EV71–mediated HFMD patients presenting with differing degrees of disease severity. Methodology Comprehensive cytokine and chemokine expression were broadly evaluated by cytokine antibody array in EV71–infected patients hospitalized for HFMD compared to Coxsackievirus A16-infected patients and age-matched healthy controls. More detailed analysis using Luminex-based cytokine bead array was performed in EV71–infected patients stratified into diverse clinic outcomes. Additionally, immune cell frequencies in peripheral blood and EV71–specific antibodies in plasma were also examined. Principal Findings Expression of several cytokines and chemokines were significantly increased in plasma from EV71–infected patients compared to healthy controls, which further indicated that: (1) GM-CSF, MIP-1?, IL-2, IL-33, and IL-23 secretion was elevated in patients who rapidly developed disease and presented with uncomplicated neurological damage; (2) G-CSF and MCP-1 were distinguishably secreted in EV71 infected very severe patients presenting with acute respiratory failure; (3) IP-10, MCP-1, IL-6, IL-8, and G-CSF levels were much higher in cerebrospinal fluid than in plasma from patients with neurological damage; (4) FACS analysis revealed that the frequency of CD19+HLADR+ mature B cells dynamically changed over time during the course of hospitalization and was accompanied by dramatically increased EV71–specific antibodies. Our data provide a panoramic view of specific immune mediator and cellular immune responses of HFMD and may provide useful immunological profiles for monitoring the progress of EV71–induced fatal neurological symptoms with acute respiratory failure.

Wang, Linghang; Yang, Fan; Hu, Yongfeng; Ren, Xianwen; Li, Guojun; Yu, Yang; Sun, Shaoxia; Li, Yufen; Chen, Xinchun; Li, Xingwang; Jin, Qi

2013-01-01

251

Activation of human monocytes by both human ?1H and C3b  

PubMed Central

We tested whether highly purified human ?1H and C3b, two proteins of the alternative pathway of complement activation, could exert an influence on the activity of human monocytes (M?). The activation process of M? was assessed by measurements of the respiratory burst in terms of nitro blue tetrazolium (NBT) reduction and by chemiluminescence (CL) tests. In NBT reduction experiments, we found a tendency for ?1H to increase NBT reduction, while C3b was found to be rather inhibitory. In CL measurements, both ?1H and C3b displayed a stimulatory effect on M?, showing different time- and dose-dependency. For ?1H, the maximum stimulation occurred after 15 min, whereas for C3b after 45 min. Zymosan particles which served as a positive control also showed the highest stimulation after 45 min. In dose—response experiments, ?1H reached a plateau ranging from 30 to 80 ?g/ml. In contrast, using C3b up to 170 ?g/ml, no plateau was reached. M?-depletion and enrichment studies suggested at M? as being responsible for the stimulatory effects found. The differences between NBT reduction and CL could possibly be explained by the measurement of only cell-bound reductive potentials by NBT reduction, while in CL measurements, products of the extracellular space are also assessed. Our results suggest that both human ?1H and C3b are appropriate stimuli for human monocytes.

Schopf, R. E.; Hammann, K. P.; Scheiner, O.; Lemmel, E.-M.; Dierich, M. P.

1982-01-01

252

Some properties of human and bovine brain cathepsin B  

Microsoft Academic Search

Cathespin B has been purified 750-fold to apparent homogeneity from human and bovine brain cortex using ammonium sulfate fractionation (30–70%), chromatography on Sephadex G-100, CM-Sephadex C-50, and concanavalin A-Sepharose. Enzyme was assayed fluorometrically at pH 4.0 with pyridoxyl-hemoglobin in the presence of 1 mM DTT and 1 mM EDTA. Properties of the enzyme from the two sources proved to be

Anahit Azaryan; Nina Barkhudaryan; Armen Galoyan

1985-01-01

253

Nucleolar Localization of Human Hepatitis B Virus Capsid Protein  

Microsoft Academic Search

Wild-type human hepatitis B virus (HBV) exhibits selective export of virions containing mature genomes. In contrast, changing an isoleucine to a leucine at amino acid 97 (I97L) of the HBV core antigen (HBcAg) causes it to release immature genomes. To elucidate the structure-function relationship of HBcAg at amino acid 97, we systematically replaced the isoleucine residue at this position with

Bo Ning; C. Shih

2004-01-01

254

Coxsackievirus B4 Infection of Human Fetal Thymus Cells  

Microsoft Academic Search

The infection of human fetal thymus organ cultures (FTOC) with coxsackievirus B4 E2 (CVB4 E2) was investigated. Both positive- and negative-strand viral RNA were detected by real-time quantitative reverse transcription-PCR (RT-PCR) in CVB4 E2-infected FTOC, which supported high yields of virus production (106 50% tissue culture infective doses\\/ml), and in flow-sorted thymocyte populations for 7 days after inoculation. Cortical CD4

Fabienne Brilot; Vincent Geenen; Didier Hober; Cheryl A. Stoddart

2004-01-01

255

B. F. Skinner's Legacy to Human Infant Behavior and Development  

Microsoft Academic Search

B. F. Skinner's legacy to human behavioral research for the study of environment–infant interactions, and indeed for the conception of development itself, is described and exemplified. The legacy is largely the practicality, the efficiency, and the comparative advantage—relative to diverse other behavioral and nonbehavioral approaches—of using the operant-learning paradigm to organize and explain many of the sequential changes in behavior

Jacob L. Gewirtz; Martha Peláez-Nogueras

1992-01-01

256

Histone Demethylases KDM4B and KDM6B Promotes Osteogenic Differentiation Of Human MSCs  

PubMed Central

SUMMARY Human bone marrow mesenchymal stem/stromal cells (MSCs) are multipotent progenitor cells with multilineage differentiation potentials including osteogenesis and adipogenesis. While significant progress has been made in understanding transcriptional controls of MSC fate, little is known about how MSC differentiation is epigenetically regulated. Here we show that the histone demethylases KDM4B and KDM6B play critical roles in osteogenic commitment of MSCs by removing H3K9me3 and H3K27me3. Depletion of KDM4B or KDM6B significantly reduced osteogenic differentiation and increased adipogenic differentiation. Mechanistically, while KDM6B controlled HOX expression by removing H3K27me3, KDM4B promoted DLX expression by removing H3K9me3. Importantly, H3K27me3- and H3K9me3-positive MSCs of bone marrow were significantly elevated in ovariectomized and aging mice in which adipogenesis was highly active. Since histone demethylases are chemically modifiable, KDM4B and KDM6B may present as novel therapeutic targets for controlling MSC fate choices, and lead to clues for new treatment in metabolic bone diseases such as osteoporosis.

Ye, Ling; Fan, Zhipeng; Yu, Bo; Chang, Jia; Hezaimi, Khalid Al; Zhou, Xuedong; Park, No-Hee; Wang, Cun-Yu

2012-01-01

257

An investigation into parametric relationships between enterovirus and faecal indicator organisms in the coastal waters of England and Wales  

Microsoft Academic Search

Previous research has resulted in conflicting conclusions concerning possible statistical relationships between bacterial indicator and enterovirus concentrations in marine waters. Results have suggested either highly significant relationships or no significant correlation. To examine these relationships further, large compliance assessment data sets for identified bathing waters on the English and Welsh coast, between 1988 and 1992, were extended to include enterovirus

Mark D. Wyer; Jay M. Fleisher; Jacqueline Gough; David Kay; Helen Merrett

1995-01-01

258

Cleavage of Poly(A)Binding Protein by Enterovirus Proteases Concurrent with Inhibition of Translation In Vitro  

Microsoft Academic Search

Many enteroviruses, members of the family Picornaviridae, cause a rapid and drastic inhibition of host cell protein synthesis during infection, a process referred to as host cell shutoff. Poliovirus, one of the best-studied enteroviruses, causes marked inhibition of host cell translation while preferentially allowing translation of its own genomic mRNA. An abundance of experimental evidence has accumulated to indicate that

MICHELLE JOACHIMS; PIETER C. VAN BREUGEL; RICHARD E. LLOYD

1999-01-01

259

Human Papillomavirus Type 31b Infection of Human Keratinocytes and the Onset of Early Transcription  

PubMed Central

Human papillomaviruses (HPVs) cause a number of human tumors and malignancies, including cervical cancers. Epithelial differentiation is required for the complete HPV life cycle and can be achieved using the organotypic (raft) culture system. The CIN-612 9E cell line maintains episomal copies of HPV type 31b (HPV31b), an HPV type associated with cervical cancers. When grown in the raft system, CIN-612 9E cells form a differentiated epithelium such that infectious virions can be synthesized. Many aspects of the later stages of the HPV31b life cycle have been investigated in CIN-612 9E raft tissues. We used a biologically contained homogenization system for efficient virion extraction from raft epithelial tissues. Purified HPV31b virions were used to infect low-passage-number human foreskin keratinocytes and a variety of epithelial cell lines. Newly synthesized, spliced HPV31b transcripts were detected by reverse transcription and PCR (RT-PCR) following HPV31b infection. HPV31b infection was most efficient and reproducible in HaCaT cells. The onset of viral transcription following infection was also investigated using RT-PCR techniques. Spliced E1?I,E2 RNAs were present as early as 4 h postinfection (p.i.), whereas the other major viral transcripts were detected by 8 to 10 h p.i. Furthermore, we characterized the structures and temporal expression of seven novel spliced early transcripts expressed following infection.

Ozbun, Michelle A.

2002-01-01

260

Detection of enteroviruses and rhinoviruses in clinical specimens by PCR and liquid-phase hybridization.  

PubMed Central

A sensitive method based on PCR followed by liquid-phase hybridization for detection of enterovirus and rhinovirus RNAs in clinical specimens and cell culture supernatants is described. RNA was extracted from stool samples, throat swabs, nasopharyngeal aspirates, cerebrospinal fluid, urine, and plasma with a commercial phenol-guanidinium-chloroform reagent and purified on a polysulfone membrane, on which the reverse transcriptase reaction was also done. Two sets of oligonucleotide primers from the 5' noncoding region of picornaviruses were selected for DNA amplification of 153-bp (enterovirus) and 120-bp (rhinovirus) regions. Double-stranded amplicons were digested into single strands with T7 gene 6 exonuclease and quantitated by an assay using a europium-labeled probe, streptavidin- and biotinylated probe-coated microtitration wells, and time-resolved fluorometry. The sensitivity of the assay was about one template molecule when purified coxsackievirus A9 RNA was used. All enterovirus prototype strains, except echoviruses 22 and 23, and clinical isolates grown in cell culture or suckling mice were strongly positive by the enterovirus PCR-hybridization, as were selected prototype strains and untyped isolates of rhinoviruses by the rhinovirus PCR-hybridization. In a series of 100 clinical specimens tested, the results for 92 agreed with virus culture results. The detection method described will be useful in etiopathogenic studies on enteroviruses and rhinoviruses.

Halonen, P; Rocha, E; Hierholzer, J; Holloway, B; Hyypia, T; Hurskainen, P; Pallansch, M

1995-01-01

261

Human Herpesvirus 6B Genome Sequence: Coding Content and Comparison with Human Herpesvirus 6A  

Microsoft Academic Search

Human herpesvirus 6 variants A and B (HHV-6A and HHV-6B) are closely related viruses that can be readily distinguished by comparison of restriction endonuclease profiles and nucleotide sequences. The viruses are similar with respect to genomic and genetic organization, and their genomes cross-hybridize extensively, but they differ in biological and epidemiologic features. Differences include infectivity of T-cell lines, patterns of

GERALDINA DOMINGUEZ; TIMOTHY R. DAMBAUGH; FELICIA R. STAMEY; STEPHEN DEWHURST; NAOKI INOUE; PHILIP E. PELLETT

1999-01-01

262

Deamidation in Human Lens ?B2-Crystallin Destabilizes the Dimer†  

PubMed Central

Two major determinants of the transparency of the lens are protein-protein interactions and stability of the crystallins, the structural proteins in the lens. ?B2 is the most abundant ?-crystallin in the human lens and is important in formation of the complex interactions of lens crystallins. ?B2 readily forms a homodimer in vitro, with interacting residues across the monomer-monomer interface conserved among ?-crystallins. Due to their long life spans, crystallins undergo an unusually large number of modifications, with deamidation being a major factor. In this study the effects of two potential deamidation sites at the monomer-monomer interface on dimer formation and stability were determined. Glutamic acid substitutions were constructed to mimic the effects of previously reported deamidations at Q162 in the C-terminal domain and at Q70, its N-terminal homologue. The mutants had a nativelike secondary structure similar to that of wild type ?B2 with differences in tertiary structure for the double mutant, Q70E/Q162E. Multiangle light scattering and quasi-elastic light scattering experiments showed that dimer formation was not interrupted. In contrast, equilibrium unfolding and refolding in urea showed destabilization of the mutants, with an inflection in the transition of unfolding for the double mutant suggesting a distinct intermediate. These results suggest that deamidation at critical sites destabilizes ?B2 and may disrupt the function of ?B2 in the lens.

Lampi, Kirsten J.; Amyx, Kencee K.; Ahmann, Petra; Steel, Eric A.

2008-01-01

263

Characterization of a rare IL-10-competent B-cell subset in humans that parallels mouse regulatory B10 cells  

PubMed Central

Regulatory B cells control inflammation and autoimmunity in mice, including the recently identified IL-10–competent B10 cell subset that represents 1% to 3% of spleen B cells. In this study, a comparable IL-10–competent B10 cell subset was characterized in human blood. B10 cells were functionally identified by their ability to express cytoplasmic IL-10 after 5 hours of ex vivo stimulation, whereas progenitor B10 (B10pro) cells required 48 hours of in vitro stimulation before they acquired the ability to express IL-10. B10 and B10pro cells represented 0.6% and approximately 5% of blood B cells, respectively. Ex vivo B10 and B10pro cells were predominantly found within the CD24hiCD27+ B-cell subpopulation that was able to negatively regulate monocyte cytokine production through IL-10–dependent pathways during in vitro functional assays. Blood B10 cells were present in 91 patients with rheumatoid arthritis, systemic lupus erythematosus, primary Sjögren syndrome, autoimmune vesiculobullous skin disease, or multiple sclerosis, and were expanded in some cases as occurs in mice with autoimmune disease. Mean B10 + B10pro-cell frequencies were also significantly higher in patients with autoimmune disease compared with healthy controls. The characterization of human B10 cells will facilitate their identification and the study of their regulatory activities during human disease.

Iwata, Yohei; Matsushita, Takashi; Horikawa, Mayuka; DiLillo, David J.; Yanaba, Koichi; Venturi, Guglielmo M.; Szabolcs, Paul M.; Bernstein, Steven H.; Magro, Cynthia M.; Williams, Armistead D.; Hall, Russell P.; St Clair, E. William

2011-01-01

264

Human parvovirus B19 infection of monocytic cell line U937 and antibody-dependent enhancement  

Microsoft Academic Search

Human parvovirus B19 (B19) infects human erythroid lineage cells. Accumulating evidence also shows that B19 is detectable in nonerythroid lineage cells in vivo, but the mechanism of infection is still not clear. In this study, we explored the mode of B19 infection of human monocytic cell line U937. An in vitro infection study demonstrated B19 binding of U937 and slow

Yasuhiko Munakata; Ichiro Kato; Takako Saito; Takao Kodera; Keiko Kumura Ishii; Takeshi Sasaki

2006-01-01

265

Human papillomavirus type 31b infection of human keratinocytes does not require heparan sulfate.  

PubMed

Oncogenic human papillomaviruses (HPVs) are difficult to study experimentally as they replicate at low levels in vivo. This has precluded the purification of high-risk HPV virions from in vivo lesions. Virus-like particles (VLPs) and pseudovirions from low- and high-risk HPV types can emulate various aspects of HPV virion attachment and infections. These studies suggest that HPV infection is mediated by alpha6-integrin and/or heparan-sulfonated receptors. However, whether VLPs and pseudovirions accurately reflect the infection process of HPV virions has not been verified. We generated infectious HPV31b virions from organotypic (raft) tissues and performed experimental infections in a variety of cells. Successful infection following viral attachment, internalization, and nuclear transport was assayed by detecting newly synthesized, spliced HPV transcripts using reverse transcription (RT)-PCR or RT-quantitative PCR. Most human epithelial cells were infected with HPV31b at a multiplicity of infection as low as 1 to 10 viral genome equivalents per cell. HPV31b infection was detected in other cell lines, including COS-7 monkey kidney cells, but higher viral multiplicities of infection were required. Heparin preparations of various molecular weights or heparinase I treatment of cells prevented HPV31b infection of COS-7 cells and C-33A human cervical cancer cells in reproducible and dose-dependent manners. However, these reagents were unable to block infection of human keratinocytes, including HaCaT and N/TERT-1 cells and low-passage human foreskin keratinocytes. These data suggest that HPV31b infection of human keratinocytes, the natural host cell for HPV infections in vivo, does not require a heparan-sulfonated receptor, whereas heparan sulfate is important for infection of some other cells. PMID:15890923

Patterson, Nicole A; Smith, Jessica L; Ozbun, Michelle A

2005-06-01

266

Human Papillomavirus Type 31b Infection of Human Keratinocytes Does Not Require Heparan Sulfate  

PubMed Central

Oncogenic human papillomaviruses (HPVs) are difficult to study experimentally as they replicate at low levels in vivo. This has precluded the purification of high-risk HPV virions from in vivo lesions. Virus-like particles (VLPs) and pseudovirions from low- and high-risk HPV types can emulate various aspects of HPV virion attachment and infections. These studies suggest that HPV infection is mediated by ?6-integrin and/or heparan-sulfonated receptors. However, whether VLPs and pseudovirions accurately reflect the infection process of HPV virions has not been verified. We generated infectious HPV31b virions from organotypic (raft) tissues and performed experimental infections in a variety of cells. Successful infection following viral attachment, internalization, and nuclear transport was assayed by detecting newly synthesized, spliced HPV transcripts using reverse transcription (RT)-PCR or RT-quantitative PCR. Most human epithelial cells were infected with HPV31b at a multiplicity of infection as low as 1 to 10 viral genome equivalents per cell. HPV31b infection was detected in other cell lines, including COS-7 monkey kidney cells, but higher viral multiplicities of infection were required. Heparin preparations of various molecular weights or heparinase I treatment of cells prevented HPV31b infection of COS-7 cells and C-33A human cervical cancer cells in reproducible and dose-dependent manners. However, these reagents were unable to block infection of human keratinocytes, including HaCaT and N/TERT-1 cells and low-passage human foreskin keratinocytes. These data suggest that HPV31b infection of human keratinocytes, the natural host cell for HPV infections in vivo, does not require a heparan-sulfonated receptor, whereas heparan sulfate is important for infection of some other cells.

Patterson, Nicole A.; Smith, Jessica L.; Ozbun, Michelle A.

2005-01-01

267

Superactive insulin: (B10-aspartic acid)insulin(human)  

SciTech Connect

The genetic basis for a case of familial hyperproinsulinemia has been elucidated recently. It involves a single point mutation in the proinsulin gene resulting in the substitution of aspartic acid for histidine-10 of the B chain of insulin. The authors have synthesized a human insulin analogue, (Asp/sup B10/) insulin, corresponding to the mutant proinsulin and evaluated its biological activity. (Asp/sup B10/) Insulin displayed a binding affinity to insulin receptors in rat liver plasma membranes that was 534 +- 146% relative to the natural hormone. In lipogenesis assays, the synthetic analogue exhibited a potency that was 435 +- 144% relative to insulin, which is statistically not different from its binding affinity. Reversed-phase HPLC indicated that the synthetic analogue is more apolar than natural insulin. They suggest that the observed properties reflect changes in the conformation of the analogue relative to natural insulin, which results in a stronger interaction with the insulin receptor. Thus, a single substitution of an amino acid residue of human insulin has resulted in a superactive hormone.

Schwartz, G.P.; Burke, G.T.; Katsoyannis, P.G.

1987-09-01

268

Molecular Evolution of Enterovirus 68 Detected in the Philippines  

PubMed Central

Background Detection of Enterovirus 68 (EV68) has recently been increased. However, underlying evolutionary mechanism of this increasing trend is not fully understood. Methods Nasopharyngeal swabs were collected from 5,240 patients with acute respiratory infections in the Philippines from June 2009 to December 2011. EV68 was detected by polymerase chain reaction (PCR) targeting for 5? untranslated region (5?UTR), viral protein 1 (VP1), and VP4/VP2. Phylogenetic trees were generated using the obtained sequences. Results Of the 5,240 tested samples, 12 EV68 positive cases were detected between August and December in 2011 (detection rate, 0.23%). The detection rate was higher among inpatients than outpatients (p<0.0001). Among VP1 sequences detected from 7 patients in 2011, 5 in lineage 2 were diverged from those detected in the Philippines in 2008, however, 2 in lineage 3 were not diverged from strains detected in the Philippines in 2008 but closely associated with strains detected in the United States. Combined with our previous report, EV68 occurrences were observed twice in the Philippines within the last four years. Conclusions EV68 detections might be occurring in cyclic patterns, and viruses might have been maintained in the community while some strains might have been newly introduced.

Imamura, Tadatsugu; Suzuki, Akira; Lupisan, Socorro; Okamoto, Michiko; Aniceto, Rapunzel; Egos, Rutchie J.; Daya, Edgardo E.; Tamaki, Raita; Saito, Mariko; Fuji, Naoko; Roy, Chandra Nath; Opinion, Jaime M.; Santo, Arlene V.; Macalalad, Noel G.; Tandoc, Amado; Sombrero, Lydia; Olveda, Remigio; Oshitani, Hitoshi

2013-01-01

269

Non-polio enteroviruses and their association with acute diarrhea in children in India.  

PubMed

A causative agent in approximately 40% of diarrheal cases still remains unidentified. Though many enteroviruses (EVs) are transmitted through fecal-oral route and replicate in the intestinal cells, their association with acute diarrhea has not so far been recognized due to lack of detailed epidemiological investigations. This long-term, detailed molecular epidemiological study aims to conclusively determine the association of non-polio enteroviruses (NPEVs) with acute diarrhea in comparison with rotavirus (RV) in children. Diarrheal stool specimens from 2161 children aged 0-2 years and 169 children between 2 and 9 years, and 1800 normal stool samples from age-matched healthy children between 0 and 9 years were examined during 2008-2012 for enterovirus (oral polio vaccine strains (OPVs) and NPEVs). Enterovirus serotypes were identified by complete VP1 gene sequence analysis. Enterovirus and rotavirus were detected in 19.01% (380/2330) and 13.82% (322/2330) diarrheal stools. During the study period, annual prevalence of EV- and RV-associated diarrhea ranged between 8% and 22%, but with contrasting seasonal prevalence with RV predominating during winter months and NPEV prevailing in other seasons. NPEVs are associated with epidemics-like outbreaks during which they are detected in up to 50% of diarrheic children, and in non-epidemic seasons in 0-10% of the patients. After subtraction of OPV-positive diarrheal cases (1.81%), while NPEVs are associated with about 17% of acute diarrhea, about 6% of healthy children showed asymptomatic NPEV excretion. Of 37 NPEV serotypes detected in diarrheal children, seven echovirus types 1, 7, 11, 13, 14, 30 and 33 are frequently observed, with E11 being more prevalent followed by E30. In conclusion, NPEVs are significantly associated with acute diarrhea, and NPEVs and rotavirus exhibit contrasting seasonal predominance. This study signifies the need for a new direction of research on enteroviruses involving systematic analysis of their contribution to diarrheal burden. PMID:23602838

Rao, Durga C; Ananda Babu, M; Raghavendra, A; Dhananjaya, D; Kumar, Sudheendra; Maiya, P P

2013-04-17

270

Genetic and Antigenic Characterization of Enterovirus 71 in Ho Chi Minh City, Vietnam, 2011  

PubMed Central

Enterovirus 71 (EV71) frequently causes fatal infections in young children in Asia. In 2011, EV71 epidemics occurred in southern Vietnam. We conducted genetic and antigenic analysis of the EV71 isolates and found that 94% of them were genotype C4a related to two lineages circulating in China and 6% were genotype C5 which have circulated in Vietnam since 2003. Antigenic variants were not detected. EV71 vaccines are being developed. Longitudinal enterovirus surveillance data are critical to formulate vaccination policy in Vietnam.

Thoa, Le Phan Kim; Chiang, Pai-Shan; Khanh, Truong Huu; Luo, Shu-Ting; Dan, Tran Ngoc Hanh; Wang, Ya-Fang; Thuong, Tang Chi; Chung, Wan-Yu; Hung, Nguyen Thanh; Wang, Jen-Ren; Nhan, Le Nguyen Thanh; Thinh, Le Quoc; Su, Ih-Jen; Dung, Than Duc; Lee, Min-Shi

2013-01-01

271

Innate immunity and human B cell clonal expansion: effects on the recirculating B2 subpopulation.  

PubMed

Foci of autoantigen-specific B lymphocytes in nonlymphoid tissues have been associated with development of autoimmune disease. To better understand the genesis of such ectopic lymphoid tissue, this study investigated whether several B cell-tropic innate immune system molecules, known to be elevated in response to inflammatory stimuli, can cooperate in fostering the T cell-independent clonal expansion of mature human B2 cells under conditions of limiting BCR engagement. Notable synergy was observed between BCR coligation with the C3dg-binding CD21/CD19 costimulatory complex, B cell-activating factor belonging to the TNF family (BAFF), and IL-4 in generating B cell progeny with sustained CD86 and DR expression. The synergy was observed over a wide range of BCR:ligand affinities and involved: 1) cooperative effects at promoting early cell cycle progression and viability; 2) BCR:CD21 coligation-promoted increases in BAFF receptors that were highly regulated by IL-4; 3) reciprocal effects of IL-4 and BAFF at dampening daughter cell apoptosis typical of stimulation by BCR:CD21 and either cytokine alone; and 4) BAFF-sustained expression of antiapoptotic Mcl-1 within replicating lymphoblasts. The results suggest that significant clonal proliferation of recirculating B2 cells occurs upon limited binding to C3dg-coated Ag in an inflammatory in vivo milieu containing both BAFF and IL-4. When rare autoantigen-presenting B cells undergo such expansions, both B cell and T cell autoimmunity may be promoted. PMID:16237111

Mongini, Patricia K A; Inman, John K; Han, Hanna; Kalled, Susan L; Fattah, Rasem J; McCormick, Steven

2005-11-01

272

Isolation of human monoclonal antibodies from peripheral blood B cells.  

PubMed

Isolation of monoclonal antibodies is an important technique for understanding the specificities and characteristics of antibodies that underlie the humoral immune response to a given antigen. Here we describe a technique for isolating monoclonal antibodies from human peripheral blood mononuclear cells. The protocol includes strategies for the isolation of switch-memory B cells from peripheral blood, the culture of B cells, the removal of the supernatant for screening and the lysis of B cells in preparation for immunoglobulin heavy-chain and light-chain amplification and cloning. We have observed that the addition of cytokines IL-2, IL-21 and irradiated 3T3-msCD40L feeder cells can successfully stimulate switch-memory B cells to produce high concentrations of IgG in the supernatant. The supernatant may then be screened by appropriate assays for binding or for other functions. This protocol can be completed in 2 weeks. It is adaptable to use in other species and enables the efficient isolation of antibodies with a desired functional characteristic without prior knowledge of specificity. PMID:24030440

Huang, Jinghe; Doria-Rose, Nicole A; Longo, Nancy S; Laub, Leo; Lin, Chien-Li; Turk, Ellen; Kang, Byong H; Migueles, Stephen A; Bailer, Robert T; Mascola, John R; Connors, Mark

2013-09-12

273

Evidence for APOBEC3B mutagenesis in multiple human cancers.  

PubMed

Thousands of somatic mutations accrue in most human cancers, and their causes are largely unknown. We recently showed that the DNA cytidine deaminase APOBEC3B accounts for up to half of the mutational load in breast carcinomas expressing this enzyme. Here we address whether APOBEC3B is broadly responsible for mutagenesis in multiple tumor types. We analyzed gene expression data and mutation patterns, distributions and loads for 19 different cancer types, with over 4,800 exomes and 1,000,000 somatic mutations. Notably, APOBEC3B is upregulated, and its preferred target sequence is frequently mutated and clustered in at least six distinct cancers: bladder, cervix, lung (adenocarcinoma and squamous cell carcinoma), head and neck, and breast. Interpreting these findings in the light of previous genetic, cellular and biochemical studies, the most parsimonious conclusion from these global analyses is that APOBEC3B-catalyzed genomic uracil lesions are responsible for a large proportion of both dispersed and clustered mutations in multiple distinct cancers. PMID:23852168

Burns, Michael B; Temiz, Nuri A; Harris, Reuben S

2013-07-14

274

Some properties of human and bovine brain cathepsin B  

Microsoft Academic Search

Cathespin B has been purified 750-fold to apparent homogeneity from human and bovine brain cortex using ammonium sulfate fractionation\\u000a (30–70%), chromatography on Sephadex G-100, CM-Sephadex C-50, and concanavalin A-Sepharose. Enzyme was assayed fluorometrically\\u000a at pH 4.0 with pyridoxyl-hemoglobin in the presence of 1 mM DTT and 1 mM EDTA. Properties of the enzyme from the two sources\\u000a proved to be

Anahit Azaryan; Nina Barkhudaryan; Armen Galoyan

1985-01-01

275

Study of human B7 homolog 1 expression in patients with hepatitis B virus infection  

PubMed Central

AIM: To further investigate the role of human B7 homolog 1 (B7-H1) in the mechanism of persistent hepatitis B virus (HBV) infection. METHODS: Peripheral and intra-hepatic B7-H1 expression were compared by ?ow cytometry and immunochemical staining between two 2 distinct groups, one being chronic HBV tolerance patients (CHB-T) and the other being acute hepatitis B patients (AHB). B7-H1 mRNA expression level was also compared by real time polymerase chain reaction between CHB-T and AHB patients. The location of intra-hepatic B7-H1 and CD40 expression were analyzed by immunofluorescence. The levels of B7-H1 and CD40 expression on cultured myeloid dendritic cells (mDCs) with or without hepatitis B surface antigen (HBsAg) treatment were analyzed dynamically by ?ow cytometry. Intracellular interferon-? (IFN-?) staining and the stimulatory capacity of mDC of cultured mDC with or without HBsAg treatment were also compared by ?ow cytometry. RESULTS: Peripheral B7-H1 expression on mDCs was increased significantly in AHB compared to CHB-T patients (P < 0.05). In the liver tissues from CHB-T patients, B7-H1 positive cells were almost absent despite a persistently elevated serum HBsAg load. In contrast, there were indeed increased B7-H1-positive cells in situ in the liver tissue from AHB. In vitro analysis showed the parallel upregulation of B7-H1 and CD40 on CD11c+ mDCs after the onset of stimulation. Addition of recombinant hepatitis B surface antigen (rHBsAg) significantly decreased CD40 expression (P < 0.05 at 16 h, 20 h and 24 h time points). B7-H1 expression was also inhibited by rHBsAg, and the inhibition rate of CD40 was greater than that of B7-H1. This preferential inhibition of CD40 expression on mDCs by rHBsAg resulted in the dysfunction of mDCs and T cells in the mixed leucocyte reaction (MLR) system. With rHBsAg pretreatment, in a carboxy?uorescein diacetate succinimidyl ester (CFSE) labeled MLR system at a ratio of 1:5 responder cell-stimulator cell (R/S), the CFSEdim percentage of T cells decreased from 85.1% to 25.4% and decreased from 30.3% to 12.0% at 1:10 R/S. IFN-? production by CD8+ T cells, in the MLR system, was reduced significantly by HBsAg pretreatment. At ratios of 1:5 R/S, the percentage of IFN-? and CD8 dual positive T cells decreased from 55.2% ± 5.3% to 15.1% ± 3.1% (P < 0.001), and decreased from 35.0% ± 5.1% to 7.3% ± 2.7% at ratios of 1:10 R/S (P < 0.001). CONCLUSION: B7-H1 is not a signature of immune dysfunction, but an inflammation marker. HBsAg regulate immune response by tipping the balance between B7-H1 and CD40.

Zhang, Wen-Jin; Xie, Hai-Yang; Duan, Xin; Wan, Yun-Le; Peng, Chuan-Hui; Shi, Shao-Hua; Su, Rong; Zheng, Zhang-Hui; Pan, Le-Lin; Zhou, Lin; Zheng, Shu-Sen

2012-01-01

276

Human Monoclonal Antibodies Broadly Neutralizing against Influenza B Virus  

PubMed Central

Influenza virus has the ability to evade host immune surveillance through rapid viral genetic drift and reassortment; therefore, it remains a continuous public health threat. The development of vaccines producing broadly reactive antibodies, as well as therapeutic strategies using human neutralizing monoclonal antibodies (HuMAbs) with global reactivity, has been gathering great interest recently. Here, three hybridoma clones producing HuMAbs against influenza B virus, designated 5A7, 3A2 and 10C4, were prepared using peripheral lymphocytes from vaccinated volunteers, and were investigated for broad cross-reactive neutralizing activity. Of these HuMAbs, 3A2 and 10C4, which recognize the readily mutable 190-helix region near the receptor binding site in the hemagglutinin (HA) protein, react only with the Yamagata lineage of influenza B virus. By contrast, HuMAb 5A7 broadly neutralizes influenza B strains that were isolated from 1985 to 2006, belonging to both Yamagata and Victoria lineages. Epitope mapping revealed that 5A7 recognizes 316G, 318C and 321W near the C terminal of HA1, a highly conserved region in influenza B virus. Indeed, no mutations in the amino acid residues of the epitope region were induced, even after the virus was passaged ten times in the presence of HuMAb 5A7. Moreover, 5A7 showed significant therapeutic efficacy in mice, even when it was administered 72 hours post-infection. These results indicate that 5A7 is a promising candidate for developing therapeutics, and provide insight for the development of a universal vaccine against influenza B virus.

Yasugi, Mayo; Kubota-Koketsu, Ritsuko; Yamashita, Akifumi; Kawashita, Norihito; Du, Anariwa; Sasaki, Tadahiro; Nishimura, Mitsuhiro; Misaki, Ryo; Kuhara, Motoki; Boonsathorn, Naphatsawan; Fujiyama, Kazuhito; Okuno, Yoshinobu; Nakaya, Takaaki; Ikuta, Kazuyoshi

2013-01-01

277

Interplay between the human TCR\\/CD3? and the B-cell antigen receptor associated Ig-? (B29)  

Microsoft Academic Search

T and B lymphocytes are characterized by the surface expression of highly variable antigen receptors called the T-cell (TCR) and B-cell (BCR) receptor. In both B and T cells, binding of antigen to their respective surface receptors results in transmembrane signaling which leads either to programmed cell death or to proliferatin and differentiation. The human and murine TCR consist of

Brigitte Müller; Laura Cooper; Cox Terhorst

1995-01-01

278

RNA-sequence analysis of human B-cells  

PubMed Central

RNA-sequencing (RNA-seq) allows quantitative measurement of expression levels of genes and their transcripts. In this study, we sequenced complementary DNA fragments of cultured human B-cells and obtained 879 million 50-bp reads comprising 44 Gb of sequence. The results allowed us to study the gene expression profile of B-cells and to determine experimental parameters for sequencing-based expression studies. We identified 20,766 genes and 67,453 of their alternatively spliced transcripts. More than 90% of the genes with multiple exons are alternatively spliced; for most genes, one isoform is predominantly expressed. We found that while chromosomes differ in gene density, the percentage of transcribed genes in each chromosome is less variable. In addition, genes involved in related biological processes are expressed at more similar levels than genes with different functions. Besides characterizing gene expression, we also used the data to investigate the effect of sequencing depth on gene expression measurements. While 100 million reads are sufficient to detect most expressed genes and transcripts, about 500 million reads are needed to measure accurately their expression levels. We provide examples in which deep sequencing is needed to determine the relative abundance of genes and their isoforms. With data from 20 individuals and about 40 million sequence reads per sample, we uncovered only 21 alternatively spliced, multi-exon genes that are not in databases; this result suggests that at this sequence coverage, we can detect most of the known genes. Results from this project are available on the UCSC Genome Browser to allow readers to study the expression and structure of genes in human B-cells.

Toung, Jonathan M.; Morley, Michael; Li, Mingyao; Cheung, Vivian G.

2011-01-01

279

Seroconversion to islet autoantibodies after enterovirus infection in early pregnancy.  

PubMed

Gestational enterovirus (EV) infections have been associated with an increased risk for type 1 diabetes in the offspring. We therefore analyzed non-diabetic mothers for EV exposure in early pregnancy in relation to type 1 diabetes HLA-DQ risk genotypes and seroconversion to islet autoantibodies during pregnancy. Non-diabetic mothers who had islet autoantibodies (n=365) against glutamic acid decarboxylase (GADA), islet antigen-2 autoantibodies (IA-2A), or insulin autoantibodies (IAA), in early pregnancy and at delivery were compared to islet autoantibody-negative mothers (n=1457) matched for age and sampling date. Mothers were genotyped for HLA-DQ and analyzed for both EV-RNA and EV-IgM. EV-IgM, but not EV-RNA, was detected during early pregnancy in 12% of islet autoantibody-positive mothers compared to 11% of the controls. In early pregnancy, mothers with HLA-DQ 2/2 or 2/X genotypes showed an increased risk for islet autoantibodies at delivery (OR 1.85; p=0.001). After adjusting for parity, maternal age, year of birth, and season of early pregnancy, early pregnancy EV-IgM combined with DQ2/2 or 2/X increased the risk for islet autoantibodies (OR 3.10; 95% CI 1; p=0.008). EV-IgM in early pregnancy increased the risk for islet autoantibodies at delivery in non-diabetic mothers with HLA-DQ 2/2 or 2/X type 1 diabetes risk genotypes. PMID:22746839

Reši? Lindehammer, Sabina; Honkanen, Hanna; Nix, William Allan; Oikarinen, Maarit; Lynch, Kristian Francis; Jönsson, Ida; Marsal, Karel; Oberste, Steven; Hyöty, Heikki; Lernmark, Åke

2012-07-02

280

Human Herpes Virus 6B: A possible Role in Epilepsy?  

PubMed Central

Nearly ubiquitous infection, including central nervous system invasion, with human herpesvirus 6 occurs in childhood. There are two variants, HHV6A and HHV6B. Usually asymptomatic, it is associated with the common, self-limited childhood illness roseola infantum, and rarely with more severe syndromes. In patients with immune compromise, reactivation of viral activity may lead to severe limbic encephalitis. HHV6 has been identified as a possible etiologic agent in multiple sclerosis. A preponderance of evidence supports an association between HHV6 and febrile seizures. An ongoing multicenter study is investigating possible links between HHV6 infection, febrile status epilepticus, and development of mesial temporal sclerosis (MTS). Investigation of temporal lobectomy specimens showed evidence of active HHV6B but not HHV6A replication in hippocampal astrocytes in about two-thirds of patients with MTS, but not other causes of epilepsy. HHV6 has been detected in similar clinical syndromes by other investigators, although less frequently, and additional viruses have been detected as well. It has been suggested that HHV6B may cause ‘excitotoxicity’ by interfering with astrocyte excitatory amino acid transport. Although conventional inflammatory changes are not found in most MTS specimens, inflammatory modulators may play a role in neuronal injury leading to MTS as well. If the link between early viral infection and later development of intractable temporal lobe epilepsy is confirmed, possibly associated with febrile seizures, new therapeutic approaches to a common intractable epilepsy syndrome may be possible.

Theodore, William H; Shinnar, Shlomo; Gaillard, William D; Wainwright, Mark S.; Epstein, Leon; Jacobson, Steven

2009-01-01

281

Protection against Enterovirus 71 with Neutralizing Epitope Incorporation within Adenovirus Type 3 Hexon  

PubMed Central

Enterovirus 71 (EV71) is responsible for hand, foot and mouth disease with high mortality among children. Various neutralizing B cell epitopes of EV71 have been identified as potential vaccine candidates. Capsid-incorporation of antigens into adenovirus (Ad) has been developed for a novel vaccine approach. We constructed Ad3-based EV71 vaccine vectors by incorporating a neutralizing epitope SP70 containing 15 amino acids derived from capsid protein VP1 of EV71 within the different surface-exposed domains of the capsid protein hexon of Ad3EGFP, a recombinant adenovirus type 3 (Ad3) expressing enhanced green fluorescence protein. Thermostability and growth kinetic assays suggested that the SP70 epitope incorporation into hypervariable region (HVR1, HVR2, or HVR7) of the hexon did not affect Ad fitness. The SP70 epitopes were thought to be exposed on all hexon-modified intact virion surfaces. Repeated administration of BALB/c mice with the modified Ads resulted in boosting of the anti-SP70 humoral immune response. Importantly, the modified Ads immunization of mother mice conferred protection in vivo to neonatal mice against the lethal EV71 challenge, and the modified Ads-immunized mice serum also conferred passive protection against the lethal challenge in newborn mice. Compared with the recombinant GST-fused SP70 protein immunization, immunization with the Ads containing SP70 in HVR1 or HVR2 elicited higher SP70-specific IgG titers, higher neutralization titers, and conferred more effective protection to neonatal mice. Thus, this study provides valuable information for hexon-modified Ad3 vector development as a promising EV71 vaccine candidate and as an epitope-delivering vehicle for other pathogens.

Tian, Xingui; Su, Xiaobo; Li, Xiao; Li, Haitao; Li, Ting; Zhou, Zhichao; Zhong, Tianhua; Zhou, Rong

2012-01-01

282

Enterovirus infection induces cytokine and chemokine expression in insulin-producing cells.  

PubMed

Despite evidence supporting an association between enterovirus (EV) infection and type 1 diabetes, the etiological mechanism(s) for EV-induced beta cell destruction is(are) not well understood. In this study, the effects of Coxsackievirus B (CVB) 1-6 on cell lysis and cytokine/chemokine expression in the insulinoma-1 (INS-1) beta cell line were investigated. Cytolysis was assessed using tissue culture infectious dose 50 (TCID(50)). Quantitative RT-PCR was used to measure viral RNA and mRNA of cytokines interferon (IFN)-?, IFN-?, IFN-?, tumor necrosis factor (TNF)-?, and chemokine (C-X-C motif) ligand 10 (CXCL10), chemokine (C-C motif) ligand 2 (CCL2), and chemokine (C-C motif) ligand 5 (CCL5) in infected INS-1 cells. CVB2, 4, 5, and 6 lysed and replicated in INS-1 cells; TCID(50) was lowest for CVB5 and highest for CVB6. IFN-?, CXCL10, and CCL5 mRNA levels all increased significantly following infection with CVB2, 4, 5, and 6 (P<0.05). CCL2 mRNA increased with CVB2, 5, and 6 (P<0.05), IFN-? mRNA increased with CVB5 infection (P<0.05), while TNF-? mRNA and IFN-? mRNA (P<0.001) increased with CVB2 infection. Dose-dependent effects of infection on cytokine mRNA levels were observed for all (P<0.01) except IFN-?. Following inoculation of INS-1 cells with CVB1 and 3, viral RNA was not detected and cytokine/chemokine mRNA levels were unchanged. In conclusion, CVB2, 4, 5, and 6 induce dose-dependent cytokine and chemokine mRNA production from INS-1 cells suggesting that pro-inflammatory cytokine and chemokine secretion by beta cells is a potential mechanism for EV-induced beta cell damage in type 1 diabetes. PMID:20872723

Nair, Sandhya; Leung, Kin-Chuen; Rawlinson, William D; Naing, Zin; Craig, Maria E

2010-11-01

283

Human antibodies for immunotherapy development generated via a human B cell hybridoma technology  

PubMed Central

Current strategies for the production of therapeutic mAbs include the use of mammalian cell systems to recombinantly produce Abs derived from mice bearing human Ig transgenes, humanization of rodent Abs, or phage libraries. Generation of hybridomas secreting human mAbs has been previously reported; however, this approach has not been fully exploited for immunotherapy development. We previously reported the use of transient regulation of cellular DNA mismatch repair processes to enhance traits (e.g., affinity and titers) of mAb-producing cell lines, including hybridomas. We reasoned that this process, named morphogenics, could be used to improve suboptimal hybridoma cells generated by means of ex vivo immunization and immortalization of antigen-specific human B cells for therapeutic Ab development. Here we present a platform process that combines hybridoma and morphogenics technologies for the generation of fully human mAbs specific for disease-associated human antigens. We were able to generate hybridoma lines secreting mAbs with high binding specificity and biological activity. One mAb with strong neutralizing activity against human granulocyte–macrophage colony-stimulating factor was identified that is now considered for preclinical development for autoimmune disease indications. Moreover, these hybridoma cells have proven suitable for genetic optimization using the morphogenics process and have shown potential for large-scale manufacturing.

Li, Jian; Sai, Tao; Berger, Marc; Chao, Qimin; Davidson, Diane; Deshmukh, Gaurav; Drozdowski, Brian; Ebel, Wolfgang; Harley, Stephen; Henry, Marianne; Jacob, Sara; Kline, Brad; Lazo, Ella; Rotella, Frank; Routhier, Eric; Rudolph, Kathryn; Sage, Jeaneen; Simon, Paul; Yao, Jun; Zhou, Yuhong; Kavuru, Mani; Bonfield, Tracey; Thomassen, Mary Jane; Sass, Philip M.; Nicolaides, Nicholas C.; Grasso, Luigi

2006-01-01

284

Maternal enterovirus infection during pregnancy as a risk factor in offspring diagnosed with type 1 diabetes between 15 and 30 years of age.  

PubMed

Maternal enterovirus infections during pregnancy may increase the risk of offspring developing type 1 diabetes during childhood. The aim of this study was to investigate whether gestational enterovirus infections increase the offspring's risk of type 1 diabetes later in life. Serum samples from 30 mothers without diabetes whose offspring developed type 1 diabetes between 15 and 25 years of age were analyzed for enterovirus-specific immunoglobulin M (IgM) antibodies and enterovirus genome (RNA), and compared to a control group. Among the index mothers, 9/30 (30%) were enterovirus IgM-positive, and none was positive for enterovirus RNA. In the control group, 14/90 (16%) were enterovirus IgM-positive, and 4/90 (4%) were positive for enterovirus RNA (n.s.). Boys of enterovirus IgM-positive mothers had approximately 5 times greater risk of developing diabetes (OR 4.63; 95% CI 1.22-17.6), as compared to boys of IgM-negative mothers (P < .025). These results suggest that gestational enterovirus infections may be related to the risk of offspring developing type 1 diabetes in adolescence and young adulthood. PMID:18670622

Elfving, Maria; Svensson, Johan; Oikarinen, Sami; Jonsson, Björn; Olofsson, Per; Sundkvist, Göran; Lindberg, Bengt; Lernmark, Ake; Hyöty, Heikki; Ivarsson, Sten-Anders

2008-01-01

285

Acute Encephalitis Caused by Intrafamilial Transmission of Enterovirus 71 in Adult  

PubMed Central

Enterovirus 71 (EV71) is a common cause of hand, foot, and mouth disease and sometimes causes severe neurologic complications, mainly in children. We report a case of adult-onset encephalitis caused by intrafamilial transmission of a subgenogroup C4 strain of EV71. This case elucidates the risk for EV71 encephalitis even in adults.

Fujisawa, Hironori; Sakai, Kenji; Okino, Soichi; Kurosaki, Naoko; Nishimura, Yorihiro; Shimizu, Hiroyuki; Yamada, Masahito

2008-01-01

286

Detection of infectious enteroviruses and adenoviruses in tap water in urban areas in Korea  

Microsoft Academic Search

We investigated the viral contamination of tap water at 11 urban sites in Korea between 1997 and 1998 over a period of 11 months. A total of 23 tap water samples were examined for infectious enteroviruses and adenoviruses by a cell culture technique followed by polymerase chain reaction (PCR) amplification. To identify the recovered viruses, sequence analysis of PCR products

Seung-Hoon Lee; Sang-Jong Kim

2002-01-01

287

EFFECT OF PARTICULATES ON DISINFECTION OF ENTEROVIRUSES IN WATER BY CHLORINE DIOXIDE  

EPA Science Inventory

The inactivation kinetics of ClO2 on two enteroviruses, poliovirus 1 (Mahoney) and coxsackie virus A9, and an enteric indicator of fecal pollution, Escherichia coli, were examined in laboratory studies. In addition, the disinfecting ability of ClO2 as affected by particulates (bo...

288

Demonstration of persistent enterovirus in the pancreas of diabetic mice by in situ polymerase chain reaction  

Microsoft Academic Search

Background: Although Enterovirus (EV) do not persist in the tissue, which is essential to maintain autoimmunity, they have been associated as the cause of chronic autoimmunity in some cases of insulin dependent diabetes mellitus (IDDM). Convincing reports, demonstrating persistent EV infections in the pancreases, are rare.Objectives: To determine the role of EV in IDDM, a mouse model was tested and

M. M Berger; D. M See; M Aymard; B Lina

1998-01-01

289

Detection of Infectious Enteroviruses by an Integrated Cell Culture-PCR Procedure  

Microsoft Academic Search

Rapid detection of infectious enteroviruses in environmental samples was made possible by utilizing an integrated cell culture-reverse transcriptase PCR approach. By this method, the presence of infectious entero- virus was confirmed within 24 h, compared with >3 days by cell culture alone. The combined methodology eliminated typical problems normally associated with direct reverse transcriptase PCR by increasing the equivalent volume

KELLY A. REYNOLDS; CHARLES P. GERBA; ANDIAN L. PEPPER

290

An outbreak of enterovirus 71 infection in Taiwan, 1998: epidemiologic and clinical manifestations  

Microsoft Academic Search

Background: An outbreak of enterovirus infections occurred throughout Taiwan in 1998. The diseases were manifectated with hand, foot, and mouth disease (HFMD), some associated with meningitis, encephalitis, or acute flaccid paralysis (AFP). Objectives: This study is aimed to characterize and analyze the epidermologic and clinical features during the outbreak. Study design: The epidemiologic information was collected from the Ministry of

Ching-Chuan Liu; Hui-Wan Tseng; Shih-Min Wang; Jen-Ren Wang; Ih-Jen Su

2000-01-01

291

Molecular identification of new picornaviruses and characterization of a proposed enterovirus 73 serotype  

Microsoft Academic Search

Enteroviruses (EV) have traditionally been identified by using serotype-specific antisera in a virus- neutralization test. Three EV strains isolated in California, USA, in 1955, 1964 and 1978, and a 1995 Oman isolate, were found to be antigenically related to one another; however, the strains were not neutralized by standard EV typing antisera, suggesting that they may represent a new EV

M. Steven Oberste; David Schnurr; Kaija Maher; Mark A. Pallansch

292

Phylogenetic Analysis of Enterovirus 71 Strains Isolated during Linked Epidemics in Malaysia, Singapore, and Western Australia  

Microsoft Academic Search

Enterovirus 71 (EV71) is a frequent cause of hand, foot, and mouth disease (HFMD) epidemics associated with severe neurological sequelae in a small proportion of cases. There has been a significant increase in EV71 epidemic activity throughout the Asia-Pacific region since 1997. Recent HFMD epidemics in this region have been associated with a severe form of brainstem encephalitis associated with

PETER MCMINN; KATIE LINDSAY; DAVID PERERA; HUNG MING CHAN; KWAI PENG CHAN; MARY JANE CARDOSA

2001-01-01

293

Enterovirus 68 in Children with Acute Respiratory Tract Infections, Osaka, Japan  

PubMed Central

Enterovirus 68 strains were detected in 14 specimens from children with respiratory tract infections and 1 specimen from a child with febrile convulsions during 2010 in Osaka, Japan. These strains had deletions in the 5? untranslated region and were genetically different from reported strains. This virus is associated with respiratory tract infections in Japan.

Kubo, Hideyuki; Sekiguchi, Jun-ichiro; Kohdera, Urara; Togawa, Masao; Shiomi, Masashi; Nishigaki, Toshinori; Iritani, Nobuhiro

2011-01-01

294

Expression of human apolipoprotein B and assembly of lipoprotein(a) in transgenic mice  

SciTech Connect

The atherogenic macromolecule lipoprotein(a) [Lp(a)] has resisted in vivo analyses partly because it is found in a limited number of experimental animals. Although transgenic mice expressing human apolipoprotein (a) [apo(a)] have previously been described, they failed to assemble Lp(a) particles because of the inability of human apo(a) to associate with mouse apolipoprotein B (apoB). The authors isolated a 90-kilobase P1 phagemid containing the human apoB gene and with this DNA generated 13 lines of transgenic mice of which 11 expressed human apoB. The human apoB transcript was expressed and edited in the liver of the transgenic mice. Plasma concentrations of human apoB, as well as low density lipoprotein (LDL), were related to transgene copy number; the transgenic line with the most copies of human apoB had a >4-fold increase in LDL cholesterol compared with nontransgenics and a lipoprotein profile similar to that of humans. When human apoB and apo(a) transgenic mice were bred together, plasma apo(a) in mice expressing both human proteins was tightly associated with lipoproteins in the LDL density region. These studies demonstrate the successful expression of human apoB and the efficient assembly of Lp(a) in mice.

Callow, M.J.; Stoltzfus, L.J.; Rubin, E.M. [Lawrence Berkeley Lab., CA (United States); Lawn, R.M. [Stanford Univ., CA (United States)

1994-03-15

295

Expression of the Aldo-Ketoreductases AKR1B1 and AKR1B10 in Human Cancers.  

PubMed

The American Cancer Society estimates that there will be more than 1.5 million new cases of cancer in 2011, underscoring the need for identification of new therapeutic targets and development of novel cancer therapies. Previous studies have implicated the human aldo-ketoreductases AKR1B1 and AKR1B10 in cancer, and therefore we examined AKR1B1 and AKR1B10 expression across all major human cancer types using the Oncomine cancer gene expression database (Compendia Biosciences, www.oncomine.com). Using this database, we found that expression of AKR1B1 and AKR1B10 varies greatly by cancer type and tissue of origin, including agreement with previous reports that AKR1B10 is significantly over-expressed in cancers of the lungs and liver. AKR1B1 is more broadly over-expressed in human cancers than AKR1B10, albeit at a generally lower magnitude. AKR1B1 over-expression was found to be associated with shortened patient survival in acute myelogenous leukemias and multiple myelomas. High AKR1B10 expression tends to predict less aggressive clinical course generally, notably within lung cancers, where it tends to be highly over-expressed compared to normal tissue. These findings suggest that AKR1B1 inhibitors in particular hold great potential as novel cancer therapeutics. PMID:22685431

Laffin, Brian; Petrash, J Mark

2012-06-06

296

Expression of the Aldo-Ketoreductases AKR1B1 and AKR1B10 in Human Cancers  

PubMed Central

The American Cancer Society estimates that there will be more than 1.5 million new cases of cancer in 2011, underscoring the need for identification of new therapeutic targets and development of novel cancer therapies. Previous studies have implicated the human aldo-ketoreductases AKR1B1 and AKR1B10 in cancer, and therefore we examined AKR1B1 and AKR1B10 expression across all major human cancer types using the Oncomine cancer gene expression database (Compendia Biosciences, www.oncomine.com). Using this database, we found that expression of AKR1B1 and AKR1B10 varies greatly by cancer type and tissue of origin, including agreement with previous reports that AKR1B10 is significantly over-expressed in cancers of the lungs and liver. AKR1B1 is more broadly over-expressed in human cancers than AKR1B10, albeit at a generally lower magnitude. AKR1B1 over-expression was found to be associated with shortened patient survival in acute myelogenous leukemias and multiple myelomas. High AKR1B10 expression tends to predict less aggressive clinical course generally, notably within lung cancers, where it tends to be highly over-expressed compared to normal tissue. These findings suggest that AKR1B1 inhibitors in particular hold great potential as novel cancer therapeutics.

Laffin, Brian; Petrash, J. Mark

2012-01-01

297

Treatment with recombinant human erythropoietin increases antibody titers after hepatitis B vaccination in dialysis patients  

Microsoft Academic Search

Treatment with recombinant human erythropoietin increases antibody titers after hepatitis B vaccination in dialysis patients. The effect of recombinant human erythropoietin (rHuEPO) on the immune system of hemodialysis patients has been studied by evaluating their response to hepatitis B (HB) vaccination. Fifty hemodialysis patients were given four doses of 20 ?g recombinant DNA hepatitis B vaccine (SKF) at an interval

Jacques J Sennesael; Patricia Van der Niepen; Dierik L Verbeelen

1991-01-01

298

Human Hepatocellular Carcinoma Cell Lines Secrete the Major Plasma Proteins and Hepatitis B Surface Antigen  

Microsoft Academic Search

Analysis of the cell culture fluid from two new human hepatoma-derived cell lines reveals that 17 of the major human plasma proteins are synthesized and secreted by these cells. One of these cell lines, Hep 3B, also produces the two major polypeptides of the hepatitis B virus surface antigen. When Hep 3B is injected into athymic mice, metastatic hepatocellular carcinomas

Barbara B. Knowles; Chin C. Howe; David P. Aden

1980-01-01

299

Comparison of heart failure in children with enterovirus 71 rhombencephalitis and cats with norepinephrine cardiotoxicity.  

PubMed

The mechanism of heart failure in patients with enterovirus 71 rhombencephalitis (brain stem encephalitis) remains unknown. Our previous reports hypothesized that a catecholamine storm induced by rhombencephalitis may account for the heart failure. The aim of this study was to develop a novel feline model of norepinephrine cardiotoxicity and compare the resulting heart failure to that in children with enterovirus 71 rhombencephalitis. Nine of 75 children (12%) with enterovirus 71 rhombencephalitis (5 boys and 4 girls; age, 4-28 months; median age, 16 months) were complicated with left ventricular hypokinesia (ejection fraction, 31 +/- 9%). Six cats (weight, 3.03 +/- 0.64 kg) were administered intravenous norepinephrine 30 microg/kg/min for 3 hours. Echocardiography assessed the left ventricular diameter and function before and after the administration of norepinephrine. Pathology studies included hematoxylin and eosin stain and in situ terminal deoxyribonucleotidyl transferase-mediated dUTP nick end-labeling assay. In the feline model, norepinephrine induced significant left ventricular dilatation (end diastolic diameter from 1.18 +/- 0.19 to 1.62 +/- 0.22 cm, p = 0.001; endsystolic diameter from 0.54 +/- 0.09 to 1.36 +/- 0.32 cm, p = < 0.001) and hypokinesia (ejection fraction from 87.5 +/- 4.1 to 35.2 +/- 16.3%, p = 0.001). Heart specimens from 4 patients and six cats showed similar pathology findings, including myocardial hemorrhage, cardiomyocyte apoptosis, and coagulative myocytolysis, which is characterized by sarcoplasmic coagulation, granulation, vacuolization, myofibrillar waving, and disruption. Both groups showed no significant inflammatory reaction. In conclusion, heart failure in patients with enterovirus 71 rhombencephalitis is similar to that in cats with norepinephrine cardiotoxicity. Norepinephrine cardiotoxicity may play a role in the pathogenesis of heart failure in enterovirus 71 rhombencephalitis. PMID:16933070

Fu, Y-C; Chi, C-S; Lin, N-N; Cheng, C-C; Jan, S-L; Hwang, B; Hsu, S-L; Gong, C-L; Chen, Y-T; Chiu, Y-T

300

Search for Coxsackievirus B3 RNA in idiopathic dilated cardiomyopathy using gene amplification by polymerase chain reaction.  

PubMed

A polymerase chain reaction (PCR) amplification assay was developed to detect Coxsackievirus B3 ribonucleic acid (RNA) in blood and myocardial tissue of explanted hearts from 40 patients who underwent cardiac transplantation and in 1 normal heart. Twenty-one patients were affected by idiopathic dilated cardiomyopathy of different duration and 19 by coronary artery disease. Coxsackievirus B3 in vitro infected Vero cells and cells infected by related human enteroviruses (Coxsackievirus B2, B4, and poliovirus 1) were used as reaction controls. PCR was performed using 4 pairs of primers homologous to Coxsackie-virus B3 sequences. Three sets were located in regions of the genome conserved at nucleotide level between several enterovirus species (replicase gene, 5' noncoding region), while one was located in a Coxsackievirus B3-specific region (VP1 gene). Total RNA was prepared by acid guanidinium isothiocyanate extraction from tissue stored frozen at -80 degrees C. One microgram of total RNA was retrotranscribed with either antisense primer or with random hexanucleotide primers and then subjected to 40 cycles of amplification. PCR products were separated by electrophoresis on a 10% polyacrylamide gel, electrotransferred to a nylon membrane and then hybridized to oligonucleotide probes specific for the coxsackievirus B3 genome radiolabeled with radioactive isotope of phosphorous. All pairs of primers yielded specific amplification products when tested on Coxsackievirus B3-infected Vero cells, with a sensitivity of 1 infected cell out of 10(5) to 10(6) cells starting from 1 microgram total RNA. Primer sets for regions of Coxsackievirus B3 genome highly conserved between related enteroviral species gave positive amplification also when challenged with RNA from cells infected by Coxsackievirus B2, B4 and poliovirus 1. PMID:1311139

Grasso, M; Arbustini, E; Silini, E; Diegoli, M; Percivalle, E; Ratti, G; Bramerio, M; Gavazzi, A; Vigano, M; Milanesi, G

1992-03-01

301

Modular Structure of the Human Lamin B2 Replicator  

PubMed Central

The cis-acting elements necessary for the activity of DNA replication origins in metazoan cells are still poorly understood. Here we report a thorough characterization of the DNA sequence requirements of the origin associated with the human lamin B2 gene. A 1.2-kb DNA segment, comprising the start site of DNA replication and located within a large protein-bound region, as well as a CpG island, displays origin activity when moved to different ectopic positions. Genomic footprinting analysis of both the endogenous and the ectopic origins indicates that the large protein complex is assembled in both cases around the replication start site. Replacement of this footprinted region with an unrelated sequence, maintaining the CpG island intact, abolishes origin activity and the interaction with hORC2, a subunit of the origin recognition complex. Conversely, the replacement of 17 bp within the protected region reduces the extension of the protection without affecting the interaction with hORC2. This substitution does not abolish the origin activity but makes it more sensitive to the integration site. Finally, the nearby CpG island positively affects the efficiency of initiation. This analysis reveals the modular structure of the lamin B2 origin and supports the idea that sequence elements close to the replication start site play an important role in origin activation.

Paixao, Sonia; Colaluca, Ivan N.; Cubells, Matthieu; Peverali, Fiorenzo A.; Destro, Annarita; Giadrossi, Sara; Giacca, Mauro; Falaschi, Arturo; Riva, Silvano; Biamonti, Giuseppe

2004-01-01

302

Human rhinoviruses.  

PubMed

Human rhinoviruses (HRVs), first discovered in the 1950s, are responsible for more than one-half of cold-like illnesses and cost billions of dollars annually in medical visits and missed days of work. Advances in molecular methods have enhanced our understanding of the genomic structure of HRV and have led to the characterization of three genetically distinct HRV groups, designated groups A, B, and C, within the genus Enterovirus and the family Picornaviridae. HRVs are traditionally associated with upper respiratory tract infection, otitis media, and sinusitis. In recent years, the increasing implementation of PCR assays for respiratory virus detection in clinical laboratories has facilitated the recognition of HRV as a lower respiratory tract pathogen, particularly in patients with asthma, infants, elderly patients, and immunocompromised hosts. Cultured isolates of HRV remain important for studies of viral characteristics and disease pathogenesis. Indeed, whether the clinical manifestations of HRV are related directly to viral pathogenicity or secondary to the host immune response is the subject of ongoing research. There are currently no approved antiviral therapies for HRVs, and treatment remains primarily supportive. This review provides a comprehensive, up-to-date assessment of the basic virology, pathogenesis, clinical epidemiology, and laboratory features of and treatment and prevention strategies for HRVs. PMID:23297263

Jacobs, Samantha E; Lamson, Daryl M; St George, Kirsten; Walsh, Thomas J

2013-01-01

303

Frequent Importation of Enterovirus 71 from Surrounding Countries into the Local Community of Yamagata, Japan, between 1998 and 2003  

Microsoft Academic Search

Phylogenetic analysis of 45 enterovirus 71 (EV71) isolates for 6 years in Yamagata, Japan, clarified that the annual outbreak of hand-foot-and-mouth disease was due to four genetically distinct subgenogroups, including a novel \\

K. Mizuta; C. Abiko; T. Murata; Y. Matsuzaki; T. Itagaki; K. Sanjoh; M. Sakamoto; S. Hongo; S. Murayama; K. Hayasaka

2005-01-01

304

Dysfunction in ankyrin-B-dependent ion channel and transporter targeting causes human sinus node disease  

PubMed Central

The identification of nearly a dozen ion channel genes involved in the genesis of human atrial and ventricular arrhythmias has been critical for the diagnosis and treatment of fatal cardiovascular diseases. In contrast, very little is known about the genetic and molecular mechanisms underlying human sinus node dysfunction (SND). Here, we report a genetic and molecular mechanism for human SND. We mapped two families with highly penetrant and severe SND to the human ANK2 (ankyrin-B/AnkB) locus. Mice heterozygous for AnkB phenocopy human SND displayed severe bradycardia and rate variability. AnkB is essential for normal membrane organization of sinoatrial node cell channels and transporters, and AnkB is required for physiological cardiac pacing. Finally, dysfunction in AnkB-based trafficking pathways causes abnormal sinoatrial node (SAN) electrical activity and SND. Together, our findings associate abnormal channel targeting with human SND and highlight the critical role of local membrane organization for sinoatrial node excitability.

Le Scouarnec, Solena; Bhasin, Naina; Vieyres, Claude; Hund, Thomas J.; Cunha, Shane R.; Koval, Olha; Marionneau, Celine; Chen, Biyi; Wu, Yuejin; Demolombe, Sophie; Song, Long-Sheng; Le Marec, Herve; Probst, Vincent; Schott, Jean-Jacques; Anderson, Mark E.; Mohler, Peter J.

2008-01-01

305

Pilot Scale Production of Highly Efficacious and Stable Enterovirus 71 Vaccine Candidates  

PubMed Central

Background Enterovirus 71 (EV71) has caused several epidemics of hand, foot and mouth diseases (HFMD) in Asia and now is being recognized as an important neurotropic virus. Effective medications and prophylactic vaccine against EV71 infection are urgently needed. Based on the success of inactivated poliovirus vaccine, a prototype chemically inactivated EV71 vaccine candidate has been developed and currently in human phase 1 clinical trial. Principal Finding In this report, we present the development of a serum-free cell-based EV71 vaccine. The optimization at each step of the manufacturing process was investigated, characterized and quantified. In the up-stream process development, different commercially available cell culture media either containing serum or serum-free was screened for cell growth and virus yield using the roller-bottle technology. VP-SFM serum-free medium was selected based on the Vero cell growth profile and EV71 virus production. After the up-stream processes (virus harvest, diafiltration and concentration), a combination of gel-filtration liquid chromatography and/or sucrose-gradient ultracentrifugation down-stream purification processes were investigated at a pilot scale of 40 liters each. Although the combination of chromatography and sucrose-gradient ultracentrifugation produced extremely pure EV71 infectious virus particles, the overall yield of vaccine was 7–10% as determined by a VP2-based quantitative ELISA. Using chromatography as the downstream purification, the virus yield was 30–43%. To retain the integrity of virus neutralization epitopes and the stability of the vaccine product, the best virus inactivation was found to be 0.025% formalin-treatment at 37°C for 3 to 6 days. Furthermore, the formalin-inactivated virion vaccine candidate was found to be stable for >18 months at 4°C and a microgram of viral proteins formulated with alum adjuvant could induce strong virus-neutralizing antibody responses in mice, rats, rabbits, and non-human primates. Conclusion These results provide valuable information supporting the current cell-based serum-free EV71 vaccine candidate going into human Phase I clinical trials.

Chang, Cheng-Peng; Guo, Meng-Shin; Hsieh, Shih-Yang; Yang, Wen-Hsueh; Chao, Hsin-Ju; Wu, Chien-Long; Huang, Ju-Lan; Lee, Min-Shi; Hu, Alan Yung-Chi; Lin, Sue-Chen; Huang, Yu-Yun; Hu, Mei-Hua; Chow, Yen-Hung; Chiang, Jen-Ron; Chang, Jui-Yuan; Chong, Pele

2012-01-01

306

Detection of naturally occurring enteroviruses in waters using direct RT-PCR and integrated cell culture-RT-PCR  

Microsoft Academic Search

Viruses detected by rapid molecular assays are not always infectious. In this study we compared enterovirus levels in natural waters using culture and reverse transcription-polymerase chain reaction (RT-PCR) techniques to determine whether molecular units of naturally occurring enteroviruses can be utilized to predict viral infectivity. Viruses were concentrated from 12 river water and effluent samples using 1MDS filter–filtration and beef

Y. C. Shieh; C. I. Wong; J. A. Krantz; F. C. Hsu

2008-01-01

307

Localization of Monoamine Oxidase A and B in Human Pancreas, Thyroid, and Adrenal Glands  

Microsoft Academic Search

We studied monoamine oxidase (MAO) A and B localization in human pancreas, thyroid gland, and adrenal gland by immunohistochemistry. The primary antibodies used were mouse monoclonal anti-human MAO-A (6G11\\/E1) and anti-human MAO-B (3F12\\/ G10\\/2E3). Samples were obtained from six routine autopsy cases and fixed in 2% paraformaldehyde. Exocrine pancreas showed a widespread distribution of MAO-A, whereas MAO-B was present only

Manuel J. Rodríguez; Josep Saura; Cheryl C. Finch; Nicole Mahy; Ellen E. Billett

2000-01-01

308

HUMAN CYTOSOLIC SULFOTRANSFERASE 2B1: ISOFORM EXPRESSION, TISSUE SPECIFICITY AND SUBCELLULAR LOCALIZATION  

PubMed Central

Sulfation is an important Phase II conjugation reaction involved in the synthesis and metabolism of steroids in humans. Two different isoforms (2B1a and 2B1b) are encoded by the sulfotransferase (SULT) 2B1 gene utilizing different start sites of transcription resulting in the incorporation of different first exons. SULT2B1a and SULT2B1b are 350 and 365 amino acids in length, respectively, and the last 342 aa are identical. Message for both SULT2B1 isoforms is present in human tissues although SULT2B1b message is generally more abundant. However, to date only SULT2B1b protein has been detected in human tissues or cell lines. SULT2B1b is localized in the cytosol and/or nuclei of human cells. A unique 3?-extension of SULT2B1b is required for nuclear localization in human BeWo placental choriocarcinoma cells. Nuclear localization is stimulated by forskolin treatment in BeWo cells and serine phosphorylation has been identified in the 3?-extension. SULT2B1b is selective for the sulfation of 3?-hydroxysteroids such as dehydroepiandrosterone and pregnenolone, and may also have a role in cholesterol sulfation in human skin. The substrate specificity, nuclear localization, and tissue localization of SULT2B1b suggest a role in regulating the responsiveness of cells to adrenal androgens via their direct inactivation or by preventing their conversion to more potent androgens and estrogens.

Falany, C.N.; He, D.; Dumas, N.; Frost, A.R.; Falany, J.L.

2007-01-01

309

B lymphopoiesis is active throughout human life, but there are developmental age-related changes  

Microsoft Academic Search

This study addressed several questions concerning age-related changes in hu- man B lymphopoiesis. The relative abun- dance of pro-B, pre-B, immature, naive, and mature B cells among the CD19 lymphocyte fraction of human bone mar- row was found not to change appreciably over the interval between 24 and 88 years of age. Moreover, proliferation of pro-B and large pre-B cells

Maria Isabel; D. Rossi; Takafumi Yokota; Kay L. Medina; Karla P. Garrett; Philip C. Comp; Arthur H. Schipul; Paul W. Kincade

2003-01-01

310

Preparation of human immortalized B-cells secreting antibodies against food allergens  

Microsoft Academic Search

An immortalized B-cell library consisting of cells secreting antibodies to food allergens has been prepared. Peripheral B-cells from seven healthy donors were transformed with Epstein-Barr virus and an immortalized human B-cell library with 2,202 ampoules of multi-clone B-lymphoblastoid cells was obtained. The B-cell library contained various types of B-lymphoblastoid cells secreting antibodies to rice, soybeans, milk, and eggs, and can

Hiroshi Shinmoto; Kazuhiko Nakahara; Masuko Kobori; Tojiro Tsushida

1998-01-01

311

Rapid and Sensitive Routine Detection of All Members of the Genus Enterovirus in Different Clinical Specimens by Real-Time PCR  

Microsoft Academic Search

We developed a rapid and sensitive method for the routine detection of all members of the enterovirus genus in different clinical specimens by using real-time TaqMan quantitative PCR. Multiple primer and probe sets were selected in the highly conserved 5-untranslated region of the enterovirus genome. Our assay detected all 60 different enterovirus species tested, whereas no reactivity was observed with

Monique Nijhuis; Noortje van Maarseveen; Rob Schuurman; Sandra Verkuijlen; Machiel de Vos; Karin Hendriksen; Anton M. van Loon

2002-01-01

312

Rapid Detection of Enterovirus RNA in Cerebrospinal Fluid Specimens with a Novel Single-Tube Real-Time Reverse Transcription-PCR Assay  

Microsoft Academic Search

A single-tube real-time reverse transcription-PCR (RT-PCR) assay for enterovirus detection in cerebrospi- nal fluid (CSF) was developed based on a fluorogenic probe and primers directed to highly conserved sequences in the 5 untranslated region of the enterovirus genome. Quantitative detection of enterovirus genome was demonstrated in a linear range spanning at least 5 logs. Endpoint titration experiments revealed that the

WALTER A. VERSTREPEN; SOFIE KUHN; MARK M. KOCKX; MARTINE E. VAN DE VYVERE; AN H. MERTENS

2001-01-01

313

Direct Ex Vivo Measurement of CD8+ T-Lymphocyte Responses to Human Parvovirus B19  

PubMed Central

Parvovirus B19 is a common human pathogen which can cause severe syndromes, including aplastic anemia and fetal hydrops. The mapping of the first parvovirus B19-derived CD8+ T-lymphocyte epitope is described. This HLA-B35-restricted peptide derives from the nonstructural (NS1) protein and is strongly immunogenic in B19 virus-seropositive donors.

Tolfvenstam, T.; Oxenius, A.; Price, D. A.; Shacklett, B. L.; Spiegel, H. M. L.; Hedman, K.; Norbeck, O.; Levi, M.; Olsen, K.; Kantzanou, M.; Nixon, D. F.; Broliden, K.; Klenerman, P.

2001-01-01

314

Interleukin1 b and Rhinovirus Sensitize Adenylyl Cyclase in Human Airway Smooth-Muscle Cells  

Microsoft Academic Search

Rhinovirus (RV) is a major cause of wheezing in asthmatics and has been reported to cause b 2 adrenergic receptor hypo- responsiveness in human airway smooth muscle (HASM) via cellular secretion of interleukin (IL)-1 b . We studied the effects of IL-1 b and RV on cyclic adenosine monophosphate (cAMP) production in HASM cells. Chronic incubation with IL-1 b or

Charlotte K. Billington; Rodolfo M. Pascual; Michael L. Hawkins; Raymond B. Penn; Ian P. Hall

315

Expression of activation-induced cytidine deaminase in human B-cell non-Hodgkin lymphomas  

Microsoft Academic Search

Activation-induced cytidine deaminase (AID) induces somatic hypermutation (SHM), class switch recombination (CSR), and immunoglobulin gene conversion in B-lymphocytes. Here we report for the first time the expression of AID in healthy human B-lymphocytes and in B-cell non- Hodgkin lymphomas (B-NHL). AID mRNA expression in humans is restricted to the CD19CD38IgD germinal center cells, namely the CD19CD38CD44 centro- blasts. After in

Jobst Greeve; Antje Philipsen; Kristina Krause; Wolfram Klapper; Klaus Heidorn; Brian E. Castle; Joe Janda; Kenneth B. Marcu; Reza Parwaresch

2003-01-01

316

CD4 T-Cell-Independent Antibody Response Reduces Enterovirus 71 Lethality in Mice by Decreasing Tissue Viral Loads  

PubMed Central

Enterovirus 71 (EV71) has induced fatal encephalitis in hundreds of thousands of infants and young children in the Asia-Pacific region since the past decade. Lymphocyte and antibody responses have been suspected to aggravate EV71-induced neurological symptoms, so anti-inflammatory agents have been used to treat patients with neurological symptoms. In the present study, we found that mice deficient in CD4+ T cells were resistant to EV71 infection as wild-type mice, whereas mice deficient in B cells were highly susceptible to viral infection. Compensation of CD4 T-cell function by other immune cells was not likely, because wild-type mice depleted of CD4+ T cells were also resistant to viral infection. Infected CD4 T-cell-deficient mice produced virus-specific neutralizing antibodies, IgM and IgG. Moreover, adoptive transfer of the virus-specific antibody produced by infected CD4 T-cell-deficient mice protected B-cell-deficient mice from infection by reducing tissue viral loads. Collectively, our results show that the CD4 T-cell-independent antibody response promotes the survival of EV71-infected mice and suggest great potential for the use of vaccines and neutralizing antibodies to reduce fatal symptoms in patients.

Wang, Li-Chiu; Kao, Chia-Min; Ling, Pin; Su, Ih-Jen; Chang, Tung-Miao; Chen, Shun-Hua

2012-01-01

317

Exogenous interleukin-6, interleukin-13, and interferon-gamma provoke pulmonary abnormality with mild edema in enterovirus 71-infected mice  

PubMed Central

Background Neonatal mice developed neurological disease and pulmonary dysfunction after an infection with a mouse-adapted human Enterovirus 71 (EV71) strain MP4. However, the hallmark of severe human EV71 infection, pulmonary edema (PE), was not evident. Methods To test whether EV71-induced PE required a proinflammatory cytokine response, exogenous pro-inflammatory cytokines were administered to EV71-infected mice during the late stage of infection. Results After intracranial infection of EV71/MP4, 7-day-old mice developed hind-limb paralysis, pulmonary dysfunction, and emphysema. A transient increase was observed in serum IL-6, IL-10, IL-13, and IFN-?, but not noradrenaline. At day 3 post infection, treatment with IL-6, IL-13, and IFN-? provoked mild PE and severe emphysema that were accompanied by pulmonary dysfunction in EV71-infected, but not herpes simplex virus-1 (HSV-1)-infected control mice. Adult mice did not develop PE after an intracerebral microinjection of EV71 into the nucleus tractus solitarii (NTS). While viral antigen accumulated in the ventral medulla and the NTS of intracerebrally injected mice, neuronal loss was observed in the ventral medulla only. Conclusions Exogenous IL-6, IL-13, and IFN-? treatment could induce mild PE and exacerbate pulmonary abnormality of EV71-infected mice. However, other factors such as over-activation of the sympathetic nervous system may also be required for the development of classic PE symptoms.

2011-01-01

318

Enterovirus 71-induced neurological disorders in young gerbils, Meriones unguiculatus: development and application of a neurological disease model.  

PubMed

A reliable disease model mimicking Enterovirus 71 (EV71) infection in humans is essential for understanding pathogenesis and for developing a safe and effective vaccine. Commonly used rodent models including mouse or rat models are not suitable for vaccine evaluation because the rodents are resistant to EV71 infection after they reach the age of 6 days. In this study, 21-day-old gerbils inoculated intraperitoneally (IP) with a non mouse-adapted EV71 strain developed neurological lesion-related signs including hind limb paralysis, slowness, ataxia and lethargy similar to those of central nervous system (CNS) infection of EV71 in humans. The infected gerbils eventually died of the neurological lesions and EV71 could be isolated from lung, liver, spleen, kidney, heart, spinal cord, brain cortex, brainstem and skeletal muscle. Significantly high virus replication was detected in spinal cord, brainstem and skeletal muscle by cellular analysis, real-time quantitative PCR (RT-PCR) and immunohistochemical staining. Histopathologic changes such as neuronal degeneration, neuronal loss and neuronophagia were observed in spinal cord, brain cortex, brainstem, and skeletal muscle along with necrotizing myositis and splenic atrophy. Gerbils that received two doses of inactive whole-virus vaccine showed no EV71-specific symptoms after challenged with EV71. In contrast, gerbils that received mock vaccination died of EV71-induced neuropathology after challenged with EV71. The result indicates that gerbils can serve as a reliable disease model for evaluating safety and efficacy of EV71 vaccine. PMID:23284845

Yao, Ping-Ping; Qian, Lei; Xia, Yong; Xu, Fang; Yang, Zhang-Nv; Xie, Rong-Hui; Li, Xiao; Liang, Wei-Feng; Huang, Xiao-Xiao; Zhu, Zhi-Yong; Zhu, Han-Ping

2012-12-21

319

Antenatal diagnosis of intrauterine infection with coxsackievirus B3 associated with live birth.  

PubMed Central

BACKGROUND: Prior reported cases of stillbirth and neonates infected with enteroviruses suggest transplacental infection. We present a case of fetal infection with coxsackievirus B3, diagnosed antenatally and resulting in live birth. CASE: A pregnant woman presented at 26 weeks with fetal tachycardia and non-immune hydrops fetalis. Coxsackievirus B3 was cultured from amniotic fluid. Maternal antibody to coxsackievirus B3 was positive at 1:512. At 32 weeks, the fetus deteriorated and was delivered. Cord blood antibody to coxsackievirus B3 was positive at a higher titer. Following neonatal death, brain and placental tissues were positive for enterovirus ribonucleic acid by polymerase chain reaction. CONCLUSION: Intrauterine infection by enteroviruses should be considered in the differential diagnosis of non-immune hydrops fetalis. Antenatal diagnosis of coxsackievirus B3 infection is associated with poor outcome.

Ouellet, Annie; Sherlock, Rebecca; Toye, Baldwin; Fung, Karen Fung Kee

2004-01-01

320

Complete Genome Sequence of a Recombinant Coxsackievirus B4 from a Patient with a Fatal Case of Hand, Foot, and Mouth Disease in Guangxi, China  

PubMed Central

The coxsackievirus B4 (CVB4) belongs to human enterovirus B species within the family Picornaviridae. Here we report a novel complete genome sequence of a recombinant CVB4 strain, CVB4/GX/10, which was isolated from a patient with a fatal case of hand, foot, and mouth disease in China. The complete genome consists of 7,293 nucleotides, excluding the 3? poly(A) tail, and has an open reading frame that maps between nucleotide positions 742 and 7293 and encodes a 2,183-amino-acid polyprotein. Phylogenetic analysis based on different genome regions reveals that CVB4/GX/10 is closest to a CVB4 strain, EPIHFMD-CLOSE CONTACT-16, in the 5? half (VP4?2B) of the genome, although it is closer to a Chinese CVB5 strain, CVB5/Henan/2010, in the 3? half (2C?3D) of the genome. Furthermore, similar bootscan analysis based on the whole genomes demonstrates that recombination has possibly occurred within the 2C domain and that CVB4/GX/10 is a possible progeny of intertypic recombination of the CVB4 strain EPIHFMD-CLOSE CONTACT-16 and CVB5/Henan/2010 that occurred during their cocirculation and evolution, which is a relatively common phenomenon in enteroviruses.

Hu, Y. F.; Du, J.; Zhao, R.; Xue, Y.

2012-01-01

321

Production of monoclonal antibody recognizing a subpopulation of human B lymphocytes producing B cell growth factor (BCGF)  

SciTech Connect

Several laboratories have recently reported production of B cell growth factors (BCGF) by a variety of human B cell lines. The authors have described production of BCGF by clones of B cell lymphoma lines and by normal Staphylococcus aureus Cowan I-activated B cells. To further evaluate whether BCGF production was the function of particular subpopulations of B cells, they immunized mice with the BCGF producing line Namalva and then developed a panel of hybridomas. Monoclonal antibodies were screened for binding to Namalva and the absence of binding to a non-BCGF producing B cell line. One monoclonal antibody, NN4, which met these criteria was also noted by fluorescence-activated cell sorter analysis to stain clones from a B cell lymphoma line producing BCGF but not clones from the same line which do not produce BCGF. The monoclonal antibody did not stain T cells or monocytes but stained 1-5% of normal human B lymphocytes from peripheral blood or tonsils. Binding of /sup 125/I-labeled NN4 to NN4/sup +/ B cells was not affected by the presence of BCGF. Therefore, NN4 recognizes a unique antigen present on B cell lines which produces BCGF; studies are in progress to determine the significance of this antigen on normal B cells.

Witzel, N.; Ambrus, J.L. Jr.; Jurgensen, C.H.; Mostowski, H.; Fauci, A.S.

1986-03-05

322

Antigen-Specific Antibody Production of Human B Cells in NOG Mice Reconstituted with the Human Immune System  

Microsoft Academic Search

Passive antibody administration shows strong potential as a new therapeutic method. In clinical applications, human-derived\\u000a antibodies with antigen specificity are more useful without putting individuals at risk. Production of human-derived antibodies\\u000a against given antigens can be obtained from animal models if the human immune system is established in the animals. In fact,\\u000a past reports revealed that human T and B

M. Ito; M. Shiina; Y. Saito; Y. Tokuda; Y. Kametani; Sonoko Habu

323

Relation between T-cell responses to glutamate decarboxylase and coxsackievirus B4 in patients with insulin-dependent diabetes mellitus  

Microsoft Academic Search

Background: the role of enteroviruses has been implicated in the etiology of insulin-dependent diabetes mellitus (IDDM). A possible connection between glutamate decarboxylase (GAD) autoimmunity and enterovirus infections in IDDM has been suggested to be based on a homology region between GAD and the non-structural protein 2C of coxsackievirus B4 (CVB4). Objectives: the aims of the study were to measure the

P Klemetti; H Hyöty; M Roivainen; J Ilonen; K Savola; M Knip; H. K Åkerblom; O Vaarala

1999-01-01

324

Lung tumorigenesis associated with erb-B-2 and erb-B-3 overexpression in human erb-B-3 transgenic mice is enhanced by methylnitrosourea.  

PubMed

Erb-B-3 overexpression is associated with poor prognosis in non-small cell lung cancer and is often overexpressed in breast cancers. MMTVhuman-erb-B-3 transgenic mice were generated to evaluate the impact of erb-B-3 overexpression on lung and mammary gland tumorigenesis. These transgenic mice developed a high incidence of lung adenocarcinomas but not mammary gland tumors. The tumors overexpressed transgenic human [h]-erb-B-3 but also overexpressed endogenous erb-B-2, indicating that the heterodimer of h-erb-B-3-erb-B-2 was required for proliferative signal transduction to the nucleus. Lung tumor latency was shorter and the incidence higher in erb-B-3 transgenic mice treated with the methylating agent, methylnitrosourea [MNU]. In MNU treated mice, K-ras activating point mutations in codon 12, synergized with h-erb-B-3 in lung tumorogenesis. In bitransgenic MMTVrat-erb-B2/MMTV-human-erb-B-3 mice, lung tumor latency was also significantly shortened. Unlike over-expression of rat-erb-B-2, overexpression of h-erb-B-3 did not alter the incidence or latency of mammary tumors. Coupled erb-B-2 and erb-B-3 overexpression as well as K-ras activation induced signaling through mitogen-activated protein kinase (MAPK). This animal model links erb-B-3 with lung cancer, suggests that erb-B-2 and erb-B-3 heterodimerization is a necessary intermediate, and documents latency shortening by methylating agent-induced mutation of K-ras. This erb-B-3 mouse lung cancer model will help dissect genetic changes in lung tumorigenesis and may be useful for chemoprevention studies. PMID:12483526

Zhou, Hang; Liu, Lili; Lee, Keunmyoung; Qin, Xiusheng; Grasso, Adam W; Kung, Hsing-Jien; Willis, Joesph E; Kern, Jeffery; Wagner, Thomas; Gerson, Stanton L

2002-12-12

325

Ribonucleoprotein complex formation by the human hepatitis B virus polymerase.  

PubMed

Human hepatitis B virus (HBV) polymerase (pol or RT), when expressed in Xenopus oocytes upon injection of synthetic minimal pol RNA (RT RNA), assembles into a higher molecular weight complex with the characteristics of a ribonuclear protein (RNP) complex. In vitro RNA competition binding data suggest that RT RNA is preferentially packaged into this complex even though it lacks the authentic viral encapsidation signal, epsilon, and viral capsid protein sequences. Consistent with this finding, the in vitro polymerase reaction performed in pol-expressing oocyte extracts generates primarily HBV-specific DNAs even when the pol template is challenged with a coinjected non-HBV competitor RNA. These results suggest that interaction between pol and its cognate RNA can be mediated by sequences other than the known packaging elements. We speculate that HBV RNP complexes containing at least polymerase and viral RNA may play a role in viral nucleocapsid assembly and may help to segregate HBV reverse transcription from the cellular milieu in vivo. PMID:8724861

Seifer, M; Standring, D N

1995-01-01

326

SURFACE MARKERS ON HUMAN T AND B LYMPHOCYTES  

PubMed Central

By using the two criteria (a) high density of immunoglobulin determinants on the cell surface and (b) presence of receptors for C'3 on the cell surface for defining bone marrow-derived lymphocytes, it is indirectly shown that all or at least a major population of human thymus-derived lymphocytes under certain conditions will form nonimmune rosettes with sheep red blood cells (SRBC). Almost all thymocytes tested from two different donors formed rosettes. The SRBC rosettes are not formed by virtue of immunoglobulin receptors and form only around living cells. Positive bivalent ions are required for rosette formation since EDTA will block rosette formation. Sodium iodoacetate will also block rosette formation demonstrating the dependence on an intact glycolytic pathway. Rosette formation is temperature dependent and will not appear at 37°C. Trypsin treatment of lymphocytes will abolish their SRBC-binding ability which cannot be restored by treating them with fresh donor serum or fetal calf serum, but which will reappear after culturing the lymphocytes. It is suggested that these rosettes are formed by a rapidly released or metabolized receptor substance on the living cell surface which behaves as a trypsin-sensitive structure produced by the cells themselves.

Jondal, M.; Holm, G.; Wigzell, H.

1972-01-01

327

The export of immunoglobulin D by human neoplastic B lymphocytes  

PubMed Central

An investigation has been made into the ability of human neoplastic B lymphocytes expressing surface IgM and IgD to export IgD in culture. Cells that expressed surface Ig of the lambda light chain type frequently exported IgD (10/12 patients), whereas cells expressing surface Ig of the kappa light chain type exported no IgD, although most (8/11 patients) were able to export IgM. It appears, therefore, that in most of the 23 cases studied, cells synthesizing IgD with lambda light chains can both express and export IgD, whereas those synthesizing IgD kappa can only insert it into the surface membrane. This finding and the known preponderance of lambda in plasma IgD imply that the possession of a lambda chain facilitates the IgD secretory pathway, a conclusion that implicates a control mechanism subsequent to the surface/secretory dichotomy arising from different splicings of heavy chain messenger RNA.

1983-01-01

328

EphB and Ephrin-B Interactions Mediate Human Mesenchymal Stem Cell Suppression of Activated T-Cells.  

PubMed

Mesenchymal stromal/stem cells (MSC) express the contact-dependent erythropoietin-producing hepatocellular (Eph) receptor tyrosine kinase family and their cognate ephrin ligands, which are known to regulate thymocyte maturation and selection, T-cell transendothelial migration, activation, co-stimulation, and proliferation. However, the contribution of Eph/ephrin molecules in mediating human MSC suppression of activated T-cells remains to be determined. In the present study, we showed that EphB2 and ephrin-B2 are expressed by ex vivo expanded MSC, while the corresponding ligands, ephrin-B1 and EphB4, respectively, are highly expressed by T-cells. Initial studies demonstrated that EphB2-Fc and ephrin-B2-Fc molecules suppressed T-cell proliferation in allogeneic mixed lymphocyte reaction (MLR) assays compared with human IgG-treated controls. While the addition of a third-party MSC population demonstrated dramatic suppression of T-cell proliferation responses in the MLR, blocking the function of EphB2 or EphB4 receptors using inhibitor binding peptides significantly increased T-cell proliferation. Consistent with these observations, shRNA EphB2 or ephrin-B2 knockdown expression in MSC reduced their ability to inhibit T-cell proliferation. Importantly, the expression of immunosuppressive factors, indoleamine 2, 3-dioxygenase, transforming growth factor-?1, and inducible nitric oxide synthase expressed by MSC, was up-regulated after stimulation with EphB4 and ephrin-B1 in the presence of interferon (IFN)-?, compared with untreated controls. Conversely, key factors involved in T-cell activation and proliferation, such as interleukin (IL)-2, IFN-?, tumor necrosis factor-?, and IL-17, were down-regulated by T-cells treated with EphB2 or ephrin-B2 compared with untreated controls. Studies utilizing signaling inhibitors revealed that inhibition of T-cell proliferation is partly mediated through EphB2-induced ephrin-B1 reverse signaling or ephrin-B2-mediated EphB4 forward signaling by activating Src, PI3Kinase, Abl, and JNK kinase pathways, activated by tyrosine phosphorylation. Taken together, these observations suggest that EphB/ephrin-B interactions play an important role in mediating human MSC inhibition of activated T cells. PMID:23711177

Nguyen, Thao M; Arthur, Agnes; Hayball, John D; Gronthos, Stan

2013-06-29

329

Coxsackievirus B5 induced apoptosis of HeLa cells: Effects on p53 and SUMO  

SciTech Connect

Coxsackievirus B5 (CVB5), a human enterovirus of the family Picornaviridae, is a frequent cause of acute and chronic human diseases. The pathogenesis of enteroviral infections is not completely understood, and the fate of the CVB5-infected cell has a pivotal role in this process. We have investigated the CVB5-induced apoptosis of HeLa cells and found that it happens by the intrinsic pathway by a mechanism dependent on the ubiquitin-proteasome system, associated with nuclear aggregation of p53. Striking redistribution of both SUMO and UBC9 was noted at 4 h post-infection, simultaneously with a reduction in the levels of the ubiquitin-ligase HDM2. Taken together, these results suggest that CVB5 infection of HeLa cells elicit the intrinsic pathway of apoptosis by MDM2 degradation and p53 activation, destabilizing protein sumoylation, by a mechanism that is dependent on a functional ubiquitin-proteasome system.

Gomes, Rogerio, E-mail: roggomes@usp.b [Department of Cell Biology, University of Sao Paulo School of Medicine at Ribeirao Preto, Av. dos Bandeirantes, 3900, 14049-900, SP (Brazil); Guerra-Sa, Renata [Federal University of Ouro Preto, MG (Brazil); Arruda, Eurico [Department of Cell Biology, University of Sao Paulo School of Medicine at Ribeirao Preto, Av. dos Bandeirantes, 3900, 14049-900, SP (Brazil); Virology Research Center, University of Sao Paulo School of Medicine at Ribeirao Preto, SP (Brazil)

2010-01-20

330

The Human Igp cDNA Sequence, a Homologue of Murine B29, Is Identical in B Cell and Plasma Cell Lines Producing All the Human Ig Isotypes  

Microsoft Academic Search

The B cell Ag receptor complex consists of at least two disulfide-linked, heterodimeric structures: the clonally restricted membrane Ig (mlg) molecule and the nonpolymorphic Ig-a:lg-\\/3 protein dimer. The latter molecule is encoded by two separate genes, mb-1 and B29. The DNA sequences of murine and human mb-1 and murine B29 have been determined previously. This study describes the sequence of

Shiori Hashimoto; Peter K. Gregersen; Nicholas Chiorazzi

331

The Prevalence and Persistence of Human Parvovirus B19 Infection in Thalassemic Patients  

Microsoft Academic Search

SUMMARY Human parvovirus B19 infection was studied in 60 thalassemic patients in Thailand. Seroprevalence, persistence of parvovirus B19 and their genotypes were identified in blood samples. Prevalence of anti-parvovirus B19 IgG and DNA found in thalassemic patients were 38% and 13%, respectively. Anti-parvovirus B19 IgM could be detected in 4% of these positive anti-parvovirus B19 IgG patients. The seroprevalence and

Sontana Siritantikorn; Noppadol Siritanaratkul; Apiradee Theamboonlers; Wannee Kantakamalakul; Chantapong Wasi

332

Proteolytic Processing of Human Cytomegalovirus Glycoprotein B Is Dispensable for Viral Growth in Culture  

Microsoft Academic Search

Glycoprotein B (gB) of human cytomegalovirus (HCMV), which is considered essential for the viral life cycle, is proteolytically processed during maturation. Since gB homologues of several other herpesviruses remain uncleaved, the relevance of this property of HCMV gB for viral infectivity is unclear. Here we report on the construction of a viral mutant in which the recognition site of gB

Tanja Strive; Eva Borst; Martin Messerle; Klaus Radsak

2002-01-01

333

A Virus Similar to Human Hepatitis B Virus Associated with Hepatitis and Hepatoma in Woodchucks  

Microsoft Academic Search

Particles with properties similar to those associated with human hepatitis B were found in serum from woodchucks with chronic hepatitis and hepatocellular carcinoma. It is suggested that woodchuck hepatitis virus is a second member of a novel class of viruses represented by the human hepatitis B virus.

Jesse Summers; Jo Marie Smolec; Robert Snyder

1978-01-01

334

B-Virus (Cercopithecine herpesvirus 1) Infection in Humans and Macaques: Potential for Zoonotic Disease  

Microsoft Academic Search

Nonhuman primates are widely used in biomedical research because of their genetic, anatomic, and physiologic similarities to humans. In this setting, human contact directly with macaques or with their tissues and fluids sometimes occurs. Cercopithecine herpesvirus 1 (B virus), an alphaherp- esvirus endemic in Asian macaques, is closely related to her- pes simplex virus (HSV). Most macaques carry B virus

Jennifer L. Huff; Peter A. Barry

2003-01-01

335

Identification of Functional Human Splenic Memory B Cells by Expression of CD148 and CD27  

Microsoft Academic Search

Summary Memory B cells isolated from human tonsils are characterized by an activated cell surface phe- notype, localization to mucosal epithelium, expression of somatically mutated immunoglobulin (Ig) variable (V) region genes, and a preferential differentiation into plasma cells in vitro. In spleens of both humans and rodents, a subset of memory B cells is believed to reside in the marginal

Stuart G. Tangye; Yong-Jun Liu; Gregorio Aversa; Joseph H. Phillips; Jan E. de Vries

2010-01-01

336

Canonical pathway of nuclear factor B activation selectively regulates proinflammatory and prothrombotic responses in human atherosclerosis  

Microsoft Academic Search

Nuclear factor B (NF-B) activation has been observed in human atherosclerotic plaques and is enhanced in unstable coronary plaques, but whether such activation has a protective or pathophysiological role remains to be determined. We addressed this question by developing a short-term culture system of cells isolated from human atherosclerotic tissue, allowing efficient gene transfer to directly investigate signaling pathways in

Claudia Monaco; Evangelos Andreakos; Serafim Kiriakidis; Claudia Mauri; Colin Bicknell; Brian Foxwell; Nicholas Cheshire; Ewa Paleolog; Marc Feldmann

2004-01-01

337

Influence of pan-caspase inhibitors on coxsackievirus B3-infected CD19+ B lymphocytes.  

PubMed

Coxsackievirus B3 (CVB3), together with other enteroviruses of the picornavirus family, is associated with a wide variety of acute and chronic forms of human diseases. Using the murine model of CVB3-caused myocarditis, this pathogen can be detected not only in solid organs but also in different types of immune cells, preferentially in B lymphocytes. Therefore, these cells could represent a non-cardiac virus reservoir and may play an important role with regard to viral dissemination in the infected host. In addition, the infection of specific immune cells might modulate the severity of tissue injury and the pattern of virus-caused pathology in susceptible or resistant individuals. In the present study it could be demonstrated that CVB3 was capable to infect productively a certain percentage of murine CD19(+) B cells. In vivo studies revealed that CVB3 invaded murine CD19(+) B cells during an acute infection. Three days p. i. approximately 0.5-1.0% of these cells were productively infected. This proportion could be decreased up to 45%, if 3 days p. i. mice were intravenously treated with the pan-caspase inhibitors Z-VAD-FMK or Q-VD-OPH. These data were compared with results obtained from CVB3-infected human Raji cells. PMID:17520192

Jarasch, Nadine; Martin, Ulrike; Zell, Roland; Wutzler, Peter; Henke, Andreas

2007-09-01

338

An epidemic of acute haemorrhagic conjunctivitis caused by enterovirus 70 in Okinawa, Japan, in 1994  

Microsoft Academic Search

· Background: Although enterovirus 70 (EV70) has been identified as the major aetiological agent of acute haemorrhagic conjunctivities\\u000a (ACH),no EV70 strain has been isolated by cell culture method since 1988. Therefore, recent clinical and epidemiological characteristics\\u000a of AHC caused by EV70 have not been clarified. · Methods: Clinical and serological studies were carried out on patients during\\u000a the AHC epidemic

Eiichi Uchio; Kenji Yamazaki; Hideo Ishikawa; Isao Matsunaga; Yoshimori Asato; Koki Aoki; Shigeaki Ohno

1999-01-01

339

Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity  

Microsoft Academic Search

Enterovirus type 70, an etiologic agent of acute hemorrhagic conjunctivitis, may bind different cellular receptors depending on cell type. To understand how EV70-receptor interaction is controlled, we studied two variants of the virus with distinct receptor utilization. EV70-Rmk, derived by passage in rhesus monkey kidney cells, replicates poorly in HeLa cells and does not cause cytopathic effects. Decay accelerating factor

Melissa Stewart Kim; Vincent R. Racaniello

2007-01-01

340

RT-PCR, nucleotide, amino acid and phylogenetic analyses of enterovirus type 71 strains from Asia  

Microsoft Academic Search

A specific and sensitive method based on RT-PCR was developed to detect enterovirus 71 (EV71) from patients with hand, foot and mouth disease, myocarditis, aseptic meningitis and acute flaccid paralysis. RT-PCR primers from conserved parts of the VP1 capsid gene were designed on the basis of good correlation with sequences of EV71 strains. These primers successfully amplified 44 strains of

Sunita Singh; Vincent T. K. Chow; K. P. Chan; A. E. Ling; C. L. Poh

2000-01-01

341

An overview of the evolution of enterovirus 71 and its clinical and public health significance  

Microsoft Academic Search

Since its discovery in 1969, enterovirus 71 (EV71) has been recognised as a frequent cause of epidemics of hand-foot-and-mouth disease (HFMD) associated with severe neurological sequelae in a small proportion of cases. There has been a significant increase in EV71 epidemic activity throughout the Asia–Pacific region since 1997. Recent HFMD epidemics in this region have been associated with a severe

Peter C McMinn

2002-01-01

342

Development of a quantitative enzyme linked immunosorbent assay for monitoring the Enterovirus 71 vaccine manufacturing process  

Microsoft Academic Search

Enterovirus 71 (EV71), the etiologic agent causes outbreaks with significant mortality in young children in Asia and currently there is no vaccine available. In this study, we report a quantitative enzyme linked immunosorbent assay (Q-ELISA) to determine the concentration of the EV71 VP2 antigen. EV71 virus-like particles (VLPs) were produced in the baculovirus expression system and used as the EV71

Chia-Chyi Liu; Hsuen-Wen Chang; Grace Yang; Jen-Ron Chiang; Yen-Hung Chow; I-Hsi Sai; Jui-Yuan Chang; Sue-Chen Lin; Charles Sia; Chia-Hsin Hsiao; Ai-Hsiang Chou; Pele Chong

2011-01-01

343

Oral immunization of mice using transgenic tomato fruit expressing VP1 protein from enterovirus 71  

Microsoft Academic Search

Enterovirus 71 (EV71) causes seasonal epidemics of hand-foot-and-mouth disease associated with fatal neurological complications in young children, and several major outbreaks have occurred recently. This study developed an effective antiviral agent by transforming the gene for VP1 protein, a previously defined epitope and also a coat protein of EV71, into tomato plant. VP1 protein was first fused with sorting signals

Hsuan-Fu Chen; Meng-Huei Chang; Bor-Luen Chiang; Shih-Tong Jeng

2006-01-01

344

Heterologous expression and epitope mapping of a human small nuclear ribonucleoprotein-associated Sm-B'/B autoantigen.  

PubMed

The U snRNP associated B'/B polypeptides are primary targets of Sm autoantibodies in patients with systemic lupus erythematosus. We have bacterially expressed a Sm-B'/B autoantigen from Raji cells as a fusion with the anthranilate synthase protein from Escherichia coli. The recombinant Sm-B'/B fusion displays comparable immunologic reactivity to the native protein when tested with both monoclonal and polyclonal antibodies. To map Sm-B'/B epitopes, we constructed a series of 12 anthranilate synthase fusions spanning different regions of Sm-B'/B and tested such fusions on immunoblots against a panel of characterized sera. In this manner, we have identified six epitopes, five of which overlap the proline-rich carboxyl-terminus of the protein. Some of these epitopes appear to be conformational. The human sera tested can be divided, according to the epitopes they recognize, into six groups. Finally, we have shown that anti-Sm recognition of the (U1)RNP-specific A protein is attributable to cross-reactivity between the Sm-B'/B and A autoantigens. PMID:1688587

Rokeach, L A; Jannatipour, M; Hoch, S O

1990-02-01

345

A role for gut-associated lymphoid tissue in shaping the human B cell repertoire.  

PubMed

We have tracked the fate of immature human B cells at a critical stage in their development when the mature B cell repertoire is shaped. We show that a major subset of bone marrow emigrant immature human B cells, the transitional 2 (T2) B cells, homes to gut-associated lymphoid tissue (GALT) and that most T2 B cells isolated from human GALT are activated. Activation in GALT is a previously unknown potential fate for immature human B cells. The process of maturation from immature transitional B cell through to mature naive B cell includes the removal of autoreactive cells from the developing repertoire, a process which is known to fail in systemic lupus erythematosus (SLE). We observe that immature B cells in SLE are poorly equipped to access the gut and that gut immune compartments are depleted in SLE. Thus, activation of immature B cells in GALT may function as a checkpoint that protects against autoimmunity. In healthy individuals, this pathway may be involved in generating the vast population of IgA plasma cells and also the enigmatic marginal zone B cell subset that is poorly understood in humans. PMID:23940259

Vossenkämper, Anna; Blair, Paul A; Safinia, Niloufar; Fraser, Louise D; Das, Lisa; Sanders, Theodore J; Stagg, Andrew J; Sanderson, Jeremy D; Taylor, Kirstin; Chang, Fuju; Choong, Lee M; D'Cruz, David P; Macdonald, Thomas T; Lombardi, Giovanna; Spencer, Jo

2013-08-12

346

High sensitivity and label-free detection of Enterovirus 71 by nanogold modified electrochemical impedance spectroscopy  

NASA Astrophysics Data System (ADS)

Enterovirus 71 (EV71), which is the most fulminant and invasive species of enterovirus, can cause children neurologic complications and death within 2-3 days after fever and rash developed. Besides, EV71 has high sequence similarity with Coxsackie A 16 (CA16) that makes differential diagnosis difficult in clinic and laboratory. Since conventional viral diagnostic method cannot diagnose EV71 quickly and EV71 can transmit at low viral titer, the patients might delay in treatment. A quick, high sensitive, and high specific test for EV71 detection is pivotal. Electrochemical impedance spectroscopy (EIS) has been applied for detecting bio-molecules as biosensors recently. In this study, we try to build a detection platform for EV71 detection by nanogold modified EIS probe. The result shows that our probe can detect 3.6 VP1/50 ?l (one EV71 particle has 60 VP1) in 3 minutes. The test can also distinguish EV71 from CA16 and lysozyme. Diagnosis of enterovirus 71 by electrochemical impedance spectroscopy has the potential to apply in clinic.

Wang, Fang-Yu; Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Yang, Jyh-Yuan; Chang, Chia-Ching

2013-03-01

347

Outbreak of vertigo in Wyoming: possible role of an enterovirus infection.  

PubMed

An epidemiologic investigation was conducted to characterize and evaluate the possibility of a viral aetiology of an outbreak of acute vertigo in Hot Springs Country, Wyoming, during autumn 1992. Case-finding identified Hot Springs County residents who sought medical attention for new onset vertigo during 1 August, 1992-31 January 1993. Thirty-five case-patients and 61 matched controls were interviewed and serum specimens were obtained during January 1993. Case-patients were more likely than controls to report symptoms (e.g. fatigue, sore throat, fever, diarrhoea) of antecedent acute illness. Case-patients did not have a significantly greater prevalence or mean titre of IgG antibodies to respiratory syncytial virus, parainfluenza viruses, Epstein-Barr virus, and cytomegalovirus than controls. Serologic evidence of recent enterovirus infection (IgM antibodies) was found for 74% of case-patients compared with 54% of controls (P < 0.05), suggesting a possible association between vertigo and enterovirus infection. Future studies are needed to define the role of enteroviruses in innerear diseases. PMID:8760963

Simonsen, L; Khan, A S; Gary, H E; Hanson, C; Pallansch, M A; Music, S; Holman, R C; Stewart, J A; Erdman, D D; Arden, N H; Arenberg, I K; Schonberger, L B

1996-08-01

348

B-less: a strain of profoundly B cell-deficient mice expressing a human lambda transgene  

PubMed Central

We have created several transgenic mouse strains that bear the human lambda light chain gene driven by its own promoter and a mouse immunoglobulin heavy chain enhancer. The transgene is expressed in many tissues, with particularly high levels of expression in the bone marrow, thymus, spleen, and lymph nodes. One of these transgenic lines, B-less, displays a dramatic phenotype characterized by an acute susceptibility to bacterial and viral infections. Analysis of this strain shows it to be profoundly deficient in both immature (pre-B) and mature B cells, as well as in circulating immunoglobulin. The pre-B and B cell defects are cell autonomous, as judged by cell culture and bone marrow graft chimeras. Despite this B cell deficiency, the T cell lineage appears grossly normal as assessed by flow cytometric analysis and by its response to mitogen stimulation. Since an independently derived transgenic strain bearing the same human lambda construct displays a partial B-less phenotype, it is likely that the B lineage deficiency is due to a dominant effect of transgene expression rather than to the insertional perturbation of an endogenous mouse gene. It is interesting that the deficiency phenotype is fully expressed in the FVB/N genetic background, but is suppressed in F1 hybrids formed between the FVB/N and C57BL/6 inbred strains. Evidently, there are one or more dominant genetic suppressors of B-less in the C57BL/6 genome.

1992-01-01

349

ErbB receptor tyrosine kinase/NF-?B signaling controls mammosphere formation in human breast cancer  

PubMed Central

Breast cancer is one of the most common cancers in humans. However, our understanding of the cellular and molecular mechanisms underlying tumorigenesis in breast tissues is limited. Here, we identified a molecular mechanism that controls the ability of breast cancer cells to form multicellular spheroids (mammospheres). We found that heregulin (HRG), a ligand for ErbB3, induced mammosphere formation of a breast cancer stem cell (BCSC)–enriched population as well as in breast cancer cell lines. HRG-induced mammosphere formation was reduced by treatment with inhibitors for phosphatidyl inositol 3-kinase (PI3K) or NF-?B and by expression of I?B?-Super Repressor (I?B?SR), a dominant-negative inhibitor for NF-?B. Moreover, the overexpression of I?B?SR in breast cancer cells inhibited tumorigenesis in NOD/SCID mice. Furthermore, we found that the expression of IL8, a regulator of self-renewal in BCSC-enriched populations, was induced by HRG through the activation of the PI3K/NF-?B pathway. These findings illustrate that HRG/ErbB3 signaling appears to maintain mammosphere formation through a PI3K/NF-?B pathway in human breast cancer.

Hinohara, Kunihiko; Kobayashi, Seiichiro; Kanauchi, Hajime; Shimizu, Seiichiro; Nishioka, Kotoe; Tsuji, Ei-ichi; Tada, Kei-ichiro; Umezawa, Kazuo; Mori, Masaki; Ogawa, Toshihisa; Inoue, Jun-ichiro; Tojo, Arinobu; Gotoh, Noriko

2012-01-01

350

Stereoselective Metabolism of Bupropion by Cytochrome P4502B6 (CYP2B6) and Human Liver Microsomes  

PubMed Central

Purpose Hydroxylation of the antidepressant and smoking deterrent drug bupropion is a clinically important bioactivation and elimination pathway. Bupropion hydroxylation is catalyzed selectively by cytochrome P4502B6 (CYP2B6). CYP2B6-catalyzed bupropion hydroxylation has been used as an in vitro and in vivo phenotypic probe for CYP2B6 activity and CYP2B6 drug interactions. Bupropion is chiral, used clinically as a racemate, and disposition is stereoselective. Nevertheless, it is unknown whether CYP2B6-catalyzed bupropion hydroxylation is stereoselective. Methods Hydroxylation of racemic bupropion by recombinant CYP2B6 and human liver microsomes was evaluated using a stereoselective assay. Results At therapeutic concentrations, hydroxylation of (S)-bupropion was 3-fold and 1.5-greater than (R)-bupropion, respectively, by recombinant CYP2B6 and human liver microsomes. In vitro intrinsic clearances were likewise different for bupropion enantiomers. Conclusions Stereoselective bupropion hydroxylation may have implications for the therapeutic efficacy of bupropion as an antidepressant or smoking cessation therapy, and for the use of bupropion as an in vivo phenotypic probe for CYP2B6 activity.

Coles, Rebecka; Kharasch, Evan D.

2013-01-01

351

The catalytic properties of human hepatitis B virus polymerase.  

PubMed

The DNA-dependent DNA polymerase (DDDP) and RNA-dependent DNA polymerase (RDDP) activities of hepadnavirus polymerases are both essential for viral replication. Human hepatitis B virus (HBV) polymerase has been successfully expressed in Escherichia coli as a fusion protein in frame with maltose-binding protein. The present study was undertaken to characterize these two activities and introduce an in vitro assay system. In situ activity gel assays show that the polymerase has both types of activities. One hundred thirty-four kilodaltons of active full-length product was proteolytically cleaved into approximately 73 kDa of active fragment by proteinase K preincubation. Mutation of conserved YMDD motif also confirms that the activities were due to the recombinant polymerase and that this motif is essential to polymerase activity. Two activities of the polymerase show their optima under conditions of 1 mM (DDDP) or 0.25 mM (RDDP) of MnCl2, 400 mM KCl, 37 degrees C (DDDP) or 24 degrees C (RDDP), and pH 7.0-7.7. Substitution of Mg2+ for Mn2+ results in reduction of processivity, which may explain why Mn2+ supports nucleotide incorporation to a higher level than Mg2+. The polymerase is resistant to aphidicolin. Actinomycin D acts selectively on DDDP activity, whereas phosphonoformic acid inhibits both activities. The in vitro HBV polymerase assay system demonstrated herein will be useful for screening potential HBV polymerase inhibitor for the development of anti-HBV drugs. PMID:8670270

Jeong, J H; Kwak, D S; Rho, H M; Jung, G

1996-06-14

352

The genomic signature of human rhinoviruses A, B and C.  

PubMed

Human rhinoviruses are single stranded positive sense RNA viruses that are presented in more than 50% of acute upper respiratory tract infections. Despite extensive studies on the genetic diversity of the virus, little is known about the forces driving it. In order to explain this diversity, many research groups have focused on protein sequence requirements for viable, functional and transmissible virus but have missed out an important aspect of viral evolution such as the genomic ontology of the virus. This study presents for the first time the genomic signature of 111 fully sequenced HRV strains from all three groups HRV-A, HRV-B and HRV-C. We observed an HRV genome tendency to eliminate CpG and UpA dinucleotides, coupling with over-representation of UpG and CpA. We propose a specific mechanism which describes how rapid changes in the HRV genomic sequence can take place under the strict control of conservation of the polypeptide backbone. Moreover, the distribution of the observed under- and over-represented dinucleotides along the HRV genome is presented. Distance matrice tables based on CpG and UpA odds ratios were constructed and viewed as heatmaps and distance trees. None of the suppressions can be attributed to codon usage or in RNA secondary structure requirements. Since viral recognition is dependent on RNA motifs rich in CpG and UpA, it is possible that the overall described genome evolution mechanism acts in order to protect the virus from host recognition. PMID:23028561

Megremis, Spyridon; Demetriou, Philippos; Makrinioti, Heidi; Manoussaki, Alkistis E; Papadopoulos, Nikolaos G

2012-09-13

353

Evidence for a wide extra-astrocytic distribution of S100B in human brain  

Microsoft Academic Search

BACKGROUND: S100B is considered an astrocytic in-situ marker and protein levels in cerebrospinal fluid (CSF) or serum are often used as biomarker for astrocytic damage or dysfunction. However, studies on S100B in the human brain are rare. Thus, the distribution of S100B was studied by immunohistochemistry in adult human brains to evaluate its cell-type specificity. RESULTS: Contrary to glial fibrillary

Johann Steiner; Hans-Gert Bernstein; Hendrik Bielau; Annika Berndt; Ralf Brisch; Christian Mawrin; Gerburg Keilhoff; Bernhard Bogerts

2007-01-01

354

Stimulatory and Inhibitory Influences of Human Immunodeficiency Virus on Normal B Lymphocytes  

Microsoft Academic Search

B-lymphocyte dysfunction is a characteristic feature of the acquired immunodeficiency syndrome (AIDS) and of the AIDS-related complex. The aim of the present study was to further examine the influences exercised by the human immunodeficiency virus (HIV; formerly called human T-lymphotropic virus type III or lymphadenopathy-associated virus, HTLV-III\\/LAV) on normal human B lymphocytes. An unfractionated protein preparation, made from HIV purified

Savita Pahwa; Rajendra Pahwa; Robert A. Good; Robert C. Gallo; Carl Saxinger

1986-01-01

355

Selective Uptake of Specifically Bound Cobalt58 Vitamin B12 by Human and Mouse Tumour Cells  

Microsoft Academic Search

MANY body fluids have been reported to contain substances that combine with vitamin B12. However, research concerning the role of these substances in the absorption of vitamin B12 by animal cells has been confined mainly to work involving intrinsic factor preparations or normal human gastric juice. A large amount of evidence has led to general acceptance that vitamin B12 bound

Bernard A. Cooper; William Paranchych

1961-01-01

356

Regulation of human B-cell activation, proliferation, and differentiation by soluble factors  

Microsoft Academic Search

This review describes a series of studies performed in our laboratory which have focused on the activation and subsequent proliferation and differentiation of human B lymphocytes. Utilizing polyclonal signals which activate B cells by interacting with their surface membrane Ig, we have examined the events in the transition of a resting B lymphocyte to an Ig-secreting cell. A major theme

Atsushi Muraguchi; John H. Kehrl; Joseph L. Butler; Anthony S. Fauci

1984-01-01

357

Epidemiology of human parvovirus B19 in children with sickle cell disease  

Microsoft Academic Search

Human parvovirus (HPV) B19 causes sig- nificant morbidity and mortality in chil- dren with sickle cell disease (SCD), but little data are published about the epidemi- ology of HPV B19 infection and its associ- ated complications in this patient popula- tion. In this study, prevalence and incidence rates of HPV B19 were deter- mined in 633 patients with SCD followed

Kim Smith-Whitley; Huaqing Zhao; Richard L. Hodinka; Janet Kwiatkowski; Renee Cecil; Tamara Cecil; Avital Cnaan; Kwaku Ohene-Frempong

2004-01-01

358

An insertion deletion polymorphism in the signal peptide of the human apolipoprotein B gene  

Microsoft Academic Search

In this communication we report the genetic properties of an insertion\\/deletion polymorphism in the signal peptide of the human apolipoprotein B (apo b) gene. There are two alleles of the apo B signal peptide; one codes for a peptide 27 amino acids in length and the other a peptide only 24 amino acids in length. Using the polymerase chain reaction

Sophia Visvikis; Lawrence Chan; Gerard Siest; Pierre Drouin; Eric Boerwinkle

1990-01-01

359

Rapid Simultaneous Detection of Enterovirus and Parechovirus RNAs in Clinical Samples by One-Step Real-Time Reverse Transcription-PCR Assay ?  

PubMed Central

Enteroviruses (EVs) are recognized as the major etiological agent in meningitis in children and young adults. The use of molecular techniques, such as PCR, has substantially improved the sensitivity of enterovirus detection compared to that of virus culture methods. PCR-based methods also can detect a much wider range of EV variants, including those within species A, as well as human parechoviruses (HPeVs) that often grow poorly in vitro and which previously have been underdiagnosed by traditional methods. To exploit these developments, we developed a real-time one-step reverse transcription-PCR (RT-PCR) for the rapid and sensitive detection of EV and HPeV in clinical specimens. Two commercially available RT-PCR kits were used (method I, Platinum one-step kit; method II, Express qPCR one-step kit) with primers and probes targeting the EV and HPeV 5?-untranslated regions (5?UTR). Amplification dynamics (threshold cycle [CT]values and efficiencies) of absolutely quantified full-length RNA transcripts representative of EV species A to D and HPeV were similar, demonstrating the effectiveness of both assays across the range of currently described human EV and HPeV variants. Probit analysis of multiple endpoint replicates demonstrated comparable sensitivities of the assays for EV and HPeV (method I, approximately 10 copies per reaction for both targets; method II, 20 copies per reaction). CT values were highly reproducible on repeat testing of positive controls within assays and between assay runs. Considering the sample turnaround time of less than 3 h, the multiplexed one-step RT-PCR method provides rapid diagnostic testing for EV and HPeV in cases of suspected central nervous system infections in a clinically relevant time frame.

Bennett, Susan; Harvala, Heli; Witteveldt, Jeroen; McWilliam Leitch, E. Carol; McLeish, Nigel; Templeton, Kate; Gunson, Rory; Carman, William F.; Simmonds, Peter

2011-01-01

360

DNMT3B7, a truncated DNMT3B isoform expressed in human tumors, disrupts embryonic development and accelerates lymphomagenesis  

PubMed Central

Epigenetic changes are among the most common alterations observed in cancer cells, yet the mechanism by which cancer cells acquire and maintain abnormal DNA methylation patterns is not understood. Cancer cells have an altered distribution of DNA methylation and express aberrant DNA methyltransferase 3B transcripts, which encode truncated proteins, some of which lack the C-terminal catalytic domain. To test if a truncated DNMT3B isoform disrupts DNA methylation in vivo, we constructed two lines of transgenic mice expressing DNMT3B7, a truncated DNMT3B isoform commonly found in cancer cells. DNMT3B7 transgenic mice exhibit altered embryonic development, including lymphopenia, craniofacial abnormalities, and cardiac defects, similar to Dnmt3b-deficient animals, but rarely develop cancer. However, when DNMT3B7 transgenic are bred with E?-Myc transgenic mice, which model aggressive B cell lymphoma, DNMT3B7 expression increases the frequency of mediastinal lymphomas in E?-Myc animals. E?-Myc/DNMT3B7 mediastinal lymphomas have more chromosomal rearrangements, increased global DNA methylation levels, and more locus-specific perturbations in DNA methylation patterns compared to E?-Myc lymphomas. These data represent the first in vivo modeling of cancer-associated DNA methylation changes and suggest that truncated DNMT3B isoforms contribute to the re-distribution of DNA methylation characterizing virtually every human tumor.

Shah, Mrinal Y.; Vasanthakumar, Aparna; Barnes, Natalie Y.; Figueroa, Maria E.; Kamp, Anna; Hendrick, Christopher; Ostler, Kelly R.; Davis, Elizabeth M.; Lin, Shang; Anastasi, John; Le Beau, Michelle M.; Moskowitz, Ivan; Melnick, Ari; Pytel, Peter; Godley, Lucy A.

2010-01-01

361

Genomic analysis of coxsackieviruses A1, A19, A22, enteroviruses 113 and 104: viruses representing two clades with distinct tropism within enterovirus C.  

PubMed

Coxsackieviruses (CV) A1, CV-A19 and CV-A22 have historically comprised a distinct phylogenetic clade within Enterovirus (EV) C. Several novel serotypes that are genetically similar to these three viruses have been recently discovered and characterized. Here, we report the coding sequence analysis of two genotypes of a previously uncharacterized serotype EV-C113 from Bangladesh and demonstrate that it is most similar to CV-A22 and EV-C116 within the capsid region. We sequenced novel genotypes of CV-A1, CV-A19 and CV-A22 from Bangladesh and observed a high rate of recombination within this group. We also report genomic analysis of the rarely reported EV-C104 circulating in the Gambia in 2009. All available EV-C104 sequences displayed a high degree of similarity within the structural genes but formed two clusters within the non-structural genes. One cluster included the recently reported EV-C117, suggesting an ancestral recombination between these two serotypes. Phylogenetic analysis of all available complete genome sequences indicated the existence of two subgroups within this distinct Enterovirus C clade: one has been exclusively recovered from gastrointestinal samples, while the other cluster has been implicated in respiratory disease. PMID:23761409

Tokarz, Rafal; Haq, Saddef; Sameroff, Stephen; Howie, Stephen R C; Lipkin, W Ian

2013-06-12

362

The signs B and B-bent in Israeli sign language according to the theory of Phonology as Human Behavior.  

PubMed

The purpose of the present research is to examine which of the two factors: (1) the iconic-semiotic factor; or (2) the human-phonetic factor is more relevant in explaining the appearance and distribution of the hand shape B-bent in Israeli Sign Language (ISL). The B-bent shape has been the subject of much attention in sign language research revolving around the question of its status as a phoneme. The arguments supporting the phonemic status of the B-bent hand shape have been primarily based on the semiotic opposition between the hand shape B and the hand shape B-bent. It has been claimed that in Italian Sign Language the hand shape B is perceptually distinct from the hand shape B-bent, i.e. in opposition to the general, neutral, unmarked meaning of the hand shape B, the iconic hand shape B-bent has a more narrow, specific and marked meaning: DELIMIT. The B-bent hand shape appears in spatial-temporal signs such as "a little before, ahead, postpone or behind". In these signs the iconic structure of the hand shape B-bent is utilized to mark borders in space and time. The arguments opposing the perceptual/phonemic distinction between these hand shapes is based on the human-phonetic factor, i.e. the need to reduce the effort on the part of the wrist joints in specific phonetic environments. We performed a quantitative and qualitative content analysis of the distribution of the basic units of 560 lexical signs taken from a stratified random sample from the ISL dictionary. The results were analyzed in the framework of the sign-oriented linguistic theory of the Columbia School including the theory of Phonology as Human Behavior. Our data revealed that the B-bent hand shape--as all the "building blocks" of the ISL--is a morpho-phonemic unit. We found that there is not only a phonemic distinction between hand shape B and hand shape B-bent in ISL (based on minimal pairs), but there is also a perceptual distinction between them. The qualitative analysis shows that the distribution of the B and the B-bent shapes in the lexicon of ISL is not random but rather motivated by a specific iconic meaning attached to each one of them. The results of our research lead to the conclusion that iconicity is the dominant explanatory factor in the Phonology of ISL. PMID:18415739

Fuks, Orit; Tobin, Yishai

363

B Cell Repertoire Analysis Identifies New Antigenic Domains on Glycoprotein B of Human Cytomegalovirus which Are Target of Neutralizing Antibodies  

PubMed Central

Human cytomegalovirus (HCMV), a herpesvirus, is a ubiquitously distributed pathogen that causes severe disease in immunosuppressed patients and infected newborns. Efforts are underway to prepare effective subunit vaccines and therapies including antiviral antibodies. However, current vaccine efforts are hampered by the lack of information on protective immune responses against HCMV. Characterizing the B-cell response in healthy infected individuals could aid in the design of optimal vaccines and therapeutic antibodies. To address this problem, we determined, for the first time, the B-cell repertoire against glycoprotein B (gB) of HCMV in different healthy HCMV seropositive individuals in an unbiased fashion. HCMV gB represents a dominant viral antigenic determinant for induction of neutralizing antibodies during infection and is also a component in several experimental HCMV vaccines currently being tested in humans. Our findings have revealed that the vast majority (>90%) of gB-specific antibodies secreted from B-cell clones do not have virus neutralizing activity. Most neutralizing antibodies were found to bind to epitopes not located within the previously characterized antigenic domains (AD) of gB. To map the target structures of these neutralizing antibodies, we generated a 3D model of HCMV gB and used it to identify surface exposed protein domains. Two protein domains were found to be targeted by the majority of neutralizing antibodies. Domain I, located between amino acids (aa) 133–343 of gB and domain II, a discontinuous domain, built from residues 121–132 and 344–438. Analysis of a larger panel of human sera from HCMV seropositive individuals revealed positivity rates of >50% against domain I and >90% against domain II, respectively. In accordance with previous nomenclature the domains were designated AD-4 (Dom II) and AD-5 (Dom I), respectively. Collectively, these data will contribute to optimal vaccine design and development of antibodies effective in passive immunization.

Wiegers, Anna-Katharina; Fisch, Tanja; Rucker, Pia; Sticht, Heinrich; Grieb, Nina; Baroti, Tina; Weisel, Florian; Stamminger, Thomas; Martin-Parras, Luis; Mach, Michael; Winkler, Thomas H.

2011-01-01

364

Cloning and chromosomal localization of the human TRK-B tyrosine kinase receptor gene (NTRK2)  

SciTech Connect

There is increasing evidence that neurotrophins and their receptors play an important role in regulating development of both the central and the peripheral nervous systems. Human TRK-A (NTRK1) and TRK-C (NTRK3) have been cloned and sequenced, but only a truncated form of human TRK-B has been published. Therefore, we isolated complementary DNAs spanning the entire coding region of both human full-length and truncated forms of TRK-B from human brain cDNA libraries. Human full-length TRK-B codes for a protein of 822 amino acid residues. The putative mature peptide sequence is 49% homologous to human TRK-A and 55% full-length human TRK-C, with 40% amino acid identity among TRK-A, -B, and -C. Nine of 13 cysteine residues, 4 of 12 N-glycosylation sites in the extracellular domain, and 10 of 13 tyrosine residues in the intracellular domain are conserved among human TRK-A, -B, and -C. There is a cluster of 10 serine residues in the juxtamembrane region of TRK-B that is absent in TRK-A. Two major sizes of TRK-B transcripts were expressed in human brain. Northern blot analysis using probes specific for the extracellular or the tyrosine kinase domain revealed that the 9.5-kb band encodes the truncated from of TRK-B mRNA. By fluorescence in situ hybridization and somatic cell hybrid mapping, the human TRK-B gene was localized to chromosome 9q22.1. 31 refs., 4 figs.

Nakagawara, A.; Liu, X.G.; Ikegaki, N. [Children`s Hospital of Philadelphia, PA (United States)] [and others

1995-01-20

365

Enrichment of cerebrospinal fluid samples on cell culture for enhancement of sensitivity of mumps and enterovirus detection by multiplex RT-PCR  

Microsoft Academic Search

It has been demonstrated that the detection of enteroviruses and mumps virus nucleic acid in cerebrospinal fluid (CSF) specimens improves the management of the patients with aseptic meningitis. To determine the effect of overnight enrichment of mumps and enteroviruses in CSF samples on cell culture for increasing the sensitivity of viral detection, we developed a multiplex reverse transcriptase-polymerase chain reaction

Masoumeh Kermanian; Hoorieh Soleimanjahi; Ehsan Arefian; Taravat Bamdad

2008-01-01

366

Simple method of detecting enteroviruses in contaminated molluscs and sewage by using polymerase chain reaction coupled with a colorimetric microwell detection assay  

Microsoft Academic Search

The methods normally used for the detection of enteroviruses in environmental [MM1]samples involve the use of cell cultures, which are expensive and time consuming. The Polymerase Chain Reaction (PCR) is a useful tool for the detection of enteroviruses in several matrixes because primary cell culture is not needed and the increased sensitivity of PCR allows detection of the low numbers

O. Gualillo; D. Biscardi; R. Di Carlo; R. De Fusco

1999-01-01

367

A REAL-TIME RT-PCR REACTION SPECIFIC FOR BOVINE ENTEROVIRUSES PROVIDES A RAPID ASSAY FOR FECAL CONTAMINATION OF THE ENVIRONMENT BY CATTLE  

Technology Transfer Automated Retrieval System (TEKTRAN)

Characterization of bovine enteroviruses found in a closed herd of cattle and the surrounding environment indicated that bovine enteroviruses (BEV) could serve as useful markers of contamination with cattle waste. BEV was found in feces from 76% of cattle, 38% of white-tail deer in the same area an...

368

An outbreak of enterovirus 71 infection in Taiwan 1998: A comprehensive pathological, virological, and molecular study on a case of fulminant encephalitis  

Microsoft Academic Search

Background: In a recent enterovirus outbreak in Taiwan, serotype 71 was the culprit of encephalitis causing rapid clinical deterioration and death among young children. Objectives: Since knowledge of enterovirus 71 (EV71) infection in the central nervous system is still limited, the purpose of the present case study was attempted to uncover the pathogenesis of the virus. Study design: We performed

Jing-Jou Yan; Jen-Ren Wang; Ching-Chuan Liu; Hsiao-Bai Yang; Ih-Jen Su

2000-01-01

369

Inhibition of Nuclear Factor-Kappa B Activation Decreases Survival of Mycobacterium tuberculosis in Human Macrophages  

PubMed Central

Nuclear factor-kappa B (NF?B) is a ubiquitous transcription factor that mediates pro-inflammatory responses required for host control of many microbial pathogens; on the other hand, NF?B has been implicated in the pathogenesis of other inflammatory and infectious diseases. Mice with genetic disruption of the p50 subunit of NF?B are more likely to succumb to Mycobacterium tuberculosis (MTB). However, the role of NF?B in host defense in humans is not fully understood. We sought to examine the role of NF?B activation in the immune response of human macrophages to MTB. Targeted pharmacologic inhibition of NF?B activation using BAY 11-7082 (BAY, an inhibitor of I?B? kinase) or an adenovirus construct with a dominant-negative I?B? significantly decreased the number of viable intracellular mycobacteria recovered from THP-1 macrophages four and eight days after infection. The results with BAY were confirmed in primary human monocyte-derived macrophages and alveolar macrophages. NF?B inhibition was associated with increased macrophage apoptosis and autophagy, which are well-established killing mechanisms of intracellular MTB. Inhibition of the executioner protease caspase-3 or of the autophagic pathway significantly abrogated the effects of BAY. We conclude that NF?B inhibition decreases viability of intracellular MTB in human macrophages via induction of apoptosis and autophagy.

Chmura, Kathryn; Ovrutsky, Alida R.; Su, Wen-Lin; Griffin, Laura; Pyeon, Dohun; McGibney, Mischa T.; Strand, Matthew J.; Numata, Mari; Murakami, Seiji; Gaido, Loretta; Honda, Jennifer R.; Kinney, William H.; Oberley-Deegan, Rebecca E.; Voelker, Dennis R.; Ordway, Diane J.; Chan, Edward D.

2013-01-01

370

Detection of enterovirus RNA in peripheral blood mononuclear cells of type 1 diabetic patients beyond the stage of acute infection.  

PubMed

Previous studies have shown that enteroviral RNA can be detected in blood at the onset of type 1 diabetes (T1D). The infection may play a role in triggering T1D and genetic host factors may contribute to this process. We investigated (1) whether enterovirus is present at the onset of T1D in peripheral blood mononuclear cells (PBMC), plasma, throat, or stool, and (2) whether enteroviral presence is linked with HLA-DR type and/or polymorphisms in melanoma differentiation-associated gene 5 (MDA5) and 2'-5' oligoadenylate synthetase 1 (OAS1), factors of antiviral immunity. To this end, PBMC, plasma, throat, and stool samples from 10 T1D patients and 20 unrelated controls were tested for the presence of enteroviruses (RT-PCR), for HLA-DR type, and polymorphisms in MDA5 and OAS1. Enterovirus RNA was detected in PBMC of 4/10 T1D patients, but none of 20 controls. Plasma was positive in 2/10 T1D patients and none of 20 controls, suggesting that enteroviruses found at the onset of T1D are mainly present in PBMC. All throat samples from positive T1D patients were virus-negative and only 1 fecal sample was positive. The negative results for all throat and most stool samples argues against acute infection. Enterovirus presence was linked with HLA-DR4, but not with polymorphisms in MDA5 or OAS1. PMID:20121407

Schulte, Barbara M; Bakkers, Judith; Lanke, Kjerstin H W; Melchers, Willem J G; Westerlaken, Ciska; Allebes, Wil; Aanstoot, Henk-Jan; Bruining, G Jan; Adema, Gosse J; Van Kuppeveld, Frank J M; Galama, Jochem M D

2010-02-01

371

Human apolipoprotein (Apo) B-48 and ApoB-100 kinetics with stable isotopes.  

PubMed

The kinetics of apolipoprotein (apo) B-100 and apoB-48 within triglyceride-rich lipoproteins (TRLs) and of apoB-100 within IDL and LDL were examined with a primed-constant infusion of (5,5,5-(2)H(3)) leucine in the fed state (hourly feeding) in 19 subjects after consumption of an average American diet (36% fat). Lipoproteins were isolated by ultracentrifugation and apolipoproteins by SDS gels, and isotope enrichment was assessed by gas chromatography/mass spectrometry. Kinetic parameters were calculated by multicompartmental modeling of the data with SAAM II. The pool sizes (PS) of TRL apoB-48, VLDL apoB-100, and LDL apoB-100 were 17+/-10, 273+/-167, and 3325+/-1146 mg, respectively. There was a trend toward a faster fractional catabolic rate (FCR) for VLDL apoB-100 than for TRL apoB-48 (6.73+/-3.48 versus 5.02+/-2.07 pools/d, respectively, P=0.06). The mean FCRs for IDL and LDL apoB-100 were 10.07+/-7.28 and 0.27+/-0.08 pools/d, respectively. The mean production rate (PR) of TRL apoB-48 was 6.5% of VLDL apoB-100 (1. 3+/-0.90 versus 20.06+/-6.53 mg. kg(-1). d(-1), P<0.0001). TRL apoB-48 PS was correlated with apoB-48 PR (r=0.780, P<0.0001) but not FCR (r=-0.1810, P=0.458). VLDL apoB-100 PS was correlated with both PR (r=0.713, P=0.0006) and FCR (r=-0.692, P=0.001) of VLDL apoB-100 and by apoB-48 PR (r=0.728, P=0.0004). LDL apoB-100 PS was correlated with FCR (r=-0.549, P=0.015). These data indicate that (1) the FCRs of TRL apoB-48 and VLDL apoB-100 are similar in the fed state, (2) TRL apoB-48 PS is correlated with TRL apoB-48 PR, (3) VLDL apoB-100 PS is correlated with both PR and FCR of VLDL apoB-100 and PR of TRL apoB-48, and (4) LDL apoB-100 PS is correlated with LDL FCR. PMID:10591677

Welty, F K; Lichtenstein, A H; Barrett, P H; Dolnikowski, G G; Schaefer, E J

1999-12-01

372

Association between human parvovirus B19 infection and arthritis  

Microsoft Academic Search

OBJECTIVE--To gain information concerning the association between parvovirus B19 infection and arthritis. METHODS--Blood or synovial fluid, or both, from a total of 77 adult patients with various arthropathies (rheumatoid arthritis 13; mechanical arthropathies 11; crystal induced arthritis 13; idiopathic mono\\/oligoarthritis 25; suspicion of viral arthritis 15) were tested for the presence of the viral genome and anti-B19 antibodies. B19 DNA

P Cassinotti; S Bas; G Siegl; T L Vischer

1995-01-01

373

Structure of Apolipoprotein B100 of Human Low Density Lipoproteins  

Microsoft Academic Search

We have analyzed low density lipoproteins (LDL) apolipoprotein (apo) B structure by direct sequence analysis of LDL apo B-100 tryptlc peptides. Native LDL were digested with trypsin, and the products were fractionated on a Sephadex G-50 column. The partially digested apo B-100 still associated with liplds was recovered In the void volume (designated trypsln-nonreleasable, TN, peptides). The released peptides (designated

Chao-yuh Yang; Zi-Wei Gu; Shi-ai Weng; Tae W. Kim; San-Hwan Chen; Henry J. Pownall; Paul M. Sharp; Shyan-Woei Liu; Wen-Hsiung Li; Antonio M. Gotto; Lawrence Chan

2009-01-01

374

Why do human B cells secrete granzyme B? Insights into a novel B-cell differentiation pathway  

PubMed Central

B cells are generally believed to operate as producers of high affinity antibodies to defend the body against microorganisms, whereas cellular cytotoxicity is considered as an exclusive prerogative of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs). In conflict with this dogma, recent studies have demonstrated that the combination of interleukin-21 (IL-21) and B-cell receptor (BCR) stimulation enables B cells to produce and secrete the active form of the cytotoxic serine protease granzyme B (GrB). Although the production of GrB by B cells is not accompanied by that of perforin as in the case of many other GrB-secreting cells, recent findings suggest GrB secretion by B cells may play a significant role in early antiviral immune responses, in the regulation of autoimmune responses, and in cancer immunosurveillance. Here, we discuss in detail how GrB-secreting B cells may influence a variety of immune processes. A better understanding of the role that GrB-secreting B cells are playing in the immune system may allow for the development and improvement of novel immunotherapeutic approaches against infectious, autoimmune and malignant diseases.

Hagn, Magdalena; Jahrsdorfer, Bernd

2012-01-01

375

Cathepsin B and cysteine proteinase inhibitors in human lung cancer cell lines  

Microsoft Academic Search

Cell lines derived from human squamous cell (EPCL), large cell (LCLC), and small cell lung cancer (SCLC) lines were investigated for the expression of cathepsin B (Cat B) and cysteine proteinase inhibitors (CPIs). The EPLC and LCLC lines expressed 5- to 50-fold more Cat B activity and contained more mature Cat B of Mr 27-29kDa (>2.5mg\\/mg total protein) than the

Hans-Heinrich Heidtmann; Ursula Salge; Magnus Abrahamson; Mojca Bencina; Lili Kastelic; Natasa Kopitar-Jerala; Vito Turk; Tamara T. Lah

1997-01-01

376

Use of recombinant human parvovirus B19 antigens in serological assays  

Microsoft Academic Search

AIMS--To compare the sensitivity, specificity, and practicality of recombinant proteins in serological tests for the detection of human parvovirus B19 IgG and IgM. METHODS--Indirect enzyme linked immunosorbent assays using B19 structural proteins expressed in Escherichia coli were developed for the detection of B19 specific IgG and IgM (rELISA-G and rELISA-M). Cells infected with baculovirus expressing B19 structural proteins were also

H A Cubie; E E Leslie; S Smith; H J ONeill; H Hart; B J Cohen; J M Inglis

1993-01-01

377

Formation of Two Different Types of Ion Channels by Amphotericin B in Human Erythrocyte Membranes  

Microsoft Academic Search

The polyene antibiotic amphotericin B (AmB) is known to form aqueous pores in lipid membranes and biological membranes. Here,\\u000a membrane potential and ion permeability measurements were used to demonstrate that AmB can form two types of selective ion\\u000a channels in human erythrocytes, differing in their interaction with cholesterol. We show that AmB induced a cation efflux\\u000a (negative membrane polarization) across

Eneida A. Romero; Elizabeth Valdivieso; B. Eleazar Cohen

2009-01-01

378

Human LilrB2 is a ?-amyloid receptor and its murine homolog PirB regulates synaptic plasticity in an Alzheimer's model.  

PubMed

Soluble ?-amyloid (A?) oligomers impair synaptic plasticity and cause synaptic loss associated with Alzheimer's disease (AD). We report that murine PirB (paired immunoglobulin-like receptor B) and its human ortholog LilrB2 (leukocyte immunoglobulin-like receptor B2), present in human brain, are receptors for A? oligomers, with nanomolar affinity. The first two extracellular immunoglobulin (Ig) domains of PirB and LilrB2 mediate this interaction, leading to enhanced cofilin signaling, also seen in human AD brains. In mice, the deleterious effect of A? oligomers on hippocampal long-term potentiation required PirB, and in a transgenic model of AD, PirB not only contributed to memory deficits present in adult mice, but also mediated loss of synaptic plasticity in juvenile visual cortex. These findings imply that LilrB2 contributes to human AD neuropathology and suggest therapeutic uses of blocking LilrB2 function. PMID:24052308

Kim, Taeho; Vidal, George S; Djurisic, Maja; William, Christopher M; Birnbaum, Michael E; Garcia, K Christopher; Hyman, Bradley T; Shatz, Carla J

2013-09-20

379

Identification of human germinal center light and dark zone cells and their relationship to human B-cell lymphomas  

PubMed Central

Germinal centers (GCs) are sites of B-cell clonal expansion, hypermutation, and selection. GCs are polarized into dark (DZ) and light zones (LZ), a distinction that is of key importance to GC selection. However, the difference between the B cells in each of these zones in humans remains unclear. We show that, as in mice, CXCR4 and CD83 can be used to distinguish human LZ and DZ cells. Using these markers, we show that LZ and DZ cells in mice and humans differ only in the expression of characteristic “activation” and “proliferation” programs, suggesting that these populations represent alternating states of a single-cell type rather than distinct differentiation stages. In addition, LZ/DZ transcriptional profiling shows that, with the exception of “molecular” Burkitt lymphomas, nearly all human B-cell malignancies closely resemble LZ cells, which has important implications for our understanding of the molecular programs of lymphomagenesis.

Dominguez-Sola, David; Holmes, Antony B.; Deroubaix, Stephanie; Dalla-Favera, Riccardo; Nussenzweig, Michel C.

2012-01-01

380

Identification of human germinal center light and dark zone cells and their relationship to human B-cell lymphomas.  

PubMed

Germinal centers (GCs) are sites of B-cell clonal expansion, hypermutation, and selection. GCs are polarized into dark (DZ) and light zones (LZ), a distinction that is of key importance to GC selection. However, the difference between the B cells in each of these zones in humans remains unclear. We show that, as in mice, CXCR4 and CD83 can be used to distinguish human LZ and DZ cells. Using these markers, we show that LZ and DZ cells in mice and humans differ only in the expression of characteristic "activation" and "proliferation" programs, suggesting that these populations represent alternating states of a single-cell type rather than distinct differentiation stages. In addition, LZ/DZ transcriptional profiling shows that, with the exception of "molecular" Burkitt lymphomas, nearly all human B-cell malignancies closely resemble LZ cells, which has important implications for our understanding of the molecular programs of lymphomagenesis. PMID:22740445

Victora, Gabriel D; Dominguez-Sola, David; Holmes, Antony B; Deroubaix, Stephanie; Dalla-Favera, Riccardo; Nussenzweig, Michel C

2012-06-26

381

Human Genome Diversity Project. Summary of planning workshop 3(B): Ethical and human-rights implications  

SciTech Connect

The third planning workshop of the Human Genome Diversity Project was held on the campus of the US National Institutes of Health in Bethesda, Maryland, from February 16 through February 18, 1993. The second day of the workshop was devoted to an exploration of the ethical and human-rights implications of the Project. This open meeting centered on three roundtables, involving 12 invited participants, and the resulting discussions among all those present. Attendees and their affiliations are listed in the attached Appendix A. The discussion was guided by a schedule and list of possible issues, distributed to all present and attached as Appendix B. This is a relatively complete, and thus lengthy, summary of the comments at the meeting. The beginning of the summary sets out as conclusions some issues on which there appeared to be widespread agreement, but those conclusions are not intended to serve as a set of detailed recommendations. The meeting organizer is distributing his recommendations in a separate memorandum; recommendations from others who attended the meeting are welcome and will be distributed by the meeting organizer to the participants and to the Project committee.

NONE

1993-12-31

382

Expression of the inhibin\\/activin subunits alpha (?), beta-A (?A) and beta-B (?B) in benign human endometrial polyps and tamoxifen-associated polyps  

Microsoft Academic Search

Background: Inhibins (INH) are dimeric glycoproteins, composed of an alpha subunit (INH-a) and one of two possible beta subunits (INH-bA or INH- bB). They have substantial roles in human reproduction and in endocrine-responsive tumours. Therefore, the aims of this study were to determine the frequency and tissue distribution of INH-a ,I NH-bA and INH-b Bi n normal human endometrium and

Ioannis Mylonas; Josef Makovitzky; Anja Fernow; Dagmar-Ulrike Richter; Udo Jeschke; Volker Briese; Bernd Gerber; Klaus Friese

2005-01-01

383

A human TRIM5? B30.2\\/SPRY domain mutant gains the ability to restrict and prematurely uncoat B-tropic murine leukemia virus  

Microsoft Academic Search

Human TRIM5? restricts N-tropic murine leukemia virus (N-MLV) but not B-tropic MLV (B-MLV) infection. Here we study B30.2\\/SPRY domain mutants of human TRIM5? that acquire the ability to inhibit B-MLV infection prior to reverse transcription without losing the ability to restrict N-MLV infection. Remarkably, these mutants gain the ability to decrease the amount of particulate B-MLV capsids in the cytosol

Felipe Diaz-Griffero; Michel Perron; Kathleen McGee-Estrada; Robert Hanna; Pierre V. Maillard; Didier Trono; Joseph Sodroski

2008-01-01

384

Comparison of splicing factor 3b inhibitors in human cells.  

PubMed

Name your splice: FR901464 analogues and herboxidiene inhibit constitutive splicing, most likely by inhibiting spliceosomal subunit SF3b. A parallel comparison of these compounds in a cell-based assay system showed meayamycin B as the most potent splicing inhibitor among these small molecules. PMID:23172726

Gao, Yang; Vogt, Andreas; Forsyth, Craig J; Koide, Kazunori

2012-11-22

385

Hyperactivated B cells in human inflammatory bowel disease  

PubMed Central

IBD is characterized by a chronic, dysregulated immune response to intestinal bacteria. Past work has focused on the role of T cells and myeloid cells in mediating chronic gastrointestinal and systemic inflammation. Here, we show that circulating and tissue B cells from CD patients demonstrate elevated basal levels of activation. CD patient B cells express surface TLR2, spontaneously secrete high levels of IL-8, and contain increased ex vivo levels of phosphorylated signaling proteins. CD clinical activity correlates directly with B cell expression of IL-8 and TLR2, suggesting a positive relationship between these B cell inflammatory mediators and disease pathogenesis. In contrast, B cells from UC patients express TLR2 but generally do not demonstrate spontaneous IL-8 secretion; however, significant IL-8 production is inducible via TLR2 stimulation. Furthermore, UC clinical activity correlates inversely with levels of circulating TLR2+ B cells, which is opposite to the association observed in CD. In conclusion, TLR2+ B cells are associated with clinical measures of disease activity and differentially associated with CD- and UC-specific patterns of inflammatory mediators, suggesting a formerly unappreciated role of B cells in the pathogenesis of IBD

Noronha, Ansu Mammen; Liang, YanMei; Hetzel, Jeremy T.; Hasturk, Hatice; Kantarci, Alpdogan; Stucchi, Arthur; Zhang, Yue; Nikolajczyk, Barbara S.; Farraye, Francis A.; Ganley-Leal, Lisa M.

2009-01-01

386

Human Serum Bactericidal Activity against Haemophilus influenzae Type b  

Microsoft Academic Search

We examined bactericidal and opsonizing activity of pooled adult 'immune' serum against Haemophilus influenzae type b with and without the addition of phagocytes. Four type b strains from cerebrospinal fluid (CSF) and three such strains from the nasopharynx (NP) of healthy children were examined. Duplicate reaction mixtures contained organisms in exponential (E) or stationary phase (S) of growth, serum, a

TERRENCE L. STULL; RICHARD F. JACOBS; JOEL E. HAAS; MARILYN C. ROBERTS; CHRISTOPHER B. WILSON; ARNOLD L. SMITH

1984-01-01

387

Efficient lentiviral transduction and transgene expression in primary human B cells.  

PubMed

Primary human B cells are an attractive target for gene-therapeutic applications, but have been found to be relatively resistant toward transduction with lentiviral vectors (LVVs), even though a number of different envelope pseudotypes were tested. Moreover, low transgene expression in primary human B cells has impeded the use of LVVs for this target cell. We investigated the transduction potential of gibbon-ape leukemia virus (GALV) Env-pseudotyped LVVs for primary human B cells. By establishing optimized transduction kinetics and multiplicities of infection, we were able to regularly obtain transduction efficiencies of more than 50% in CD40L-activated B cells. Noteworthy, with the use of GALV-pseudotyped LVVs we could achieve a more than 10-fold higher yield of transduced activated B cells in direct comparison with LVVs pseudotyped with measles virus glycoproteins. Phenotyping of transduced primary B cells revealed a majority of memory B cells, a long-lived phenotype, presumed to be well suited for enduring therapeutic interventions. Finally, by combining the enhancer (E?) and the matrix/scaffold-attachment regions (MARs) of the human immunoglobulin heavy chain with the promoter of spleen focus-forming virus (SFFV) we aimed at generating a novel LVV particularly suitable for B cell transgenesis. We show that the optimized vector facilitated significantly higher transgene expression in various B cell lines and, more importantly, primary human B cells (mean factor of three). In summary, we have established a novel protocol for the efficient lentiviral transduction of primary human B cells and have improved transgene expression in B cells by a specific vector modification. PMID:23240650

Mock, Ulrike; Thiele, Regine; Uhde, Almut; Fehse, Boris; Horn, Stefan

2012-12-01

388

Human Exposure to Herpesvirus B-Seropositive Macaques, Bali, Indonesia  

PubMed Central

Herpesvirus B (Cercopithecine herpesvirus 1) has been implicated as the cause of approximately 40 cases of meningoencephalitis affecting persons in direct or indirect contact with laboratory macaques. However, the threat of herpesvirus B in nonlaboratory settings worldwide remains to be addressed. We investigated the potential for exposure to herpesvirus B in workers at a “monkey forest” (a temple that has become a tourist attraction because of its monkeys) in Bali, Indonesia. In July 2000, 105 workers at the Sangeh Monkey Forest in Central Bali were surveyed about contact with macaques (Macaca fascicularis). Nearly half of those interviewed had either been bitten or scratched by a macaque. Prevalence of injury was higher in those who fed macaques. Serum from 31 of 38 Sangeh macaques contained antibodies to herpesvirus B. We conclude that workers coming into contact with macaques at the Sangeh Monkey Forest are at risk for exposure to herpesvirus B.

Engel, Gregory A.; Schillaci, Michael A.; Suaryana, Komang Gde; Putra, Artha; Fuentes, Agustin; Henkel, Richard

2002-01-01

389

Metabolism and cytotoxicity of aflatoxin b1 in cytochrome p-450-expressing human lung cells.  

PubMed

The mycotoxin aflatoxin B(1) (AFB(1)) is a hepatocarcinogen in many animal models and probably a human carcinogen. Besides being a dietary carcinogen, AFB(1) has been detected in dusts generated in the processing and transportation of AFB(1)-contaminated products. Inhalation of grain dusts contaminated with AFB(1) may be a risk factor in human lung cancer. Aflatoxin B(1) requires cytochrome P-450 (CYP)-mediated activation to form cytotoxic and DNA-reactive intermediates, and this activation in human liver is mediated by the CYP 1A2 and 3A4 isoforms. Which isoforms are important in AFB(1) activation in human lung is not well understood. To investigate whether these CYPs can activate AFB(1) at low, environmentally relevant concentrations in human lung cells, SV40 immortalized human bronchial epithelial cells (BEAS-2B) that were transfected with cDNA for CYPs 3A4 (B3A4) or 1A2 (B-CMV1A2) were used. B-CMV1A2 cultured in 15 nM AFB(1) produced the AFB(1)-glutathione conjugate (AFB(1)-GSH) and aflatoxin M(1) (AFM(1)), while B3A4 cells produced only aflatoxin Q(1) (AFQ(1)) at 0.15 microM AFB(1). Nontransfected BEAS-2B cells produced no metabolites, even at 1.5 mM AFB(1). Microsomes prepared from B-CMV1A2 and B3A4 cells activated AFB(1) to AFB(1) 8,9-epoxide (AFBO), while those from BEAS-2B cells did not produce AFBO. Cytosol from all three cell types was ineffective at glutathione S-transferase (GST)-mediated trapping of enzymatically generated AFB(1) 8,9-epoxide. B-CMV1A2 cells were 100-fold more sensitive to AFB(1) compared to B3A4 cells, and were 6000-fold more sensitive than control BEAS-2B cells. Western immunoblots confirmed that only B-CMV1A2 cells expressed CYP 1A2 protein, while CYP 3A4 was only in B3A4 cells. B-CMV1A2 cells were the most sensitive to AFB(1), followed by B3A4 cells. CYP 3A4, which has been predicted to activate AFB(1) primarily at higher AFB(1) concentrations, was also responsible for significant AFB(1) toxicity at low concentrations. These data indicate that human lung cells expressing these CYP isoforms are capable of activating AFB(1), even at environmentally relevant concentrations. PMID:12079611

Van Vleet, Terry R; Klein, Patrick J; Coulombe, Roger A

2002-06-28

390

Antiviral vaccines license T cell responses by suppressing granzyme B levels in human plasmacytoid dendritic cells.  

PubMed

Human plasmacytoid dendritic cells (pDC) are important modulators of adaptive T cell responses during viral infections. Recently, we found that human pDC produce the serine protease granzyme B (GrB), thereby regulating T cell proliferation in a GrB-dependent manner. In this study, we demonstrate that intrinsic GrB production by pDC is significantly inhibited in vitro and in vivo by clinically used vaccines against viral infections such as tick-borne encephalitis. We show that pDC GrB levels inversely correlate with the proliferative response of coincubated T cells and that GrB suppression by a specific Ab or a GrB substrate inhibitor results in enhanced T cell proliferation, suggesting a predominant role of GrB in pDC-dependent T cell licensing. Functionally, we demonstrate that GrB(high) but not GrB(low) pDC transfer GrB to T cells and may degrade the ?-chain of the TCR in a GrB-dependent fashion, thereby providing a possible explanation for the observed T cell suppression by GrB-expressing pDC. Modulation of pDC-derived GrB activity represents a previously unknown mechanism by which both antiviral and vaccine-induced T cell responses may be regulated in vivo. Our results provide novel insights into pDC biology during vaccinations and may contribute to an improvement of prophylactic and therapeutic vaccines. PMID:23785122

Fabricius, Dorit; Nußbaum, Benedikt; Busch, Daniel; Panitz, Verena; Mandel, Birgit; Vollmer, Angelika; Westhoff, Mike-Andrew; Kaltenmeier, Christof; Lunov, Oleg; Tron, Kyrylo; Nienhaus, G Ulrich; Jahrsdörfer, Bernd; Debatin, Klaus-Michael

2013-06-19

391

Enterovirus RNA in Peripheral Blood May Be Associated with the Variants of rs1990760, a Common Type 1 Diabetes Associated Polymorphism in IFIH1  

PubMed Central

Objective Polymorphisms in the IFIH1 (common rs1990760 and four rare rs35667974, rs35337543, rs35744605, rs35732034) have been convincingly associated with type 1 diabetes. The encoded protein (interferon-induced helicase C domain-containing protein 1) senses double-stranded RNA during replication of Picornavirales, including Enterovirus, a genus suspected in the etiology of type 1 diabetes. We therefore investigated whether the polymorphisms are associated with differences in the frequency of enterovirus RNA in blood. Research Design and Methods The study included 1001 blood samples, each from a child participating in the Norwegian ‘Environmental Triggers of Type 1 Diabetes: the MIDIA study’. The enterovirus RNA was tested using qualitative semi-nested real-time reverse transcriptase PCR on RNA extracted from frozen cell packs after removal of plasma. Stool samples previously analyzed for enterovirus RNA were available in 417 children. Results The genotypes of IFIH1 rs1990760 were associated with different frequencies of enterovirus RNA in blood (7.0%, 14.4% and 9.5% bloods were enterovirus positive among children carrying the Ala/Ala, Ala/Thr and Thr/Thr genotypes, respectively, p?=?0.012). This association remained essentially unchanged after adjustment for age and calendar year. The presence of enterovirus in the concomitantly sampled stool further increased the likelihood of enterovirus RNA in blood (odds ratio 2.40, CI 95% 1.13–4.70), but did not affect the association with IFIH1 rs1990760. The rare polymorphisms (individually, or pooled) were not significantly associated with enterovirus RNA in blood. Conclusions The common IFIH1 SNP may modify the frequency of enterovirus RNA in blood of healthy children. This effect can help explain the association of IFIH1 with type 1 diabetes.

Cinek, Ondrej; Tapia, German; Wits?, Elisabet; Kramna, Lenka; Holkova, Katerina; Rasmussen, Trond; Stene, Lars C.; R?nningen, Kjersti S.

2012-01-01

392

Human immunodeficiency virus/human parvovirus B19 co-infection in blood donors and AIDS patients in Sichuan, China  

PubMed Central

Background Human parvovirus B19 (B19) is a common pathogen which causes a variety of diseases. Persistent B19 infection is related to the degree of host immunodeficiency in patients with human immunodeficiency virus (HIV) infection. However, the existence, loading, virus evolution and distribution of B19 in Chinese HIV-positive patients have not been determined. Materials and methods. We investigated 573 HIV-positive blood donors and AIDS patients in Sichuan, China in the last two decades. Bl9-specific serology and quantitative polymerase chain reaction were used to determine the prevalence of B19/HIV co-infection. Viral genome fragments were subjected to phylogeny and haplotype analysis. Results B19 genomic DNA was found in 26 of 573 (4.5%) HIV-positive individuals, a higher prevalence than in blood donors. DNA levels ranged from 5.3×102–1.1×105 copies/mL. The seroprevalence of IgG was significantly lower in HIV-positive samples than in HIV-negative blood donors, indicating deficient production of B19-specific IgG in the former. The B19 isolates were genotype-1 subtype B19-1A which formed a monophyletic group; seven distinct haplotypes were discovered with 60% of the B19/HIV co-infected variants sharing one central haplotype. Discussion. This study on the prevalence, phylogeny and distribution of human parvovirus B19 in Sichuan, China, demonstrates the persistence of B19 in the circulation of both immunocompetent and immunocompromised subjects, with implications for blood safety.

He, Miao; Zhu, Jiang; Yin, Huimin; Ke, Ling; Gao, Lei; Pan, Zhihong; Yang, Xiuhua; Li, Wuping

2012-01-01

393

NF?B Is Persistently Activated in Continuously Stimulated Human Neutrophils  

PubMed Central

Increased activation of the transcription factor NF?B in the neutrophils has been associated with the pathogenesis of sepsis, acute lung injury (ALI), bronchopulmonary dysplasia (BPD), and other neutrophil-mediated inflammatory disorders. Despite recent progress in analyzing early NF?B activation in human neutrophils, activation of NF?B in persistently stimulated neutrophils has not been previously studied. Because it is the persistent NF?B activation that is thought to be involved in the host response to sepsis and the pathogenesis of ALI and BPD, we hypothesized that continuously stimulated human neutrophils may exhibit a late phase of NF?B activity. The goal of this study was to analyze the NF?B activation and expression of I?B and NF?B proteins during neutrophil stimulation with inflammatory signals for prolonged times. We demonstrate that neutrophil stimulation with lipopolysaccharide (LPS) and tumor necrosis factor-? (TNF?) induces, in addition to the early activation at 30–60 min, a previously unrecognized late phase of NF?B activation. In LPS-stimulated neutrophils, this NF?B activity typically had a biphasic character, whereas TNF?-stimulated neutrophils exhibited a continuous NF?B activity peaking around 9 h after stimulation. In contrast to the early NF?B activation that inversely correlates to the nuclear levels of I?B?, however, in continuously stimulated neutrophils, NF?B is persistently activated despite considerable levels of I?B? present in the nucleus. Our data suggest that NF?B is persistently activated in human neutrophils during neutrophil-mediated inflammatory disorders, and this persistent NF?B activity may represent one of the underlying mechanisms for the continuous production of proinflammatory mediators.

Miskolci, Veronika; Rollins, Janet; Vu, Hai Yen; Ghosh, Chandra C; Davidson, Dennis; Vancurova, Ivana

2007-01-01

394

Effect of acrolein on human alveolar macrophage NF-kappaB activity.  

PubMed

Acrolein is an environmental pollutant that is known to suppress respiratory host defense against infections; however, the mechanism of the decrease in host defense is not yet clear. We have previously reported that acrolein inhibited endotoxin-induced cytokine release and induced apoptosis in human alveolar macrophages, suggesting that the inhibition of cytokine release and/or cytotoxicity to alveolar macrophages may, in part, be responsible for acrolein-induced immunosuppression in the lung. Because nuclear factor-kappaB (NF-kappaB) is an important transcription factor for a number of cytokine genes and is also an important regulator of apoptosis, the effect of acrolein on NF-kappaB activity was examined by electrophoresis mobility shift assay. Acrolein caused a dose-dependent inhibition of endotoxin-induced NF-kappaB activation as well as an inhibition of basal level NF-kappaB activity. Because IkappaB is a principal regulator of NF-kappaB activity in the nucleus, changes in IkappaB were determined by Western blotting. Acrolein-inhibited IkappaB phosphorylation leads to an increase in cellular IkappaB levels preventing NF-kappaB nuclear translocation and is likely the mechanism of acrolein-induced inhibition of NF-kappaB activity. The role of basal level NF-kappaB in acrolein-induced apoptosis was also examined. An NF-kappaB inhibitor (MG-132) also induced apoptosis in human alveolar macrophages, suggesting that a certain basal level NF-kappaB activity may be required for macrophage cell survival. Taken together, our results suggest that the acrolein-inhibited endotoxin-induced NF-kappaB activation decreased the basal level NF-kappaB activity, which may be responsible for the inhibition of cytokine release and the induction of apoptosis in human alveolar macrophages. PMID:10484462

Li, L; Hamilton, R F; Holian, A

1999-09-01

395

Targeting granzyme B to tumor cells using a yoked human chorionic gonadotropin  

Microsoft Academic Search

Purpose  Luteinizing hormone receptor (LHR) is found in abundance on human ovarian, breast, endometrial and prostate carcinomas but\\u000a at only low levels on non-gonadal tissues. To selectively kill LHR-expressing tumors, granzyme B (GrB) was linked to a protein\\u000a in which both chains of human chorionic gonadotropin were yoked together (YCG).\\u000a \\u000a \\u000a \\u000a \\u000a Methods  GrB-YCG was expressed and secreted from insect Sf9 cells. Its GrB

Isao Kanatani; Xinjian Lin; Xiaoqin Yuan; Gerald Manorek; Xiying Shang; Lawrence H. Cheung; Michael G. Rosenblum; Stephen B. Howell

396

The effect of recombinant human interferon ? B D compared to interferon ? 2b on SIV infection in rhesus macaques  

Microsoft Academic Search

The model of simian immunodeficiency virus (SIV) infection in rhesus macaques was used to evaluate the effects of recombinant human interferon ?, Hu IFN-? 2b and Hu IFN-? BD, at two doses. Administration began 1 day prior to infection and was continued for 90 days postinfection. Both interferons suppressed SIV antigenemia during the treatment period. Following treatment animals were monitored

H. Schellekens; H. Niphuis; L. Buijs; P. Douw van der Krap; H. K. Hochkeppel; J. L. Heeney

1996-01-01

397

Development of a human T-T cell hybridoma secreting B cell growth factor  

PubMed Central

The success of long-term culture of normal human and murine B cells has been hampered by the limited availability of soluble factors capable of maintaining proliferation of activated B lymphocytes. Previous experiments using various culture-derived supernatants in a human system were unable to separate the activities of B cell growth factor (BCGF) and interleukin 2 (IL-2) by immunochemical means. Thus, purified factors with BCGF activity in the absence of IL-2 activity have not been available for study. In the present study, normal human peripheral blood T cells were fused with the hypoxanthine/aminopterin/thymidine- sensitive human T-leukemic cell line, CEM-6. Supernatants from the resulting hybrid cells were tested for the ability to maintain proliferation of normal human B cells in a recently described assay system for human BCGF. Hybrids demonstrating BCGF activity were cloned by limiting dilution. One hybrid clone, 2B11, continued to support proliferation of B cells in both long-term cultures and 6-d assays at a level significantly above that seen with conventionally produced growth factors. No IL-2 activity was found in the supernatant from hybrid 2B11. The hybridoma supernatant was fractionated by gel filtration, and maximum proliferation of B cells was supported by the 18-20,000 mol wt protein fraction. Thus, a human T-T cell hybridoma that has BCGF activity in the absence of any demonstrable IL-2 activity has been developed. Human T-T cell hybridomas secreting discrete immunoregulatory factors should prove to be powerful tools in dissecting the mechanisms of immunoregulation of human lymphocyte function.

1983-01-01

398

Evaluation of five commercial tests for detection of immunoglobulin M antibodies to human parvovirus B19.  

PubMed Central

The following commercial tests for detection of immunoglobulin M antibodies to human parvovirus B19 were evaluated: Ideia Parvovirus B19-IgM, MRL Diagnostics Human Parvovirus B19 IgM ELISA, Parvoscan-B19, and Biotrin Parvo B19 IgM EIA and IF. A total of 203 serum specimens from patients who probably have current B19 infections or have other viral infections and sera with rheumatoid factor were investigated. Between 75 and 79 of 102 serum samples from patients thought to have current B19 infections yielded positive results with the different tests. Ideia had the highest specificity (94.8%), while Parvoscan showed a specificity of only 70.1%. Our evaluation results show that Ideia, MRL, and Biotrin EIA and IF can be recommended for diagnostic purposes.

Bruu, A L; Nordb?, S A

1995-01-01

399

3C Protease of Enterovirus 68: Structure-Based Design of Michael Acceptor Inhibitors and Their Broad-Spectrum Antiviral Effects against Picornaviruses  

PubMed Central

We have determined the cleavage specificity and the crystal structure of the 3C protease of enterovirus 68 (EV68 3Cpro). The protease exhibits a typical chymotrypsin fold with a Cys...His...Glu catalytic triad; its three-dimensional structure is closely related to that of the 3Cpro of rhinovirus 2, as well as to that of poliovirus. The phylogenetic position of the EV68 3Cpro between the corresponding enzymes of rhinoviruses on the one hand and classical enteroviruses on the other prompted us to use the crystal structure for the design of irreversible inhibitors, with the goal of discovering broad-spectrum antiviral compounds. We synthesized a series of peptidic ?,?-unsaturated ethyl esters of increasing length and for each inhibitor candidate, we determined a crystal structure of its complex with the EV68 3Cpro, which served as the basis for the next design round. To exhibit inhibitory activity, compounds must span at least P3 to P1?; the most potent inhibitors comprise P4 to P1?. Inhibitory activities were found against the purified 3C protease of EV68, as well as with replicons for poliovirus and EV71 (50% effective concentration [EC50] = 0.5 ?M for the best compound). Antiviral activities were determined using cell cultures infected with EV71, poliovirus, echovirus 11, and various rhinovirus serotypes. The most potent inhibitor, SG85, exhibited activity with EC50s of ?180 nM against EV71 and ?60 nM against human rhinovirus 14 in a live virus–cell-based assay. Even the shorter SG75, spanning only P3 to P1?, displayed significant activity (EC50 = 2 to 5 ?M) against various rhinoviruses.

Tan, Jinzhi; George, Shyla; Kusov, Yuri; Perbandt, Markus; Anemuller, Stefan; Mesters, Jeroen R.; Norder, Helene; Coutard, Bruno; Lacroix, Celine; Leyssen, Pieter; Neyts, Johan

2013-01-01

400

3C protease of enterovirus 68: structure-based design of Michael acceptor inhibitors and their broad-spectrum antiviral effects against picornaviruses.  

PubMed

We have determined the cleavage specificity and the crystal structure of the 3C protease of enterovirus 68 (EV68 3C(pro)). The protease exhibits a typical chymotrypsin fold with a Cys...His...Glu catalytic triad; its three-dimensional structure is closely related to that of the 3C(pro) of rhinovirus 2, as well as to that of poliovirus. The phylogenetic position of the EV68 3C(pro) between the corresponding enzymes of rhinoviruses on the one hand and classical enteroviruses on the other prompted us to use the crystal structure for the design of irreversible inhibitors, with the goal of discovering broad-spectrum antiviral compounds. We synthesized a series of peptidic ?,?-unsaturated ethyl esters of increasing length and for each inhibitor candidate, we determined a crystal structure of its complex with the EV68 3C(pro), which served as the basis for the next design round. To exhibit inhibitory activity, compounds must span at least P3 to P1'; the most potent inhibitors comprise P4 to P1'. Inhibitory activities were found against the purified 3C protease of EV68, as well as with replicons for poliovirus and EV71 (50% effective concentration [EC(50)] = 0.5 ?M for the best compound). Antiviral activities were determined using cell cultures infected with EV71, poliovirus, echovirus 11, and various rhinovirus serotypes. The most potent inhibitor, SG85, exhibited activity with EC(50)s of ?180 nM against EV71 and ?60 nM against human rhinovirus 14 in a live virus-cell-based assay. Even the shorter SG75, spanning only P3 to P1', displayed significant activity (EC(50) = 2 to 5 ?M) against various rhinoviruses. PMID:23388726

Tan, Jinzhi; George, Shyla; Kusov, Yuri; Perbandt, Markus; Anemüller, Stefan; Mesters, Jeroen R; Norder, Helene; Coutard, Bruno; Lacroix, Céline; Leyssen, Pieter; Neyts, Johan; Hilgenfeld, Rolf

2013-02-06

401

CD27+ B cells in human lymphatic organs: re-evaluating the splenic marginal zone  

PubMed Central

The marginal zone of human spleens is regarded as an organ-specific region harbouring sessile memory B cells. This opinion has arisen by extrapolating from results obtained in mice and rats. Detection of CD27+ B cells in situ now revealed similarities among the most superficial region of B-cell follicles in human spleens, reactive lymph nodes, inflamed appendices, tonsils and terminal ilea. The follicular surface in these organs consists of small naïve immunoglobulin D (IgD)+ CD27– B cells predominating in an inner area and larger IgD+/– CD27+ B cells prevailing in a more superficial position. CD27+ B cells may, however, also occupy the entire follicular periphery around the germinal centre. Together with additional peculiarities this distribution indicates a fundamental microanatomical difference among the human and rodent splenic white pulp. We hypothesize that the follicular periphery represents a recirculation compartment both for naïve and memory/natural reactive B cells in all human secondary lymphatic organs. This assumption implies a difference in recirculation behaviour among human and rodent B memory cells.

Steiniger, Birte; Timphus, Eva-Maria; Jacob, Ralf; Barth, Peter J

2005-01-01

402

Treatment with ephrin B2 positively impacts the abnormal metabolism of human osteoarthritic chondrocytes  

PubMed Central

Introduction Members of the ephrin system, the ephrin receptor erythropoietin-producing hepatocellular B4 (EphB4) and its specific ligand, ephrin B2, appear to be involved in the bone remodelling process. We recently showed that their interaction inhibits the resorptive activity of human osteoarthritic (OA) subchondral bone osteoblasts. Hence, we further investigated the possible implication of these ephrin members on the catabolic/anabolic activities of human OA chondrocytes. Methods EphB4 receptor and ephrin B2 levels were determined by quantitative PCR and immunohistochemistry, and the effects of ephrin B2 on the expression/production of factors involved in the OA process. Results EphB4 receptors and ephrin B2 ligands are expressed and produced by human normal and OA chondrocytes. Ephrin B2 protein was found at similar levels in both cartilage types, whereas EphB4 receptor expression (P < 0.0001) and production (P < 0.01) levels were significantly increased in OA chondrocytes/cartilage. Ephrin B2 treatment significantly inhibited the interleukin (IL)-1beta, IL-6, matrix metalloproteinase-1 (MMP-1), MMP-9, MMP-13, and proteinase-activated receptor-2 (PAR-2) gene expression levels, whereas MMP-2 was unaffected, and significantly increased collagen type II, a cartilage specific macromolecule. It also inhibited the IL-1beta stimulated protein production of IL-6, MMP-1 and MMP-13. Conclusions Our study is the first to provide data on the presence and role of ephrin B2/EphB4 receptors in human chondrocytes/cartilage. Data showed that ephrin B2 treatment positively impacts the abnormal metabolism of OA cartilage by inhibiting important catabolic factors involved in this disease at the same time as increasing anabolic activity.

Kwan Tat, Steeve; Pelletier, Jean-Pierre; Amiable, Nathalie; Boileau, Christelle; Lavigne, Martin; Martel-Pelletier, Johanne

2009-01-01

403

Inhibition of human acid-sensing ion channel 1b by zinc  

PubMed Central

Acid-sensing ion channel 1b (ASIC1b) is expressed in peripheral sensory neurons and has been implicated in nociception. Understanding the modulation of ASIC1b will provide important insight into how ASIC1b contributes to pain sensation. In our previous study, we showed that zinc, an important modulator of pain sensation, reduces rat ASIC1b current. However, rat ASIC1b shows several important differences from its recently identified human homolog. Most noticeably, human ASIC1b (hASIC1b) has a sustained component, which may play a role in persistent pain. Therefore, we tested here the hypothesis that zinc modulates the current properties of hASIC1b. Bath application of zinc suppressed the peak amplitude of hASIC1b currents, with a half-maximum inhibitory concentration of 37 ?M. However, zinc did not affect the sustained component of hASIC1b currents. The effect of zinc was independent of pH-dependent activation, steady-state desensitization, and extracellular Ca2+, suggesting noncompetitive mechanisms. Further, we found that extracellular site(s) of the hASIC1b subunit is important for the effect of zinc. Mutating cysteine 196, but not cysteine 309, in the extracellular domain of the hASIC1b abolished the zinc inhibition. These results suggest that, through modulating cysteine196, zinc may have a modulatory role in acute pain.

Jiang, Qian; Zha, Xiang-Ming; Chu, Xiang-Ping

2012-01-01

404

REPSIM-B: A Computer Simulation Model of Human Reproduction.  

National Technical Information Service (NTIS)

REPSIM-B simulates the detailed reproductive history of a hypothetical cohort of woman. The computer program provides for a woman's marrying, dying, becoming sterile, becoming pregnant and for varying outcomes of pregnancy, including induced abortion. Pro...

R. Geremia J. C. Ridley

1979-01-01

405

Human Immunodeficiency Virus Types 1 and 2 (E. coli, B. ...  

Center for Biologics Evaluation and Research (CBER)

Text Version... subtype B, while the predominant strains in India - are subtype C.1113 Group O is found primarily in Cameroon and west central Africa, but also has ... More results from www.fda.gov/downloads/biologicsbloodvaccines/bloodbloodproducts

406

FGF8 isoform b expression in human prostate cancer  

Microsoft Academic Search

Overexpression of fibroblast growth factor 8 (FGF8) mRNA has been previously described in prostate cancer. Of its four isoforms, FGF8b is thought to be the most important in carcinogenesis. We hypothesised that immunodetection of FGF8b in archival prostate cancer specimens is of potential prognostic value. Using a selected cohort of prostate tumours from transurethral (n=30) and radical prostatectomies (n=59), an

V J Gnanapragasam; M C Robinson; C Marsh; C N Robson; F C Hamdy; H Y Leung

2003-01-01

407

CENP-B box is required for de novo centromere chromatin assembly on human alphoid DNA  

PubMed Central

Centromere protein (CENP) B boxes, recognition sequences of CENP-B, appear at regular intervals in human centromeric ?-satellite DNA (alphoid DNA). In this study, to determine whether information carried by the primary sequence of alphoid DNA is involved in assembly of functional human centromeres, we created four kinds of synthetic repetitive sequences: modified alphoid DNA with point mutations in all CENP-B boxes, resulting in loss of all CENP-B binding activity; unmodified alphoid DNA containing functional CENP-B boxes; and nonalphoid repetitive DNA sequences with or without functional CENP-B boxes. These four synthetic repetitive DNAs were introduced into cultured human cells (HT1080), and de novo centromere assembly was assessed using the mammalian artificial chromosome (MAC) formation assay. We found that both the CENP-B box and the alphoid DNA sequence are required for de novo MAC formation and assembly of functional centromere components such as CENP-A, CENP-C, and CENP-E. Using the chromatin immunoprecipitation assay, we found that direct assembly of CENP-A and CENP-B in cells with synthetic alphoid DNA required functional CENP-B boxes. To the best of our knowledge, this is the first reported evidence of a functional molecular link between a centromere-specific DNA sequence and centromeric chromatin assembly in humans.

Ohzeki, Jun-ichirou; Nakano, Megumi; Okada, Teruaki; Masumoto, Hiroshi

2002-01-01

408

Improved crystallization of the coxsackievirus B3 RNA-dependent RNA polymerase  

PubMed Central

The Picornaviridae virus family contains a large number of human pathogens such as poliovirus, hepatitis A virus and rhinoviruses. Amongst the viruses belonging to the genus Enterovirus, several serotypes of coxsackievirus coexist for which neither vaccine nor therapy is available. Coxsackievirus B3 is involved in the development of acute myocarditis and dilated cardiomyopathy and is thought to be an important cause of sudden death in young adults. Here, the first crystal of a coxsackievirus RNA-dependent RNA polymerase is reported. Standard crystallization methods yielded crystals that were poorly suited to X-­ray diffraction studies, with one axis being completely disordered. Crystallization was improved by testing crystallization solutions from commercial screens as additives. This approach yielded crystals that diffracted to 2.1?Å resolution and that were suitable for structure determination.

Jabafi, Ilham; Selisko, Barbara; Coutard, Bruno; De Palma, Armando M.; Neyts, Johan; Egloff, Marie-Pierre; Grisel, Sacha; Dalle, Karen; Campanacci, Valerie; Spinelli, Silvia; Cambillau, Christian; Canard, Bruno; Gruez, Arnaud

2007-01-01

409

Improved crystallization of the coxsackievirus B3 RNA-dependent RNA polymerase.  

PubMed

The Picornaviridae virus family contains a large number of human pathogens such as poliovirus, hepatitis A virus and rhinoviruses. Amongst the viruses belonging to the genus Enterovirus, several serotypes of coxsackievirus coexist for which neither vaccine nor therapy is available. Coxsackievirus B3 is involved in the development of acute myocarditis and dilated cardiomyopathy and is thought to be an important cause of sudden death in young adults. Here, the first crystal of a coxsackievirus RNA-dependent RNA polymerase is reported. Standard crystallization methods yielded crystals that were poorly suited to X-ray diffraction studies, with one axis being completely disordered. Crystallization was improved by testing crystallization solutions from commercial screens as additives. This approach yielded crystals that diffracted to 2.1 A resolution and that were suitable for structure determination. PMID:17554171

Jabafi, Ilham; Selisko, Barbara; Coutard, Bruno; De Palma, Armando M; Neyts, Johan; Egloff, Marie Pierre; Grisel, Sacha; Dalle, Karen; Campanacci, Valerie; Spinelli, Silvia; Cambillau, Christian; Canard, Bruno; Gruez, Arnaud

2007-05-12

410

Comparative Gene Expression Profiling Identifies Common Molecular Signatures of NF-?B Activation in Canine and Human Diffuse Large B Cell Lymphoma (DLBCL).  

PubMed

We present the first comparison of global transcriptional changes in canine and human diffuse large B-cell lymphoma (DLBCL), with particular reference to the nuclear factor-kappa B (NF-?B) pathway. Microarray data generated from canine DLBCL and normal lymph nodes were used for differential expression, co-expression and pathway analyses, and compared with analysis of microarray data from human healthy and DLBCL lymph nodes. The comparisons at gene level were performed by mapping the probesets in canine microarrays to orthologous genes in humans and vice versa. A considerable number of differentially expressed genes between canine lymphoma and healthy lymph node samples were also found differentially expressed between human DLBCL and healthy lymph node samples. Principal component analysis using a l