Sample records for enterovirus b human

  1. Biological Significance of a Human Enterovirus B-Specific RNA Element in the 3? Nontranslated Region

    PubMed Central

    Merkle, Ingrid; van Ooij, Mark J. M.; van Kuppeveld, Frank J. M.; Glaudemans, Dirk H. R. F.; Galama, Jochem M. D.; Henke, Andreas; Zell, Roland; Melchers, Willem J. G.

    2002-01-01

    The secondary structures predicted for the enteroviral 3? nontranslated region (3?NTR) all seem to indicate a conformation consisting of two (X and Y) hairpin structures. The higher-order RNA structure of the 3?NTR appears to exist as an intramolecular kissing interaction between the loops of these two hairpin structures. The enterovirus B-like subgroup possesses an additional stem-loop structure, domain Z, which is not present in the poliovirus-like enteroviruses. It has been suggested that the Z domain originated from a burst of short sequence repetitions (E. V. Pilipenko, S. V. Maslova, A. N. Sinyakov, and V. I. Agol, Nucleic Acids Res. 20:1739-1745, 1992). However, no functional features have yet been ascribed to this enterovirus B-like-specific RNA element in the 3?NTR. In this study, we tested the functional characteristics and biological significance of domain Z. A mutant of the cardiovirulent coxsackievirus group B3 strain Nancy which completely lacked the Z domain and which therefore acquired enterovirus C-like secondary structures exhibited a wild-type growth phenotype, as determined by single-cycle growth analysis with BGM cells. This result proves that the Z domain is virtually dispensable for viral growth in tissue cultures. Partial distortion of the Z domain structure resulted in a disabled virus with reduced growth kinetics, probably due to alternative conformations of the overall structure of the domain. Infection of mice showed that the recombinant coxsackievirus group B3 mutant which completely lacked the Z domain was less virulent. Pancreatic tissues from mice infected with wild-type virus and recombinant virus were equally affected. However, the heart tissue from mice infected with the recombinant virus showed only slight signs of myocarditis. These results suggest that the enterovirus B-like-specific Z domain plays a role in coxsackievirus-induced pathogenesis. PMID:12208967

  2. Genetic Relationship between Cocirculating Human Enteroviruses Species C

    E-print Network

    Paris-Sud XI, Université de

    Genetic Relationship between Cocirculating Human Enteroviruses Species C Mae¨l Bessaud1, Razafindratsimandresy R, Delpeyroux F (2011) Genetic Relationship between Cocirculating Human Enteroviruses Species C Recombination events between human enteroviruses (HEV) are known to occur frequently and to participate

  3. Molecular strategy for 'serotyping' of human enteroviruses

    Microsoft Academic Search

    Sophie Guillot; Francis Delpeyroux; Radu Crainic

    2001-01-01

    To explore further the phylogenetic relationships between human enteroviruses and to develop new diagnostic approaches, we designed a pair of generic primers in order to study a 1452 bp genomic fragment (relative to the poliovirus Mahoney genome), including the 3« end of the VP1- coding region, the 2A- and 2B-coding regions, and the 5« moiety of the 2C-coding region. Fifty-

  4. Epidemiology and seroepidemiology of human enterovirus 71 among Thai populations

    PubMed Central

    2014-01-01

    Background Human enterovirus 71 (EV71) is an important pathogen caused large outbreaks in Asian-Pacific region with severe neurological complications and may lead to death in young children. Understanding of the etiological spectrum and epidemic changes of enterovirus and population’s immunity against EV71 are crucial for the implementation of future therapeutic and prophylactic intervention. Results A total of 1,182 patients who presented with the symptoms of hand foot and mouth disease (67.3%) or herpangina (HA) (16.7%) and admitted to the hospitals during 2008-2013 were tested for enterovirus using pan-enterovirus PCR targeting 5?-untranslated region and specific PCR for viral capsid protein 1 gene. Overall, 59.7% were pan-enterovirus positive comprising 9.1% EV71 and 31.2% coxsackievirus species A (CV-A) including 70.5% CV-A6, 27.6% CV-A16, 1.1% CV-A10, and 0.8% CV-A5. HFMD and HA occurred endemically during 2008-2011. The number of cases increased dramatically in June 2012 with the percentage of the recently emerged CV-A6 significantly rose to 28.4%. Co-circulation between different EV71 genotypes was observed during the outbreak. Total of 161 sera obtained from healthy individuals were tested for neutralizing antibodies (NAb) against EV71 subgenotype B5 (EV71-B5) using microneutralization assay. The seropositive rate of EV71-B5 was 65.8%. The age-adjusted seroprevalence for individuals was found to be lowest in children aged >6 months to 2 years (42.5%). The seropositive rate remained relatively low in preschool children aged?>?2 years to 6 years (48.3%) and thereafter increased sharply to more than 80% in individuals aged?>?6 years. Conclusions This study describes longitudinal data reflecting changing patterns of enterovirus prevalence over 6 years and demonstrates high seroprevalences of EV71-B5 NAb among Thai individuals. The rate of EV71 seropositive increased with age but without gender-specific significant difference. We identified that relative lower EV71 seropositive rate in early 2012 may demonstrate widely presented of EV71-B5 in the population before account for a large outbreak scale epidemic occurred in 2012 with due to a relatively high susceptibility of the younger population. PMID:24548776

  5. Prevalence of human enteroviruses among apparently healthy nursery school children in Accra

    PubMed Central

    Attoh, Juliana; Obodai, Evangeline; Adiku, Theophilus; Odoom, John Kofi

    2014-01-01

    Introduction Human enteroviruses are common in children causing asymptomatic infections ranging from mild to severe illnesses. In Ghana, information on the prevalence of non-polio enterovirus causing acute flaccid paralysis is available but data on surveillance of these viruses in school children is scanty. Here, the prevalence of human enteroviruses among apparently healthy children in selected school in Accra was studied. Methods Stool samples from 273 apparently healthy children less than eight years of age in 9 selected nursery schools were collected between December 2010 and March 2011and processed for human enteroviruses on L20B, RD and Hep-2 cell lines. Positive Isolates were characterized by microneutralisation assay with antisera pools from RIVM, the Netherlands according to standard methods recommended by WHO. Results Of the 273 samples processed, 66 (24.2%) non-polio enteroviruses were isolated. More growth was seen on Hep-2C (46%) only than RD (18%) only and on both cell lines (34%). No growth was seen on L20B even after blind passage. Excretion of non-polio enteroviruses was found in all the schools with majority in BD school. Serotyping of the isolates yielded predominantly Coxsackie B viruses followed by echoviruses 13 and 7. More than half of the isolates could not be typed by the antisera pools. Conclusion The study detected 13 different serotypes of non-polio enteroviruses in circulation but no poliovirus was found. BD school was found to have the highest prevalence of NPEV. Complete identification through molecular methods is essential to establish the full range of NPEVs in circulation in these schools. PMID:25400833

  6. Human enterovirus 71 epidemics: what's next?

    PubMed Central

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

    2013-01-01

    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

  7. Cleavage of eukaryotic initiation factor eIF5B by enterovirus 3C proteases

    PubMed Central

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

    2008-01-01

    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. PMID:18572216

  8. Cleavage of eukaryotic initiation factor eIF5B by enterovirus 3C proteases.

    PubMed

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

    2008-08-15

    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 (VVEQG, equivalent to VMEQG479 in 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 shutoff of translation that occurs in enterovirus-infected cells. PMID:18572216

  9. Epidemics and Frequent Recombination within Species in Outbreaks of Human Enterovirus B-Associated Hand, Foot and Mouth Disease in Shandong China in 2010 and 2011.

    PubMed

    Zhang, Ting; Du, Jiang; Xue, Ying; Su, Haoxiang; Yang, Fan; Jin, Qi

    2013-01-01

    The epidemiology and molecular characteristics of human enterovirus B (HEV-B) associated with hand, foot and mouth disease (HFMD) outbreaks in China are not well known. In the present study, we tested 201 HEV isolates from 233 clinical specimens from patients with severe HFMD during 2010-2011 in Linyi, Shandong, China. Of the 201 isolates, 189 were fully typed and 18 corresponded to HEV-B species (six serotypes CVA9, CVB1, CVB4, Echo 6, Echo 25 and Echo 30) using sensitive semi-nested polymerase chain reaction analysis of VP1 gene sequences. Phylogenetic analysis based on the VP1 region showed that eight E30SD belonged to a novel sub-genogroup D2; E25SD belonged to a novel sub-genogroup D6; E6SD belonged to sub-lineage C6 and five CVB1SD belonged to subgroup 4C; and B4SD belonged sub-lineage D2. The full viral genomes of the CVB1SD, E6SD, E25SD and E30SD isolates were sequenced. Analysis of phylogenetic and similarity plots indicated that E25SD recombined with E25-HN-2, E30FDJS03 and E4AUS250 at noncontiguous P2A-P3D regions, while E30SD, E30FDJ03, E25-HN-2 and E9 DM had shared sequences in discrete regions of P2 and P3. Both E6SD and B1SD shared sequences with E1-HN, B4/GX/10, B5-HN, and A9-Alberta in contiguous regions of most of P2 and P3. Genetic algorithm recombination detection analysis further confirmed the existence of multiple potential recombination points. In conclusion, analysis of the complete genomes of E25SD, E30SD, CVB1SD and E6SD isolated from HFMD patients revealed that they formed novel subgenogroup. Given the prevalence and recombination of these viruses in outbreaks of HFMD, persistent surveillance of HFMD-associated HEV-B pathogens is required to predict potential emerging viruses and related disease outbreaks. PMID:23840610

  10. Enterovirus 71 induces apoptosis of SH-SY5Y human neuroblastoma cells through stimulation of endogenous microRNA let-7b expression

    PubMed Central

    DU, XILING; WANG, HAIPENG; XU, FUHUI; HUANG, YONGYI; LIU, ZHIXUE; LIU, TE

    2015-01-01

    Enterovirus 71 (EV71) is a pathogenic microorganism that causes hand, foot and mouth disease. However, the epigenetic mechanisms behind how EV71 regulates host cell proliferation and apoptosis are unclear. In the present study, the ability of EV71 to induce apoptosis was analyzed in the SH-SY5Y human neuroblastoma cell line and the effect of this virus on the mRNA expression levels of various apoptotic markers, miRNA let-7b and cyclin D1 (CCND1), was also investigated. The results demonstrated that EV71 induced SH-SY5Y cell apoptosis. An MTT assay revealed a significant inhibitory effect of EV71 on cell proliferation between 12–72 h post injection, compared with the control group. Furthermore, quantitative polymerase chain reaction and western blot analyses demonstrated that expression level of the apoptosis inhibitor Bcl-2 was markedly reduced, but the expression levels of the apoptosis-promoting factors Bax, caspase-7, caspase-3 and active caspase-3 were markedly higher in the SH-SY5Y cells 12–48 h after EV71 infection, compared with the non-infected cells. In addition, flow cytometric assays revealed that EV71 arrested the cell cycle of host SH-SY5Y cells. Northern blot analysis revealed a marked miRNA let-7b hybridization signal in the EV71 virus-infected group compared with the non-infected group. Furthermore, western blotting confirmed that the CCND1 protein expression levels were significantly reduced in EV71-infected SH-SY5Y cells. EV71-inhibited SH-SY5Y proliferation was abrogated using let-7b specific 2?-O-Methyl-RNA, which inhibited endogenous miRNA let-7b expression. Thus, EV71 regulated the host SH-SY5Y cell cycle and cell proliferation via stimulating endogenous miRNA let-7b and directly targeting CCND1, therefore EV71 is a potential candidate for antiviral therapy. PMID:25779425

  11. Genome Characterisation of Enteroviruses 117 and 118: A New Group within Human Enterovirus Species C

    PubMed Central

    Scala, Alessia; Greenberg, David; Usonis, Vytautas; Principi, Nicola; Baldanti, Fausto; Esposito, Susanna

    2013-01-01

    The more than 120 genotypes of human enteroviruses (HEVs) reflect a wide range of evolutionary divergence, and there are 23 currently classified as human enterovirus C species (HEV-C). Two new HEV-C (EV-C117 and EV-C118) were identified in the Community-Acquired Pneumonia Pediatric Research Initiative (CAP-PRI) study, and the present paper describes the characterisation of the complete genome of one EV-C117 strain (LIT22) and two EV-C118 (ISR38 and ISR10) strains. The EV-C117 and EV-C118 5?UTR sequences were related to those of EV-C104, EV-C105 and EV-C109, and were slightly shorter than those of other HEV A-D species. Similarity plot analyses showed that EV-C117 and EV-C118 have a P1 region that is highly divergent from that of the other HEV-C, and phylogenetic analyses highly supported a monophyletic group consisting of EV-C117, EV-C118, EV-C104, EV-C105 and EV-C109 strains. Phylogenetic, Simplot and Bootscan analyses indicated that recombination was not the main mechanism of EV-C117 and EV-C118 evolution, thus strengthening the hypothesis of the monophyletic origin of the coding regions, as in the case of other HEV-C. Phylogenetic analysis also revealed the emergence of a new group within HEV-C that is divided into two subgroups. Nucleotide and amino acid identity in VP1 sequences have been established as useful criteria for assigning new HEV types, but analysis of the complete P1 region improves resolution. PMID:23565264

  12. [Enterovirus infections in new disguise].

    PubMed

    Fohlman, J; Friman, G; Tuvemo, T

    1997-07-01

    Enteroviruses (Coxsackie A and B, echovirus, poliovirus) belong to a group of small RNA-viruses, picomavirus, which are widespread in nature. Enteroviruses cause a number of wellknown diseases and symptoms in humans, from subclinical infections and the common cold to poliomyelitis with paralysis. The development of polio vaccines is the greatest accomplishment within the field of enterovirus research and the background work was awarded the Nobel prize in 1954. New knowledge implies that enteroviruses play a more important part in the morbidity panorama than was previously thought. Chronic (persistent) enteroviruses were formerly unknown. Serologic and molecular biology techniques have now demonstrated that enteroviral genomes, in certain situations, persist after the primary infection (which is often silent). Persistent enteroviral infection or recurrent infections and/or virus-stimulated autoimmunity might contribute to the development of diseases with hitherto unexplained pathogenesis, such as post polio syndrome, dilated cardiomyopathy, juvenile (type 1) diabetes and possibly some cases of chronic fatigue syndrome. PMID:9254324

  13. Genetic Relationship between Cocirculating Human Enteroviruses Species C

    PubMed Central

    Holmblat, Barbara; Razafindratsimandresy, Richter; Delpeyroux, Francis

    2011-01-01

    Recombination events between human enteroviruses (HEV) are known to occur frequently and to participate in the evolution of these viruses. In a previous study, we reported the isolation of a panel of viruses belonging to the Human enterovirus species C (HEV-C) that had been cocirculating in a small geographic area of Madagascar in 2002. This panel included type 2 vaccine-derived polioviruses (PV) that had caused several cases of acute flaccid paralysis in humans. Previous partial sequencing of the genome of these HEV-C isolates revealed considerable genetic diversity, mostly due to recombination. In the work presented herein, we carried out a more detailed characterization of the genomes of viruses from this collection. First, we determined the full VP1 sequence of 41 of these isolates of different types. These sequences were compared with those of HEV-C isolates obtained from other countries or in other contexts. The sequences of the Madagascan isolates of a given type formed specific clusters clearly differentiated from those formed by other strains of the same type isolated elsewhere. Second, we sequenced the entire genome of 10 viruses representing most of the lineages present in this panel. All but one of the genomes appeared to be mosaic assemblies of different genomic fragments generated by intra- and intertypic recombination. The location of the breakpoints suggested potential preferred genomic regions for recombination. Our results also suggest that recombination between type HEV-99 and other HEV-C may be quite rare. This first exhaustive genomic analysis of a panel of non-PV HEV-C cocirculating in a small human population highlights the high frequency of inter and intra-typic genetic recombination, constituting a widespread mechanism of genetic plasticity and continually shifting the HEV-C biodiversity. PMID:21931857

  14. Molecular epidemiology of enterovirus B77 isolated from non polio acute flaccid paralytic patients in Pakistan during 2013.

    PubMed

    Angez, Mehar; Shaukat, Shahzad; Zahra, Rabaab; Khurshid, Adnan; Sharif, Salmaan; Alam, Muhammad Masroor; Zaidi, Syed Sohail Zahoor

    2015-01-01

    Human enteroviruses are associated with various clinical syndromes and severe neurological disorders. The aim of this study was to determine the molecular epidemiology of non polio enteroviruses and their correlation with acute flaccid paralysis (AFP) patients living in Khyber Pakhtunkhwa (KP) and Federally Administered Tribal Areas (FATA) of Pakistan. The stool samples collected from these patients were used for isolation of non polio enteroviruses (NPEVs). Out of 38 samples, 29 (76.3%) were successfully typed by microneutralization assay into eleven serotypes including echovirus (E)-3 (5.3%), E-7 (2.6%), E-11 (13.2%), E-12 (7.9%), E-13 (10.5%), E-20 (7.9%), E-27 (5.3%), E-29 (10.5%), E-30 (7.9%), E-33 (2.6%), coxsackievirus (CV) B5 (2.6%) and nine isolates (23.7%) remained untyped which were confirmed as NPEVs by real time RT-PCR. Complete VP1 genetic sequencing data characterized untypeable isolates into enterovirus B77 (EV-B77). Moreover, molecular phylogenetic analysis classified these viruses into two new genotypes having high genetic diversity (at least 17.7%) with prototype. This study provides valuable information on extensive genetic diversity of EV-B77 genotypes. Although, its association with neurological disorder has not yet been known but isolation of nine EV-B77 viruses from AFP cases highlights the fact that they may have a contributing role in the etiology of AFP. In addition, it is needed to establish enterovirus surveillance system and laboratory diagnostic facilities for early detection of NPEVs that may cause poliomyelitis like paralysis especially in the situation when we are at the verge of polio eradication. PMID:25433133

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

    PubMed Central

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

    2013-01-01

    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-Ĺ-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. PMID:23509286

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

    PubMed

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

    2011-10-01

    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 Ĺ, 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

  17. Structural Basis for Antiviral Inhibition of the Main Protease, 3C, from Human Enterovirus 93 ?

    PubMed Central

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

    2011-01-01

    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 (3Cpro), which has narrow, conserved substrate specificity. These cleavages are essential for virus replication, making 3Cpro an excellent target for antivirus drug development. In this study, we report the first determination of the crystal structure of 3Cpro 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 Ĺ, respectively. The EV-93 3Cpro 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 3Cpro 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

  18. Characterization of the genome of human enteroviruses: design of generic primers for amplification and sequencing of different regions of the viral genome

    E-print Network

    Boyer, Edmond

    human enteroviruses into 5 species (Hyypia et al., 1997): HEV-A (at least 17 serotypes), HEV-B (at least 56 serotypes), HEV-C (at least 13 serotypes), HEV-D (at least 3 serotypes) and the 3 serotypes of PV

  19. Survey of human enterovirus occurrence in fresh and marine surface waters on Long Island

    Microsoft Academic Search

    J. M. Vaughn; E. F. Landry; M. Z. Thomas; T. J. Vicale; W. F. Penello

    1979-01-01

    A variety of surface water systems, including a lake, a creek, and two marine embayments, were analyzed on a monthly basis for indigenous human enteroviruses and coliform bacteria. Findings are discussed in terms of the probable pollution sources to each system and their relationship to data from previous studies.

  20. Identification of site-specific adaptations conferring increased neural cell tropism during human enterovirus 71 infection.

    PubMed

    Cordey, Samuel; Petty, Tom J; Schibler, Manuel; Martinez, Yannick; Gerlach, Daniel; van Belle, Sandra; Turin, Lara; Zdobnov, Evgeny; Kaiser, Laurent; Tapparel, Caroline

    2012-01-01

    Enterovirus 71 (EV71) is one of the most virulent enteroviruses, but the specific molecular features that enhance its ability to disseminate in humans remain unknown. We analyzed the genomic features of EV71 in an immunocompromised host with disseminated disease according to the different sites of infection. Comparison of five full-length genomes sequenced directly from respiratory, gastrointestinal, nervous system, and blood specimens revealed three nucleotide changes that occurred within a five-day period: a non-conservative amino acid change in VP1 located within the BC loop (L97R), a region considered as an immunogenic site and possibly important in poliovirus host adaptation; a conservative amino acid substitution in protein 2B (A38V); and a silent mutation in protein 3D (L175). Infectious clones were constructed using both BrCr (lineage A) and the clinical strain (lineage C) backgrounds containing either one or both non-synonymous mutations. In vitro cell tropism and competition assays revealed that the VP1?? Leu to Arg substitution within the BC loop conferred a replicative advantage in SH-SY5Y cells of neuroblastoma origin. Interestingly, this mutation was frequently associated in vitro with a second non-conservative mutation (E167G or E167A) in the VP1 EF loop in neuroblastoma cells. Comparative models of these EV71 VP1 variants were built to determine how the substitutions might affect VP1 structure and/or interactions with host cells and suggest that, while no significant structural changes were observed, the substitutions may alter interactions with host cell receptors. Taken together, our results show that the VP1 BC loop region of EV71 plays a critical role in cell tropism independent of EV71 lineage and, thus, may have contributed to dissemination and neurotropism in the immunocompromised patient. PMID:22910880

  1. [Detection of human enteroviruses with real-time PCR assay using TaqMan fluorescent probe].

    PubMed

    Le?, Katarzyna; Przybylski, Maciej; Dzieciatkowski, Tomasz; M?ynarczyk, Grazyna

    2010-01-01

    Infections with human enteroviruses are common worldwide and cause a wide range of signs and symptoms. Nowadays in current diagnostics procedures older virological methods, such virus isolation in a cell cultures and seroneutralisation assay, are replaced with molecular biology tests. The aim of the study was development of real-time PCR assay for detection of human adenoviruses. DNA isolated from MK2 cell line infected with nineteen different enterovirus strains was used for development of a qualitative real-time PCR assay using primers targeting a conserved region of the 5'UTR region and a specific TaqMan probe. The analytical sensitivity of real-time PCR assay was tested using serial dilutions of Coxackie A9 cDNA in range between 10 degrees and 10(-8). For comparison typical end-point detected RT-PCR for enterovirus detection with the same cDNA dilutions was made. The sensitivity of novel method was about ten thousand-fold higher than older one. The conclusion is that real-time PCR is very advisable in diagnostics of diseases caused with enteroviruses. The high level of sensitivity, specificity, accuracy, and rapidity provided by this assay are favorable for the use in the detection of enteroviral RNA in clinical specimens, especially from neuroinfections. PMID:21114017

  2. Viral kinetics of Enterovirus 71 in human abdomyosarcoma cells

    PubMed Central

    Lu, Jing; He, Ya-Qing; Yi, Li-Na; Zan, Hong; Kung, Hsiang-Fu; He, Ming-Liang

    2011-01-01

    AIM: To characterise the viral kinetics of enterovirus 71 (EV71). METHODS: In this study, human rhabdomyosarcoma (RD) cells were infected with EV71 at different multiplicity of infection (MOI). After infection, the cytopathic effect (CPE) was monitored and recorded using a phase contrast microscope associated with a CCD camera at different time points post viral infection (0, 6, 12, 24 h post infection). Cell growth and viability were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in both EV71 infected and mock infected cells at each time point. EV71 replication kinetics in RD cells was determined by measuring the total intracellular viral RNA with real-time reverse-transcription polymerase chain reaction (qRT-PCR). Also, the intracellular and extracellular virion RNA was isolated and quantified at different time points to analyze the viral package and secretion. The expression of viral protein was determined by analyze the levels of viral structure protein VP1 with Western blotting. RESULTS: EV71 infection induced a significant CPE as early as 6 h post infection (p.i.) in both RD cells infected with high ratio of virus (MOI 10) and low ratio of virus (MOI 1). In EV71 infected cells, the cell growth was inhibited and the number of viable cells was rapidly decreased in the later phase of infection. EV71 virions were uncoated immediately after entry. The intracellular viral RNA began to increase at as early as 3 h p.i. and the exponential increase was found between 3 h to 6 h p.i. in both infected groups. For viral structure protein synthesis, results from western-blot showed that intracellular viral protein VP1 could not be detected until 6 h p.i. in the cells infected at either MOI 1 or MOI 10; and reached the peak at 9 h p.i. in the cells infected with EV71 at both MOI 1 and MOI 10. Simultaneously, the viral package and secretion were also actively processed as the virus underwent rapid replication. The viral package kinetics was comparable for both MOI 1 and MOI 10 infected groups. It was observed that at 3 h p.i, the intracellular virions obviously decreased, thereafter, the intracellular virions began to increase and enter into the exponential phase until 12 h p.i. The total amounts of intracellular virons were decreased from 12 to 24 h p.i. Consistent with this result, the increase of virus secretion occurred during 6 to 12 h p.i. CONCLUSION: The viral kinetics of EV71 were established by analyzing viral replication, package and secretion in RD cells. PMID:22039330

  3. Molecular Characterization of Human Enteroviruses in the Central African Republic: Uncovering Wide Diversity and Identification of a New Human Enterovirus A71 Genogroup

    PubMed Central

    Pillet, Sylvie; Ibrahim, Wafa; Joffret, Marie-Line; Pozzetto, Bruno; Gouandjika-Vasilache, Ionela

    2012-01-01

    Human enteroviruses (HEV) are among the most common viruses infecting humans. Their circulation has been widely studied in most parts of the world but not in sub-Saharan Africa, where poliomyelitis remains prevalent. We report here the molecular characterization of 98 nonpoliovirus (non-PV) HEV strains isolated from 93 randomly selected cell culture-positive supernatants from stool samples collected from 1997 through 2006 from children with acute flaccid paralysis living in the Central African Republic (CAR). The isolates were typed by sequencing the VP1 coding region and sequenced further in the VP2 coding region, and phylogenetic studies were carried out. Among the 98 VP1 sequences, 3, 74, 18, and 3 were found to belong to the HEV-A, -B, -C, and -D species, respectively. Overall, 42 types were detected. In most cases, the VP2 type was correlated with that of the VP1 region. Some of the isolates belonged to lineages that also contain viruses isolated in distant countries, while others belonged to lineages containing viruses isolated only in Africa. In particular, one isolate (type EV-A71) did not fall into any of the genogroups already described, indicating the existence of a previously unknown genogroup for this type. These results illustrate the considerable diversity of HEV isolates from the stools of paralyzed children in the CAR. The presence of diverse HEV-C types makes recombination between poliovirus and other HEV-C species possible and could promote the emergence of recombinant vaccine-derived polioviruses similar to those that have been implicated in repeated poliomyelitis outbreaks in several developing countries. PMID:22337981

  4. Human enteroviruses in oysters and their overlying waters.

    PubMed

    Goyal, S M; Gerba, C P; Melnick, J L

    1979-03-01

    The presence of enteroviruses in oysters and oyster-harvesting waters of the Texas Gulf coast was monitored over a period of 10 months. Viruses were detected in water and oyster samples obtained from areas both open and closed to shellfish harvesting. Viruses were detected periodically in waters that met current bacteriological standards for shellfish harvesting. No significant statistical relationship was demonstrated between virus concentration in oysters and the bacteriological and physiochemical quality of water and shellfish. Viruses in water were, however, moderately correlated with total coliforms in water and oysters and with fecal coliforms in oysters. Total coliforms in water were realted to total coliforms in sediment were related only to total coliforms in sediment. Among the physiochemical characteristics of water, turbidity was related statistically to the organic matter content of water and to fecal coliforms in water. There was a marked effect of rainfall on the bacteriological quality of water. Of a total of 44 water samples, 26 yielded virus in concentrations from 4 to 167 plaque-forming units per 100-gallon (ca. 378.5-liter) sample. Of a total of 40 pools of 10 to 12 oysters each, virus was found in 14 pools at a concentration of 6 to 224 plaque-forming units per 100 g of oyster meat. On five occasions, virus was found in water samples when no virus could be detected in oysters harvested from the same sites. This study indicates that current bacteriological standards for determining the safety of shellfish and shellfish-growing waters do no reflect the occurrence of enteroviruses. PMID:222210

  5. Molecular Evolution of the Human Enteroviruses: Correlation of Serotype with VP1 Sequence and Application to Picornavirus Classification

    PubMed Central

    Oberste, M. Steven; Maher, Kaija; Kilpatrick, David R.; Pallansch, Mark A.

    1999-01-01

    Sixty-six human enterovirus serotypes have been identified by serum neutralization, but the molecular determinants of the serotypes are unknown. Since the picornavirus VP1 protein contains a number of neutralization domains, we hypothesized that the VP1 sequence should correspond with neutralization (serotype) and, hence, with phylogenetic lineage. To test this hypothesis and to analyze the phylogenetic relationships among the human enteroviruses, we determined the complete VP1 sequences of the prototype strains of 47 human enterovirus serotypes and 10 antigenic variants. Our sequences, together with those available from GenBank, comprise a database of complete VP1 sequences for all 66 human enterovirus serotypes plus additional strains of seven serotypes. Phylogenetic trees constructed from complete VP1 sequences produced the same four major clusters as published trees based on partial VP2 sequences; in contrast to the VP2 trees, however, in the VP1 trees strains of the same serotype were always monophyletic. In pairwise comparisons of complete VP1 sequences, enteroviruses of the same serotype were clearly distinguished from those of heterologous serotypes, and the limits of intraserotypic divergence appeared to be about 25% nucleotide sequence difference or 12% amino acid sequence difference. Pairwise comparisons suggested that coxsackie A11 and A15 viruses should be classified as strains of the same serotype, as should coxsackie A13 and A18 viruses. Pairwise identity scores also distinguished between enteroviruses of different clusters and enteroviruses from picornaviruses of different genera. The data suggest that VP1 sequence comparisons may be valuable in enterovirus typing and in picornavirus taxonomy by assisting in the genus assignment of unclassified picornaviruses. PMID:9971773

  6. Activity of Pleconaril against Enteroviruses

    PubMed Central

    Pevear, Daniel C.; Tull, Tina M.; Seipel, Martin E.; Groarke, James M.

    1999-01-01

    The activity of pleconaril in cell culture against prototypic enterovirus strains and 215 clinical isolates of the most commonly isolated enterovirus serotypes was examined. The latter viruses were isolated by the Centers for Disease Control and Prevention during the 1970s and 1980s from clinically ill subjects. Pleconaril at a concentration of ?0.03 ?M inhibited the replication of 50% of all clinical isolates tested. Ninety percent of the isolates were inhibited at a drug concentration of ?0.18 ?M. The most sensitive serotype, echovirus serotype 11, was also the most prevalent enterovirus in the United States from 1970 to 1983. Pleconaril was further tested for oral activity in three animal models of lethal enterovirus infection: coxsackievirus serotype A9 infection in suckling mice, coxsackievirus serotype A21 strain Kenny infection in weanling mice, and coxsackievirus serotype B3 strain M infection in adult mice. Treatment with pleconaril increased the survival rate in all three models for both prophylactic and therapeutic dosing regimens. Moreover, pleconaril dramatically reduced virus levels in target tissues of coxsackievirus serotype B3 strain M-infected animals. Pleconaril represents a promising new drug candidate for potential use in the treatment of human enteroviral infections. PMID:10471549

  7. A neonatal gnotobiotic pig model of human enterovirus 71 infection and associated immune responses

    PubMed Central

    Yang, Xingdong; Li, Guohua; Wen, Ke; Bui, Tammy; Liu, Fangning; Kocher, Jacob; Jortner, Bernard S; Vonck, Marlice; Pelzer, Kevin; Deng, Jie; Zhu, Runan; Li, Yuyun; Qian, Yuan; Yuan, Lijuan

    2014-01-01

    Vaccine development and pathogenesis studies for human enterovirus 71 are limited by a lack of suitable animal models. Here, we report the development of a novel neonatal gnotobiotic pig model using the non-pig-adapted neurovirulent human enterovirus 71 strain BJ110, which has a C4 genotype. Porcine small intestinal epithelial cells, peripheral blood mononuclear cells and neural cells were infected in vitro. Oral and combined oral–nasal infection of 5-day-old neonatal gnotobiotic pigs with 5×108 fluorescence forming units (FFU) resulted in shedding up to 18 days post-infection, with viral titers in rectal swab samples peaking at 2.22×108 viral RNA copies/mL. Viral capsid proteins were detected in enterocytes within the small intestines on post-infection days (PIDs) 7 and 14. Additionally, viral RNA was detected in intestinal and extra-intestinal tissues, including the central nervous system, the lung and cardiac muscle. The infected neonatal gnotobiotic pigs developed fever, forelimb weakness, rapid breathing and some hand, foot and mouth disease symptoms. Flow cytometry analysis revealed increased frequencies of both CD4+ and CD8+ IFN-?-producing T cells in the brain and the blood on PID 14, but reduced frequencies were observed in the lung. Furthermore, high titers of serum virus-neutralizing antibodies were generated in both orally and combined oral–nasally infected pigs on PIDs 7, 14, 21 and 28. Together, these results demonstrate that neonatal gnotobiotic pigs represent a novel animal model for evaluating vaccines for human enterovirus 71 and for understanding the pathogenesis of this virus and the associated immune responses.

  8. Molecular epidemiology of severe respiratory disease by human rhinoviruses and enteroviruses at a tertiary paediatric hospital in Barcelona, Spain.

    PubMed

    Launes, C; Armero, G; Anton, A; Hernandez, L; Gimferrer, L; Cisneros, C; Jordan, I; Muńoz-Almagro, C

    2015-08-01

    In order to describe the molecular epidemiology of human rhinovirus (HRV) and enterovirus (EV) infection in severely ill children, we studied all episodes of bronchospasm/bronchopneumonia in 6-month-old to 18-year-old patients from January 2010 to May 2012 who required mechanical ventilation. HRV/EVs were detected in 55 (57.3%) of 96 patients, of which 50 (91%) were HRV (HRV-A, 16; HRV-B, 1; HRV-C, 18) and 5 (9%) were EVs (EV-D68, 3). No significant differences in epidemiologic and clinical characteristics were found between different types. In six of the 13 patients who required invasive mechanical ventilation, HRV was the only pathogen detected. PMID:25964153

  9. Factors affecting the detection of enteroviruses in cerebrospinal fluid with coxsackievirus B3 and poliovirus 1 cDNA probes.

    PubMed Central

    Rotbart, H A; Levin, M J; Villarreal, L P; Tracy, S M; Semler, B L; Wimmer, E

    1985-01-01

    Enteroviruses are common pathogens of meningitis and encephalitis, and infections are often difficult to distinguish clinically from bacterial and herpetic infections of the central nervous system. An array of enteroviruses added to cerebrospinal fluid in reconstruction experiments were detected by a dot hybridization assay. Optimal handling and processing conditions for infected cerebrospinal fluid were established, and the effect on the hybridization reaction of humoral and cellular components of the inflammatory response was determined. Six hybridization probes, derived from poliovirus 1 and coxsackievirus B3, were then tested, singly and in combinations, to optimize the sensitivity and spectrum of the assay. Implications for enteroviral taxonomy based on these experiments are discussed. Images PMID:2993351

  10. Evidence of Recombination among Enteroviruses

    Microsoft Academic Search

    JUHANA SANTTI; TIMO HYYPIA; LEENA KINNUNEN; MIKA SALMINEN

    1999-01-01

    Human enteroviruses consist of more than 60 serotypes, reflecting a wide range of evolutionary divergence. They have been genetically classified into four clusters on the basis of sequence homology in the coding region of the single-stranded RNA genome. To explore further the genetic relationships between human enteroviruses and to characterize the evolutionary mechanisms responsible for variation, previously sequenced genomes were

  11. Recombination in Circulating Enteroviruses

    PubMed Central

    Lukashev, Alexander N.; Lashkevich, Vasilii A.; Ivanova, Olga E.; Koroleva, Galina A.; Hinkkanen, Ari E.; Ilonen, Jorma

    2003-01-01

    Recombination is a well-known phenomenon for enteroviruses. However, the actual extent of recombination in circulating nonpoliovirus enteroviruses is not known. We have analyzed the phylogenetic relationships in four genome regions, VP1, 2A, 3D, and the 5? nontranslated region (NTR), of 40 enterovirus B strains (coxsackie B viruses and echoviruses) representing 11 serotypes and isolated in 1981 to 2002 in the former Soviet Union states. In the VP1 region, strains of the same serotype expectedly grouped with their prototype strain. However, as early as the 2A region, phylogenetic grouping differed significantly from that in the VP1 region and indicated recombination within the 2A region. Moreover, in the 5? NTR and 3D region, only 1 strain of 40 grouped with its prototype strain. Instead, we observed a major group in both the 5? NTR and the 3D region that united most (in the 5? NTR) or all (in the 3D region) of the strains studied, regardless of the serotype. Subdivision within that major group in the 3D region correlated with the time of virus isolation but not with the serotype. Therefore, we conclude that a majority, if not all, circulating enterovirus B strains are recombinants relative to the prototype strains, isolated mostly in the 1950s. Moreover, the ubiquitous recombination has allowed different regions of the enterovirus genome to evolve independently. Thus, a novel model of enterovirus genetics is proposed: the enterovirus genome is a stable symbiosis of genes, and enterovirus species consist of a finite set of capsid genes responsible for different serotypes and a continuum of nonstructural protein genes that seem to evolve in a relatively independent manner. PMID:12970427

  12. Evolutionary Genetics of Human Enterovirus 71: Origin, Population Dynamics, Natural Selection, and Seasonal Periodicity of the VP1 Gene

    Microsoft Academic Search

    Kok Keng Tee; Tommy Tsan-Yuk Lam; Yoke Fun Chan; Jon M. Bible; Adeeba Kamarulzaman; C. Y. William Tong; Yutaka Takebe; Oliver G. Pybus

    2010-01-01

    Human enterovirus 71 (EV-71) is one of the major etiologic causes of hand, foot, and mouth disease (HFMD) among young children worldwide, with fatal instances of neurological complications becoming increasingly common. Global VP1 capsid sequences (n = 628) sampled over 4 decades were collected and subjected to comprehensive evolutionary analysis using a suite of phylogenetic and population genetic methods. We

  13. Stability of human enteroviruses in estuarine and marine waters.

    PubMed Central

    Lo, S; Gilbert, J; Hetrick, F

    1976-01-01

    Studies of the effects of temperature and salinity on the survival of three enteric viruses (poliomyelitis type 1, echovirus-6, and coxsackievirus B-5) under controlled laboratory conditions and in situ indicate that temperature rather than salinity is the critical factor affecting their stability, in that the higher the temperature the more rapid was the loss of viral infectivity. In the laboratory studies, all three viruses were quite stable at 4 degrees C, with infectious virus still detectable after 46 weeks of incubation. In situ studies on virus survival in free-flowing estuarine or marine waters showed that, although the viruses were more labile in natural waters than in the laboratory studies, they persisted for several months, in some cases during the winter months. At all temperatures and salinities, coxsackievirus B-5 was the most stable, echovirus-6 was intermediate, and poliovirus 1 was the least stable of the viruses tested. PMID:184736

  14. Identification of Enteroviruses in Naturally Infected Captive Primates

    Microsoft Academic Search

    W. Allan Nix; Baoming Jiang; Kaija Maher; Elizabeth Strobert; M. Steven Oberste

    2008-01-01

    In a recent study, we investigated cases of diarrheal disease among monkeys at a U.S. primate center. In that study, enteroviruses were detected in a high proportion of the fecal specimens tested. To determine whether the enterovirus detections represented the circulation of one or more simian enteroviruses within the colony or the transmission of human enteroviruses from animal handlers, we

  15. Differential Susceptibility and Response of Primary Human Myeloid BDCA1+ Dendritic Cells to Infection with Different Enteroviruses

    PubMed Central

    Schulte, Barbara M.; Kers-Rebel, Esther D.; Prosser, Amy C.; Galama, Jochem M. D.; van Kuppeveld, Frank J. M.; Adema, Gosse J.

    2013-01-01

    Coxsackie B viruses (CVBs) and echoviruses (EVs) form the Human Enterovirus-B (HEV-B) species within the family Picornaviridae. HEV-B infections are widespread and generally cause mild disease; however, severe infections occur and HEV-B are associated with various chronic diseases such as cardiomyopathy and type 1 diabetes. Dendritic cells (DCs) are the professional antigen-presenting cells of our immune system and initiate and control immune responses to invading pathogens, yet also maintain tolerance to self-antigens. We previously reported that EVs, but not CVBs, can productively infect in vitro generated monocyte-derived DCs. The interactions between HEV-B and human myeloid DCs (mDCs) freshly isolated from blood, however, remain unknown. Here, we studied the susceptibility and responses of BDCA1+ mDC to HEV-B species and found that these mDC are susceptible to EV, but not CVB infection. Productive EV7 infection resulted in massive, rapid cell death without DC activation. Contrary, EV1 infection, which resulted in lower virus input at the same MOI, resulted in DC activation as observed by production of type I interferon-stimulated genes (ISGs), upregulation of co-stimulatory and co-inhibitory molecules (CD80, CD86, PDL1) and production of IL-6 and TNF-?, with a relative moderate decrease in cell viability. EV1-induced ISG expression depended on virus replication. CVB infection did not affect DC viability and resulted in poor induction of ISGs and CD80 induction in part of the donors. These data show for the first time the interaction between HEV-B species and BDCA1+ mDCs isolated freshly from blood. Our data indicate that different HEV-B species can influence DC homeostasis in various ways, possibly contributing to HEV-B associated pathology. PMID:23638101

  16. The interaction between human enteroviruses and type I IFN signaling pathway.

    PubMed

    Lu, Jing; Yi, Lina; Ke, Changwen; Zhang, Yonghui; Liu, Ren; Chen, Jinfei; Kung, Hsiang-Fu; He, Ming-Liang

    2015-06-01

    Human enteroviruses (HEV), very common and important human pathogens, cause infections in diverse ways. Recently, the large epidemic of HFMD caused by HEV infection became a growing threat to public health in China. As the first line of immune response, the type I interferon (IFN-?/?) pathway plays an essential role in antiviral infection, particularly in limiting both the early and late stages of infection. Because of co-evolution with the host, the viruses have evolved multiple strategies to evade or subvert the host immunity to ensure their survival. In this paper, we systematically reviewed and summarized the interaction between HEV infections and host type I IFN responses. We firstly described the recent findings of HEV recognition and IFN induction, specifically on host pattern-recognition receptors (PRRs) in HEV infection. Then we discussed the antiviral effect of IFN in HEV infection. Finally, we timely summarized the mechanisms of HEV to circumvent the IFN responses. Clarification of the complexity in this battle may provide us new strategies for prevention and antiviral treatment. PMID:23919297

  17. Molecular epidemiology and evolution of human enterovirus 71 and hand, foot and mouth disease.

    PubMed

    Zhifang, Liu; Juanjuan, Gui; Qihang, Hua; Changzheng, Dong

    2015-05-01

    Human enterovirus 71(EV71), one of the major pathogens of the hand, foot and mouth disease (HFMD), causes skin rashes in palms, feet and mouth ulcers and complication in the central nervous system such as aseptic meningitis and acute flaccid paralysis that may lead to death. EV71 infection has been reported to be associated with many outbreaks of HFMD worldwide, especially the great outbreaks that occurred in the Asia-Pacific region and caused numerous death since 1997. The studies of molecular epidemiology and evolution of EV71 are important for the prevention and control of HFMD since no vaccines and antiviral drugs have been developed except symptomatic treatment for HFMD. In this review, we summarize genotype classification, temporal and spatial distribution, evolutionary characteristics and modes of EV71 as well as typical EV71 epidemics. Further studies on EV71 and HFMD may lead to better understanding of pathological mechanisms of EV71, development of antiviral drugs and prevention and control of HFMD. PMID:25998430

  18. Evidence of Recombination among Enteroviruses

    PubMed Central

    Santti, Juhana; Hyypiä, Timo; Kinnunen, Leena; Salminen, Mika

    1999-01-01

    Human enteroviruses consist of more than 60 serotypes, reflecting a wide range of evolutionary divergence. They have been genetically classified into four clusters on the basis of sequence homology in the coding region of the single-stranded RNA genome. To explore further the genetic relationships between human enteroviruses and to characterize the evolutionary mechanisms responsible for variation, previously sequenced genomes were subjected to detailed comparison. Bootstrap and genetic similarity analyses were used to systematically scan the alignments of complete genomic sequences. Bootstrap analysis provided evidence from an early recombination event at the junction of the 5? noncoding and coding regions of the progenitors of the current clusters. Analysis within the genetic clusters indicated that enterovirus prototype strains include intraspecies recombinants. Recombination breakpoints were detected in all genomic regions except the capsid protein coding region. Our results suggest that recombination is a significant and relatively frequent mechanism in the evolution of enterovirus genomes. PMID:10482628

  19. Excessive proinflammatory cytokine and chemokine responses of human monocyte-derived macrophages to enterovirus 71 infection

    PubMed Central

    2012-01-01

    Background The levels of proinflammatory cytokine or chemokine in blood and cerebrospinal fluid are thought to be one of predictors for clinical severity of enterovirus 71 (EV71) infection, yet the cellular sources or signalling mechanism remain undefined. Here, we focused on the response of human primary monocyte-derived macrophages (MDMs) to EV71 virus and its possible mechanisms. Methods Human primary MDMs were infected by EV71 virus in vitro. Infectivity and viral replication were assayed, and cytokine responses were determined by Cytometric Bead Array(CBA) analysis. The relative changes of Toll-like receptors, retinoic acid-inducible gene I (RIG-I) and melamoma differentiation associated gene 5 (MDA5) mRNA expression were detected by real-time RT-PCR. Results Effective infection and viral replication were detected in EV71-infected MDMs. The titters of progeny virus released from EV71-infected MDMs gradually increased from 6-h to 48-h point of infection (POI.). Proinflammatory cytokines: IL-1, IL-6, TNF-? but not IFN-? and ? were induced in MDMs by EV71. EV71 infection significantly increased the release of IL-8, IP-10 and RANTES at 12-h or 24-h POI. Upregulation of TLR2, TLR7 and TLR8 mRNA expression rather than TLR3, TLR4, TLR6, TLR9, TLR10, RIG-I, MDA5 were found at different time points in EV71-infected MDMs. Conclusions Our findings suggested that macrophages are not only the important target cells but also the effectors during EV71 infection, and they may play an important role in the pathogenesis of EV71 infection. And the proinflammatory cytokine and chemokine responses in EV71-infected MDMs may be mediated by the activation of differential pattern of TLRs. PMID:22994237

  20. Genetic and phylogenetic clustering of enteroviruses

    Microsoft Academic Search

    T. Poyry; L. Kinnunen; T. Hyypia; B. Brown; C. Horsnell; T. Hovi; G. Stanway

    1996-01-01

    Genetic and phylogenetic analysis of enteroviruses showed that in the 5'NCR enteroviruses formed three clusters: polioviruses (PVs), coxsackievirus A type 21 (CAV21), CAV24 and enterovirus type 70 (ENV70) formed one cluster; coxsackievirus B isolates (CBVs), CAV9, CAV16, ENV71, echovirus type 11 (EV11), EV12 and all partially sequenced echoviruses and swine vesicular disease virus (SVDV) belonged to another cluster and bovine

  1. Antiviral Activity of Hederasaponin B from Hedera helix against Enterovirus 71 Subgenotypes C3 and C4a

    PubMed Central

    Song, JaeHyoung; Yeo, Sang-Gu; Hong, Eun-Hye; Lee, Bo-Ra; Kim, Jin-Won; Kim, JeongHoon; Jeong, HyeonGun; Kwon, YongSoo; Kim, HyunPyo; Lee, SangWon; Park, Jae-Hak; Ko, Hyun-Jeong

    2014-01-01

    Enterovirus 71 (EV71) is the predominant cause of hand, foot and mouth disease (HFMD). The antiviral activity of hederasaponin B from Hedera helix against EV71 subgenotypes C3 and C4a was evaluated in vero cells. In the current study, the antiviral activity of hederasaponin B against EV71 C3 and C4a was determined by cytopathic effect (CPE) reduction method and western blot assay. Our results demonstrated that hederasaponin B and 30% ethanol extract of Hedera helix containing hederasaponin B showed significant antiviral activity against EV71 subgenotypes C3 and C4a by reducing the formation of a visible CPE. Hederasaponin B also inhibited the viral VP2 protein expression, suggesting the inhibition of viral capsid protein synthesis.These results suggest that hederasaponin B and Hedera helix extract containing hederasaponin B can be novel drug candidates with broad-spectrum antiviral activity against various subgenotypes of EV71. PMID:24596620

  2. Characterization of Enteroviruses from Non-Human Primates in Cameroon Revealed Virus Types Widespread in Humans along with Candidate New Types and Species

    PubMed Central

    Sadeuh-Mba, Serge Alain; Bessaud, Maël; Joffret, Marie-Line; Endegue Zanga, Marie-Claire; Balanant, Jean; Mpoudi Ngole, Eitel; Njouom, Richard; Reynes, Jean-Marc; Delpeyroux, Francis; Rousset, Dominique

    2014-01-01

    Enteroviruses (EVs) infecting African Non-Human Primates (NHP) are still poorly documented. This study was designed to characterize the genetic diversity of EVs among captive and wild NHP in Cameroon and to compare this diversity with that found in humans. Stool specimens were collected in April 2008 in NHP housed in sanctuaries in Yaounde and neighborhoods. Moreover, stool specimens collected from wild NHP from June 2006 to October 2008 in the southern rain forest of Cameroon were considered. RNAs purified directly from stool samples were screened for EVs using a sensitive RT-nested PCR targeting the VP1 capsid coding gene whose nucleotide sequence was used for molecular typing. Captive chimpanzees (Pan troglodytes) and gorillas (Gorilla gorilla) were primarily infected by EV types already reported in humans in Cameroon and elsewhere: Coxsackievirus A13 and A24, Echovirus 15 and 29, and EV-B82. Moreover EV-A119, a novel virus type recently described in humans in central and west Africa, was also found in a captive Chimpanzee. EV-A76, which is a widespread virus in humans, was identified in wild chimpanzees, thus suggesting its adaptation and parallel circulation in human and NHP populations in Cameroon. Interestingly, some EVs harbored by wild NHP were genetically distinct from all existing types and were thus assigned as new types. One chimpanzee-derived virus was tentatively assigned as EV-J121 in the EV-J species. In addition, two EVs from wild monkeys provisionally registered as EV-122 and EV-123 were found to belong to a candidate new species. Overall, this study indicates that the genetic diversity of EVs among NHP is more important than previously known and could be the source of future new emerging human viral diseases. PMID:25079078

  3. Prevalence and Characterization of Enterovirus Infections among Pediatric Patients with Hand Foot Mouth Disease, Herpangina and Influenza Like Illness in Thailand, 2012

    PubMed Central

    Puenpa, Jiratchaya; Mauleekoonphairoj, John; Linsuwanon, Piyada; Suwannakarn, Kamol; Chieochansin, Thaweesak; Korkong, Sumeth; Theamboonlers, Apiradee; Poovorawan, Yong

    2014-01-01

    Hand, foot, and mouth disease (HFMD) and herpangina are common infectious diseases caused by several genotypes of human enterovirus species A and frequently occurring in young children. This study was aimed at analyzing enteroviruses from patients with these diseases in Thailand in 2012. Detection and genotype determination of enteroviruses were accomplished by reverse transcription-polymerase chain reaction and sequencing of the VP1 region. Enterovirus-positive samples were differentiated into 17 genotypes (coxsackievirus A4 (CAV4), A5, A6, A8, A9, A10, A12, A16, A21, B1, B2, B4, B5, echovirus 7, 16, 25 and Enterovirus 71). The result showed CAV6 (33.5%), followed by CAV16 (9.4%) and EV71 (8.8%) as the most frequent genotypes in HFMD, CAV8 (19.3%) in herpangina and CAV6 (1.5%) in influenza like illness. Enterovirus infections were most prevalent during July with 34.4% in HFMD, 39.8% in herpangina and 1.6% in ILI. The higher enterovirus infection associated with HFMD and herpangina occurred in infants over one year-old. This represents the first report describing the circulation of multiple enteroviruses in Thailand. PMID:24887237

  4. New PCR Test That Recognizes All Human Prototypes of Enterovirus: Application for Clinical Diagnosis

    PubMed Central

    Bourlet, Thomas; Caro, Valerie; Minjolle, Sophie; Jusselin, Isabelle; Pozzetto, Bruno; Crainic, Radu; Colimon, Ronald

    2003-01-01

    We describe a new PCR test (Penter RT-PCR) that recognizes all 64 prototypes of enterovirus. Sixty clinical samples were analyzed in parallel with this Penter RT-PCR and previously described PCR tests: 34 and 32 samples tested positive, respectively. This assay is suitable for use in clinical diagnosis, and its ability to amplify all known serotypes makes it more useful than other consensus PCR tests. PMID:12682177

  5. Chemical modification of the plant isoprenoid cytokinin N(6)-isopentenyladenosine yields a selective inhibitor of human enterovirus 71 replication.

    PubMed

    Tararov, Vitali I; Tijsma, Aloys; Kolyachkina, Svetlana V; Oslovsky, Vladimir E; Neyts, Johan; Drenichev, Mikhail S; Leyssen, Pieter; Mikhailov, Sergey N

    2015-01-27

    In this study, we demonstrate that N(6)-isopentenyladenosine, which essentially is a plant cytokinin-like compound, exerts a potent and selective antiviral effect on the replication of human enterovirus 71 with an EC50 of 1.0 ± 0.2 ?M and a selectivity index (SI) of 5.7. The synthesis of analogs with modification of the N(6)-position did not result in a lower EC50 value. However, in particular with the synthesis of N(6)-(5-hexene-2-yne-1-yl)adenosine (EC50 = 4.3 ± 1.5 ?M), the selectivity index was significantly increased: because of a reduction in the adverse effect of this compound on the host cells, an SI > 101 could be calculated. With this study, we for the first time provide proof that a compound class that is based on the plant cytokinin skeleton offers an interesting starting point for the development of novel antivirals against mammalian viruses, in the present context in particular against enterovirus 71. PMID:25461889

  6. Receptors for enterovirus 71

    PubMed Central

    Yamayoshi, Seiya; Fujii, Ken; Koike, Satoshi

    2014-01-01

    Enterovirus 71 (EV71) is one of the major causative agents of hand, foot and mouth disease (HFMD). Occasionally, EV71 infection is associated with severe neurological diseases, such as acute encephalitis, acute flaccid paralysis and cardiopulmonary failure. Several molecules act as cell surface receptors that stimulate EV71 infection, including scavenger receptor B2 (SCARB2), P-selectin glycoprotein ligand-1 (PSGL-1), sialylated glycan, heparan sulfate and annexin II (Anx2). SCARB2 plays critical roles in attachment, viral entry and uncoating, and it can facilitate efficient EV71 infection. The three-dimensional structures of the mature EV71 virion, procapsid and empty capsid, as well as the exofacial domain of SCARB2, have been elucidated. This structural information has greatly increased our understanding of the early steps of EV71 infection. Furthermore, SCARB2 plays essential roles in the development of EV71 neurological disease in vivo. Adult mice are not susceptible to infection by EV71, but transgenic mice that express human SCARB2 become susceptible to EV71 infection and develop similar neurological diseases to those found in humans. This mouse model facilitates the in vivo investigation of many issues related to EV71. PSGL-1, sialylated glycan, heparan sulfate and Anx2 are attachment receptors, which enhance viral infection by retaining the virus on the cell surface. These molecules also contribute to viral infection in vitro either by interacting with SCARB2 or independently of SCARB2. However, the cooperative effects of these receptors, and their contribution to EV71 pathogenicity in vivo, remain to be elucidated. PMID:26038749

  7. High-titred neutralizing antibodies to human enterovirus 71 preferentially bind to the N-terminal portion of the capsid protein VP1

    Microsoft Academic Search

    C.-S. Tan; M. J. Cardosa

    2007-01-01

    Summary  Human enterovirus 71 has emerged as an important pathogen of children in the Asia Pacific region, and it may be important\\u000a to consider the development of a vaccine against this virus. Human cord serum was used as a source of neutralizing antibodies\\u000a to determine whether the N- or C-terminal half of the VP1 capsid protein was more likely to harbour

  8. Epidemic 2014 Enterovirus D68 Cross-Reacts with Human Rhinovirus on a Respiratory Molecular Diagnostic Platform

    PubMed Central

    McAllister, Shane C.; Schleiss, Mark R.; Arbefeville, Sophie; Steiner, Marie E.; Hanson, Ryan S.; Pollock, Catherine; Ferrieri, Patricia

    2015-01-01

    Enterovirus D68 (EV-D68) is an emerging virus known to cause sporadic disease and occasional epidemics of severe lower respiratory tract infection. However, the true prevalence of infection with EV-D68 is unknown, due in part to the lack of a rapid and specific nucleic acid amplification test as well as the infrequency with which respiratory samples are analyzed by enterovirus surveillance programs. During the 2014 EV-D68 epidemic in the United States, we noted an increased frequency of “low-positive” results for human rhinovirus (HRV) detected in respiratory tract samples using the GenMark Diagnostics eSensor respiratory viral panel, a multiplex PCR assay able to detect 14 known respiratory viruses but not enteroviruses. We simultaneously noted markedly increased admissions to our Pediatric Intensive Care Unit for severe lower respiratory tract infections in patients both with and without a history of reactive airway disease. Accordingly, we hypothesized that these “low-positive” RVP results were due to EV-D68 rather than rhinovirus infection. Sequencing of the picornavirus 5’ untranslated region (5’-UTR) of 49 samples positive for HRV by the GenMark RVP revealed that 33 (67.3%) were in fact EV-D68. Notably, the mean intensity of the HRV RVP result was significantly lower in the sequence-identified EV-D68 samples (20.3 nA) compared to HRV (129.7 nA). Using a cut-off of 40 nA for the differentiation of EV-D68 from HRV resulted in 94% sensitivity and 88% specificity. The robust diagnostic characteristics of our data suggest that the cross-reactivity of EV-D68 and HRV on the GenMark Diagnostics eSensor RVP platform may be an important factor to consider in making accurate molecular diagnosis of EV-D68 at institutions utilizing this system or other molecular respiratory platforms that may also cross-react. PMID:25799541

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

    PubMed

    Brinkman, Nichole E; Haffler, Tyler D; Cashdollar, Jennifer L; Rhodes, Eric R

    2013-10-01

    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, making wastewater a source of a potentially diverse group of enteric viruses. In this study, two procedures were evaluated to concentrate noroviruses, adenoviruses and enteroviruses from primary effluent of wastewater. In the first procedure, indigenous enteroviruses, noroviruses and adenoviruses were concentrated using celite (diatomaceous earth) followed by centrifugation through a 30K MWCO filter and nucleic acid extraction. The second procedure used celite concentration followed by nucleic acid extraction only. Virus quantities were measured using qPCR. A second set of primary effluent samples were seeded with Coxsackievirus A7, Coxsackievirus B1, poliovirus 1 or enterovirus 70 before concentration and processed through both procedures for recovery evaluation of enterovirus species representatives. The pairing of the single step extraction procedure with the celite concentration process resulted in 47-98% recovery of examined viruses, while the celite concentration process plus additional centrifugal concentration before nucleic acid extraction showed reduced recovery (14-47%). The celite concentration process followed by a large volume nucleic acid extraction technique proved to be an effective procedure for recovering these important human pathogens from wastewater. PMID:23727118

  10. THERMAL AND WATER SOURCE EFFECTS UPON THE STABILITY OF ENTEROVIRUSES IN SURFACE FRESHWATERS

    EPA Science Inventory

    The long-term survival of three human enterovirus serotypes, coxsackievirus B3, echovirus 7, and poliovirus 1 was examined in samples of surface freshwater collected from five sites of physically different character. hese were an artificial lake created by damming a creek, a smal...

  11. The Association of Recombination Events in the Founding and Emergence of Subgenogroup Evolutionary Lineages of Human Enterovirus 71

    PubMed Central

    McWilliam Leitch, E. C.; Cabrerizo, M.; Cardosa, J.; Harvala, H.; Ivanova, O. E.; Koike, S.; Kroes, A. C. M.; Lukashev, A.; Perera, D.; Roivainen, M.; Susi, P.; Trallero, G.; Evans, D. J.

    2012-01-01

    Enterovirus 71 (EV71) is responsible for frequent large-scale outbreaks of hand, foot, and mouth disease worldwide and represent a major etiological agent of severe, sometimes fatal neurological disease. EV71 variants have been classified into three genogroups (GgA, GgB, and GgC), and the latter two are further subdivided into subgenogroups B1 to B5 and C1 to C5. To investigate the dual roles of recombination and evolution in the epidemiology and transmission of EV71 worldwide, we performed a large-scale genetic analysis of isolates (n = 308) collected from 19 countries worldwide over a 40-year period. A series of recombination events occurred over this period, which have been identified through incongruities in sequence grouping between the VP1 and 3Dpol regions. Eleven 3Dpol clades were identified, each specific to EV71 and associated with specific subgenogroups but interspersed phylogenetically with clades of coxsackievirus A16 and other EV species A serotypes. The likelihood of recombination increased with VP1 sequence divergence; mean half-lives for EV71 recombinant forms (RFs) of 6 and 9 years for GgB and GgC overlapped with those observed for the EV-B serotypes, echovirus 9 (E9), E30, and E11, respectively (1.3 to 9.8 years). Furthermore, within genogroups, sporadic recombination events occurred, such as the linkage of two B4 variants to RF-W instead of RF-A and of two C4 variants to RF-H. Intriguingly, recombination events occurred as a founding event of most subgenogroups immediately preceding their lineage expansion and global emergence. The possibility that recombination contributed to their subsequent spread through improved fitness requires further biological and immunological characterization. PMID:22205739

  12. Occurrence of enteroviruses in community swimming pools.

    PubMed

    Keswick, B H; Gerba, C P; Goyal, S M

    1981-09-01

    Municipal swimming pools and wading pools were examined for the presence of human enteric viruses using a portable virus concentrator at the site to concentrate viruses from 100-gallon to 500-gallon samples. Ten of 14 samples contained viruses; three of these were positive for virus in the presence of residual free chlorine. Enteroviruses were isolated from two pools which exceeded the 0.4 ppm free residual chlorine standard. This study appears to be supportive of recent evidence that indicates a higher incidence of enterovirus infection among bathers. All seven wading pool samples contained virus. Coxsackieviruses B3 and B4, poliovirus 1, and echovirus 7 were isolated. Total coliform bacteria were not adequate indicators of the presence of virus, as six of the samples were positive for virus but negative for coliforms. Total plate counts appeared to provide a better indication of the sanitary quality of the pool water, but viruses could still be detected in samples that met currently recommended bacterial levels. It is possible that swimming and wading pools may serve as a means of transmission of enteroviral disease, especially in children, during summer months. PMID:6267950

  13. Enterovirus D68

    MedlinePLUS

    ... Related Links Water-related Hygiene Viral Conjunctivitis Hand, Foot, and Mouth Disease Viral Meningitis What is ... D68 Enterovirus D68 (EV-D68) is one of more than 100 non-polio enteroviruses. This virus was first identified in California in 1962. What ...

  14. Anti-enterovirus activity and structure–activity relationship of a series of 2,6-dihalophenyl-substituted 1 H,3 H-thiazolo[3,4- a]benzimidazoles

    Microsoft Academic Search

    Armando M. De Palma; Ward Heggermont; Pieter Leyssen; Gerhard Pürstinger; Eva Wimmer; Erik De Clercq; Angela Rao; Anna-Maria Monforte; Alba Chimirri; Johan Neyts

    2007-01-01

    Despite the fact that enteroviruses are implicated in a variety of human diseases, there is no approved therapy for the treatment of enteroviral infections. Here, a series of 2,6-dihalophenyl-substituted 1H,3H-thiazolo[3,4-a]benzimidazoles with anti-enterovirus activity is reported. The compounds elicit potent activity against coxsackievirus A9, echovirus 9 and 11 and all six strains of coxsackievirus B. A structure–activity relationship analysis revealed that

  15. Enteroviruses, hygiene and type 1 diabetes: toward a preventive vaccine.

    PubMed

    Drescher, Kristen M; von Herrath, Matthias; Tracy, Steven

    2015-01-01

    Enteroviruses and humans have long co-existed. Although recognized in ancient times, poliomyelitis and type 1 diabetes (T1D) were exceptionally rare and not epidemic, due in large part to poor sanitation and personal hygiene which resulted in repeated exposure to fecal-oral transmitted viruses and other infectious agents and viruses and the generation of a broad protective immunity. As a function of a growing acceptance of the benefits of hygienic practices and microbiologically clean(er) water supplies, the likelihood of exposure to diverse infectious agents and viruses declined. The effort to vaccinate against poliomyelitis demonstrated that enteroviral diseases are preventable by vaccination and led to understanding how to successfully attenuate enteroviruses. Type 1 diabetes onset has been convincingly linked to infection by numerous enteroviruses including the group B coxsackieviruses (CVB), while studies of CVB infections in NOD mice have demonstrated not only a clear link between disease onset but an ability to reduce the incidence of T1D as well: CVB infections can suppress naturally occurring autoimmune T1D. We propose here that if we can harness and develop the capacity to use attenuated enteroviral strains to induce regulatory T cell populations in the host through vaccination, then a vaccine could be considered that should function to protect against both autoimmune as well as virus-triggered T1D. Such a vaccine would not only specifically protect from certain enterovirus types but more importantly, also reset the organism's regulatory rheostat making the further development of pathogenic autoimmunity less likely. PMID:25430610

  16. Detection of enterovirus specific IgG and IgM antibodies in humans by an indirect solid phase radioimmunoassay

    Microsoft Academic Search

    R. Dörries; V. Meulen

    1980-01-01

    The development of a solid phase radioimmunoassay which is able to detect virus-specific IgG and IgM antibodies in serum specimens from patients with enterovirus infections is described. Viral antigen partially purified from infected tissue culture fluid was adsorbed passively to individual polystyrene microtiter wells. Dilutions of sera were incubated on these antigens and bound anti-viral antibodies were monitored by the

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

    Microsoft Academic Search

    Christina Baboonian; Tom Treasure

    1997-01-01

    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

  18. Real-time monitoring of human enterovirus (HEV)-infected cells and anti-HEV 3C protease potency by fluorescence resonance energy transfer.

    PubMed

    Tsai, Meng-Tian; Cheng, Yun-Hsiang; Liu, Yu-Ning; Liao, Nien-Chien; Lu, Wen-Wen; Kung, Szu-Hao

    2009-02-01

    A real-time assay system that allows monitoring of intracellular human enterovirus (HEV) protease activity was established using the principle of fluorescence resonance energy transfer (FRET). It was accomplished by engineering cells to constitutively express a genetically encoded FRET probe. The FRET-based probe was designed to contain an enterovirus 71 3C protease (3C(pro)) cleavage motif flanked by the FRET pair composed of green fluorescent protein 2 and red fluorescent protein 2 (DsRed2). Efficient FRET from the stable line was detected in a real-time manner by fluorescence microscopy, and the disruption of FRET was readily monitored upon HEV infection. The level of the repressed FRET was proportional to the input virus titer and the infection duration as measured by the fluorometric method. The FRET biosensor cell line was also responsive to other related HEV serotypes, but not to the phylogenetically distant herpes simplex virus, which was confirmed by Western blot analysis. The FRET biosensor was then utilized to develop a format for the determination of antiviral susceptibility, as the reduced FRET appeared to reflect viral replication. Evaluations of the FRET biosensor system with representative HEV serotypes demonstrated that their susceptibilities to a 3C(pro) inhibitor, rupintrivir, were all accurately determined. In summary, this novel FRET-based system is a means for rapid detection, quantification, and drug susceptibility testing for HEVs, with potential for the development of a high-throughput screening assay. PMID:19015331

  19. Real-Time Monitoring of Human Enterovirus (HEV)-Infected Cells and Anti-HEV 3C Protease Potency by Fluorescence Resonance Energy Transfer? †

    PubMed Central

    Tsai, Meng-Tian; Cheng, Yun-Hsiang; Liu, Yu-Ning; Liao, Nien-Chien; Lu, Wen-Wen; Kung, Szu-Hao

    2009-01-01

    A real-time assay system that allows monitoring of intracellular human enterovirus (HEV) protease activity was established using the principle of fluorescence resonance energy transfer (FRET). It was accomplished by engineering cells to constitutively express a genetically encoded FRET probe. The FRET-based probe was designed to contain an enterovirus 71 3C protease (3Cpro) cleavage motif flanked by the FRET pair composed of green fluorescent protein 2 and red fluorescent protein 2 (DsRed2). Efficient FRET from the stable line was detected in a real-time manner by fluorescence microscopy, and the disruption of FRET was readily monitored upon HEV infection. The level of the repressed FRET was proportional to the input virus titer and the infection duration as measured by the fluorometric method. The FRET biosensor cell line was also responsive to other related HEV serotypes, but not to the phylogenetically distant herpes simplex virus, which was confirmed by Western blot analysis. The FRET biosensor was then utilized to develop a format for the determination of antiviral susceptibility, as the reduced FRET appeared to reflect viral replication. Evaluations of the FRET biosensor system with representative HEV serotypes demonstrated that their susceptibilities to a 3Cpro inhibitor, rupintrivir, were all accurately determined. In summary, this novel FRET-based system is a means for rapid detection, quantification, and drug susceptibility testing for HEVs, with potential for the development of a high-throughput screening assay. PMID:19015331

  20. A serological classification of bovine enteroviruses

    Microsoft Academic Search

    N. J. Knowles; I. T. R. Barnett

    1985-01-01

    Summary Cross virus neutralization (VN), complement fixation (CF) and immunoprecipitation (IP) tests were employed to compare the seven currently recognized bovine enterovirus (BEV) serotypes with seven serologically distinct strains previously isolated in Great Britain and two other BEV from the United Kingdom. Based on criteria used to differentiate other human and animal picornavirus serotypes, it was discovered that BEV types

  1. Molecular Typing of Enteroviruses: Current Status and Future Requirements

    PubMed Central

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

    1998-01-01

    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. PMID:9457433

  2. Epidemiology and clinical characteristics of infants with human parechovirus or human herpes virus-6 detected in cerebrospinal fluid tested for enterovirus or herpes simplex virus.

    PubMed

    Messacar, Kevin; Breazeale, Garrett; Wei, Qi; Robinson, Christine C; Dominguez, Samuel R

    2015-05-01

    Human parechovirus (HPeV) and human herpes virus-6 (HHV-6) are acquired commonly in infancy and associated with central nervous system infection. The prevalence of HPeV and HHV-6 in the cerebrospinal fluid (CSF) of infants tested for enterovirus (EV) and herpes-simplex virus (HSV) is unknown. All stored CSF samples from EV or HSV testing in infants less than 6 months of age at Children's Hospital Colorado between January 1, 2010 and December 31, 2011 were tested for HPeV, HHV-6, EV, and HSV by PCR. Clinical characteristics and epidemiological data were collected using retrospective electronic chart review. Of 239 infants tested, 29 cases of EV (12.1%), 7 cases of HPeV (2.9%), 5 cases of HHV-6 (2.1%), and 5 cases of HSV (2.1%) were identified with no bacterial co-infections. HPeV cases occurred between July and October in infants with median age of 24 days. Infants with HPeV had a median maximum temperature of 39?°C, median fever duration of 3 days and median peripheral white blood cell count of 5.2?×?10(3)/?L. HHV-6 cases occurred in infants with median age of 61 days without seasonality. Five percent of infants less than 6 months of age undergoing testing for EV or HSV have HPeV or HHV-6 in the CSF. Targeting testing of HPeV towards febrile infants less than 2 months of age with leukopenia in the late summer to early fall, and HHV-6 towards older infants may increase diagnostic yield. The clinical and fiscal impact of testing infants for HPeV and HHV-6 needs to be determined. PMID:25650069

  3. Genetic and phylogenetic clustering of enteroviruses.

    PubMed

    Pöyry, T; Kinnunen, L; Hyypiä, T; Brown, B; Horsnell, C; Hovi, T; Stanway, G

    1996-08-01

    Genetic and phylogenetic analysis of enteroviruses showed that in the 5'NCR enteroviruses formed three clusters: polioviruses (PVs), coxsackievirus A type 21 (CAV21), CAV24 and enterovirus type 70 (ENV70) formed one cluster; coxsackievirus B isolates (CBVs), CAV9, CAV16, ENV71, echovirus type 11 (EV11), EV12 and all partially sequenced echoviruses and swine vesicular disease virus (SVDV) belonged to another cluster and bovine enteroviruses (BEVs) formed the third cluster. In the capsid coding region five clusters were seen: PVs, CAV21 and CAV24 formed one cluster (PV-like); ENV70 formed a cluster of its own; all CBVs, CAV9, EV11, EV12 and SVDV formed the third cluster (CBV-like); CAV16, CAV2 and ENV71 belonged to the fourth cluster (CAV16-like) and BEVs formed their own cluster (BEV-like). In the 3'NCR the same clusters were seen as in the coding region suggesting a close association of the 3'NCR with viral proteins while the cellular environment may be more important in the evolution of the 5'NCR. Secondary structures were predicted in the 3'NCR, which showed two different patterns among the five clusters. A potential pseudoknot region common in all five clusters was identified. Although the BEV-like viruses formed a separate cluster in all genomic regions, in the coding region they seem to be phylogenetically related to the CAV16-like viruses. PMID:8760417

  4. Detection of human adenovirus, rotavirus and enterovirus in water samples collected on dairy farms from Tenente Portela, Northwest of Rio Grande do Sul, Brazil

    PubMed Central

    Spilki, Fernando Rosado; da Luz, Roger Bordin; Fabres, Rafael Bandeira; Soliman, Mayra Cristina; Kluge, Mariana; Fleck, Juliane Deise; Rodrigues, Manoela Tressoldi; Comerlato, Juliana; Cenci, Alexander; Cerva, Cristine; Dasso, Maurício Gautério; Roehe, Paulo Michel

    2013-01-01

    Viral gastroenteritis and other waterborne diseases are a major concern for health in Brazil. A number of studies were conducted about the presence of viruses on water samples from Brazilian areas. However, the knowledge about the occurrence of viral contamination of drinking water sources in rural settings of the country is insufficient. On the present work, 15 samples from 5 dairy farms located at the municipality of Tenente Portela were collected and analysed for the presence of human adenoviruses (HAdV), as well as human enteroviruses (EV) and rotaviruses (RV). HAdV was present on 66.66% of the water samples, and have been found in all samples from artesian wells and springs, which are used as sources of drinking water for the individuals inhabiting those farms. EV and RV found only in one sample each. The detection rates of HAdV on the water from these dairy farms are alarming and point towards a situation of elevated environmental contamination by fecal microorganisms of human origin and poor basic sanitation conditions. PMID:24516464

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

    Microsoft Academic Search

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

    1998-01-01

    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

  6. MOLECULAR EPIDEMIOLOGY OF ANIMAL ENTEROVIRUSES: IMPLICATIONS FOR THEIR USE AS MARKERS FOR ENVIRONMENTAL FECAL CONTAMINATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enteroviruses are the most common viruses, infecting a wide variety of mammals. It is generally accepted that only a small number of entero viruses are known. Every year several new isolates are identified and named. Human enteroviruses have been found as environmental contaminants, and contaminatio...

  7. [Research methods in protein-protein and protein-nucleic acid interactions and application in the study of human enterovirus A71].

    PubMed

    Zhang, Zhi-Xiao; Zheng, Ying; Hu, Ting-Song; Li, Qi-Han; Fan, Quan-Shui

    2014-09-01

    Methods for analyses of protein-protein interactions include: yeast two hybrid (Y2H), phage dis- play (PD), co-immunoprecipitation (Co-IP), glutathione S-transferase pull-down (GST pull-down), cellular co-localization, far-western blotting, virus overlay protein binding assay (VOPBA), surface plasmon resonance (SPR), and fluorescence resonance energy transfer (FRET). Technologies for the detection of protein-nucleic acid interactions include: yeast one hybrid (Y1H), chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA), Southwestern blotting, reporter gene, Co-IP, GST pull-down, and PD. These methods are often used in the study of the human enterovirus A71 (EV-A71) by our research team. Reviews in the Chinese literature in this field are lacking, so we reviewed applications of these methods in the study of EV-A71. This review may impart important knowledge in the research of other viruses with regard to protein-protein and protein-nucleic acid interactions. PMID:25562971

  8. Identification of a Common Epitope between Enterovirus 71 and Human MED25 Proteins Which May Explain Virus-Associated Neurological Disease

    PubMed Central

    Fan, Peihu; Li, Xiaojun; Sun, Shiyang; Su, Weiheng; An, Dong; Gao, Feng; Kong, Wei; Jiang, Chunlai

    2015-01-01

    Enterovirus 71 (EV71) is a major causative pathogen of hand, foot and mouth disease with especially severe neurologic complications, which mainly account for fatalities from this disease. To date, the pathogenesis of EV71 in the central neurons system has remained unclear. Cytokine-mediated immunopathogenesis and nervous tissue damage by virus proliferation are two widely speculated causes of the neurological disease. To further study the pathogenesis, we identified a common epitope (co-epitope) between EV71 VP1 and human mediator complex subunit 25 (MED25) highly expressed in brain stem. A monoclonal antibody (2H2) against the co-epitope was prepared, and its interaction with MED25 was examined by ELISA, immunofluorescence assay and Western blot in vitro and by live small animal imaging in vivo. Additionally, 2H2 could bind to both VP1 and MED25 with the affinity constant (Kd) of 10?7 M as determined by the ForteBio Octet System. Intravenously injected 2H2 was distributed in brain stem of mice after seven days of EV71 infection. Interestingly, 2H2-like antibodies were detected in the serum of EV71-infected patients. These findings suggest that EV71 infection induces the production of antibodies that can bind to autoantigens expressed in nervous tissue and maybe further trigger autoimmune reactions resulting in neurological disease. PMID:25826188

  9. Identification of a common epitope between enterovirus 71 and human MED25 proteins which may explain virus-associated neurological disease.

    PubMed

    Fan, Peihu; Li, Xiaojun; Sun, Shiyang; Su, Weiheng; An, Dong; Gao, Feng; Kong, Wei; Jiang, Chunlai

    2015-04-01

    Enterovirus 71 (EV71) is a major causative pathogen of hand, foot and mouth disease with especially severe neurologic complications, which mainly account for fatalities from this disease. To date, the pathogenesis of EV71 in the central neurons system has remained unclear. Cytokine-mediated immunopathogenesis and nervous tissue damage by virus proliferation are two widely speculated causes of the neurological disease. To further study the pathogenesis, we identified a common epitope (co-epitope) between EV71 VP1 and human mediator complex subunit 25 (MED25) highly expressed in brain stem. A monoclonal antibody (2H2) against the co-epitope was prepared, and its interaction with MED25 was examined by ELISA, immunofluorescence assay and Western blot in vitro and by live small animal imaging in vivo. Additionally, 2H2 could bind to both VP1 and MED25 with the affinity constant (Kd) of 10-7 M as determined by the ForteBio Octet System. Intravenously injected 2H2 was distributed in brain stem of mice after seven days of EV71 infection. Interestingly, 2H2-like antibodies were detected in the serum of EV71-infected patients. These findings suggest that EV71 infection induces the production of antibodies that can bind to autoantigens expressed in nervous tissue and maybe further trigger autoimmune reactions resulting in neurological disease. PMID:25826188

  10. Assessment of the risks for human health of adenoviruses, hepatitis A virus, rotaviruses and enteroviruses in the Buffalo River and three source water dams in the Eastern Cape.

    PubMed

    Chigor, Vincent N; Sibanda, Timothy; Okoh, Anthony I

    2014-06-01

    Buffalo River is an important water resource in the Eastern Cape Province of South Africa. The potential risks of infection constituted by exposure to human enteric viruses in the Buffalo River and three source water dams along its course were assessed using mean values and static quantitative microbial risk assessment (QMRA). The daily risks of infection determined by the exponential model [for human adenovirus (HAdV) and enterovirus (EnV)] and the beta-Poisson model (for hepatitis A virus (HAV) and rotavirus (RoV)) varied with sites and exposure scenario. The estimated daily risks of infection values at the sites where the respective viruses were detected, ranged from 7.31 × 10(-3) to 1 (for HAdV), 4.23 × 10(-2) to 6.54 × 10(-1) (RoV), 2.32 × 10(-4) to 1.73 × 10(-1) (HAV) and 1.32 × 10(-4) to 5.70 × 10(-2) (EnV). The yearly risks of infection in individuals exposed to the river/dam water via drinking, recreational, domestic or irrigational activities were unacceptably high, exceeding the acceptable risk of 0.01% (10(-4) infection/person/year), and the guideline value used as by several nations for drinking water. The risks of illness and death from infection ranged from 6.58 × 10(-5) to 5.0 × 10(-1) and 6.58 × 10(-9) to 5.0 × 10(-5), respectively. The threats here are heightened by the high mortality rates for HAV, and its endemicity in South Africa. Therefore, we conclude that the Buffalo River and its source water dams are a public health hazard. The QMRA presented here is the first of its kinds in the Eastern Cape Province and provides the building block for a quantitatively oriented local guideline for water quality management in the Province. PMID:24676673

  11. Binding of Glutathione to Enterovirus Capsids Is Essential for Virion Morphogenesis

    PubMed Central

    Thibaut, Hendrik Jan; Thys, Bert; Canela, María-Dolores; Aguado, Leire; Wimmer, Eckard; Paul, Aniko; Pérez-Pérez, María-Jesús; van Kuppeveld, Frank J. M.; Neyts, Johan

    2014-01-01

    Enteroviruses (family of the Picornaviridae) cover a large group of medically important human pathogens for which no antiviral treatment is approved. Although these viruses have been extensively studied, some aspects of the viral life cycle, in particular morphogenesis, are yet poorly understood. We report the discovery of TP219 as a novel inhibitor of the replication of several enteroviruses, including coxsackievirus and poliovirus. We show that TP219 binds directly glutathione (GSH), thereby rapidly depleting intracellular GSH levels and that this interferes with virus morphogenesis without affecting viral RNA replication. The inhibitory effect on assembly was shown not to depend on an altered reducing environment. Using TP219, we show that GSH is an essential stabilizing cofactor during the transition of protomeric particles into pentameric particles. Sequential passaging of coxsackievirus B3 in the presence of low GSH-levels selected for GSH-independent mutants that harbored a surface-exposed methionine in VP1 at the interface between two protomers. In line with this observation, enteroviruses that already contained this surface-exposed methionine, such as EV71, did not rely on GSH for virus morphogenesis. Biochemical and microscopical analysis provided strong evidence for a direct interaction between GSH and wildtype VP1 and a role for this interaction in localizing assembly intermediates to replication sites. Consistently, the interaction between GSH and mutant VP1 was abolished resulting in a relocalization of the assembly intermediates to replication sites independent from GSH. This study thus reveals GSH as a novel stabilizing host factor essential for the production of infectious enterovirus progeny and provides new insights into the poorly understood process of morphogenesis. PMID:24722756

  12. Enterovirus and type 1 diabetes: What is the matter?

    PubMed Central

    Bergamin, Carla Sanchez; Dib, Sergio Atala

    2015-01-01

    A complex interaction of genetic and environmental factors can trigger the immune-mediated mechanism responsible for type 1 diabetes mellitus (T1DM) establishment. Environmental factors may initiate and possibly sustain, accelerate, or retard damage to ?-cells. The role of environmental factors in this process has been exhaustive studied and viruses are among the most probable ones, especially enteroviruses. Improvements in enterovirus detection methods and randomized studies with patient follow-up have confirmed the importance of human enterovirus in the pathogenesis of T1DM. The genetic risk of T1DM and particular innate and acquired immune responses to enterovirus infection contribute to a tolerance to T1DM-related autoantigens. However, the frequency, mechanisms, and pathways of virally induced autoimmunity and ?-cell destruction in T1DM remain to be determined. It is difficult to investigate the role of enterovirus infection in T1DM because of several concomitant mechanisms by which the virus damages pancreatic ?-cells, which, consequently, may lead to T1DM establishment. Advances in molecular and genomic studies may facilitate the identification of pathways at earlier stages of autoimmunity when preventive and therapeutic approaches may be more effective. PMID:26131324

  13. Development of a Transcription-Reverse Transcription Concerted Reaction Method for Specific Detection of Human Enterovirus 71 from Clinical Specimens

    PubMed Central

    Nakajima, Naoto; Kitamori, Yuka; Ohnaka, Satoru; Mitoma, Yasutami; Mizuta, Katsumi; Wakita, Takaji; Shimizu, Hiroyuki

    2012-01-01

    A transcription-reverse transcription (RT) concerted reaction (TRCR) method was developed for rapid and specific detection of EV71 from clinical specimens. This method was validated with EV71 strains from all of the known genotypes (genotypes A, B1 to B5, and C1 to C5), with detection limits of 10 to 103 copies, and was useful for identification of EV71 from throat swabs of patients with hand, foot, and mouth disease (HFMD). PMID:22357494

  14. Real-Time Monitoring of Human Enterovirus (HEV)Infected Cells and Anti-HEV 3C Protease Potency by Fluorescence Resonance Energy Transfer

    Microsoft Academic Search

    Meng-Tian Tsai; Yun-Hsiang Cheng; Yu-Ning Liu; Nien-Chien Liao; Wen-Wen Lu; Szu-Hao Kung

    2009-01-01

    designed to contain an enterovirus 71 3C protease (3Cpro) cleavage motif flanked by the FRET pair composed of green fluorescent protein 2 and red fluorescent protein 2 (DsRed2). Efficient FRET from the stable line was detected in a real-time manner by fluorescence microscopy, and the disruption of FRET was readily monitored upon HEV infection. The level of the repressed FRET

  15. Identification of a Series of Compounds with Potent Antiviral Activity for the Treatment of Enterovirus Infections

    PubMed Central

    2013-01-01

    Rhinovirus (genus enterovirus) infections are responsible for many of the severe exacerbations of asthma and chronic obstructive pulmonary disease. Other members of the genus can cause life-threatening acute neurological infections. There is currently no antiviral drug approved for the treatment of such infections. We have identified a series of potent, broad-spectrum antiviral compounds that inhibit the replication of the human rhinovirus, Coxsackie virus, poliovirus, and enterovirus-71. The mechanism of action of the compounds has been established as inhibition of a lipid kinase, PI4KIII?. Inhibition of hepatitis C replication in a replicon assay correlated with enterovirus inhibition. PMID:24900715

  16. Identification of a series of compounds with potent antiviral activity for the treatment of enterovirus infections.

    PubMed

    MacLeod, Angus M; Mitchell, Dale R; Palmer, Nicholas J; Van de Poël, Hervé; Conrath, Katja; Andrews, Martin; Leyssen, Pieter; Neyts, Johan

    2013-07-11

    Rhinovirus (genus enterovirus) infections are responsible for many of the severe exacerbations of asthma and chronic obstructive pulmonary disease. Other members of the genus can cause life-threatening acute neurological infections. There is currently no antiviral drug approved for the treatment of such infections. We have identified a series of potent, broad-spectrum antiviral compounds that inhibit the replication of the human rhinovirus, Coxsackie virus, poliovirus, and enterovirus-71. The mechanism of action of the compounds has been established as inhibition of a lipid kinase, PI4KIII?. Inhibition of hepatitis C replication in a replicon assay correlated with enterovirus inhibition. PMID:24900715

  17. The role of Misshapen NCK-related kinase (MINK), a novel Ste20 family kinase, in the IRES-mediated protein translation of human enterovirus 71.

    PubMed

    Leong, Shi Yun; Ong, Bryan Kit Teck; Chu, Justin Jang Hann

    2015-03-01

    Human Enterovirus 71 (EV71) commonly causes Hand, Foot and Mouth Disease in young children, and occasional occurrences of neurological complications can be fatal. In this study, a high-throughput cell-based screening on the serine/threonine kinase siRNA library was performed to identify potential antiviral agents against EV71 replication. Among the hits, Misshapen/NIKs-related kinase (MINK) was selected for detailed analysis due to its strong inhibitory profile and novelty. In the investigation of the stage at which MINK is involved in EV71 replication, virus RNA transfection in MINK siRNA-treated cells continued to cause virus inhibition despite bypassing the normal entry pathway, suggesting its involvement at the post-entry stage. We have also shown that viral RNA and protein expression level was significantly reduced upon MINK silencing, suggesting its involvement in viral protein synthesis which feeds into viral RNA replication process. Through proteomic analysis and infection inhibition assay, we found that the activation of MINK was triggered by early replication events, instead of the binding and entry of the virus. Proteomic analysis on the activation profile of p38 Mitogen-activated Protein Kinase (MAPK) indicated that the phosphorylation of p38 MAPK was stimulated by EV71 infection upon MINK activation. Luciferase reporter assay further revealed that the translation efficiency of the EV71 internal ribosomal entry site (IRES) was reduced after blocking the MINK/p38 MAPK pathway. Further investigation on the effect of MINK silencing on heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) localisation demonstrated that cytoplasmic relocalisation of hnRNP A1 upon EV71 infection may be facilitated via the MINK/p38 MAPK pathway which then positively regulates the translation of viral RNA transcripts. These novel findings hence suggest that MINK plays a functional role in the IRES-mediated translation of EV71 viral RNA and may provide a potential target for the development of specific antiviral strategies against EV71 infection. PMID:25747578

  18. The Role of Misshapen NCK-related kinase (MINK), a Novel Ste20 Family Kinase, in the IRES-Mediated Protein Translation of Human Enterovirus 71

    PubMed Central

    Leong, Shi Yun; Ong, Bryan Kit Teck; Chu, Justin Jang Hann

    2015-01-01

    Human Enterovirus 71 (EV71) commonly causes Hand, Foot and Mouth Disease in young children, and occasional occurrences of neurological complications can be fatal. In this study, a high-throughput cell-based screening on the serine/threonine kinase siRNA library was performed to identify potential antiviral agents against EV71 replication. Among the hits, Misshapen/NIKs-related kinase (MINK) was selected for detailed analysis due to its strong inhibitory profile and novelty. In the investigation of the stage at which MINK is involved in EV71 replication, virus RNA transfection in MINK siRNA-treated cells continued to cause virus inhibition despite bypassing the normal entry pathway, suggesting its involvement at the post-entry stage. We have also shown that viral RNA and protein expression level was significantly reduced upon MINK silencing, suggesting its involvement in viral protein synthesis which feeds into viral RNA replication process. Through proteomic analysis and infection inhibition assay, we found that the activation of MINK was triggered by early replication events, instead of the binding and entry of the virus. Proteomic analysis on the activation profile of p38 Mitogen-activated Protein Kinase (MAPK) indicated that the phosphorylation of p38 MAPK was stimulated by EV71 infection upon MINK activation. Luciferase reporter assay further revealed that the translation efficiency of the EV71 internal ribosomal entry site (IRES) was reduced after blocking the MINK/p38 MAPK pathway. Further investigation on the effect of MINK silencing on heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) localisation demonstrated that cytoplasmic relocalisation of hnRNP A1 upon EV71 infection may be facilitated via the MINK/p38 MAPK pathway which then positively regulates the translation of viral RNA transcripts. These novel findings hence suggest that MINK plays a functional role in the IRES-mediated translation of EV71 viral RNA and may provide a potential target for the development of specific antiviral strategies against EV71 infection. PMID:25747578

  19. Proteomic Analysis of Human Brain Microvascular Endothelial Cells Reveals Differential Protein Expression in Response to Enterovirus 71 Infection

    PubMed Central

    Luo, Wenying; Zhong, Jiayu; Zhao, Wei; Liu, Jianjun; Zhang, Renli; Peng, Liang; Hong, Wenxu; Huang, Sheng He; Cao, Hong

    2015-01-01

    2D DIGE technology was employed on proteins prepared from human brain microvascular endothelial cells (HBMEC), to study the differentially expressed proteins in cells at 0?h, 1?h, 16?h, and 24?h after infection. Proteins found to be differentially expressed were identified with matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDITOF/TOF MS) analysis. We identified 43 spots showing changes of at least 2.5 fold up- or downregulated expressions in EV71-infected cells at different time when comparing to control, and 28 proteins could be successfully identified by MALDI TOF/TOF mass spectrometry analysis. 4 proteins were significantly upregulated, and 6 proteins were downregulated, another 18 proteins were different expression at different incubation time. We identified changes in the expression of 12 cellular metabolism-related proteins, 5 molecules involved in cytoskeleton, 3 molecules involved in energy metabolism, 2 molecules involved in signal transduction, 1 molecule involved in the ubiquitin-proteasome pathway, 1 molecule involved in cell cycle, 1 molecule involved in apoptosis-related protein, 1 molecular chaperone, and 2 unknown proteins. These findings build up a comprehensive profile of the HBMEC proteome and provide a useful basis for further analysis of the pathogenic mechanism that underlies EV71 infections to induce severe neural complications. PMID:25821824

  20. COPI Is Required for Enterovirus 71 Replication

    PubMed Central

    Wang, Jianmin; Wu, Zhiqiang; Jin, Qi

    2012-01-01

    Enterovirus 71 (EV71), a member of the Picornaviridae family, is found in Asian countries where it causes a wide range of human diseases. No effective therapy is available for the treatment of these infections. Picornaviruses undergo RNA replication in association with membranes of infected cells. COPI and COPII have been shown to be involved in the formation of picornavirus-induced vesicles. Replication of several picornaviruses, including poliovirus and Echovirus 11 (EV11), is dependent on COPI or COPII. Here, we report that COPI, but not COPII, is required for EV71 replication. Replication of EV71 was inhibited by brefeldin A and golgicide A, inhibitors of COPI activity. Furthermore, we found EV71 2C protein interacted with COPI subunits by co-immunoprecipitation and GST pull-down assay, indicating that COPI coatomer might be directed to the viral replication complex through viral 2C protein. Additionally, because the pathway is conserved among different species of enteroviruses, it may represent a novel target for antiviral therapies. PMID:22662263

  1. Reemergence of Enterovirus 71 Epidemic in Northern Taiwan, 2012

    PubMed Central

    Chung, Wan-Yu; Chia, Min-Yuan; Tsao, Kuo-Chien; Wang, Ying-Hsiang; Lin, Tzou-Yien; Lee, Min-Shi

    2015-01-01

    Background Enterovirus 71 (EV71) belongs to picornavirus family and could be classified phylogenetically into three major genogroups (A, B and C) including 11 genotypes (A, B1-B5 and C1-C5). Since 1997, EV71 has caused large-scale of epidemics with neurological complications in Asian children. In Taiwan, nationwide EV71 epidemics with different predominant genotypes have occurred cyclically since 1998. A nationwide EV71 epidemic occurred again in 2012. We conducted genetic and antigenic characterizations of the 2012 epidemic. Methods Chang Gung Memorial Hospital (CGMH) is a medical center in northern Taiwan. In CGMH, specimens were collected from pediatric inpatients with suspected enterovirus infections for virus isolation. Enterovirus isolates were serotyped and genotyped and sera from EV71 inpatients were collected for measuring neutralizing antibody titers. Results There were 10, 16 and 99 EV71 inpatients identified in 2010, 2011 and 2012, respectively. There were 82 EV71 isolates genotyped, which identified 17 genotype C4a viruses and 65 genotype B5 viruses. The genotype B5 viruses were not detected until November 2011 and caused epidemics in 2012. Interestingly, the B5-2011 viruses were genetically distinguishable from the B5 viruses causing the 2008 epidemic and are likely introduced from China or Southeastern Asia. Based on antigenic analysis, minor antigenic variations were detected among the B5-2008, B5-2011, C4a-2008 and C4a-2012 viruses but these viruses antigenically differed from genotype A. Conclusions Genotype B5 and C4a viruses antigenically differ from genotype A viruses which have disappeared globally for 30 years but have been detected in China since 2008. Enterovirus surveillance should monitor genetic and antigenic variations of EV71. PMID:25774888

  2. Coxsackie B viruses that use human DAF as a receptor infect pig cells via pig CAR and do not use pig DAF

    Microsoft Academic Search

    O. Brad Spiller; Ian G. Goodfellow; David J. Evans; Stewart J. Hinchliffe; B. Paul Morgan

    Coxsackie B viruses (CVB) are enteroviruses belonging to the family Picornaviridae. Serotypes 1, 3 and 5 of CVB bind to the human membrane complement regulator decay-accelerating factor (DAF) and the coxsackievirus\\/adenovirus receptor (CAR), using either or both as receptors. These viruses are known to infect pig cell lines, but the receptor(s) involved has not been identified. We have recently characterized

  3. Conformational Plasticity of the 2A Proteinase from Enterovirus 71

    PubMed Central

    Cai, Qixu; Yameen, Muhammad; Liu, Weihua; Gao, Zhenting; Li, Yaozong; Peng, Xuanjia; Cai, Yaxian; Wu, Caiming; Zheng, Qian

    2013-01-01

    The 2A proteinase (2Apro) is an enterovirally encoded cysteine protease that plays essential roles in both the processing of viral precursor polyprotein and the hijacking of host cell translation and other processes in the virus life cycle. Crystallographic studies of 2Apro from enterovirus 71 (EV71) and its interaction with the substrate are reported here. EV71 2Apro was comprised of an N-terminal domain of a four-stranded antiparallel ? sheet and a C-terminal domain of a six-stranded antiparallel ? barrel with a tightly bound zinc atom. Unlike in other 2Apro structures, there is an open cleft across the surface of the protein in an open conformation. As demonstrated by the crystallographic studies and modeling of the complex structure, the open cleft could be fitted with the substrate. On comparison 2Apro of EV71 to those of the human rhinovirus 2 and coxsackievirus B4, the open conformation could be closed with a hinge motion in the bII2 and cII ? strands. This was supported by molecular dynamic simulation. The structural variation among different 2Apro structures indicates a conformational flexibility in the substrate-binding cleft. The open structure provides an accessible framework for the design and development of therapeutics against the viral target. PMID:23616646

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

    PubMed

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

    2008-01-01

    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

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

    PubMed Central

    McLeish, Nigel J.; Williams, Çi?dem H.; Kaloudas, Dimitrios; Roivainen, Merja M.

    2012-01-01

    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. PMID:22855495

  6. Failure of indicator bacteria to reflect the occurrence of enteroviruses in marine waters.

    PubMed Central

    Gerba, C P; Goyal, S M; LaBelle, R L; Cech, I; Bodgan, G F

    1979-01-01

    The results of several studies conducted along the upper Texas Gulf coast, where a substantial amount of quantitative virological data were collected, are compared to bacteriological indicators and other environmental factors on a statistical basis. Variables common to all these studies were anlayzed by multivariate regression. Although multivariate analysis indicated that the number of viruses detected in water was related to rainfall, salinity, and total coliforms in the water, the amount of variation in the number of viruses accounted for by these factors was not large enough to make them good predictors. Enteroviruses were detected 43 per cent of the time in recreational waters considered acceptable as judged by coliform standards, and 44 per cent of the time when judged by fecal coliform standards. Enteroviruses were detected 35 per cent of the time in waters which met acceptable standards for shellfish-harvesting. Our failure to correlate the occurrence of enteroviruses in marine waters with indicator bacteria, and the frequent occurrence of enteroviruses in water which met current bacteriological standards, indicates that these standards do not reflect the occurrence of enteroviruses, and perhaps other human pathogenic viruses, in marine waters. PMID:228561

  7. Strategies to develop antivirals against enterovirus 71

    PubMed Central

    2013-01-01

    Enterovirus 71 (EV71) is an important human pathogen which may cause severe neurological complications and death in children. The virus caused several outbreaks in the Asia-Pacific region during the past two decades and has been considered a significant public health problem in the post-poliovirus eradication era. Unlike poliovirus, there is no effective vaccine or approved antivirals against EV71. To explore anti-EV71 agents therefore is of vital importance. Several strategies have been employed to develop antivirals based on the molecular characteristics of the virus. Among these, some small molecules that were developed against human rhinoviruses and poliovirus are under evaluation. In this review, we discuss the recent development of such small molecules against EV71, known drug resistance and possible solutions to it, and animal models for evaluating the efficacy of these antivirals. Although further investigation is required for clinical applications of the existing candidates, the molecular mechanisms revealed for the inhibition of EV71 replication can be used for designing new molecules against this virus in the future. PMID:23339605

  8. Oncolysis of human ovarian cancers by echovirus type 1

    Microsoft Academic Search

    Darren R. Shafren; Dianne Sylvester; E. Susanne Johansson; Ian G. Campbell; Richard D. Barry

    2005-01-01

    A small number of enteroviruses possess the capacity to induce rapid and marked lytic infections in cells of various human malig- nancies. During screening of representative human enteroviruses for their oncolytic capacity, we observed that echovirus type 1 (EV1) displayed a high level of tropism for human ovarian cancer cells. EV1 is an enterovirus which largely causes asymptomatic infections in

  9. ADSORPTION OF ENTEROVIRUSES TO SOIL CORES AND THEIR SUBSEQUENT ELUTION BY ARTIFICIAL RAINWATER

    EPA Science Inventory

    The adsorption and elution of a variety of human enteroviruses in a highly permeable, sandy soil was studied by using cores (43 by 125 mm) collected from an operating recharge basin on Long Island. Viruses studied included field and reference strains of polioviruses types 1 and 3...

  10. QUANTIFICATION OF ENTEROVIRUS AND HEPATITIS A VIRUSES IN WELLS AND SPRINGS IN EAST TENNESSEE USING REAL-TIME REVERSE TRANSCIPTION PCR

    EPA Science Inventory

    This project involves development, validation testing and application of a fast, efficient method of quantitatively measuring occurrence and concentration of common human viral pathogens, enterovirus and hepatitis A virus, in ground water samples using real-time reverse transcrip...

  11. High Seroprevalence of Enterovirus Infections in Apes and Old World Monkeys

    PubMed Central

    McIntyre, Chloe L.; Imai, Natsuko; Clasper, Lucy; Djoko, Cyrille F.; LeBreton, Matthew; Vermeulen, Marion; Saville, Andrew; Mutapi, Francisca; Tamoufé, Ubald; Kiyang, John; Biblia, Tafon G.; Midzi, Nicholas; Mduluza, Takafira; Pépin, Jacques; Njoum, Richard; Smura, Teemu; Fair, Joseph N.; Wolfe, Nathan D.; Roivainen, Merja; Simmonds, Peter

    2012-01-01

    To estimate population exposure of apes and Old World monkeys in Africa to enteroviruses (EVs), we conducted a seroepidemiologic study of serotype-specific neutralizing antibodies against 3 EV types. Detection of species A, B, and D EVs infecting wild chimpanzees demonstrates their potential widespread circulation in primates. PMID:22305156

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

    PubMed Central

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

    1994-01-01

    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 PMID:8085832

  13. New Approaches for Enhanced Detection of Enteroviruses from Hawaiian Environmental Waters

    PubMed Central

    Connell, Christina; Tong, Hsin-I; Wang, Zi; Allmann, Erin; Lu, Yuanan

    2012-01-01

    Health risks associated with sewage-contaminated recreational waters are of important public health concern. Reliable water monitoring systems are therefore crucial. Current recreational water quality criteria rely predominantly on the enumeration of bacterial indicators, while potentially dangerous viral pathogens often remain undetected. Human enteric viruses have been proposed as alternative indicators; however, their detection is often hindered by low viral concentrations present in the environment. Reported here are novel and effective laboratory protocols for viral concentration and highly sensitive and optimized RT-PCR for the efficient detection of enteroviruses, an important enteric virus subset, in Hawaiian environmental waters. Eighteen published enterovirus primer pairs were comparatively evaluated for detection sensitivity. The primer set exhibiting the lowest detection limit under optimized conditions, EQ-1/EQ-2, was validated in a field survey of 22 recreational bodies of water located around the island of Oahu, Hawaii. Eleven sites tested positive for enterovirus, indicating fecal contamination at these locations. As an additional means of viral concentration, shellfish were collected from 9 sample sites and subjected to dissection, RNA extraction, and subsequent RT-PCR. Shellfish tissue from 6 of 9 sites tested positive for enterovirus. The techniques implemented here are valuable resources to aid accurate reflection of microbial contamination in Hawaii’s environmental waters. PMID:22567083

  14. The cross-reactivity of the enterovirus 71 to human brain tissue and identification of the cross-reactivity related fragments

    Microsoft Academic Search

    Chun Shi Jia; Jiang Ning Liu; Wan Bo Li; Chun Mei Ma; Shu Zhu Lin; Yi Hao; Xue Zhong Gao; Xiao Lin Liu; Yan Feng Xu; Lian Feng Zhang; Chuan Qin

    2010-01-01

    BACKGROUND: EV71 occasionally cause a series of severe neurological symptoms, including aseptic meningitis, encephalitis, and poliomyelitis-like paralysis. However, the neurological destruction mechanism was remained to be clarified. This study described the cross reaction between EV71 induced IgG and human brain tissue. RESULTS: Cross reaction of the IgG from 30 EV71 infected patients' sera to human tissues of cerebra was observed,

  15. Characterization of full-length enterovirus 71 strains from severe and mild disease patients in northeastern China.

    PubMed

    Wang, Xiaomei; Zhu, Chunfeng; Bao, Wanguo; Zhao, Ke; Niu, Junqi; Yu, Xiao-Fang; Zhang, Wenyan

    2012-01-01

    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. PMID:22479324

  16. Molecular modeling, organ culture and reverse genetics for a newly identified human rhinovirus C

    E-print Network

    51 89 74 87 81 80 52 80 44 81 72 100 05101520 p-distance (% change) HRV-B HRV-A HRV-C HEV HRV-A A7-distance). Human enteroviruses (HEV) are included as an outgroup. The tree was generated by Mega 4.1 software . 2

  17. [Characteristics of Coxsackie A enteroviruses circulating in children in Azerbaijan].

    PubMed

    Rustamova, L I; Aliev, K N; Tagizade, F D; Mamedova, M N

    2008-01-01

    Spectrum of serotypes of Coxsackie A virus and several others markers of virions, which cause enterovirus infection in children in Azerbaijan, was determined. Symptom of myotropism was used as a clinical sign. It was shown that enteroviruses were most frequently detected (41.1%) in children aged registered as enterovirus monoinfection. All isolated strains were characterized by such markers as cytopathic effect, delayed formation of plaques under the layer of agar with low content of sodium bicarbonate. Both attenuated and avirulent virions of Coxsackie A enteroviruses were detected. PMID:19186556

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

    PubMed

    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

    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

  19. Autism spectrum disorder secondary to enterovirus encephalitis.

    PubMed

    Marques, Filipa; Brito, Maria Joăo; Conde, Marta; Pinto, Mónica; Moreira, Ana

    2014-05-01

    Millions of children are infected by enteroviruses each year, usually exhibiting only mild symptoms. Nevertheless, these viruses are also associated with severe and life-threatening infections, such as meningitis and encephalitis. We describe a 32-month-old patient with enteroviral encephalitis confirmed by polymerase chain reaction in cerebrospinal fluid, with unfavorable clinical course with marked developmental regression, autistic features, persistent stereotypes and aphasia. She experienced slow clinical improvement, with mild residual neurologic and developmental deficits at follow-up. Viral central nervous system infections in early childhood have been associated with autism spectrum disorders but the underlying mechanisms are still poorly understood. This case report is significant in presenting a case of developmental regression with autistic features and loss of language improving on follow-up. To our knowledge, this is the first published report of enterovirus encephalitis leading to an autism spectrum disorder. PMID:24782421

  20. Discovery of a Bovine Enterovirus in Alpaca

    PubMed Central

    McClenahan, Shasta D.; Scherba, Gail; Borst, Luke; Fredrickson, Richard L.; Krause, Philip R.; Uhlenhaut, Christine

    2013-01-01

    A cytopathic virus was isolated using Madin-Darby bovine kidney (MDBK) cells from lung tissue of alpaca that died of a severe respiratory infection. To identify the virus, the infected cell culture supernatant was enriched for virus particles and a generic, PCR-based method was used to amplify potential viral sequences. Genomic sequence data of the alpaca isolate was obtained and compared with sequences of known viruses. The new alpaca virus sequence was most similar to recently designated Enterovirus species F, previously bovine enterovirus (BEVs), viruses that are globally prevalent in cattle, although they appear not to cause significant disease. Because bovine enteroviruses have not been previously reported in U.S. alpaca, we suspect that this type of infection is fairly rare, and in this case appeared not to spread beyond the original outbreak. The capsid sequence of the detected virus had greatest homology to Enterovirus F type 1 (indicating that the virus should be considered a member of serotype 1), but the virus had greater homology in 2A protease sequence to type 3, suggesting that it may have been a recombinant. Identifying pathogens that infect a new host species for the first time can be challenging. As the disease in a new host species may be quite different from that in the original or natural host, the pathogen may not be suspected based on the clinical presentation, delaying diagnosis. Although this virus replicated in MDBK cells, existing standard culture and molecular methods could not identify it. In this case, a highly sensitive generic PCR-based pathogen-detection method was used to identify this pathogen. PMID:23950875

  1. [Serotype distribution of enteroviruses isolated from paediatric cases prediagnosed as aseptic meningitis between 2001-2004 period].

    PubMed

    Ozkaya, Etem; Uysal, Gülnar; Atak, Tunca; Alkan, Mehmet

    2005-01-01

    Enteroviruses have major clinical and public health importance and are one of the leading causes of aseptic meningitis. There are many diseases with similar clinical symptoms and cerebrospinal fluid (CSF) findings of aseptic meningitis, thus virus isolation and identification is crucial for definitive diagnosis. Virological diagnosis is nonetheless important to distinguish between induced meningitis and other treatable causes of disease with a similar clinical picture. A total of 249 samples obtained from 246 cases (age range: 0-15 years), prediagnosed as aseptic meningitis, were sent to Virology Laboratory of Refik Saydam Hygiene Center. The patients were followed at Department of Pediatric Infectious Diseases in the Social Security Hospital, Ankara, Turkey, between 2001 and 2004. Stool (n: 180), CSF (n: 54) and throat swab (n: 15) samples have been inoculated to RD (rhabdomyosarcoma), Hep-2 (human epithelioma) and L20B (transgenic mice) cell lines, and followed up for the presence of cytopathic effects. A total of 95 enterovirus strains were isolated from 85 (34.6%) cases, and serotyped by using RIVM (National Institute of Public and the Environment, Nederlands) antisera with microneutralization method. As a result, the most frequently isolated types were found as echovirus type 30 (n: 24) and coxsackievirus type B (n: 19), which were most frequently isolated between July to October. This is the first report from Turkey for aseptic meningitis cases due to echovirus type 25 (n:3), 18 (n:2), 14 (n:1), 13 (n:4), 11 (n:6), 9 (n:1), 6 (n:9), 5 (n:1), 4 (n:1) and coxsackievirus type A9 (n:1). PMID:15900836

  2. Isolation of naturally occurring enteroviruses from a variety of shellfish species residing in Long Island and New Jersey marine embayments

    Microsoft Academic Search

    J. M. Vaughn; E. F. Landry; T. J. Vicale; M. C. Dahl

    1980-01-01

    Shellfish and shellfish-raising waters from a variety of Long Island and New Jersey marine embayments were examined for the presence of human enteroviruses. Little difference in virological quality was noted between areas designated as being open or closed to shellfishing. Viral isolations could not be correlated with coliform counts from identical samples, indicating the need to re-evaluate the use of

  3. Enter at your own risk: how enteroviruses navigate the dangerous world of pattern recognition receptor signaling.

    PubMed

    Harris, Katharine G; Coyne, Carolyn B

    2013-09-01

    Enteroviruses are the most common human viral pathogens worldwide. This genus of small, non-enveloped, single stranded RNA viruses includes coxsackievirus, rhinovirus, echovirus, and poliovirus species. Infection with these viruses can induce mild symptoms that resemble the common cold, but can also be associated with more severe syndromes such as poliomyelitis, neurological diseases including aseptic meningitis and encephalitis, myocarditis, and the onset of type I diabetes. In humans, polarized epithelial cells lining the respiratory and/or digestive tracts represent the initial sites of infection by enteroviruses. Control of infection in the host is initiated through the engagement of a variety of pattern recognition receptors (PRRs). PRRs act as the sentinels of the innate immune system and serve to alert the host to the presence of a viral invader. This review assembles the available data annotating the role of PRRs in the response to enteroviral infection as well as the myriad ways by which enteroviruses both interrupt and manipulate PRR signaling to enhance their own replication, thereby inducing human disease. PMID:23764548

  4. Detection of enterovirus D68 in Canadian laboratories.

    PubMed

    Hatchette, Todd F; Drews, Steven J; Grudeski, Elsie; Booth, Tim; Martineau, Christine; Dust, Kerry; Garceau, Richard; Gubbay, Jonathan; Karnauchow, Tim; Krajden, Mel; Levett, Paul N; Mazzulli, Tony; McDonald, Ryan R; McNabb, Alan; Mubareka, Samira; Needle, Robert; Petrich, Astrid; Richardson, Susan; Rutherford, Candy; Smieja, Marek; Tellier, Raymond; Tipples, Graham; LeBlanc, Jason J

    2015-05-01

    The recent emergence of a severe respiratory disease caused by enterovirus D68 prompted investigation into whether Canadian hospital and provincial laboratories can detect this virus using commercial and laboratory-developed assays. This study demonstrated analytical sensitivity differences between commercial and laboratory-developed assays for the detection of enterovirus D68. PMID:25740765

  5. Role of sediment in the persistence of enteroviruses in the estuarine environment.

    PubMed Central

    Smith, E M; Gerba, C P; Melnick, J L

    1978-01-01

    The survival of four enteroviruses commonly found in sewage effluents was examined when the viruses were adsorped to marine sediments in estuarine water and compared with virus survival in estuarine water alone. Echovirus 1, coxsackieviruses B3 and A9, and poliovirus 1 survived longer when associated with marine sediment. When the estuarine water was polluted with secondarily treated sewage effluent, virus survived for prolonged periods in sediments, but not in the overlaying estuarine water. PMID:206204

  6. Rapid diagnosis of enterovirus infection by magnetic bead extraction and polymerase chain reaction detection of enterovirus RNA in clinical specimens.

    PubMed Central

    Muir, P; Nicholson, F; Jhetam, M; Neogi, S; Banatvala, J E

    1993-01-01

    We describe a rapid method for extraction and detection of enterovirus RNA in clinical samples. By using magnetic bead technology, enterovirus RNA was efficiently and rapidly extracted from cerebrospinal fluid, stool, saliva, blood, pericardial fluid, urine, and cryopreserved or formalin-fixed solid tissue. Enterovirus RNA was then detected by reverse transcription followed by polymerase chain reaction amplification with primers designed to allow detection of most enterovirus serotypes. For detection of enteroviruses in specimens from patients with acute enteroviral disease, the overall sensitivity of enzymatic RNA amplification was greater than that of cell culture isolation, especially in blood specimens and in stool specimens from patients with acute cardiac disease. Enterovirus RNA was also detected in cryopreserved and archival formalin-fixed myocardial tissue from patients with acute myocarditis and chronic dilated cardiomyopathy. The ability to study archival specimens is of particular value in conducting retrospective investigation. The RNA extraction procedure used was considerably faster than extraction methods using organic reagents, used less hazardous reagents, and was of similar sensitivity. This detection protocol may therefore be useful both for the diagnosis of enterovirus infection and in studying the pathogenesis of acute and chronic enterovirus-induced disease. Images PMID:8380182

  7. Complete genome sequence analysis of two human coxsackievirus A9 strains isolated in Yunnan, China, in 2009.

    PubMed

    Liu, Jiansheng; Zhu, Yanju; Pan, Yue; Liu, Zhengling; Guo, Chen; Ma, Shaohui

    2015-06-01

    Human coxsackievirus A9 (CVA9) is a member of Enterovirus B species and may cause aseptic meningitis. The complete genome analyses of two strains CVA9 A242/YN/CHN/2009 and A108/YN/CHN/2009 isolated from aseptic meningitis cases in Yunnan Province, China, in 2009 were performed. These two strains shared 81.3 and 80.7, 81.0 and 81.1 % nucleotide similarity with prototype strain Griggs in the VP1-encoding sequence and the complete genome sequence, respectively. Through phylogenetic analysis and homogeneity analysis for twenty-eight VP1-encoding sequences, CVA9 strains could be divided into four genotypes and the Chinese strains might belong to genotype D. Similarity plot and bootscanning analyses showed evidence of recombination with other EVB viruses. In conclusion, persistent surveillance of circulating enterovirus might help understand the enterovirus evolution. PMID:25680342

  8. Bovine enteroviruses: classification and serological characterization 

    E-print Network

    Samal, Siba Kumar

    1981-01-01

    Committee: Dr. Stewart McConnel I Seventeen and 26 strains of bovine enterovirus were isolated from nasal and fecal samples of 18 cattle respectively. These viruses were serotyped using rabbit immune sera prepared aga1nst the 7 1nternational reference... of the virus and is inconsistent. Plaque assay revealed a mult1plic1ty of plaque sizes wi th1n selected samples. The plaque size and morphology however is not the controlling factor in determining serotype. Three cross reacting isolates were triple plaque...

  9. Detection, quantitation and identification of enteroviruses from surface waters and sponge tissue from the Florida Keys using real-time RT-PCR

    USGS Publications Warehouse

    Donaldson, K.A.; Griffin, Dale W.; Paul, J.H.

    2002-01-01

    A method was developed for the quantitative detection of pathogenic human enteroviruses from surface waters in the Florida Keys using Taqman (R) one-step Reverse transcription (RT)-PCR with the Model 7700 ABI Prism (R) Sequence Detection System. Viruses were directly extracted from unconcentrated grab samples of seawater, from seawater concentrated by vortex flow filtration using a 100kD filter and from sponge tissue. Total RNA was extracted from the samples, purified and concentrated using spin-column chromatography. A 192-196 base pair portion of the 5??? untranscribed region was amplified from these extracts. Enterovirus concentrations were estimated using real-time RT-PCR technology. Nine of 15 sample sites or 60% were positive for the presence of pathogenic human enteroviruses. Considering only near-shore sites, 69% were positive with viral concentrations ranging from 9.3viruses/ml to 83viruses/g of sponge tissue (uncorrected for extraction efficiency). Certain amplicons were selected for cloning and sequencing for identification. Three strains of waterborne enteroviruses were identified as Coxsackievirus A9, Coxsackievirus A16, and Poliovirus Sabin type 1. Time and cost efficiency of this one-step real-time RT-PCR methodology makes this an ideal technique to detect, quantitate and identify pathogenic enteroviruses in recreational waters. Copyright ?? 2002 Elsevier Science Ltd.

  10. Site-specific targeting of enterovirus capsid by functionalized monodisperse gold nanoclusters

    PubMed Central

    Marjomäki, Varpu; Lahtinen, Tanja; Martikainen, Mari; Koivisto, Jaakko; Malola, Sami; Salorinne, Kirsi; Pettersson, Mika; Häkkinen, Hannu

    2014-01-01

    Development of precise protocols for accurate site-specific conjugation of monodisperse inorganic nanoparticles to biological material is one of the challenges in contemporary bionanoscience and nanomedicine. We report here a successful site-specific covalent conjugation of functionalized atomically monodisperse gold clusters with 1.5-nm metal cores to viral surfaces. Water-soluble Au102(para-mercaptobenzoic acid)44 clusters, functionalized by maleimide linkers to target cysteines of viral capsid proteins, were synthesized and conjugated to enteroviruses echovirus 1 and coxsackievirus B3. Quantitative analysis of transmission electron microscopy images and the known virus structures showed high affinity and mutual ordering of the bound gold clusters on the viral surface and a clear correlation between the clusters and the targeted cysteine sites close to the viral surface. Infectivity of the viruses was not compromised by loading of several tens of gold clusters per virus. These advances allow for future investigations of the structure?function relations of enteroviruses and enterovirus-related virus-like particles, including their entry mechanisms into cells and uncoating in cellular endosomes. PMID:24474748

  11. Methods for Detecting Food-borne Enteroviruses

    PubMed Central

    Herrmann, John E.; Cliver, Dean O.

    1968-01-01

    A method previously reported for detecting virus in a model system composed of cottage cheese contaminated with coxsackievirus type A9 has been adapted to detecting selected strains of enteroviruses in a variety of foods. Bentonite is omitted and serum is added for extracting virus from low-protein foods. Samples of foods, usually 25 g, must contain at least 3 to 4 plaque-forming units for a 50% probability of detecting virus. Sensitivity in detecting echovirus type 6 was lower than that for the other viruses used. After extraction from potato salad, poliovirus type 2 was completely reactivated if it had been neutralized with coproantibody, but it was only partially reactivated if neutralized with hyperimmune rabbit serum. Images Fig. 1 PMID:4300896

  12. Comparative genetic analysis of VP4, VP1 and 3D gene regions of enterovirus 71 and coxsackievirus A16 circulating in Malaysia between 1997-2008.

    PubMed

    Chan, Y F; Wee, K L; Chiam, C W; Khor, C S; Chan, S Y; Amalina W, M Z; Sam, I C

    2012-09-01

    Three genomic regions, VP4 capsid, VP1 capsid and 3D RNA polymerase of human enterovirus 71 (EV-71) and coxsackievirus A16 (CV-A16) were sequenced to understand the evolution of these viruses in Malaysia. A total of 42 EV-71 and 36 CV-A16 isolates from 1997- 2008 were sequenced. Despite the presence of many EV-71 subgenotypes worldwide, only subgenotypes B3, B4, B5, C1 and C2 were present in Malaysia. Importation of other subgenotypes such as C3, C4/D and C5 from other countries was infrequent. For CV-A16, the earlier subgenotype B1 was replaced by subgenotypes B2a and the recent B2c. Subgenotype B2a was present throughout the study while B2c only emerged in 2005. No genetic signatures could be attributed to viral virulence suggesting that host factors have a major role in determining the outcome of infection. Only three EV-71 B3 isolates showed non-consistent phylogeny in the 3D RNA polymerase region which indicated occurrence of recombination in EV-71. High genetic diversity was observed in the Malaysian EV-71 but Malaysian CV-A16 showed low genetic diversity in the three genomic regions sequenced. EV-71 showed strong purifying selection, but that occurred to a lesser extent in CV-A16. PMID:23018509

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

    PubMed Central

    Park, Kwi Sung; Choi, Young Jin

    2012-01-01

    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

  14. Short Communication: New Recognition Of Enterovirus Infections In Bottlenose Dolphins (Tursiops Truncatus)

    PubMed Central

    Nollens, Hendrik H.; Rivera, Rebecca; Palacios, Gustavo; Wellehan, James F. X.; Saliki, Jeremiah T.; Caseltine, Shannon L.; Smith, Cynthia R.; Jensen, Eric D.; Hui, Jeffrey; Lipkin, W. Ian; Yochem, Pamela K.; Wells, Randall S.; St. Leger, Judy; Venn-Watson, Stephanie

    2014-01-01

    An enterovirus was cultured from an erosive tongue lesion of a bottlenose dolphin (Tursiops truncatus). The morphology of virions on negative staining electron microscopy was consistent with those of enteroviruses. Analysis of 2613 bp of the polyprotein gene identified the isolate as a novel enterovirus strain, tentatively named bottlenose dolphin enterovirus (BDEV), that nests within the species Bovine enterovirus. Serologic evidence of exposure to enteroviruses was common in both free ranging and managed collection dolphins. Managed collection dolphins were more likely to have high antibody levels, although the highest levels were reported in free ranging populations. Associations between enterovirus antibody levels, and age, sex, complete blood counts, and clinical serum biochemistries were explored. Dolphins with higher antibody levels were more likely to be hyperproteinemic and hyperglobulinemic. PMID:19581059

  15. Long-Term Immunogenicity Studies of Formalin-Inactivated Enterovirus 71 Whole-Virion Vaccine in Macaques

    PubMed Central

    Liu, Chia-Chyi; Hwang, Chyi-Sing; Yang, Wun-Syue; Tsai, Dan-Chin; Wu, Sze-Hsien; Chou, Ai-Hsiang; Chow, Yen-Hung; Wu, Suh-Chin; Wang, Jen-Ren; Chiang, Jen-Ron; Huang, Chin-Cheng; Pan, Chien-Hsiung; Chong, Pele

    2014-01-01

    Enterovirus 71 (EV71) has caused epidemics of hand, foot and mouth diseases in Asia during the past decades and no vaccine is available. A formalin-inactivated EV71 candidate vaccine (EV71vac) based on B4 subgenotype has previously been developed and found to elicit strong neutralizing antibody responses in mice and humans. In this study, we evaluated the long-term immunogenicity and safety of this EV71vac in a non-human primate model. Juvenile macaques were immunized at 0, 3 and 6 weeks either with 10 or 5 µg doses of EV71vac formulated with AlPO4 adjuvant, or PBS as control. During the 56 weeks of studies, no fever nor local redness and swelling at sites of injections was observed in the immunized macaques. After single immunization, 100% seroconversion based on 4-fold increased in neutralization titer (Nt) was detected in EV71vac immunized monkeys but not PBS controls. A dose-dependent IgG antibody response was observed in monkeys receiving EV71vac immunization. The Nt of EV71vac immunized macaques had reached the peak after 3 vaccinations, then decreased gradually; however, the GMT of neutralizing antibody in the EV71vac immunized macaques were still above 100 at the end of the study. Correspondingly, both dose- and time-dependent interferon-? and CD4+ T cell responses were detected in monkeys receiving EV71vac. Interestingly, similar to human responses, the dominant T cell epitopes of macaques were identified mainly in VP2 and VP3 regions. In addition, strong cross-neutralizing antibodies against most EV71 subgenotypes except some C2 and C4b strains, and Coxsackievirus A16 were observed. In summary, our results indicate that EV71vac elicits dose-dependent T-cell and antibody responses in macaques that could be a good animal model for evaluating the long-term immune responses elicited by EV71 vaccines. PMID:25197967

  16. Rapid one-step quantitative reverse transcriptase PCR assay with competitive internal positive control for detection of enteroviruses in environmental samples.

    PubMed

    Gregory, Jason B; Litaker, R Wayne; Noble, Rachel T

    2006-06-01

    Human enteroviruses can serve as a more accurate indicator of human fecal contamination than conventional bacteriological fecal indicators. We describe here a quantitative reverse transcriptase PCR (qRT-PCR) assay specifically tailored to detect these viruses in environmental waters. The assay included a competitive internal positive control (CIPC) that allowed the inhibition of qRT-PCRs to be quantitatively assessed. Coamplification of the CIPC with enteroviral genetic material did not affect the sensitivity, specificity, or reproducibility of the enteroviral qRT-PCR assay. The assay is rapid (less than 5 h from sample to result), has a wide dynamic range (>3 logs), and is capable of detecting as few as 25 enteroviral genomes with an average amplification efficiency of 0.91. In samples with low or moderate inhibition, the delay in CIPC amplification was used to adjust enterovirus qRT-PCR concentrations to account for losses due to inhibition. Samples exhibiting significant inhibition were not corrected but instead diluted twofold and immediately assayed again. Using significantly inhibited samples, it was found that dilution relieved inhibition in 93% (25 of 27) of the samples. In addition, 15% (4 of 27) of these previously negative samples contained enteroviral genomes. The high-throughput format of the assay compared to conventional culture-based methods offers a fast, reliable, and specific method for detecting enteroviruses in environmental water samples. The ability of the assay to identify false negatives and provide improved quantitative assessments of enterovirus concentrations will facilitate the tracking of human fecal contamination and the assessment of potential public health risk due to enteroviruses in recreational and shellfish harvesting waters. PMID:16751503

  17. Detection of enteroviruses in untreated and treated drinking water supplies in South Africa.

    PubMed

    Ehlers, M M; Grabow, W O K; Pavlov, D N

    2005-06-01

    Enteric viruses have been detected in many drinking water supplies all over the world. A meaningful number of these supplies were treated and disinfected according to internationally acceptable methods. In addition, counts of bacterial indicators (coliform bacteria and heterotrophic plate count organisms) in these water supplies were within limits generally recommended for treated drinking water and these findings have been supported by epidemiological data on infections associated with drinking water. The shortcomings of conventional treatment methods and indicator organisms to confirm the absence of enteric viruses from drinking water, was generally ascribed to the exceptional resistance of these viruses. In this study, the prevalence of enteroviruses detected from July 2000 to June 2002 in sewage, river-, borehole-, spring- and dam water as well as drinking water supplies treated and disinfected according to international specifications for the production of safe drinking water was analysed. A glass wool adsorption-elution technique was used to recover viruses from 10--20 l of sewage as well as environmental water samples, in the case of drinking water from more than 100 l. Recovered enteroviruses were inoculated onto two cell culture types (BGM and PLC/PRF/5 cells) for amplification of viral RNA with nested-PCR being used to detect the amplified viral RNA. Results from the study demonstrated the presence of enteroviruses in 42.5% of sewage and in 18.7% of treated drinking water samples. Furthermore, enteroviruses were detected in 28.5% of river water, in 26.7% of dam/spring water and in 25.3% of borehole water samples. The high prevalence of coxsackie B viruses found in this study suggested, that a potential health risk and a burden of disease constituted by these viruses might be meaningful. These findings indicated that strategies, other than end-point analysis of treated and disinfected drinking water supplies, may be required to ensure the production of drinking water that does not exceed acceptable health risks. More reliable approaches to ensure acceptable safety of drinking water supplies may be based on control by multiple-barrier principles from catchment to tap using hazard assessment and critical control point (HACCP) principles. PMID:15919105

  18. Coxsackie B4 virus infection of ? cells and natural killer cell insulitis in recent-onset type 1 diabetic patients

    PubMed Central

    Dotta, Francesco; Censini, Stefano; van Halteren, Astrid G. S.; Marselli, Lorella; Masini, Matilde; Dionisi, Sabrina; Mosca, Franco; Boggi, Ugo; Muda, Andrea Onetti; Prato, Stefano Del; Elliott, John F.; Covacci, Antonello; Rappuoli, Rino; Roep, Bart O.; Marchetti, Piero

    2007-01-01

    Type 1 diabetes is characterized by T cell-mediated autoimmune destruction of pancreatic ? cells. Several studies have suggested an association between Coxsackie enterovirus seroconversion and onset of disease. However, a direct link between ? cell viral infection and islet inflammation has not been established. We analyzed pancreatic tissue from six type 1 diabetic and 26 control organ donors. Immunohistochemical, electron microscopy, whole-genome ex vivo nucleotide sequencing, cell culture, and immunological studies demonstrated Coxsackie B4 enterovirus in specimens from three of the six diabetic patients. Infection was specific of ? cells, which showed nondestructive islet inflammation mediated mainly by natural killer cells. Islets from enterovirus-positive samples displayed reduced insulin secretion in response to glucose and other secretagogues. In addition, virus extracted from positive islets was able to infect ? cells from human islets of nondiabetic donors, causing viral inclusions and signs of pyknosis. None of the control organ donors showed signs of viral infection. These studies provide direct evidence that enterovirus can infect ? cells in patients with type 1 diabetes and that infection is associated with inflammation and functional impairment. PMID:17360338

  19. Enterovirus replication: go with the (counter)flow.

    PubMed

    Nchoutmboube, Jules; Ford-Siltz, Lauren A; Belov, George A

    2015-04-01

    All (+)RNA viruses replicate on distinct membranous domains; however, how they induce and maintain their unique lipid composition is largely unknown. Two recent studies reveal that enteroviruses harness the PI4P-cholestrol exchange cycle driven by OSBP1 protein and PI4 kinase(s), and that blocking the dynamic lipid flow inhibits virus replication. PMID:25748799

  20. Coexistence of two clades of enterovirus D68 in pediatric Swedish patients in the summer and fall of 2014.

    PubMed

    Dyrdak, Robert; Rotzén-Östlund, Maria; Samuelson, Agneta; Eriksson, Margareta; Albert, Jan

    2015-10-01

    In 2014, an outbreak of enterovirus D68 (EV-D68) was observed in North America, with cases of severe respiratory illness and a possible etiological link to cases of acute flaccid paralysis. EV-D68 has also been reported from European countries, but no data from Sweden are available. This study investigated respiratory specimens collected during July-October 2014 from 30 Swedish children aged 0-9 years who were positive for enterovirus and/or rhinovirus in routine clinical PCR. Seven samples were typed as EV-D68 by VP4/VP2 sequencing. Two genetically distinct EV-D68 variants coexisted. Six viruses belonged to clade B, the variant involved in the North American outbreak, and one virus belonged to clade A. Respiratory illness was the major symptom among EV-D68 infected patients and all fully recovered. This is the first report of EV-D68 in Sweden. Considering the current epidemiological situation, genotyping and specific EV-D68 testing should be considered in patients with severe respiratory illness who test positive for enterovirus or rhinovirus in routine diagnostics. PMID:25972105

  1. Team 2: Human Resources 2.B Human Resources (Leist/Defa)

    E-print Network

    Lawrence, Rick L.

    Team 2: Human Resources Areas: · 2.B Human Resources (Leist/Defa) · Policies and Procedures: Human. Policies and Procedures Human Resources 2.A.18 The institution maintains and publishes its human resources of human resources records. 2.B ­ Human Resources 2.B.1 The institution employs a sufficient number

  2. Yakammaoto inhibits enterovirus 71 infection by reducing viral attachment, internalization, replication, and translation.

    PubMed

    Yeh, Chia-Feng; Wang, Kuo-Chih; Lu, Chi-Yu; Chiang, Lien-Chai; Shieh, Den-En; Yen, Ming-Hong; Chang, Jung-San

    2015-06-01

    Enterovirus 71 (EV71) can cause central nervous system infections with mortality and neurologic sequelae. At present, there is no effective therapeutic modality for EV71 infection. The infection is more common in families with poor socioeconomic status. Therefore, finding a readily available, cost-effective therapeutic modality would be very helpful to these socioeconomically disadvantaged families. Yakammaoto is a cheap and readily available traditional prescription that is proven to have antiviral activity against coxsackievirus B4 (CVB4). CVB4 and EV71 are enteroviruses. In this study, we evaluated the antiviral activity of hot water extract of yakammaoto against EV71. The results of plaque reduction assay and flow cytometry demonstrated that yakammaoto dose dependently inhibited EV71 infection. In addition, reverse transcription-polymerase chain reaction (RT-PCR) and quantitative RT-PCR results showed that yakammaoto reduced viral replication. Western blotting analysis showed that yakammaoto can inhibit viral protein production. Thus, our results suggest that yakammaoto should be considered to manage EV71 infection in the future. PMID:26043408

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

    PubMed

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

    1999-01-01

    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

  4. Advances in Human B Cell Phenotypic Profiling

    PubMed Central

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

    2012-01-01

    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. PMID:23087687

  5. Preclinical Evaluation of the Immunogenicity and Safety of an Inactivated Enterovirus 71 Candidate Vaccine

    PubMed Central

    Hwa, Shi-Hsia; Lee, Yock Ann; Brewoo, Joseph N.; Partidos, Charalambos D.; Osorio, Jorge E.; Santangelo, Joseph D.

    2013-01-01

    Human enterovirus 71 (EV71) is a significant cause of morbidity and mortality from Hand, Foot and Mouth Disease (HFMD) and neurological complications, particularly in young children in the Asia-Pacific region. There are no vaccines or antiviral therapies currently available for prevention or treatment of HFMD caused by EV71. Therefore, the development of therapeutic and preventive strategies against HFMD is of growing importance. We report the immunogenic and safety profile of inactivated, purified EV71 preparations formulated with aluminum hydroxide adjuvant in preclinical studies in mice and rabbits. In mice, the candidate vaccine formulations elicited high neutralizing antibody responses. A toxicology study of the vaccine formulations planned for human use performed in rabbits showed no vaccine-related pathological changes and all animals remained healthy. Based on these preclinical studies, Phase 1 clinical testing of the EV71 inactivated vaccine was initiated. PMID:24244774

  6. Pathogenesis of murine enterovirus myocarditis: virus dissemination and immune cell targets.

    PubMed Central

    Klingel, K; Stephan, S; Sauter, M; Zell, R; McManus, B M; Bültmann, B; Kandolf, R

    1996-01-01

    In order to identify organ and cellular targets of persistent enterovirus infection in vivo, immunocompetent mice (SWR/J, H-2q) were inoculated intraperitoneally with coxsackievirus B3 (CVB3). By use of in situ hybridization for the detection of enteroviral RNA, we show that CVB3 is capable of inducing a multiorgan disease. During acute infection, viral RNA was visualized at high levels in the heart muscle, pancreas, spleen, and lymph nodes and at comparably low levels in the central nervous system, thymus, lung, and liver. At later stages of the disease, the presence of enteroviral RNA was found to be restricted to the myocardium, spleen, and lymph nodes. To characterize infected lymphoid cells during the course of the disease, enteroviral RNA and cell-specific surface antigens were visualized simultaneously in situ in spleen tissue sections. In acute infection, the majority of infected spleen cells, which are located primarily at the periphery of lymph follicles, were found to express the CD45R/B220+ phenotype of pre-B and B cells. Whereas viral RNA was also detected in certain CD4+ helper T cells and Mac-1+ macrophages, no enteroviral genomes were identified in CD8+ cytotoxic/suppressor T cells. Later in disease, the localization of enteroviral RNA revealed a persistent type of infection of B cells within the germinal centers of secondary follicles. In addition, detection of the replicative viral minus-strand RNA intermediate provided evidence for virus replication in lymphoid cells of the spleen during the course of the disease. These data indicate that immune cells are important targets of CVB3 infection, providing a noncardiac reservoir for viral RNA during acute and persistent myocardial enterovirus infection. PMID:8971018

  7. 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...

  8. Reciprocal Regulation between Enterovirus 71 and the NLRP3 Inflammasome.

    PubMed

    Wang, Hongbin; Lei, Xiaobo; Xiao, Xia; Yang, Chunfu; Lu, Wenli; Huang, Zhong; Leng, Qibin; Jin, Qi; He, Bin; Meng, Guangxun; Wang, Jianwei

    2015-07-01

    Enterovirus 71 (EV71) is the major etiological agent of hand, foot, and mouth disease (HFMD). Early studies showed that EV71-infected patients with severe complications exhibited elevated plasma levels of IL-1?, indicating that EV71 may activate inflammasomes. Our current study demonstrates that the NLRP3 inflammasome plays a protective role against EV71 infection of mice in vivo. EV71 replication in myeloid cells results in the activation of the NLRP3 inflammasome and secretion of IL-1?. Conversely, EV71 counteracts inflammasome activation through cleavage of NLRP3 by viral proteases 2A and 3C, which cleave NLRP3 protein at the G493-L494 or Q225-G226 junction, respectively. Moreover, EV71 3C interacts with NLRP3 and inhibits IL-1? secretion when expressed in mammalian cells. These results thus reveal a set of reciprocal regulations between enterovirus 71 and the NLRP3 inflammasome. PMID:26119741

  9. Ultrastructural changes of motoneurons in monkeys infected with enterovirus 71

    Microsoft Academic Search

    I. Hashimoto; A. Hagiwara; I. Uchino

    1985-01-01

    Summary Tissues of the central nervous system (CNS) of cynomolgus monkeys were examined by electron microscopy after intraspinal inoculation of enterovirus 71 (E71). A characteristic finding was the appearance of numerous membrane-bound vesicles (Mbvs) in affected motoneurons. Similar Mbvs were also present in E71-infected cynomolgus monkey kidney (CMK) cells in culture. Virus-like particles were found within or around Mbvs in

  10. Exposure to enteroviruses and hepatitis A virus among divers in environmental waters in France, first biological and serological survey of a controlled cohort.

    PubMed Central

    Garin, D.; Fuchs, F.; Crance, J. M.; Rouby, Y.; Chapalain, J. C.; Lamarque, D.; Gounot, A. M.; Aymard, M.

    1994-01-01

    An epidemiological study of hepatitis A and enteroviruses was conducted in a military diving training school, by evaluating the viral contamination of water using an ultrafiltration concentration technique, and assessing seroconversion and the presence of virus in stool specimens obtained from 109 divers and 48 controls. Three of 29 water specimens were positive for enterovirus by cell culture and 9 by molecular hybridization. There was little or no risk of virus infection during the training course (49 h exposure) because there was no significant difference between divers and controls for both viral isolation and seroconversion. However, a higher percentage of coxsackievirus B4 and B5 seropositive divers suggests that these were more exposed during previous water training. No hepatitis A virus (HAV) detection and no seroconversion to HAV was observed. The rate of HAV seropositive subjects was 17% in this 24.5-year-old population. PMID:7995363

  11. The capsid binder Vapendavir and the novel protease inhibitor SG85 inhibit enterovirus 71 replication.

    PubMed

    Tijsma, Aloys; Franco, David; Tucker, Simon; Hilgenfeld, Rolf; Froeyen, Mathy; Leyssen, Pieter; Neyts, Johan

    2014-11-01

    Antivirals against enterovirus 71 (EV71) are urgently needed. We demonstrate that the novel enteroviral protease inhibitor (PI) SG85 and capsid binder (CB) vapendavir efficiently inhibit the in vitro replication of 21 EV71 strains/isolates that are representative of the different genogroups A, B, and C. The PI rupintrivir, the CB pirodavir, and the host-targeting compound enviroxime, which were included as reference compounds, also inhibited the replication of all isolates. Remarkably, the CB compound pleconaril was devoid of any anti-EV71 activity. An in silico docking study revealed that pleconaril-unlike vapendavir and pirodavir-lacks essential binding interactions with the viral capsid. Vapendavir and SG85 (or analogues) should be further explored for the treatment of EV71 infections. The data presented here may serve as a reference when developing yet-novel inhibitors. PMID:25199773

  12. The Capsid Binder Vapendavir and the Novel Protease Inhibitor SG85 Inhibit Enterovirus 71 Replication

    PubMed Central

    Tijsma, Aloys; Franco, David; Tucker, Simon; Hilgenfeld, Rolf; Froeyen, Mathy; Leyssen, Pieter

    2014-01-01

    Antivirals against enterovirus 71 (EV71) are urgently needed. We demonstrate that the novel enteroviral protease inhibitor (PI) SG85 and capsid binder (CB) vapendavir efficiently inhibit the in vitro replication of 21 EV71 strains/isolates that are representative of the different genogroups A, B, and C. The PI rupintrivir, the CB pirodavir, and the host-targeting compound enviroxime, which were included as reference compounds, also inhibited the replication of all isolates. Remarkably, the CB compound pleconaril was devoid of any anti-EV71 activity. An in silico docking study revealed that pleconaril—unlike vapendavir and pirodavir—lacks essential binding interactions with the viral capsid. Vapendavir and SG85 (or analogues) should be further explored for the treatment of EV71 infections. The data presented here may serve as a reference when developing yet-novel inhibitors. PMID:25199773

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

    Microsoft Academic Search

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

    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

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

    Microsoft Academic Search

    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

    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

  15. Enterovirus 68 3C Protease Cleaves TRIF To Attenuate Antiviral Responses Mediated by Toll-Like Receptor 3

    PubMed Central

    Xiang, Zichun; Li, Linlin; Lei, Xiaobo; Zhou, Hongli; Zhou, Zhuo

    2014-01-01

    ABSTRACT Human enterovirus 68 (EV68) is a member of the EV-D species, which belongs to the EV genus of the Picornaviridae family. Over the past several years, there have been increasingly documented outbreaks of respiratory disease associated with EV68. As a globally emerging pathogen, EV68 infects both adults and children. However, the molecular basis of EV68 pathogenesis is unknown. Here we report that EV68 inhibits Toll-like receptor 3 (TLR3)-mediated innate immune responses by targeting the TIR domain-containing adaptor inducing beta interferon (TRIF). In infected HeLa cells, EV68 inhibits poly(I·C)-induced interferon regulatory factor 3 (IRF3) activation and beta interferon (IFN-?) expression. Further investigations revealed that TRIF, a critical adaptor downstream of TLR3, is targeted by EV68. When expressed alone, 3Cpro, an EV68-encoded protease, cleaves TRIF. 3Cpro mediates TRIF cleavage at Q312 and Q653, which are sites in the amino- and carboxyl-terminal domains, respectively. This cleavage relies on 3Cpro's cysteine protease activity. Cleavage of TRIF abolishes the capacity of TRIF to activate NF-?B and IFN-? signaling. These results suggest that control of TRIF by 3Cpro may be a mechanism by which EV68 subverts host innate immune responses. IMPORTANCE EV68 is a globally emerging pathogen, but the molecular basis of EV68 pathogenesis is unclear. Here we report that EV68 inhibits TLR3-mediated innate immune responses by targeting TRIF. Further investigations revealed that TRIF is cleaved by 3Cpro. These results suggest that control of TRIF by 3Cpro may be a mechanism by which EV68 impairs type I IFN production in response to TLR3 activation. PMID:24672048

  16. UWMadison, Office of Human Resources Human Resource System B91002.20101229

    E-print Network

    Wisconsin at Madison, University of

    UW­Madison, Office of Human Resources Human Resource System B91002.20101229 State Group Health Information Submit completed checklist and attachments to: Office of Human Resources, Benefit Services, 21 N: __________________________________________________________________________________ ____________________________________________________________________________________________ ____________________________________________________________________________________________ mm/dd/yyyy mm/dd/yyyy #12;UW­Madison, Office of Human Resources Human Resource System B91002

  17. Replication strategy of human hepatitis B virus

    SciTech Connect

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

    1987-03-01

    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.

  18. Differentiation and Characterization of Enteroviruses by Computer-Assisted Viral Protein Fingerprinting

    PubMed Central

    Holland, Diane T.; Senne, Jill; Peter, C. R.; Urmeneta, Connie; Connor, J. D.

    1998-01-01

    We have developed and standardized a computerized method for the typing and characterization of enteroviruses with radiolabeled viral protein fingerprints. Enteroviral proteins were radiolabeled with [35S]methionine during growth in cell culture and were then separated by polyacrylamide gel electrophoresis. The dried gel was scanned, and from the resulting computer image (which resembled an autoradiogram) protein patterns were computer extracted and stored in a database. The enterovirus database contained community and prototype strains belonging to 20 different enteroviral serotypes. Each serotype has a discrete protein pattern, and the most important pattern differences for determining each type are in the region of the viral capsid proteins VP1, VP2, and VP3. When the database was challenged with 148 clinical enterovirus strains, 144 (97%) were correctly identified by using the correlation coefficient as a quantitative measure of relatedness between two patterns. This method can identify a type in a single test and represents a practical alternative to virus neutralization because it is less expensive, is much faster (3 rather than 10 days), and does not rely on any virus-specific reagents. The results also show that most of the strains currently isolated from the community have protein patterns different from those of their older prototype strains. Viral protein fingerprinting is an evolving, dynamic system for the typing and characterization of enteroviruses. The method is appropriate for use in clinical virology and reference laboratories for the typing of enteroviruses, for the study of the epidemiology of enteroviruses, and for surveillance of enteroviruses. PMID:9620382

  19. A Novel Universal Neutralizing Monoclonal Antibody against Enterovirus 71 That Targets the Highly Conserved “Knob” Region of VP3 Protein

    PubMed Central

    Meng, Tao; Chow, Vincent Tak Kwong; Kwang, Jimmy

    2014-01-01

    Hand, foot and mouth disease caused by enterovirus 71(EV71) leads to the majority of neurological complications and death in young children. While putative inactivated vaccines are only now undergoing clinical trials, no specific treatment options exist yet. Ideally, EV71 specific intravenous immunoglobulins could be developed for targeted treatment of severe cases. To date, only a single universally neutralizing monoclonal antibody against a conserved linear epitope of VP1 has been identified. Other enteroviruses have been shown to possess major conformational neutralizing epitopes on both the VP2 and VP3 capsid proteins. Hence, we attempted to isolate such neutralizing antibodies against conformational epitopes for their potential in the treatment of infection as well as differential diagnosis and vaccine optimization. Here we describe a universal neutralizing monoclonal antibody that recognizes a conserved conformational epitope of EV71 which was mapped using escape mutants. Eight escape mutants from different subgenogroups (A, B2, B4, C2, C4) were rescued; they harbored three essential mutations either at amino acid positions 59, 62 or 67 of the VP3 protein which are all situated in the “knob” region. The escape mutant phenotype could be mimicked by incorporating these mutations into reverse genetically engineered viruses showing that P59L, A62D, A62P and E67D abolish both monoclonal antibody binding and neutralization activity. This is the first conformational neutralization epitope mapped on VP3 for EV71. PMID:24875055

  20. Towards identification of cis-acting elements involved in the replication of enterovirus and rhinovirus RNAs: a proposal for the existence of tRNA-like terminal structures.

    PubMed Central

    Pilipenko, E V; Maslova, S V; Sinyakov, A N; Agol, V I

    1992-01-01

    On the basis of a comparative analysis of published sequences, models for the secondary structure of the 3'-terminal [poly(A)-preceding] untranslated region of the entero- and rhinovirus RNAs were worked out. The models for all these viruses share a common core element, but there are an extra enterovirus-specific element and still an additional element characteristic of a subset of enterovirus RNAs. The two latter models were verified for poliovirus and coxsackievirus B genomes by testing with single-strand and double-strand specific enzymatic and chemical probes. A tRNA-like tertiary structure model for the 3'-terminal folding of enterovirus RNAs was proposed. A similar folding was proposed for the 3' termini of the negative RNA strands as well as for the 5' termini of the positive strand of all entero- and rhinovirus RNAs. Implications of these data for template recognition during negative and positive RNA strands synthesis and for the evolution of the picornavirus genomes are discussed. PMID:1315956

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

    PubMed Central

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

    1994-01-01

    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 PMID:7521997

  2. Evaluation of an enterovirus group-specific anti-VP1 monoclonal antibody, 5-D8/1, in comparison with neutralization and PCR for rapid identification of enteroviruses in cell culture.

    PubMed Central

    Trabelsi, A; Grattard, F; Nejmeddine, M; Aouni, M; Bourlet, T; Pozzetto, B

    1995-01-01

    We evaluated the usefulness of a commercially available monoclonal antibody (MAb) directed against a group-specific epitope of the capsid protein VP1 of enteroviruses for the rapid identification of these viruses in cell culture. The MAb was assayed in an indirect immunofluorescence test with cultured cells infected by various serotypes of enterovirus; all 39 serotypes tested, including echoviruses 22 and 23, which are considered atypical enteroviruses, were reactive. The MAb was also tested with 61 strains recovered from clinical specimens inoculated into cell cultures in comparison with seroneutralization with intersecting pools of hyperimmune sera and PCR with primers from the 5' untranslated region of enteroviruses. There was total agreement between the results obtained with the MAb and those obtained by PCR, even for those strains of enteroviruses which were found to be untypeable with polyclonal antisera. These data demonstrate the usefulness of the MAb for rapid identification of enteroviruses in cell culture. PMID:7494045

  3. Peptidyl aldehyde NK-1.8k suppresses enterovirus 71 and enterovirus 68 infection by targeting protease 3C.

    PubMed

    Wang, Yaxin; Yang, Ben; Zhai, Yangyang; Yin, Zheng; Sun, Yuna; Rao, Zihe

    2015-05-01

    Enterovirus (EV) is one of the major causative agents of hand, foot, and mouth disease in the Pacific-Asia region. In particular, EV71 causes severe central nervous system infections, and the fatality rates from EV71 infection are high. Moreover, an outbreak of respiratory illnesses caused by an emerging EV, EV68, recently occurred among over 1,000 young children in the United States and was also associated with neurological infections. Although enterovirus has emerged as a considerable global public health threat, no antiviral drug for clinical use is available. In the present work, we screened our compound library for agents targeting viral protease and identified a peptidyl aldehyde, NK-1.8k, that inhibits the proliferation of different EV71 strains and one EV68 strain and that had a 50% effective concentration of 90 nM. Low cytotoxicity (50% cytotoxic concentration, >200 ?M) indicated a high selective index of over 2,000. We further characterized a single amino acid substitution inside protease 3C (3C(pro)), N69S, which conferred EV71 resistance to NK-1.8k, possibly by increasing the flexibility of the substrate binding pocket of 3C(pro). The combination of NK-1.8k and an EV71 RNA-dependent RNA polymerase inhibitor or entry inhibitor exhibited a strong synergistic anti-EV71 effect. Our findings suggest that NK-1.8k could potentially be developed for anti-EV therapy. PMID:25691647

  4. Effect of coincident enterovirus infection and cows' milk exposure on immunisation to insulin in early infancy

    Microsoft Academic Search

    O. Vaarala; P. Klemetti; S. Juhela; O. Simell; H. Hyöty; J. Ilonen

    2002-01-01

    Aims\\/hypothesis. Insulin autoantibodies appear often as the first autoantibody in children who develop islet-cell autoimmunity. Our recent\\u000a studies indicate that primary immunisation to insulin is induced in early infancy by exposure to dietary bovine insulin present\\u000a in cows' milk formulas. As gut-associated lymphoid tissue is also the primary replication site of enteroviruses, we tested\\u000a whether enterovirus infections could modify the

  5. Diversity of enterovirus sequences detected in oysters by RT-heminested PCR

    Microsoft Academic Search

    Eric Dubois; Ghislaine Merle; Catherine Roquier; Aurélien Trompette; Françoise Le Guyader; Catherine Crucičre; Jean-Jacques Chomel

    2004-01-01

    Oysters harvested in western France, from five sites associated with outbreaks of food-borne norovirus gastroenteritis between February 2000 and March 2001, were assayed for enterovirus RNA by reverse transcriptase-heminested polymerase chain reaction (RT-heminested PCR). Forty percent (21\\/52) of shellfish samples (pool of seven oysters) were contaminated by enteroviruses. Infectious coxsackieviruses serotype A21 were isolated from three of these positive samples.

  6. Low-level Circulation of Enterovirus D68–Associated Acute Respiratory Infections, Germany, 2014

    PubMed Central

    Reiche, Janine; Böttcher, Sindy; Diedrich, Sabine; Buchholz, Udo; Buda, Silke; Haas, Walter; Schweiger, Brunhilde

    2015-01-01

    We used physician sentinel surveillance to identify 25 (7.7%) mild to severe infections with enterovirus D68 (EV-D68) in children and adults among 325 outpatients with acute respiratory infections in Germany during August–October 2014. Results suggested low-level circulation of enterovirus D68 in Germany. Viruses were characterized by sequencing viral protein (VP) 1 and VP4/VP2 genomic regions. PMID:25898320

  7. New complete genome sequences of human rhinoviruses shed light on their phylogeny and genomic features

    PubMed Central

    Tapparel, Caroline; Junier, Thomas; Gerlach, Daniel; Cordey, Samuel; Van Belle, Sandra; Perrin, Luc; Zdobnov, Evgeny M; Kaiser, Laurent

    2007-01-01

    Background Human rhinoviruses (HRV), the most frequent cause of respiratory infections, include 99 different serotypes segregating into two species, A and B. Rhinoviruses share extensive genomic sequence similarity with enteroviruses and both are part of the picornavirus family. Nevertheless they differ significantly at the phenotypic level. The lack of HRV full-length genome sequences and the absence of analysis comparing picornaviruses at the whole genome level limit our knowledge of the genomic features supporting these differences. Results Here we report complete genome sequences of 12 HRV-A and HRV-B serotypes, more than doubling the current number of available HRV sequences. The whole-genome maximum-likelihood phylogenetic analysis suggests that HRV-B and human enteroviruses (HEV) diverged from the last common ancestor after their separation from HRV-A. On the other hand, compared to HEV, HRV-B are more related to HRV-A in the capsid and 3B-C regions. We also identified the presence of a 2C cis-acting replication element (cre) in HRV-B that is not present in HRV-A, and that had been previously characterized only in HEV. In contrast to HEV viruses, HRV-A and HRV-B share also markedly lower GC content along the whole genome length. Conclusion Our findings provide basis to speculate about both the biological similarities and the differences (e.g. tissue tropism, temperature adaptation or acid lability) of these three groups of viruses. PMID:17623054

  8. Enterovirus Infections of the Central Nervous System Review

    PubMed Central

    Rhoades, Ross E.; Tabor-Godwin, Jenna M.; Tsueng, Ginger; Feuer, Ralph

    2011-01-01

    Enteroviruses (EV) frequently infect the central nervous system (CNS) and induce neurological diseases. Although the CNS is composed of many different cell types, the spectrum of tropism for each EV is considerable. These viruses have the ability to completely shut down host translational machinery and are considered highly cytolytic, thereby causing cytopathic effects. Hence, CNS dysfunction following EV infection of neuronal or glial cells might be expected. Perhaps unexpectedly given their cytolytic nature, EVs may establish a persistent infection within the CNS, and the lasting effects on the host might be significant with unanticipated consequences. This review will describe the clinical aspects of EV-mediated disease, mechanisms of disease, determinants of tropism, immune activation within the CNS, and potential treatment regimes. PMID:21251690

  9. Proactive approach to containment of enterovirus infection in the nursery.

    PubMed

    Fuchs, Inbal; Golan, Agneta; Borer, Abraham; Shemer-Avni, Yonat; Dagan, Ron; Greenberg, David

    2013-07-01

    Administration of prophylactic intravenous immunoglobulins to contacts of infants actively shedding enterovirus during a hospital nursery outbreak may attenuate severity of disease in those contacts and aid in containment of the outbreak. Four cases of neonatal enteroviral disease were treated in our hospital nursery in July and August 2011; 3 were presumed or proven vertical transmission cases and 1 was a presumed horizontal transmission. We aimed to prevent development of severe illness in contacts of affected neonates following a ministry of health advisory during the summer of 2011 warning of increased neonatal enteroviral morbidity and mortality in Israel. Strict infection control measures were implemented, including meticulous decontamination of the nursery environment and administration of intravenous immunoglobulin prophylaxis to contacts. No further horizontal transmission occurred after infection control interventions. Immunoglobulin prophylaxis to control enteroviral infection in the nursery should be considered as an auxiliary infection control intervention during a nursery outbreak. PMID:23572447

  10. Protective Efficacy of VP1-Specific Neutralizing Antibody Associated with a Reduction of Viral Load and Pro-Inflammatory Cytokines in Human SCARB2-Transgenic Mice

    PubMed Central

    Chang, Hsuen-Wen; Lin, Yi-Wen; Ho, Hui-Min; Lin, Min-Han; Liu, Chia-Chyi; Shao, Hsiao-Yun; Chong, Pele; Sia, Charles; Chow, Yen-Hung

    2013-01-01

    Hand-foot-mouth diseases (HFMD) caused by enterovirus 71 (EV71) and coxsackievirus 16 (CVA16) in children have now become a severe public health issue in the Asian-Pacific region. Recently we have successfully developed transgenic mice expressing human scavenger receptor class B member 2 (hSCARB2, a receptor of EV71 and CVA16) as an animal model for evaluating the pathogenesis of enterovirus infections. In this study, hSCARB2-transgenic mice were used to investigate the efficacy conferred by a previously described EV71 neutralizing antibody, N3. A single injection of N3 effectively inhibited the HFMD-like skin scurfs in mice pre-infected with clinical isolate of EV71 E59 (B4 genotype) or prevented severe limb paralysis and death in mice pre-inoculated with 5746 (C2 genotype). This protection was correlated with remarkable reduction of viral loads in the brain, spinal cord and limb muscles. Accumulated viral loads and the associated pro-inflammatory cytokines were all reduced. The protective efficacy of N3 was not observed in animals challenged with CVA16. This could be due to dissimilarity sequences of the neutralizing epitope found in CVA16. These results indicate N3 could be useful in treating severe EV71 infections and the hSCARB2-transgenic mouse could be used to evaluate the protective efficacy of potential anti-enterovirus agent candidates. PMID:23936115

  11. Protection against Lethal Enterovirus 71 Challenge in Mice by a Recombinant Vaccine Candidate Containing a Broadly Cross-Neutralizing Epitope within the VP2 EF Loop

    PubMed Central

    Xu, Longfa; He, Delei; Li, Zhiqun; Zheng, Jun; Yang, Lisheng; Yu, Miao; Yu, Hai; Chen, Yixin; Que, Yuqiong; Shih, James Wai Kuo; Liu, Gang; Zhang, Jun; Zhao, Qinjian; Cheng, Tong; Xia, Ningshao

    2014-01-01

    Human enterovirus 71 (EV71) is the main causative agent of hand, foot, and mouth disease (HFMD) and is associated with several severe neurological complications in the Asia-Pacific region. Here, we evaluated that while passive transfer of neutralizing monoclonal antibody (nMAb) against the VP2 protein protect against lethal EV71 infection in BALB/c mice. Protective nMAb were mapped to residues 141-155 of VP2 by peptide ELISA. High-resolution structural analysis showed that the epitope is part of the VP2 EF loop, which is the “puff” region that forms the “southern rim” of the canyon. Moreover, a three-dimensional structural characterization for the puff region with prior neutralizing epitopes and receptor-binding sites that can serve to inform vaccine strategies. Interestingly, using hepatitis B virus core protein (HBc) as a carrier, we demonstrated that the cross-neutralizing EV71 antibodies were induced, and the VP2 epitope immunized mice serum also conferred 100% in vivo passive protection. The mechanism of in vivo protection conferred by VP2 nMAb is in part attributed to the in vitro neutralizing titer and ability to bind authentic viral particles. Importantly, the anti-VP2(aa141-155) antibodies could inhibit the binding of human serum to EV71 virions showed that the VP2 epitope is immunodominant. Collectively, our results suggest that a broad-spectrum vaccine strategy targeting the high-affinity epitope of VP2 EF loop may elicits effective immune responses against EV71 infection. PMID:24669278

  12. “Eczema Coxsackium” and Unusual Cutaneous Findings in an Enterovirus Outbreak

    PubMed Central

    Oza, Vikash; Frieden, Ilona J.; Cordoro, Kelly M.; Yagi, Shigeo; Howard, Renee; Kristal, Leonard; Ginocchio, Christine C.; Schaffer, Julie; Maguiness, Sheilagh; Bayliss, Susan; Lara-Corrales, Irene; Garcia-Romero, Maria Teresa; Kelly, Dan; Salas, Maria; Oberste, M. Steven; Nix, W. Allan; Glaser, Carol; Antaya, Richard

    2013-01-01

    OBJECTIVE: To characterize the atypical cutaneous presentations in the coxsackievirus A6 (CVA6)–associated North American enterovirus outbreak of 2011–2012. METHODS: We performed a retrospective case series of pediatric patients who presented with atypical cases of hand, foot, and mouth disease (HFMD) from July 2011 to June 2012 at 7 academic pediatric dermatology centers. Patients were included if they tested positive for CVA6 or if they met clinical criteria for atypical HFMD (an enanthem or exanthem characteristic of HFMD with unusual morphology or extent of cutaneous findings). We collected demographic, epidemiologic, and clinical data including history of skin conditions, morphology and extent of exanthem, systemic symptoms, and diagnostic test results. RESULTS: Eighty patients were included in this study (median age 1.5 years, range 4 months–16 years). Seventeen patients were CVA6-positive, and 63 met clinical inclusion criteria. Ninety-nine percent of patients exhibited a vesiculobullous and erosive eruption; 61% of patients had rash involving >10% body surface area. The exanthem had a perioral, extremity, and truncal distribution in addition to involving classic HFMD areas such as palms, soles, and buttocks. In 55% of patients, the eruption was accentuated in areas of eczematous dermatitis, termed “eczema coxsackium.” Other morphologies included Gianotti-Crosti–like (37%), petechial/purpuric (17%) eruptions, and delayed onychomadesis and palm and sole desquamation. There were no patients with serious systemic complications. CONCLUSIONS: The CVA6-associated enterovirus outbreak was responsible for an exanthem potentially more widespread, severe, and varied than classic HFMD that could be confused with bullous impetigo, eczema herpeticum, vasculitis, and primary immunobullous disease. PMID:23776120

  13. Enteroviruses in Spain: virological and epidemiological studies over 10 years (1988-97).

    PubMed Central

    Trallero, G.; Casas, I.; Tenorio, A.; Echevarria, J. E.; Castellanos, A.; Lozano, A.; Breńa, P. P.

    2000-01-01

    A total of 15,662 clinical samples were analysed for enterovirus (EV) isolation in cell cultures during a 10-year period (1988-97). Furthermore, 210 isolates of EV obtained in primary laboratories within Spain from patients with meningitis were characterized. The total number of EV typed was 758, including 727 non-polio EV and 31 Sabin-like (SL) polioviruses. Twenty-eight EV serotypes were represented. Echoviruses comprised 90% (653/727) of fully typed non-polio EV. The four most prevalent serotypes were echovirus 30, echovirus 9, echovirus 6 and echovirus 4. Echovirus 30 was the main serotype associated with meningitis. Echovirus 9 was the aetiological agent in 20 outbreaks of meningitis while the occurrence of echovirus 6 was localized in 1 year (1997). Coxsackieviruses A and B occurred in 3 and 7% of the non-polio EV respectively. Coxsackievirus B5 presented the relative greater abundance. This paper examines the epidemiology of EV in Spain to serotype level over a 10-year period with special attention to non-polio EV associated with meningitis. PMID:10982074

  14. Generation of Functional Human Pancreatic b Cells In Vitro

    E-print Network

    Blackwell, Keith

    Resource Generation of Functional Human Pancreatic b Cells In Vitro Felicia W. Pagliuca,1,3 Jeffrey in diabetes. However, insu- lin-producing cells previously generated from human pluripotent stem cells (h, and transplantation of these cells ame- liorates hyperglycemia in diabetic mice. INTRODUCTION The discovery of human

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

    PubMed Central

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

    2010-01-01

    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. PMID:20066739

  16. Crystal structures of enterovirus 71 (EV71) recombinant virus particles provide insights into vaccine design.

    PubMed

    Lyu, Ke; Wang, Guang-Chuan; He, Ya-Ling; Han, Jian-Feng; Ye, Qing; Qin, Cheng-Feng; Chen, Rong

    2015-02-01

    Hand-foot-and-mouth disease (HFMD) remains a major health concern in the Asia-Pacific regions, and its major causative agents include human enterovirus 71 (EV71) and coxsackievirus A16. A desirable vaccine against HFMD would be multivalent and able to elicit protective responses against multiple HFMD causative agents. Previously, we have demonstrated that a thermostable recombinant EV71 vaccine candidate can be produced by the insertion of a foreign peptide into the BC loop of VP1 without affecting viral replication. Here we present crystal structures of two different naturally occurring empty particles, one from a clinical C4 strain EV71 and the other from its recombinant virus containing an insertion in the VP1 BC loop. Crystal structure analysis demonstrated that the inserted foreign peptide is well exposed on the particle surface without significant structural changes in the capsid. Importantly, such insertions do not seem to affect the virus uncoating process as illustrated by the conformational similarity between an uncoating intermediate of another recombinant virus and that of EV71. Especially, at least 18 residues from the N terminus of VP1 are transiently externalized. Altogether, our study provides insights into vaccine development against HFMD. PMID:25492868

  17. Structures of Enterovirus 71 3C proteinase (strain E2004104-TW-CDC) and its complex with rupintrivir.

    PubMed

    Wu, Caiming; Cai, Qixu; Chen, Chen; Li, Ning; Peng, Xuanjia; Cai, Yaxian; Yin, Ke; Chen, Xinsheng; Wang, Xiaolong; Zhang, Rongfu; Liu, Lijie; Chen, Shuhui; Li, Jian; Lin, Tianwei

    2013-05-01

    The crystal structure of 3C proteinase (3C(pro)) from Enterovirus 71 (EV71) was determined in space group C2221 to 2.2?Ĺ resolution. The fold was similar to that of 3C(pro) from other picornaviruses, but the difference in the ?-ribbon reported in a previous structure was not observed. This ?-ribbon was folded over the substrate-binding cleft and constituted part of the essential binding sites for interaction with the substrate. The structure of its complex with rupintrivir (AG7088), a peptidomimetic inhibitor, was also characterized in space group P212121 to 1.96?Ĺ resolution. The inhibitor was accommodated without any spatial hindrance despite the more constricted binding site; this was confirmed by functional assays, in which the inhibitor showed comparable potency towards EV71 3C(pro) and human rhinovirus 3C(pro), which is the target that rupintrivir was designed against. PMID:23633597

  18. First isolation and genomic characterization of enterovirus A71 and coxsackievirus A16 from hand foot and mouth disease patients in the Lao PDR

    PubMed Central

    Nguyen, V H; Sibounheuang, B; Phommasone, K; Vongsouvath, M; Newton, P N; Piorkowski, G; Baronti, C; de Lamballerie, X; Dubot-Pérčs, A

    2014-01-01

    Enterovirus A71 (EV-A71) and coxsackievirus A16 (CV-A16) are major aetiological agents of hand, foot and mouth disease in Asia. We established the first genomic characterization of strains isolated in 2011 from Lao patients. Isolates were related to EV-A71 genotype C4 and CV-A16 genotype B1a that circulated in neighbouring countries during the same period. This confirms the regional character of hand, foot and mouth disease epidemiology and makes plausible the occurrence of severe disease in the Lao population. PMID:25566395

  19. New insights in the regulation of human B cell differentiation

    PubMed Central

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

    2009-01-01

    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 naďve 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. PMID:19447676

  20. Apolipoprotein B synthesis in humans: liver synthesizes only apolipoprotein B-100

    SciTech Connect

    Edge, S.B.; Hoeg, J.M.; Schneider, P.D.; Brewer, H.B. Jr.

    1985-08-01

    Apolipoprotein (apo) B-100 and B-48 are prominent apolipoproteins in VLDL, IDL, and chylomicrons. Organ cultures of normal adult human liver were established to ascertain the form of apo B synthesized by hepatocytes in humans. Human liver was minced and incubated in 15 mL methionine-free RPMI-1640 medium with 10% dialyzed fetal calf serum plus 250 microCi /sup 35/S-methionine for eight hours at 37 degrees C. Lipoproteins secreted by the liver were isolated by ultracentrifugation and the content of newly synthesized apo B determined by quantitation of radioactivity in the apoB-100 and apoB-48 bands after separation by 3% NaDodSO/sub 4/ gel electrophoresis. In the eight-hour period, 2.5% to 3.2% of added /sup 35/S-methionine was secreted in TCA-precipitable protein of which 0.34% was apo B. Ninety-nine percent of the apo B in VLDL, IDL, and LDL was in the apo B-100 electrophoretic band. No significant radioactivity was detected in the apo B-48 electrophoretic band. Eighty-nine percent of the total radioactivity of apo B-100 was in VLDL with 3% and 8% in IDL and LDL, respectively. These results establish that adult human liver in organ culture synthesizes apo B-100 but not apo B-48.

  1. Interleukin 7 independent development of human B cells.

    PubMed Central

    Prieyl, J A; LeBien, T W

    1996-01-01

    Mammalian hematopoietic stem cell (HSC) commitment and differentiation into lymphoid lineage cells proceed through a series of developmentally restricted progenitor compartments. A complete understanding of this process, and how it differs from HSC commitment and differentiation into cells of the myeloid/erythroid lineages, requires the development of model systems that support HSC commitment to the lymphoid lineages. We now describe a human bone marrow stromal cell culture that preferentially supports commitment and differentiation of human HSC to CD19+ B-lineage cells. Fluorescence activated cell sorterpurified CD34++/lineage-cells were isolated from fetal bone marrow and cultured on human fetal bone marrow stromal cells in serum-free conditions containing no exogenous cytokines. Over a period of 3 weeks, CD34++/lineage- cells underwent commitment, differentiation, and expansion into the B lineage. Progressive changes included: loss of CD34, acquisition of and graded increases in the level of cell surface CD19, and appearance of immature B cells expressing mu/kappa or mu/lambda cell surface Ig receptors. The tempo and phenotype of B-cell development was not influenced by the addition of IL-7 (10 ng/ml), or by the addition of goat anti-IL-7 neutralizing antibody. These results indicate a profound difference between mouse and human in the requirement for IL-7 in normal B-cell development, and provide an experimental system to identify and characterize human bone marrow stromal cell-derived molecules crucial for human B lymphopoiesis. PMID:8816803

  2. Human-Automation Interaction By Thomas B. Sheridan & Raja Parasuraman

    E-print Network

    Parasuraman, Raja

    89 CHAPTER 2 Human-Automation Interaction By Thomas B. Sheridan & Raja Parasuraman Automation does with automation in complex and typically large-scale systems, including aircraft and air traffic control, nuclear-free task for either the system designer or the human operator/automation supervisor, especially as computer

  3. John B. Watson and the study of human sexual behavior

    Microsoft Academic Search

    H. W. Magoun

    1981-01-01

    This article reviews the steps in John B. Watson's career from his initial experiments in rat learning, through studies of reflexes in newborn and infant children, to his exploration of adult human sexual behavior. The latter began with questionnaire studies of World War I films on sex hygiene. Watson was next reported to have engaged in laboratory studies of human

  4. A generic assay for whole-genome amplification and deep sequencing of enterovirus A71

    PubMed Central

    Tan, Le Van; Tuyen, Nguyen Thi Kim; Thanh, Tran Tan; Ngan, Tran Thuy; Van, Hoang Minh Tu; Sabanathan, Saraswathy; Van, Tran Thi My; Thanh, Le Thi My; Nguyet, Lam Anh; Geoghegan, Jemma L.; Ong, Kien Chai; Perera, David; Hang, Vu Thi Ty; Ny, Nguyen Thi Han; Anh, Nguyen To; Ha, Do Quang; Qui, Phan Tu; Viet, Do Chau; Tuan, Ha Manh; Wong, Kum Thong; Holmes, Edward C.; Chau, Nguyen Van Vinh; Thwaites, Guy; van Doorn, H. Rogier

    2015-01-01

    Enterovirus A71 (EV-A71) has emerged as the most important cause of large outbreaks of severe and sometimes fatal hand, foot and mouth disease (HFMD) across the Asia-Pacific region. EV-A71 outbreaks have been associated with (sub)genogroup switches, sometimes accompanied by recombination events. Understanding EV-A71 population dynamics is therefore essential for understanding this emerging infection, and may provide pivotal information for vaccine development. Despite the public health burden of EV-A71, relatively few EV-A71 complete-genome sequences are available for analysis and from limited geographical localities. The availability of an efficient procedure for whole-genome sequencing would stimulate effort to generate more viral sequence data. Herein, we report for the first time the development of a next-generation sequencing based protocol for whole-genome sequencing of EV-A71 directly from clinical specimens. We were able to sequence viruses of subgenogroup C4 and B5, while RNA from culture materials of diverse EV-A71 subgenogroups belonging to both genogroup B and C was successfully amplified. The nature of intra-host genetic diversity was explored in 22 clinical samples, revealing 107 positions carrying minor variants (ranging from 0 to 15 variants per sample). Our analysis of EV-A71 strains sampled in 2013 showed that they all belonged to subgenogroup B5, representing the first report of this subgenogroup in Vietnam. In conclusion, we have successfully developed a high-throughput next-generation sequencing-based assay for whole-genome sequencing of EV-A71 from clinical samples. PMID:25704598

  5. A generic assay for whole-genome amplification and deep sequencing of enterovirus A71.

    PubMed

    Tan, Le Van; Tuyen, Nguyen Thi Kim; Thanh, Tran Tan; Ngan, Tran Thuy; Van, Hoang Minh Tu; Sabanathan, Saraswathy; Van, Tran Thi My; Thanh, Le Thi My; Nguyet, Lam Anh; Geoghegan, Jemma L; Ong, Kien Chai; Perera, David; Hang, Vu Thi Ty; Ny, Nguyen Thi Han; Anh, Nguyen To; Ha, Do Quang; Qui, Phan Tu; Viet, Do Chau; Tuan, Ha Manh; Wong, Kum Thong; Holmes, Edward C; Chau, Nguyen Van Vinh; Thwaites, Guy; van Doorn, H Rogier

    2015-04-01

    Enterovirus A71 (EV-A71) has emerged as the most important cause of large outbreaks of severe and sometimes fatal hand, foot and mouth disease (HFMD) across the Asia-Pacific region. EV-A71 outbreaks have been associated with (sub)genogroup switches, sometimes accompanied by recombination events. Understanding EV-A71 population dynamics is therefore essential for understanding this emerging infection, and may provide pivotal information for vaccine development. Despite the public health burden of EV-A71, relatively few EV-A71 complete-genome sequences are available for analysis and from limited geographical localities. The availability of an efficient procedure for whole-genome sequencing would stimulate effort to generate more viral sequence data. Herein, we report for the first time the development of a next-generation sequencing based protocol for whole-genome sequencing of EV-A71 directly from clinical specimens. We were able to sequence viruses of subgenogroup C4 and B5, while RNA from culture materials of diverse EV-A71 subgenogroups belonging to both genogroup B and C was successfully amplified. The nature of intra-host genetic diversity was explored in 22 clinical samples, revealing 107 positions carrying minor variants (ranging from 0 to 15 variants per sample). Our analysis of EV-A71 strains sampled in 2013 showed that they all belonged to subgenogroup B5, representing the first report of this subgenogroup in Vietnam. In conclusion, we have successfully developed a high-throughput next-generation sequencing-based assay for whole-genome sequencing of EV-A71 from clinical samples. PMID:25704598

  6. Characterization of a novel porcine enterovirus in wild boars in Hungary

    PubMed Central

    Boros, Ákos; Nemes, Csaba; Pankovics, Péter; Bíró, Hunor; Kapusinszky, Beatrix; Delwart, Eric; Reuter, Gábor

    2012-01-01

    Porcine enteroviruses (PEVs) are members of the family Picornaviridae, genus Enterovirus. Until now, only three different PEV genotypes (PEV-9 and -10, and PEV-3H/PEV-14) have been detected in domestic pigs, and there is no information about the presence of PEVs in wild animals. Here, we identify and characterize the complete genomes of PEV originated from 5 of 10 (50%) of wild boar (Sus scrofa) piglets by RT-PCR and pyrosequencing. Wild boar/WBD/2011/HUN (JN807387) PEV showed only 67% amino acid identity in VP1 compared to the most closely related prototype PEV-3H/PEV-14. Wild boar enterovirus represents a novel PEV genotype, provisionally called PEV-15. PMID:22350652

  7. Cell Surface Vimentin Is an Attachment Receptor for Enterovirus 71

    PubMed Central

    Du, Ning; Cong, Haolong; Tian, Hongchao; Zhang, Hua; Zhang, Wenliang; Song, Lei

    2014-01-01

    ABSTRACT Enterovirus 71 (EV71) is a highly transmissible pathogenic agent that causes severe central nervous system diseases in infected infants and young children. Here, we reported that EV71 VP1 protein could bind to vimentin intermediate filaments expressed on the host cell surface. Soluble vimentin or an antibody against vimentin could inhibit the binding of EV71 to host cells. Accompanied with the reduction of vimentin expression on the cell surface, the binding of EV71 to cells was remarkably decreased. Further evidence showed that the N terminus of vimentin is responsible for the interaction between EV71 and vimentin. These results indicated that vimentin on the host cell surface may serve as an attachment site that mediated the initial binding and subsequently increased the infectivity of EV71. IMPORTANCE This study delivers important findings on the roles of vimentin filaments in relation to EV71 infection and provides information that not only improves our understanding of EV71 pathogenesis but also presents us with potentially new strategies for the treatment of diseases caused by EV71 infections. PMID:24623428

  8. Antiviral effect of epigallocatechin gallate on enterovirus 71.

    PubMed

    Ho, Hung-Yao; Cheng, Mei-Ling; Weng, Shiue-Fen; Leu, Yann-Lii; Chiu, Daniel Tsun-Yee

    2009-07-22

    Oxidative stress is known to be a determinant of a host's susceptibility to pathogens. Natural compounds with antioxidant activity may provide a preventive measure against infection. Tea polyphenols were evaluated for their ability to inhibit enterovirus 71 (EV71) replication in Vero cell culture. Among the polyphenolic compounds tested, epigallocatechin gallate (EGCG) and gallocatechin gallate (GCG) potently inhibited replication of EV71. EGCG and GCG reduced the titer of infectious progeny virus by 95%. Quantitative RT-PCR analysis also revealed that EGCG suppressed replication of genomic RNA. It was accompanied by an increased cytoprotective effect. EGCG and GCG caused 5-fold increase in the viability of EV71-infected cells. The viral inhibitory effect correlated well with the antioxidant capacity of polyphenol. Mechanistically, EV71 infection led to increased oxidative stress, as shown by increased dichlorofluorescein and MitoSOX Red fluorescence. Upon EGCG treatment, reactive oxygen species (ROS) generation was significantly reduced. Consistent with this, EV71 replication was enhanced in glucose-6-phosphate dehydrogenase deficient cells, and such enhancement was largely reversed by EGCG. These findings suggest that EGCG may suppress viral replication via modulation of cellular redox milieu. PMID:19537794

  9. Very low density lipoprotein apolipoprotein B metabolism in humans

    Microsoft Academic Search

    Th. Demant; J. Shepherd; C. J. Packard

    1988-01-01

    Summary The human plasma lipoproteins encompass a broad spectrum of particles of widely varying physical and chemical properties whose metabolism is directed by their protein components. Apolipoprotein B100 (apo B100) is the major structural protein resident in particles within the Svedberg flotation range 0–400. The largest of these, the very low density lipoprotein (VLDL), rich in triglyceride, are metabolised by

  10. miRNA and tropism of human parvovirus B19.

    PubMed

    Berillo, Olga; Khailenko, Vladimir; Ivashchenko, Anatoly; Perlmuter-Shoshany, Lior; Bolshoy, Alexander

    2012-10-01

    Parvovirus B19 has an extreme tropism for human erythroid progenitors. Here we propose the hypothesis explaining the tropism of human parvovirus B19. Our speculations are based on experimental results related to the capsid proteins VP1 and VP2. These proteins were not detectable in nonpermissive cells in course of these experiments, although the corresponding mRNAs were synthesized. Our interpretation of these results is an inhibition of translation in nonpermissive cells by human miRNAs. We bring support to our hypothesis and propose detailed experimental procedure to test it. PMID:22902951

  11. The Transformation of Enterovirus Replication Structures: a Three-Dimensional Study of Single- and Double-Membrane Compartments

    PubMed Central

    Limpens, Ronald W. A. L.; van der Schaar, Hilde M.; Kumar, Darshan; Koster, Abraham J.; Snijder, Eric J.; van Kuppeveld, Frank J. M.; Bárcena, Montserrat

    2011-01-01

    ABSTRACT All positive-strand RNA viruses induce membrane structures in their host cells which are thought to serve as suitable microenvironments for viral RNA synthesis. The structures induced by enteroviruses, which are members of the family Picornaviridae, have so far been described as either single- or double-membrane vesicles (DMVs). Aside from the number of delimiting membranes, their exact architecture has also remained elusive due to the limitations of conventional electron microscopy. In this study, we used electron tomography (ET) to solve the three-dimensional (3-D) ultrastructure of these compartments. At different time points postinfection, coxsackievirus B3-infected cells were high-pressure frozen and freeze-substituted for ET analysis. The tomograms showed that during the exponential phase of viral RNA synthesis, closed smooth single-membrane tubules constituted the predominant virus-induced membrane structure, with a minor proportion of DMVs that were either closed or connected to the cytosol in a vase-like configuration. As infection progressed, the DMV number steadily increased, while the tubular single-membrane structures gradually disappeared. Late in infection, complex multilamellar structures, previously unreported, became apparent in the cytoplasm. Serial tomography disclosed that their basic unit is a DMV, which is enwrapped by one or multiple cisternae. ET also revealed striking intermediate structures that strongly support the conversion of single-membrane tubules into double-membrane and multilamellar structures by a process of membrane apposition, enwrapping, and fusion. Collectively, our work unravels the sequential appearance of distinct enterovirus-induced replication structures, elucidates their detailed 3-D architecture, and provides the basis for a model for their transformation during the course of infection. PMID:21972238

  12. Use of subgenomic poliovirus DNA hybridization probes to detect the major subgroups of enteroviruses.

    PubMed Central

    Rotbart, H A; Levin, M J; Villarreal, L P

    1984-01-01

    Three nucleic acid hybridization probes were derived from DNA clones of the poliovirus type 1 genome. Used in dot hybridization experiments, the probes successfully detected members of each of the major enteroviral subgroups. The hybridization patterns obtained with the three probes suggested that a highly conserved nucleotide sequence existed among the enteroviruses tested, mapping between bases 220 and 1809 in the poliovirus genome. Two new antiviral agents capable of inhibiting enterovirus replication in tissue culture were used to demonstrate the specificity of the probes for viral RNA. Images PMID:6097599

  13. Human blood IgM "memory" B cells are circulating splenic marginal zone B cells harboring a prediversified immunoglobulin repertoire

    E-print Network

    Boyer, Edmond

    Human blood IgM "memory" B cells are circulating splenic marginal zone B cells harboring authorship Abstract The human peripheral B cell compartment displays a large population of IgM IgD CD27 The human peripheral B cell compartment displays, in contrast to the mouse, a large population of CD27

  14. Recombinant porcine lactoferrin expressed in the milk of transgenic mice protects neonatal mice from a lethal challenge with enterovirus type 71.

    PubMed

    Chen, Hsiao-Ling; Wang, Li-Chung; Chang, Chi-Hsuan; Yen, Chih-Ching; Cheng, Winston T K; Wu, Shinn-Chih; Hung, Che-Ming; Kuo, Meng-Fu; Chen, Chuan-Mu

    2008-02-13

    The human Enterovirus genus of the piconavirus family causes most of the febrile illnesses that affect children during the summer season in Taiwan. Enterovirus type 71 (EV71) plays a key role in patients with hand-foot-and-mouth disease (HFMD) combined with severe paralysis or encephalitis. It is important to find a method for preventing infection with EV71 since there is no antiviral agent or vaccine for humans. In this study, we developed a transgenic mouse model for demonstrating the protective effects of recombinant lactoferrin (LF) against EV71 infection. Transgenic mice carrying alpha-lactalbumin-porcine lactoferrin (alphaLA-pLF) and BALB/c wild-type mice were subjected to EV71 inoculation. First, we analyzed the expression efficiencies of recombinant pLF (rpLF) in hemizygous and homozygous transgenic mice. Following EV71 inoculation on the 4th day of life, pups ingesting transgenic milk showed the significantly higher survival rate and heavier body weight compared to wild-type mice. RT-PCR analysis for EV71 viral RNA showed that the recombinant pLF had a blocking effect on EV71 infection. Our data suggest that oral intake of pLF-enriched milk exhibited the ability to prevent infection with EV71. The study also provides an animal model for validating the protective effects of pLF. PMID:18207613

  15. Human Factors Analysis of Predator B Crash

    E-print Network

    Carrigan, Geoff

    2008-06-10

    The 2006 crash of a Predator B in Arizona has prompted a great amount of scrutiny into Unmanned Arial System (UAS) operations. The direct cause of the Predator crash can be tied to an initial failure of the displays and a ...

  16. Co-Circulation and Genomic Recombination of Coxsackievirus A16 and Enterovirus 71 during a Large Outbreak of Hand, Foot, and Mouth Disease in Central China

    PubMed Central

    Liu, Weiyong; Wu, Shimin; Xiong, Ying; Li, Tongya; Wen, Zhou; Yan, Mingzhe; Qin, Kai; Liu, Yingle; Wu, Jianguo

    2014-01-01

    A total of 1844 patients with hand, foot, and mouth disease (HFMD), most of them were children of age 1–3-year-old, in Central China were hospitalized from 2011 to 2012. Among them, 422 were infected with coxsackievirus A16 (CVA16), 334 were infected with enterovirus 71 (EV71), 38 were co-infected with EV71 and CVA16, and 35 were infected with other enteroviruses. Molecular epidemiology analysis revealed that EV71 and CVA16 were detected year-round, but EV71 circulated mainly in July and CVA16 circulated predominantly in November, and incidence of HFMD was reduced in January and February and increased in March. Clinical data showed that hyperglycemia and neurologic complications were significantly higher in EV71-infected patients, while upper respiratory tract infection and C-reactive protein were significantly higher in CVA16-associated patients. 124 EV71 and 80 CVA16 strains were isolated, among them 56 and 68 EV71 strains were C4a and C4b, while 25 and 55 CVA16 strains were B1a and B1b, respectively. Similarity plots and bootscan analyses based on entire genomic sequences revealed that the three C4a sub-genotype EV71 strains were recombinant with C4b sub-genotype EV71 in 2B–2C region, and the three CVA16 strains were recombinant with EV71 in 2A–2B region. Thus, CVA16 and EV71 were the major causative agents in a large HFMD outbreak in Central China. HFMD incidence was high for children among household contact and was detected year-round, but outbreak was seasonal dependent. CVA16 B1b and EV71 C4b reemerged and caused a large epidemic in China after a quiet period of many years. Moreover, EV71 and CVA16 were co-circulated during the outbreak, which may have contributed to the genomic recombination between the pathogens. It should gain more attention as there may be an upward trend in co-circulation of the two pathogens globally and the new role recombination plays in the emergence of new enterovirus variants. PMID:24776922

  17. Structures of the Procapsid and Mature Virion of Enterovirus 71 Strain 1095

    PubMed Central

    Cifuente, Javier O.; Lee, Hyunwook; Yoder, Joshua D.; Shingler, Kristin L.; Carnegie, Michael S.; Yoder, Jennifer L.; Ashley, Robert E.; Makhov, Alexander M.; Conway, James F.

    2013-01-01

    Enterovirus 71 (EV71) is an important emerging human pathogen with a global distribution and presents a disease pattern resembling poliomyelitis with seasonal epidemics that include cases of severe neurological complications, such as acute flaccid paralysis. EV71 is a member of the Picornaviridae family, which consists of icosahedral, nonenveloped, single-stranded RNA viruses. Here we report structures derived from X-ray crystallography and cryoelectron microscopy (cryo-EM) for the 1095 strain of EV71, including a putative precursor in virus assembly, the procapsid, and the mature virus capsid. The cryo-EM map of the procapsid provides new structural information on portions of the capsid proteins VP0 and VP1 that are disordered in the higher-resolution crystal structures. Our structures solved from virus particles in solution are largely in agreement with those from prior X-ray crystallographic studies; however, we observe small but significant structural differences for the 1095 procapsid compared to a structure solved in a previous study (X. Wang, W. Peng, J. Ren, Z. Hu, J. Xu, Z. Lou, X. Li, W. Yin, X. Shen, C. Porta, T. S. Walter, G. Evans, D. Axford, R. Owen, D. J. Rowlands, J. Wang, D. I. Stuart, E. E. Fry, and Z. Rao, Nat. Struct. Mol. Biol. 19:424–429, 2012) for a different strain of EV71. For both EV71 strains, the procapsid is significantly larger in diameter than the mature capsid, unlike in any other picornavirus. Nonetheless, our results demonstrate that picornavirus capsid expansion is possible without RNA encapsidation and that picornavirus assembly may involve an inward radial collapse of the procapsid to yield the native virion. PMID:23637404

  18. A Mouse-Adapted Enterovirus 71 Strain Causes Neurological Disease in Mice after Oral Infection

    PubMed Central

    Wang, Ya-Fang; Chou, Chun-Ting; Lei, Huan-Yao; Liu, Ching-Chuan; Wang, Shih-Min; Yan, Jing-Jou; Su, Ih-Jen; Wang, Jen-Reng; Yeh, Trai-Ming; Chen, Shun-Hua; Yu, Chun-Keung

    2004-01-01

    A mouse-adapted enterovirus 71 (EV71) strain with increased virulence in mice, MP4, was generated after four serial passages of the parental EV71 strain 4643 in mice. Strain MP4 exhibited a larger plaque size, grew more rapidly, and was more cytotoxic in vitro than strain 4643. Although strains 4643 and MP4 both induced apoptosis of SK-N-SH human neuroblastoma cells, MP4 was more virulent than 4643 in 1-day-old mice (50% lethal doses, 102 and 104 PFU/mouse, respectively). Strain MP4 (5 × 106 PFU/mouse), but not 4643, could orally infect 7-day-old mice, resulting in rear-limb paralysis followed by death 5 to 9 days after inoculation with the virus. Histopathologically, neuronal loss and apoptosis were evident in the spinal cords as well as the brain stems of the infected mice. The limb muscles displayed massive necrosis. There was early and transient virus replication in the intestines, whereas the spinal cord, brain, and muscle became the sites of viral replication during the late phase of the infection. Virus transmission occurred among infected and noninfected cagemates, as demonstrated by the occurrence of seroconversion and the presence of viable viruses in the stool samples of the latter. Protection against EV71 challenge was demonstrated following administration of hyperimmune serum 1 day after inoculation with the virus. Nucleotide sequence analysis of the genome of EV71 strain MP4 revealed four nucleotide changes on the 5? untranslated region, three on the VP2 region, and eight on the 2C region, resulting in one and four amino acid substitutions in the VP2 and 2C proteins, respectively. PMID:15254164

  19. 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...

  20. [Enteroviruses and bacteriophages elimination from wastewater on buildings of Bortnichi aeration station].

    PubMed

    Poniatovs'ky?, V A; Bobyr, V V; Shyrobokov, V P

    2014-01-01

    The research contains data about virologic and molecular-genetic evaluation of sewage samples, obtained in 2010-2011. The efficacy of Bortnichi aeration station in an eliminating of viral agents has been evaluated. An efficiency of PCR as a method of sewage sample analysis has been proved, that results an enchanced evaluation of enteroviruses contamination in different environmental objects. PMID:25000731

  1. Molecular epidemiology of enterovirus d68 from 2013 to 2014 in Philippines.

    PubMed

    Furuse, Yuki; Chaimongkol, Natthawan; Okamoto, Michiko; Imamura, Tadatsugu; Saito, Mariko; Tamaki, Raita; Saito, Mayuko; Lupisan, Socorro P; Oshitani, Hitoshi

    2015-03-01

    Enterovirus D68 (EV-D68) has been recognized as an important cause of acute respiratory infections. Here we report the molecular epidemiology of EV-D68 in Philippines from 2013 to 2014; we found cases in areas affected by Typhoon Haiyan and found new strains in the country. PMID:25568441

  2. Sequence analysis of a porcine enterovirus serotype 1 isolate: relationships with other picornaviruses

    Microsoft Academic Search

    Michelle Doherty; Daniel Todd; Neil McFerran; Elizabeth M. Hoey

    1999-01-01

    The majority of the genomic sequence of a porcine enterovirus serotype 1 (PEV-1) isolate was determined. The genome was found to contain a large open reading frame which encoded a leader protein prior to the capsid protein region. This showed no sequence identity to other picornavirus leader regions and the sequence data suggested that it does not possess proteolytic activity.

  3. RAPID PCR-BASED MONITORING OF INFECTIOUS ENTEROVIRUSES IN DRINKING WATER. (R824756)

    EPA Science Inventory

    Abstract Currently, the standard method for the detection of enteroviruses and hepatitis A virus in water involves cell culture assay which is expensive and time consuming. Direct RT-PCR offers a rapid and sensitive alternative to virus detection but sensitivity is oft...

  4. Development of a quantitative method for the detection of enteroviruses in soil.

    PubMed Central

    Hurst, C J; Gerba, C P

    1979-01-01

    A method is described for efficiently concentrating enteroviruses from soil. Viruses were eluted from soil by mechanical agitation in high pH glycine buffer containing ethylenediaminetetraacetic acid. The eluted viruses were concentrated on a floc that formed de novo upon adjustment of the soil eluate to 0.06 M aluminum chloride and pH 3.5. Viruses not pelleted with the floc were concentrated by adsorption to and elution from membrane filters. This method yielded an average efficiency of 66% recovery from loamy sand soil for four enteroviruses. Virus recovery from soil was consistently high, with samples ranging in size from 25 to 500 g. The method was used successfully to isolate naturally occurring viruses from soil beneath a wastewater land treatment site. Recovery of enteroviruses by this method form different types of soil was dependent on percentage of clay, surface area, and cation exchange capacity. Recovery was not dependent on soil saturation pH or on percentage of organic matter. This method should prove useful for studying enterovirus migration and survival during the land application of domestic sewage. PMID:36845

  5. Broad-range inhibition of enterovirus replication by OSW-1, a natural compound targeting OSBP.

    PubMed

    Albulescu, Lucian; Strating, Jeroen R P M; Thibaut, Hendrik Jan; van der Linden, Lonneke; Shair, Matthew D; Neyts, Johan; van Kuppeveld, Frank J M

    2015-05-01

    Enteroviruses, e.g., polio-, coxsackie- and rhinoviruses, constitute a large genus within the Picornaviridae family of positive-strand RNA viruses and include many important pathogens linked to a variety of acute and chronic diseases. Despite their huge medical and economic impact, no approved antiviral therapy is yet available. Recently, the oxysterol-binding protein (OSBP) was implicated as a host factor for enterovirus replication. Here, we investigated the antiviral activity of the natural compound OSW-1, a ligand of OSBP that is under investigation as an anti-cancer drug. OSW-1 potently inhibited the replication of all enteroviruses tested, with IC50 values in the low nanomolar range, acted at the genome replication stage and was effective in all tested cell types of three different species. Importantly, OSBP overexpression rescued viral replication, demonstrating that the antiviral effect of OSW-1 is due to targeting OSBP. Together, we here report the anti-enterovirus activity of the natural anti-cancer compound OSW-1. PMID:25752737

  6. Nuclear compartmentation of glycoprotein B of human cytomegalovirus.

    PubMed

    Radsak, K; Brücher, K H; Britt, W; Shiou, H; Schneider, D; Kollert, A

    1990-08-01

    Maturation of glycoprotein B (gB) of human cytomegalovirus (HCMV) includes a series of sequential glycosylation steps followed by proteolytic cleavage of the precursor protein. Inhibitors of glycosylation and glycoprotein processing, including tunicamycin, monensin, and bromoconduritol, were used to define further the processing pathway of HCMV gB. The results of these studies indicated that cotranslational glycosylation and intracellular transport are essential for subsequent cleavage of the precursor; early trimming in the endoplasmic reticulum is not a prerequisite but facilitates this processing event. Analysis of purified nuclei with gB-specific monoclonal antibody suggested that the mannose-rich gB-precursor intermediate(s) is (are) compartmentalized in the nuclear fraction. Immunoelectron microscopy revealed that HCMV gB was localized in the outer as well as in the inner nuclear membranes of HCMV-infected fibroblasts. PMID:2115221

  7. Exploiting Human Memory B Cell Heterogeneity for Improved Vaccine Efficacy

    PubMed Central

    Pauli, Noel T.; Henry Dunand, Carole J.; Wilson, Patrick C.

    2011-01-01

    The major goal in vaccination is establishment of long-term, prophylactic humoral memory to a pathogen. Two major components to long-lived humoral memory are plasma cells for the production of specific immunoglobulin and memory B cells that survey for their specific antigen in the periphery for later affinity maturation, proliferation, and differentiation. The study of human B cell memory has been aided by the discovery of a general marker for B cell memory, expression of CD27; however, new data suggests the existence of CD27? memory B cells as well. These recently described non-canonical memory populations have increasingly pointed to the heterogeneity of the memory compartment. The novel B memory subsets in humans appear to have unique origins, localization, and functions compared to what was considered to be a “classical” memory B cell. In this article, we review the known B cell memory subsets, the establishment of B cell memory in vaccination and infection, and how understanding these newly described subsets can inform vaccine design and disease treatment. PMID:22566866

  8. Differentiation of normal human pre-B cells in vitro

    PubMed Central

    1990-01-01

    The differentiation of surface Ig- pre-B cells into surface Ig+ B cells is a critical transition in mammalian B cell ontogeny. Elucidation of the growth factor requirements and differentiative potential of human pre-B cells has been hampered by the absence of a reproducible culture system that supports differentiation. Fluorescence-activated cell sorting and magnetic bead depletion were used to purify fetal bone marrow CD10+/surface mu- cells, which contain 60-70% cytoplasmic mu+ pre-B cells. CD10+/surface mu- cells cultured for 2 d were observed to differentiate into surface mu+ cells. Analysis by Southern blotting provided direct evidence that rearrangement of kappa light chain genes occurs in culture, and flow cytometric analysis revealed the appearance of surface Ig+ B cells expressing mu/kappa or mu/lambda. Unexpectedly, the kappa/lambda ratio in differentiated cells was the inverse of what is normally observed in adult peripheral blood. Differentiation occurs in the absence of exogenous growth factors or cytokines, suggesting that a stimulus-independent differentiative inertia might characterize pre-B cells in vivo. Future use of this model will facilitate our understanding of normal and abnormal human pre-B cell differentiation. PMID:2141629

  9. Expression of Cholesterol Sulfotransferase (SULT2B1b) in Human Skin and Primary Cultures of Human Epidermal Keratinocytes

    Microsoft Academic Search

    Yuko Higashi; Hirotoshi Fuda; Hidekatsu Yanai; Young Lee; Tomoko Fukushige; Tamotsu Kanzaki; Charles A. Strott

    2004-01-01

    Cholesterol sulfate is a highly amphipathic molecule that is present in a relatively high concentration in the epidermis of human skin, particularly in the granular layer. The physiologic significance of this finding, however, is not well-understood. Therefore, we examined expression of the gene encoding for the enzyme that sulfonates cholesterol (SULT2B1b). Of the three enzymes known to sulfonate steroids\\/sterols, only

  10. Characterization of the human B cell stimulatory factor 1 receptor

    PubMed Central

    1987-01-01

    125I-labeled recombinant human B cell stimulatory factor 1 (BSF-1) was used to characterize receptors specific for this lymphokine on in vitro cell lines representing human B, T, and hematopoietic lineages, as well as on adherent cell lines of epithelial and endothelial origin, and on primary human gingival fibroblasts. BSF-1 binding was extremely rapid and saturable at both 4 and 37 degrees C, with a slow dissociation rate. On all human cell types examined, BSF-1 bound to a single class of high-affinity receptor (less than 3,000 receptors per cell) with a Ka of 0.5-1.0 X 10(10)/M. Human BSF-1 also bound to cell lines of simian but not murine origin. Comparison of kinetic characteristics obtained with a yeast-derived hyperglycosylated form of BSF-1 (Mr 60,000) and N-glycanase-treated, sugar-free BSF-1 (Mr 15,000) showed no significant differences. Among a panel of lymphokines and growth hormones, only unlabeled human BSF-1 was able to compete for the binding of 125I-labeled human BSF-1. Affinity crosslinking experiments resulted in the identification on both Raji cells and on primary human gingival fibroblasts of a receptor subunit with an average Mr of 139,000. These studies show that the BSF-1 receptor on human cells has an extremely broad cellular distribution, while further supporting the notion that the ability of BSF-1 to mediate a spectrum of biological activities cannot be accounted for by overt differences in the receptor for this lymphokine on different cell lineages. PMID:3496417

  11. Human cytochrome b5 reductase: structure, function, and potential applications.

    PubMed

    Elahian, Fatemeh; Sepehrizadeh, Zargham; Moghimi, Bahareh; Mirzaei, Seyed Abbas

    2014-06-01

    Cytochrome b5 reductase is a flavoprotein that is produced as two different isoforms that have different localizations. The amphipathic microsomal isoform, found in all cell types with the exception of erythrocytes, consists of one hydrophobic membrane-anchoring domain and a larger hydrophilic flavin catalytic domain. The soluble cytochrome b5 reductase isoform, found in human erythrocytes, is a truncated protein that is encoded by an alternative transcript and consists of the larger domain only. Cytochrome b5 reductase is involved in the transfer of reducing equivalents from the physiological electron donor, NADH, via an FAD domain to the small molecules of cytochrome b5. This protein has received much attention from researchers due to its involvement in many oxidation and reduction reactions, such as the reduction of methemoglobin to hemoglobin. Autosomal cytochrome b5 reductase gene deficiency manifests with the accumulation of oxidized Fe+3 and recessive congenital methemoglobinemia in humans. In this article, we provide a comprehensive overview of the structure and function of cytochrome b5 reductase from different eukaryotic sources and its potential use in the food industry, biosensor, and diagnostic areas. PMID:23113554

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

    PubMed

    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

    2012-01-01

    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

  13. Enterovirus 71 3C Inhibits Cytokine Expression through Cleavage of the TAK1/TAB1/TAB2/TAB3 Complex

    PubMed Central

    Lei, Xiaobo; Han, Ning; Xiao, Xia; Jin, Qi

    2014-01-01

    ABSTRACT Enterovirus 71 (EV71) causes hand, foot, and mouth disease in young children and infants. Severe infection with EV71 can lead to various neurological complications or fatal diseases. However, the mechanism of EV71 pathogenesis is poorly understood. Emerging evidence suggests that EV71 modulates type I interferon (IFN) and cytokine responses. Here, we show that EV71 disables components of the TAB2 complex through the 3C protein. When expressed in mammalian cells, EV71 3C interacts with TAB2 and TAK1, which inhibits NF-?B activation. Furthermore, 3C mediates cleavage of TAB2 and its partners, which requires the protease activity. H40D or C147S substitution in the 3C active sites abolishes its activity, whereas R84Q or V154S substitution in the RNA binding domain has no effect. The 3C protein targets TAB2 at Q113-S114, TAK1 at Q360-S361, TAB1 both at Q414-G415 and Q451-S452, and TAB3 at Q173-G174 and Q343-G344. Importantly, overexpression of TAB2 inhibits EV71 replication, whereas addition of cleaved fragments has no effect. Thus, an equilibrium between the TAB2 complex and EV71 3C represents a control point of viral infection. These results suggest that TAK1/TAB1/TAB2/TAB3 cleavage mediated by EV71 may be a mechanism to interfere with inflammatory responses. IMPORTANCE The TAK1 complex plays a critical role in the activation of NF-?B and cytokine production. However, little is known about its connection to enterovirus 71 (EV71). We demonstrate that EV71 3C suppresses cytokine expression via cleavage of the TAK1 complex proteins. EV71 3C interacts with TAB2 and TAK1. Furthermore, overexpression of TAB2 inhibits EV71 replication, whereas addition of cleaved fragment has no effect. These results suggest that the interplay of EV71 and the TAK1 complex influences the outcome of viral infection. PMID:24942571

  14. Lymphoproliferative responses after infection with human parvovirus B19.

    PubMed Central

    von Poblotzki, A; Gerdes, C; Reischl, U; Wolf, H; Modrow, S

    1996-01-01

    Immunity after infection with the parvovirus B19 is assumed to be conferred by a humoral immune response with development of neutralizing antibody. In contrast, little is known about the nature of T-cell-mediated responses to parvovirus B19 infection in humans. We used recombinant proteins VP1, VP2, and NS1, as well as a recombinant VP1-specific amino-terminal sequence, to test the proliferative responses of peripheral blood mononuclear cells after infection of otherwise healthy individuals with parvovirus B19. These proteins were used as antigens for the stimulation of freshly isolated cells. The results show that a B19 virus-specific cellular immunity develops that is directed against the capsid proteins VP1 and VP2. We also demonstrate that viral determinants are presented to CD4+ T cells by HLA class II molecules. PMID:8794392

  15. The variations of VP1 protein might be associated with nervous system symptoms caused by enterovirus 71 infection

    PubMed Central

    2014-01-01

    Background The VP1 protein of enterovirus 71 (EV71) is an important immunodominant protein which is responsible for host-receptor binding. Nevertheless, the relationship between VP1 and neurovirulence is still poorly understood. In this study, we investigated the relationship between mutation of VP1 and neurovirulent phenotype of EV71 infection. Methods One hundred and eighty-seven strains from Genbank were included, with a clear clinical background. They were divided into two groups, one with nervous system symptoms and one with no nervous system symptoms. After alignment, the significance of amino acid variation was determined by using the ?2 test and a phylogenetic tree was constructed with MEGA software (version 5.1). Results We showed no significant difference in neurovirulence between genotype B and C. Interestingly, we found that variations of E145G/Q, E164D/K and T292N/K were associated with nervous system infection in genotype B. In the case of genotype C, the N31D mutation increased the risk for nervous complications, whereas I262V mutation decreased the risk of nervous complications. We used a 3D model of VP1 to demonstrate the potential molecular basis for EV71 nervous system tropism. Conclusions Distinct variations are shown to be associated with neurovirulent phenotype in the different genotype. Detection of variation in genotypes and subtypes may be important for the prediction of clinical outcomes. PMID:24886383

  16. Seroepidemiology of human parvovirus B19 in Taiwan.

    PubMed

    Lin, K H; You, S L; Chen, C J; Wang, C F; Yang, C S; Yamazaki, S

    1999-02-01

    In order to determine the prevalence and risk factors of human parvovirus B19 (B19) infection in Taiwan, a seroepidemiological study was carried out in 19 townships. Serum samples were collected from 862 healthy residents, who were selected by stratified random sampling from various study areas. They were chosen from four different ethnic groups including aborigines, Fukien Taiwanese, Hakka Taiwanese, and mainland Chinese. Serum samples were screened for B19 IgG antibody by indirect antibody capture enzyme-linked immunosorbent assay (ELISA) and B19 IgM by IgM antibody capture (MAC)-ELISA, respectively. The overall prevalence of anti-B19 IgG and anti-B19 IgM was 32.8% and 0.35%, respectively. The anti-B19 seropositive rate in females was significantly higher than that of males (36.4% vs. 29.4%, P < .001). The age-sex-adjusted seropositive rate in urban townships (39.9%) was higher than that in aboriginal townships (30.5%, P < .001). The seropositive rate increased significantly with age showing a dose-response relationship (P=0.0001 based on a trend test). Blood transfusion was found to be associated with an increased seropositive rate showing a multivariate-adjusted odds ratios of 1.6. PMID:9892403

  17. Characterization of the human b-globin downstream promoter region

    Microsoft Academic Search

    Kelly M. Leach; Karen F. Vieira; Sung-Hae Lee Kang; Ara Aslanian; Martin Teichmann; Robert G. Roeder; Jorg Bungert

    2003-01-01

    The human b-globin gene is abundantly expressed specifically in adult erythroid cells. Stage-specific transcription is regulated principally by promoter proximal cis-regulatory elements. The basal pro- moter contains a non-canonical TATA-like motif as well as an initiator element. These two elements have been shown to interact with the TFII-D com- plex. Here we show that in addition to the TATA and

  18. Human B cells differentiate into granzyme B-secreting cytotoxic B lymphocytes upon incomplete T-cell help.

    PubMed

    Hagn, Magdalena; Sontheimer, Kai; Dahlke, Karen; Brueggemann, Sabine; Kaltenmeier, Christof; Beyer, Thamara; Hofmann, Stefanie; Lunov, Oleg; Barth, Thomas F E; Fabricius, Dorit; Tron, Kyrylo; Nienhaus, Gerd Ulrich; Simmet, Thomas; Schrezenmeier, Hubert; Jahrsdörfer, Bernd

    2012-04-01

    Recently, CD4(+) T helper cells were shown to induce differentiation of human B cells into plasma cells by expressing interleukin (IL-)21 and CD40 ligand (CD40L). In the present study we show, that in the absence of CD40L, CD4(+) T cell-derived IL-21 induces differentiation of B cells into granzyme B (GzmB)-secreting cytotoxic cells. Using fluorescence-activated cell sorting (FACS) analysis, ELISpot and confocal microscopy, we demonstrate that CD4(+) T cells, activated via their T-cell receptor without co-stimulation, can produce IL-21, but do not express CD40L and rapidly induce GzmB in co-cultured B cells in an IL-21 receptor-dependent manner. Of note, we confirmed these results with recombinant reagents, highlighting that CD40L suppresses IL-21-induced GzmB induction in B cells in a dose-dependent manner. Surprisingly, although GzmB-secreting B cells did not express perforin, they were able to transfer active GzmB to tumor cell lines, thereby effectively inducing apoptosis. In contrast, no cytotoxic effects were found when effector B cells were activated with IL-2 instead of IL-21 or when target cells were cultured with IL-21 alone. Our findings suggest GzmB(+) cytotoxic B cells may have a role in early cellular immune responses including tumor immunosurveillance, before fully activated, antigen-specific cytotoxic T cells are on the spot. CD40 ligand determines whether IL-21 induces differentiation of B cells into plasma cells or into granzyme B-secreting cytotoxic cells. PMID:21808264

  19. Detection of naturally occurring enteroviruses in waters by reverse transcription, polymerase chain reaction, and hybridization.

    PubMed Central

    Kopecka, H; Dubrou, S; Prevot, J; Marechal, J; López-Pila, J M

    1993-01-01

    Comparison in virus-seeded mineral water of three detection methods for enteroviruses, direct hybridization, cell culture, and reverse transcription into cDNA followed by polymerase chain reaction and hybridization, showed that the last procedure was 10 to 1,000 times more sensitive than detection by cell culture and 10(5) to 10(7) times more sensitive than direct hybridization. The presence of naturally occurring enteroviruses was also demonstrated in activated sludge and in concentrated and non-concentrated surface water samples by reverse transcription-polymerase chain reaction-hybridization. However, in activated sludge and in concentrated surface waters, enzymatic amplification was sometimes inhibited by contaminants. Images PMID:7683857

  20. First Identification and Characterization of Porcine Enterovirus G in the United States

    PubMed Central

    Anbalagan, Srivishnupriya; Hesse, Richard A.; Hause, Ben M.

    2014-01-01

    Porcine enterovirus G (EV-G) is a member of the family Picornavirdae, genus Enterovirus. To date, eleven EV-G types (EV-G1 through EV-G11) have been identified in pigs from Asia and Europe however they have never been reported in North America. In this study, we isolated and characterized the complete genome of NP/2013/USA, an EV-G from a porcine diarrhea sample from the United States. The complete genome consists of 7,390 nucleotides excluding the 3? poly(A) tail, and has an open reading frame that encodes a 2,169 amino acid polyprotein. NP/2013/USA was most similar at the nucleotide (84%) and amino acid (95%) level to the HM131607, an EV-G1 type isolated from China in 2012. PMID:24824640

  1. A helix-turn-helix structure unit in human centromere protein B (CENP-B).

    PubMed Central

    Iwahara, J; Kigawa, T; Kitagawa, K; Masumoto, H; Okazaki, T; Yokoyama, S

    1998-01-01

    CENP-B has been suggested to organize arrays of centromere satellite DNA into a higher order structure which then directs centromere formation and kinetochore assembly in mammalian chromosomes. The N-terminal portion of CENP-B is a 15 kDa DNA binding domain (DBD) consisting of two repeating units, RP1 and RP2. The DBD specifically binds to the CENP-B box sequence (17 bp) in centromere DNA. We determined the solution structure of human CENP-B DBD RP1 by multi-dimensional 1H, 13C and 15N NMR methods. The CENP-B DBD RP1 structure consists of four helices and has a helix-turn-helix structure. The overall folding is similar to those of some other eukaryotic DBDs, although significant sequence homology with these proteins was not found. The DBD of yeast RAP1, a telomere binding protein, is most similar to CENP-B DBD RP1. We studied the interaction between CENP-B DBD RP1 and the CENP-B box by the use of NMR chemical shift perturbation. The results suggest that CENP-B DBD RP1 interacts with one of the essential regions of the CENP-B box DNA, mainly at the N-terminal basic region, the N-terminal portion of helix 2 and helix 3. PMID:9451007

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

    PubMed Central

    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.

    2013-01-01

    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

  3. Enterovirus isolation from children with acute respiratory infections and presumptive identification by a modified microplate method

    Microsoft Academic Search

    Katsumi Mizuta; Chieko Abiko; Hiroko Goto; Toshio Murata; Shoko Murayama

    2003-01-01

    Objective: To evaluate a modified microplate method, utilizing HEF, HEp-2, Vero, MDCK and newly introduced RD-18S and GMK cell lines, for virus isolation.Methods: From June to October 2001, 723 throat swab specimens taken from children with acute respiratory infections (ARIs) were inoculated onto these cells. To analyze cell sensitivity, we also inoculated 20 serotypes of stocked enteroviruses.Results: During the period,

  4. [Preparation of human parvovirus B19 virus-like particles].

    PubMed

    Zou, Xiaohui; Dong, Liuxin; Song, Jingdong; Qu, Jianguo; Yu, Xiuping; Lu, Zhuozhuang; Hong, Tao

    2009-04-01

    The baculovirus expression system was employed to prepare the virus-like particles (VLPs) of human parvovirus B19. The synthesized VP2 gene of B19 was inserted into the multi-cloning site (MCS) of pFastBac1 vector; the resulting plasmid was transferred to the Escherichia coli DH10Bac competent cells, which contain a baculovirus shuttle vector (Bacmid), to generate Bacmid-VP2 by site-specific transposition. Recombinant baculovirus carrying VP2 gene (rBac-VP2) was then rescued from Bacmid-VP2-transfected Sf9 cells. Indirect immunofluorescence and Western blotting were used to identify the VP2 protein in rBac-VP2-infected Sf9 cells, and the VLPs were observed under transmission electron microscope after being enriched by ultracentrifugation. The B19 VLPs were successfully produced in insect cells with baculovirus expression system, which will facilitate the development of diagnostic reagents to detect the antibody against B19 virus in human serum. PMID:19637634

  5. The contrasting phylodynamics of human influenza B viruses.

    PubMed

    Vijaykrishna, Dhanasekaran; Holmes, Edward C; Joseph, Udayan; Fourment, Mathieu; Su, Yvonne C F; Halpin, Rebecca; Lee, Raphael T C; Deng, Yi-Mo; Gunalan, Vithiagaran; Lin, Xudong; Stockwell, Timothy B; Fedorova, Nadia B; Zhou, Bin; Spirason, Natalie; Kühnert, Denise; Bošková, Veronika; Stadler, Tanja; Costa, Anna-Maria; Dwyer, Dominic E; Huang, Q Sue; Jennings, Lance C; Rawlinson, William; Sullivan, Sheena G; Hurt, Aeron C; Maurer-Stroh, Sebastian; Wentworth, David E; Smith, Gavin J D; Barr, Ian G

    2015-01-01

    A complex interplay of viral, host, and ecological factors shapes the spatio-temporal incidence and evolution of human influenza viruses. Although considerable attention has been paid to influenza A viruses, a lack of equivalent data means that an integrated evolutionary and epidemiological framework has until now not been available for influenza B viruses, despite their significant disease burden. Through the analysis of over 900 full genomes from an epidemiological collection of more than 26,000 strains from Australia and New Zealand, we reveal fundamental differences in the phylodynamics of the two co-circulating lineages of influenza B virus (Victoria and Yamagata), showing that their individual dynamics are determined by a complex relationship between virus transmission, age of infection, and receptor binding preference. In sum, this work identifies new factors that are important determinants of influenza B evolution and epidemiology. PMID:25594904

  6. Concentration and purification of enterovirus 71 using a weak anion-exchange monolithic column

    PubMed Central

    2014-01-01

    Background Enterovirus 71 (EV-71) is a neurotropic virus causing Hand, Foot and Mouth Disease (HFMD) in infants and children under the age of five. It is a major concern for public health issues across Asia-Pacific region. The most effective way to control the disease caused by EV-71 is by vaccination thus a novel vaccine is urgently needed. Inactivated EV-71 induces a strong, virus-neutralizing antibody response in animal models, protecting them against a lethal EV-71 challenge and it has been shown to elicit cross-neutralizing antibodies in human trials. Hence, the large-scale production of purified EV-71 is required for vaccine development, diagnosis and clinical trials. Methods CIM® Monolith columns are single-piece columns made up of poly(glycidyl methacrylate co-ethylene dimethacrylate) as support matrix. They are designed as porous channels rather than beads with different chemistries for different requirements. As monolithic columns have a high binding capacity, flow rate and resolution, a CIM® DEAE-8f tube monolithic column was selected for purification in this study. The EV-71 infected Rhabdomyosarcoma (RD) cell supernatant was concentrated using 8% PEG 8000 in the presence of 400 mM sodium chloride. The concentrated virus was purified by weak anion exchange column using 50 mM HEPES?+?1 M sodium chloride as elution buffer. Results Highly pure viral particles were obtained at a concentration of 350 mM sodium chloride as confirmed by SDS-PAGE and electron microscopy. Presence of viral proteins VP1, VP2 and VP3 was validated by western blotting. The overall process achieved a recovery of 55%. Conclusions EV-71 viral particles of up to 95% purity can be recovered by a single step ion-exchange chromatography using CIM-DEAE monolithic columns and 1 M sodium chloride as elution buffer. Moreover, this method is scalable to purify several litres of virus-containing supernatant, using industrial monolithic columns with a capacity of up to 8 L such as CIM® cGMP tube monolithic columns. PMID:24884895

  7. High sensitivity and label-free detection of Enterovirus 71 by nanogold modified electrochemical impedance spectroscopy

    NASA Astrophysics Data System (ADS)

    Wang, Fang-Yu; Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Yang, Jyh-Yuan; Chang, Chia-Ching

    2013-03-01

    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.

  8. Recombinant adeno-vaccine expressing enterovirus 71-like particles against hand, foot, and mouth disease.

    PubMed

    Tsou, Yueh-Liang; Lin, Yi-Wen; Shao, Hsiao-Yun; Yu, Shu-Ling; Wu, Shang-Rung; Lin, Hsiao-Yu; Liu, Chia-Chyi; Huang, Chieh; Chong, Pele; Chow, Yen-Hung

    2015-04-01

    Enterovirus 71 (EV71) and coxsackieviruses (CV) are the major causative agents of hand, foot and mouth disease (HFMD). There is not currently a vaccine available against HFMD, even though a newly developed formalin-inactivated EV71 (FI-EV71) vaccine has been tested in clinical trial and has shown efficacy against EV71. We have designed and genetically engineered a recombinant adenovirus Ad-EVVLP with the EV71 P1 and 3CD genes inserted into the E1/E3-deleted adenoviral genome. Ad-EVVLP were produced in HEK-293A cells. In addition to Ad-EVVLP particles, virus-like particles (VLPs) formed from the physical association of EV71 capsid proteins, VP0, VP1, and VP3 expressed from P1 gene products. They were digested by 3CD protease and confirmed to be produced by Ad-EVVLP-producing cells, as determined using transmission electron microscopy and western blotting. Mouse immunogenicity studies showed that Ad-EVVLP-immunized antisera neutralized the EV71 B4 and C2 genotypes. Activation of VLP-specific CD4+ and CD8+/IFN-? T cells associated with Th1/Th2-balanced IFN-?, IL-17, IL-4, and IL-13 was induced; in contrast, FI-EV71 induced only Th2-mediated neutralizing antibody against EV71 and low VLP-specific CD4+ and CD8+ T cell responses. The antiviral immunity against EV71 was clearly demonstrated in mice vaccinated with Ad-EVVLP in a hSCARB2 transgenic (hSCARB2-Tg) mouse challenge model. Ad-EVVLP-vaccinated mice were 100% protected and demonstrated reduced viral load in both the CNS and muscle tissues. Ad-EVVLP successfully induced anti-CVA16 immunities. Although antisera had no neutralizing activity against CVA16, the 3C-specific CD4+ and CD8+/IFN-? T cells were identified, which could mediate protection against CVA16 challenge. FI-EV71 did not induce 3C-mediated immunity and had no efficacy against the CVA16 challenge. These results suggest that Ad-EVVLP can enhance neutralizing antibody and protective cellular immune responses to prevent EV71 infection and cellular immune responses against CV infection. PMID:25855976

  9. Interactive Domains in the Molecular Chaperone Human aB Crystallin Modulate Microtubule Assembly and

    E-print Network

    Clark, John

    Interactive Domains in the Molecular Chaperone Human aB Crystallin Modulate Microtubule Assembly. Interactive sequences on the surface of human aB crystallin collectively modulate microtubule assembly through Chaperone Human aB Crystallin Modulate Microtubule Assembly and Disassembly. PLoS ONE 2(6): e498. doi:10

  10. Novel human recombinant antibodies against Mycobacterium tuberculosis antigen 85B

    PubMed Central

    2014-01-01

    Background Tuberculosis is the leading cause of death due to bacterial infections worldwide, mainly caused by Mycobacterium tuberculosis. The antigen 85 complex comprises a set of major secreted proteins of M. tuberculosis, which are potential biomarkers for diagnostic. Results In this work, the first human single chain fragment variable (scFv) antibodies specific for the tuberculosis biomarker 85 B were selected by phage display from naďve antibody gene libraries (HAL7/8). Produced as scFv-Fc in mammalian cells, these antibodies were further characterized and analysed for specificity and applicability in different tuberculosis antigen detection assays. Sandwich detection of recombinant 85 B was successful in enzyme linked immunosorbent assay (ELISA), lateral flow immunoassay and immunoblot. Whereas detection of M. tuberculosis cell extracts and culture filtrates was only possible in direct ELISA and immunoblot assays. It was found that the conformation of 85 B, depending on sample treatment, influenced antigen detection. Conclusions Recombinant antibodies, selected by phage display, may be applicable for 85 B detection in various assays. These antibodies are candidates for the development of future point of care tuberculosis diagnostic kits. Using 85 B as a biomarker, the antigen conformation influenced by sample treatment is important. PMID:25033887

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

    PubMed Central

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

    2008-01-01

    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. PMID:16519509

  12. Hepatitis B and human immunodeficiency virus co-infection.

    PubMed

    Phung, Bao-Chau; Sogni, Philippe; Launay, Odile

    2014-12-14

    Hepatitis B and human immunodeficiency virus (HBV and HIV) infection share transmission patterns and risk factors, which explains high prevalence of chronic HBV infection in HIV infected patients. The natural course of HBV disease is altered by the HIV infection with less chance to clear acute HBV infection, faster progression to cirrhosis and higher risk of liver-related death in HIV-HBV co-infected patients than in HBV mono-infected ones. HIV infected patients with chronic hepatitis B should be counseled for liver damage and surveillance of chronic hepatitis B should be performed to screen early hepatocellular carcinoma. Noninvasive tools are now available to evaluate liver fibrosis. Isolated hepatitis B core antibodies (anti-HBc) are a good predictive marker of occult HBV infection. Still the prevalence and significance of occult HBV infection is controversial, but its screening may be important in the management of antiretroviral therapy. Vaccination against HBV infection is recommended in non-immune HIV patients. The optimal treatment for almost all HIV-HBV co-infected patients should contain tenofovir plus lamivudine or emtricitabine and treatment should not be stopped to avoid HBV reactivation. Long term tenofovir therapy may lead to significant decline in hepatitis B surface Antigen. The emergence of resistant HBV strains may compromise the HBV therapy and vaccine therapy. PMID:25516647

  13. Histone Demethylases KDM4B and KDM6B Promotes Osteogenic Differentiation Of Human MSCs

    PubMed Central

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

    2012-01-01

    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. PMID:22770241

  14. Human Health Countermeasures (HHC) Element Management Plan: Human Research Program. Revision B

    NASA Technical Reports Server (NTRS)

    Norsk, Peter; Baumann, David

    2012-01-01

    NASA s Human Research Program (HRP) is an applied research and technology program within the Human Exploration and Operations Mission Directorate (HEOMD) that addresses human health and performance risk mitigation strategies in support of exploration missions. The HRP research and technology development is focused on the highest priority risks to crew health and safety with the goal of ensuring mission success and maintaining long-term crew health. Crew health and performance standards, defined by the NASA Chief Health and Medical Officer (CHMO), set the acceptable risk level for exploration missions. The HRP conducts research to inform these standards as well as provide deliverables, such as countermeasures, that ensure standards can be met to maximize human performance and mission success. The Human Health Countermeasures (HHC) Element was formed as part of the HRP to develop a scientifically-based, integrated approach to understanding and mitigating the health risks associated with human spaceflight. These health risks have been organized into four research portfolios that group similar or related risks. A fifth portfolio exists for managing technology developments and infrastructure projects. The HHC Element portfolios consist of: a) Vision and Cardiovascular; b) Exercise and Performance; c) Multisystem; d) Bone; and e) Technology and Infrastructure. The HHC identifies gaps associated with the health risks and plans human physiology research that will result in knowledge required to more fully understand risks and will result in validated countermeasures to mitigate risks.

  15. Human parvovirus PARV4 DNA in tissues from adult individuals: a comparison with human parvovirus B19 (B19V)

    PubMed Central

    2010-01-01

    Background PARV4 is a new member of the Parvoviridae family not closely related to any of the known human parvoviruses. Viremia seems to be a hallmark of PARV4 infection and viral DNA persistence has been demonstrated in a few tissues. Till now, PARV4 has not been associated with any disease and its prevalence in human population has not been clearly established. This study was aimed to assess the tissue distribution and the ability to persist of PARV4 in comparison to parvovirus B19 (B19V). Results PARV4 and B19V DNA detection was carried out in various tissues of individuals without suspect of acute viral infection, by a real time PCR and a nested PCR, targeting the ORF2 and the ORF1 respectively. Low amount of PARV4 DNA was found frequently (>40%) in heart and liver of adults individuals, less frequently in lungs and kidneys (23,5 and 18% respectively) and was rare in bone marrow, skin and synovium samples (5,5%, 4% and 5%, respectively). By comparison, B19V DNA sequences were present in the same tissues with a higher frequency (significantly higher in myocardium, skin and bone marrow) except than in liver where the frequency was the same of PARV4 DNA and in plasma samples where B19V frequency was significantly lower than that of PARV4 Conclusions The particular tropism of PARV4 for liver and heart, here emerged, suggests to focus further studies on these tissues as possible target for viral replication and on the possible role of PARV4 infection in liver and heart diseases. Neither bone marrow nor kidney seem to be a common target of viral replication. PMID:20950445

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

    PubMed Central

    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

    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. PMID:20962324

  17. High-affinity interaction of hnRNP A1 with conserved RNA structural elements is required for translation and replication of enterovirus 71.

    PubMed

    Levengood, Jeffrey D; Tolbert, Michele; Li, Mei-Ling; Tolbert, Blanton S

    2013-07-01

    Human Enterovirus 71 (EV71) is an emerging pathogen of infectious disease and a serious threat to public health. Currently, there are no antivirals or vaccines to slow down or prevent EV71 infections, thus underscoring the urgency to better understand mechanisms of host-enterovirus interactions. EV71 uses a type I internal ribosome entry site (IRES) to recruit the 40S ribosomal subunit via a pathway that requires the cytoplasmic localization of hnRNP A1, which acts as an IRES trans-activating factor. The mechanism of how hnRNP A1 trans activates EV71 RNA translation is unknown, however. Here, we report that the UP1 domain of hnRNP A1 interacts specifically with stem loop II (SLII) of the IRES, via a thermodynamically well-defined biphasic transition that involves conserved bulge 5'-AYAGY-3' and hairpin 5'-RY(U/A)CCA-3' loops. Calorimetric titrations of wild-type and mutant SLII constructs reveal these structural elements are essential to form a high-affinity UP1-SLII complex. Mutations that alter the bulge and hairpin primary or secondary structures abrogate the biphasic transition and destabilize the complex. Notably, mutations within the bulge that destabilize the complex correlate with a large reduction in IRES-dependent translational activity and impair EV71 replication. Taken together, this study shows that a conserved SLII structure is necessary to form a functional hnRNP A1-IRES complex, suggesting that small molecules that target this stem loop may have novel antiviral properties. PMID:23727900

  18. Development of human B cells and antibodies following human hematopoietic stem cell transplantation to Rag2(-/-)?c(-/-) mice.

    PubMed

    Tanner, Anne; Hallam, Steven J; Nielsen, Stanton J; Cuadra, German I; Berges, Bradford K

    2015-06-01

    Humanized mice represent a valuable model system to study the development and functionality of the human immune system. In the RAG-hu mouse model highly immunodeficient Rag2(-/-)?c(-/-) mice are transplanted with human CD34(+) hematopoietic stem cells, resulting in human hematopoiesis and a predominant production of B and T lymphocytes. Human adaptive immune responses have been detected towards a variety of antigens in humanized mice but both cellular and humoral immune responses tend to be weak and sporadically detected. The underlying mechanisms for inconsistent responses are poorly understood. Here, we analyzed the kinetics of human B cell development and antibody production in RAG-hu mice to better understand the lack of effective antibody responses. We found that T cell levels in blood did not significantly change from 8 to 28weeks post-engraftment, while B cells reached a peak at 14weeks. Concentrations of 3 antibody classes (IgM, IgG, IgA) were found to be at levels about 0.1% or less of normal human levels, but human antibodies were still detected up to 32weeks after engraftment. Human IgM was detected in 92.5% of animals while IgG and IgA were detected in about half of animals. We performed flow cytometric analysis of human B cells in bone marrow, spleen, and blood to examine the presence of precursor B cells, immature B cells, naďve B cells, and plasma B cells. We detected high levels of surface IgM(+) B cells (immature and naďve B cells) and low levels of plasma B cells in these organs, suggesting that B cells do not mature properly in this model. Low levels of human T cells in the spleen were observed, and we suggest that the lack of T cell help may explain poor B cell development and antibody responses. We conclude that human B cells that develop in humanized mice do not receive the signals necessary to undergo class-switching or to secrete antibody effectively, and we discuss strategies to potentially overcome these barriers. PMID:25843523

  19. Possible Role of Nuclear Factor-?B Activity in Germline C? Transcription in a Human Burkitt Lymphoma B Cell Line

    Microsoft Academic Search

    Yukiyoshi Yanagihara; Yuji Basaki; Koichi Ikizawa; Keiichi Kajiwara

    1997-01-01

    Nuclear factor-?B (NF-?B) plays a broad role in gene regulation, but it is not evident whether NF-?B acts as a messenger system for germline C? transcription. We report here that the signaling cascade triggered by interleukin-4 (IL-4) or anti-CD40 monoclonal antibody (mAb) participates in NF-?B activation responsible for germline C? transcription in a human Burkitt lymphoma B cell line, DND39.

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

    SciTech Connect

    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

    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.

  1. Infectivity assays of human rhinovirus-A and -B serotypes.

    PubMed

    Lee, Wai-Ming; Chen, Yin; Wang, Wensheng; Mosser, Anne

    2015-01-01

    Infectivity is a fundamental property of viral pathogens such as human rhinoviruses (HRVs). This chapter describes two methods for measuring the infectivity of HRV-A and -B serotypes: end point dilution (TCID50) assay and plaque assay. End point dilution assay is a quantal, not quantitative, assay that determines the dilution of the sample at which 50 % of the aliquots have infectious virus. It can be used for all the HRV-A and -B serotypes and related clinical isolates that grow in cell culture and induce cytopathic effect (CPE), degenerative changes in cells that are visible under a microscope. Plaque assay is a quantitative assay that determines the number of infectious units of a virus in a sample. After an infectious unit of virus infects one cell, the infected cell produces progeny viruses that then infect and kill a circle of adjacent cells. This circle of dead cells detaches from the dish and thus leaves a clear hole in a cell monolayer. Plaque assay works only for HeLa-adapted HRV-A and -B serotypes that can make visible plaques on the cell monolayer. Currently the end point dilution assay and plaque assay have not been developed for the newly discovered HRV-C. PMID:25261308

  2. Group B Streptococcal Infection and Activation of Human Astrocytes

    PubMed Central

    Stoner, Terri D.; Weston, Thomas A.; Trejo, JoAnn; Doran, Kelly S.

    2015-01-01

    Background Streptococcus agalactiae (Group B Streptococcus, GBS) is the leading cause of life-threatening meningitis in human newborns in industrialized countries. Meningitis results from neonatal infection that occurs when GBS leaves the bloodstream (bacteremia), crosses the blood-brain barrier (BBB), and enters the central nervous system (CNS), where the bacteria contact the meninges. Although GBS is known to invade the BBB, subsequent interaction with astrocytes that physically associate with brain endothelium has not been well studied. Methodology/Principal Findings We hypothesize that human astrocytes play a unique role in GBS infection and contribute to the development of meningitis. To address this, we used a well- characterized human fetal astrocyte cell line, SVG-A, and examined GBS infection in vitro. We observed that all GBS strains of representative clinically dominant serotypes (Ia, Ib, III, and V) were able to adhere to and invade astrocytes. Cellular invasion was dependent on host actin cytoskeleton rearrangements, and was specific to GBS as Streptococcus gordonii failed to enter astrocytes. Analysis of isogenic mutant GBS strains deficient in various cell surface organelles showed that anchored LTA, serine-rich repeat protein (Srr1) and fibronectin binding (SfbA) proteins all contribute to host cell internalization. Wild-type GBS also displayed an ability to persist and survive within an intracellular compartment for at least 12 h following invasion. Moreover, GBS infection resulted in increased astrocyte transcription of interleukin (IL)-1?, IL-6 and VEGF. Conclusions/Significance This study has further characterized the interaction of GBS with human astrocytes, and has identified the importance of specific virulence factors in these interactions. Understanding the role of astrocytes during GBS infection will provide important information regarding BBB disruption and the development of neonatal meningitis. PMID:26030618

  3. Sporadic isolation of sabin-like polioviruses and high-level detection of non-polio enteroviruses during sewage surveillance in seven Italian cities, after several years of inactivated poliovirus vaccination.

    PubMed

    Battistone, A; Buttinelli, G; Fiore, S; Amato, C; Bonomo, P; Patti, A M; Vulcano, A; Barbi, M; Binda, S; Pellegrinelli, L; Tanzi, M L; Affanni, P; Castiglia, P; Germinario, C; Mercurio, P; Cicala, A; Triassi, M; Pennino, F; Fiore, L

    2014-08-01

    Sewage surveillance in seven Italian cities between 2005 and 2008, after the introduction of inactivated poliovirus vaccination (IPV) in 2002, showed rare polioviruses, none that were wild-type or circulating vaccine-derived poliovirus (cVDPV), and many other enteroviruses among 1,392 samples analyzed. Two of five polioviruses (PV) detected were Sabin-like PV2 and three PV3, based on enzyme-linked immunosorbent assay (ELISA) and PCR results. Neurovirulence-related mutations were found in the 5'noncoding region (5'NCR) of all strains and, for a PV2, also in VP1 region 143 (Ile>Thr). Intertypic recombination in the 3D region was detected in a second PV2 (Sabin 2/Sabin 1) and a PV3 (Sabin 3/Sabin 2). The low mutation rate in VP1 for all PVs suggests limited interhuman virus passages, consistent with efficient polio immunization in Italy. Nonetheless, these findings highlight the risk of wild or Sabin poliovirus reintroduction from abroad. Non-polio enteroviruses (NPEVs) were detected, 448 of which were coxsackievirus B (CVB) and 294 of which were echoviruses (Echo). Fifty-six NPEVs failing serological typing were characterized by sequencing the VP1 region (nucleotides [nt] 2628 to 2976). A total of 448 CVB and 294 Echo strains were identified; among those strains, CVB2, CVB5, and Echo 11 predominated. Environmental CVB5 and CVB2 strains from this study showed high sequence identity with GenBank global strains. The high similarity between environmental NPEVs and clinical strains from the same areas of Italy and the same periods indicates that environmental strains reflect the viruses circulating in the population and highlights the potential risk of inefficient wastewater treatments. This study confirmed that sewage surveillance can be more sensitive than acute flaccid paralysis (AFP) surveillance in monitoring silent poliovirus circulation in the population as well as the suitability of molecular approaches to enterovirus typing. PMID:24814793

  4. Molecular modeling of the human sperm associated antigen 11 B (SPAG11B) proteins.

    PubMed

    Narmadha, Ganapathy; Yenugu, Suresh

    2015-04-01

    Antimicrobial proteins and peptides are ubiquitous in nature with diverse structural and biological properties. Among them, the human beta-defensins are known to contribute to the innate immune response. Besides the defensins, a number of defensin-like proteins and peptides are expressed in many organ systems including the male reproductive system. Some of the protein isoforms encoded by the sperm associated antigen 11B (SPAG11) gene in humans are beta-defensin-like and exhibit structure dependent and salt tolerant antimicrobial activity, besides contributing to sperm maturation. Though some of the functional roles of these proteins are reported, the structural and molecular features that contribute to their antimicrobial activity is not yet reported. In this study, using in silico tools, we report the three dimensional structure of the human SPAG11B proteins and their C-terminal peptides. web-based hydropathy, amphipathicity, and topology (WHAT) analyses and grand average of hydropathy (GRAVY) indices show that these proteins and peptides are amphipathic and highly hydrophilic. Self-optimized prediction method with alignment (SOPMA) analyses and circular dichroism data suggest that the secondary structure of these proteins and peptides primarily contain beta-sheet and random coil structure and alpha-helix to a lesser extent. Ramachandran plots show that majority of the amino acids in these proteins and peptides fall in the permissible regions, thus indicating stable structures. The secondary structure of SPAG11B isoforms and their peptides were not perturbed with increasing NaCl concentration (0-300?mM) and at different pH (3, 7, and 10), thus reinforcing our previously reported observation that their antimicrobial activity is salt tolerant. To the best of our knowledge, for the first time, results of our study provide vital information on the structural features of SPAG11B protein isoforms and their contribution to antimicrobial activity. PMID:25573789

  5. Generation of Neutralizing Human Monoclonal Antibodies against Parvovirus B19 Proteins

    Microsoft Academic Search

    ANDREAS GIGLER; SIMONE DORSCH; ANDREA HEMAUER; CONSTANCE WILLIAMS; SONNIE KIM; NEAL S. YOUNG; SUSAN ZOLLA-PAZNER; HANS WOLF; MIROSLAW K. GORNY; SUSANNE MODROW; Hematology Branch

    1974-01-01

    Infections caused by human parvovirus B19 are known to be controlled mainly by neutralizing antibodies. To analyze the immune reaction against parvovirus B19 proteins, four cell lines secreting human immuno- globulin G monoclonal antibodies (MAbs) were generated from two healthy donors and one human immuno- deficiency virus type 1-seropositive individual with high serum titers against parvovirus. One MAb is specific

  6. Molecular Cell Human LSD2/KDM1b/AOF1 Regulates

    E-print Network

    Yuan, Guo-Cheng "GC"

    Molecular Cell Article Human LSD2/KDM1b/AOF1 Regulates Gene Transcription by Modulating Intragenic to the regulation of transcriptional initiation. We now show that human LSD2/KDM1b/AOF1, the human homolog of LSD1 regions of its target genes. Genome-wide mapping reveals that LSD2 associates predominantly with the gene

  7. 60 FR 28616 - Drug Export; Antibody to Hepatitis B Surface Antigen (Human) OrthoSUPTM Antibody to HBsAg Elisa...

    Federal Register 2010, 2011, 2012, 2013, 2014

    1995-06-01

    ...95N-0137] Drug Export; Antibody to Hepatitis B Surface Antigen (Human) Ortho TM...human biological product Antibody to Hepatitis B Surface Antigen (human) ORTHO TM...human biological product Antibody to Hepatitis B Surface Antigen (human) ORTHO...

  8. Gangliosides GD1b, GT1b, and GQ1b Suppress the Growth of Human Melanoma by Inhibiting Interleukin8 Production: the Inhibition of Adenylate Cyclase1

    Microsoft Academic Search

    Naoko Kanda; Kenji Nakai; Shinichi Watanabe

    2001-01-01

    We studied the effects of various gangliosides on in vitro growth of human metastatic melanoma WM266-4. GD1b, GT1b, and GQ1b inhibited 3H-thymidine uptake and growth rate of WM266-4 whereas the other gangliosides were ineffective. The growth inhibition by GD1b, GT1b, and GQ1b was counteracted by interleukin-8 but not by the other growth factors. The growth inhibition by gangliosides was not

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

    Microsoft Academic Search

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

    1980-01-01

    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

  10. The use of MR B+1 imaging for validation of FDTD electromagnetic simulations of human anatomies

    Microsoft Academic Search

    Cornelis A. T. Van den Berg; Lambertus W. Bartels; Bob van den Bergen; Hugo Kroeze; Astrid A. C. de Leeuw; Jeroen B. Van de Kamer; Jan J. W. Lagendijk

    2006-01-01

    In this study, MR B+1 imaging is employed to experimentally verify the validity of FDTD simulations of electromagnetic field patterns in human anatomies. Measurements and FDTD simulations of the B+1 field induced by a 3 T MR body coil in a human corpse were performed. It was found that MR B+1 imaging is a sensitive method to measure the radiofrequency

  11. Electrostatic Regulation of Genome Packaging in Human Hepatitis B Virus Zhen-Gang Wang,

    E-print Network

    Wu, Jianzhong

    Electrostatic Regulation of Genome Packaging in Human Hepatitis B Virus Tao Jiang, Zhen-Gang Wang, California ABSTRACT Hepatitis B virus (HBV) is a contagious human pathogen causing liver diseases-termini. INTRODUCTION Hepatitis B virus (HBV) is a pararetrovirus responsible for up to 80% of all cases

  12. The Immune Response to Hepatitis B Vaccine in Humans: Inheritance Patterns in Families

    E-print Network

    Alper, Chester A.

    The Immune Response to Hepatitis B Vaccine in Humans: Inheritance Patterns in Families By Margot S haverecently shown that the human antibody response to the hepatitis B virus surface antigen (HBs immunized with the hepatitis B vaccine, including sevenfamilies where at least one member bore the haplotype

  13. Immunobiology Human blood IgM "memory" B cells are circulating splenic marginal zone B cells harboring

    E-print Network

    Paris-Sud XI, Université de

    1 Immunobiology Human blood IgM "memory" B cells are circulating splenic marginal zone B cells des Hopitaux de Paris, Kremlin- Bicętre (France) 10 Institute of Anatomy and Cell Biology, University compartment displays a large population of IgM+ IgD+ CD27+ "memory" B cell carrying a mutated Ig receptor. We

  14. Characterization of pharmacologically active compounds that inhibit poliovirus and enterovirus 71 infectivity.

    PubMed

    Arita, Minetaro; Wakita, Takaji; Shimizu, Hiroyuki

    2008-10-01

    Poliovirus (PV) and enterovirus 71 (EV71) cause severe neurological symptoms in their infections of the central nervous system. To identify compounds with anti-PV and anti-EV71 activities that would not allow the emergence of resistant mutants, we performed drug screening by utilizing a pharmacologically active compound library targeting cellular factors with PV and EV71 pseudoviruses that encapsidated luciferase-encoding replicons. We have found that metrifudil (N-[2-methylphenyl]methyl)-adenosine) (an A2 adenosine receptor agonist), N(6)-benzyladenosine (an A1 adenosine receptor agonist) and NF449 (4,4',4'',4'''-[carbonylbis[imino-5,1,3-benzenetriyl bis(carbonyl-imino)

  15. Human cytosolic sulfotransferase 2B1: Isoform expression, tissue specificity and subcellular localization

    Microsoft Academic Search

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

    2006-01-01

    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

  16. Human B cell activating factor (BCAF): production by a human T cell tumor line.

    PubMed

    Fevrier, M; Diu, A; Mollier, P; Abadie, A; Olive, D; Mawas, C; Theze, J

    1989-01-01

    In a previous study, we demonstrated that supernatants from human T cell clones stimulated by a pair of anti-CD2 monoclonal antibodies cause resting human B cells to become activated and to proliferate in the absence of any other signals. The activity responsible for these effects was shown to be different from already characterized lymphokines and in particular from IL-2 and IL-4, and was named B Cell Activating Factor or BCAF. In this paper, we describe the production of BCAF by a human T cell tumor line T687 after phorbol myristate acetate (PMA) stimulation; this production can be potentiated by phytohemagglutinin (PHA). We further show that the stimulatory phase can be separated from the secretory phase thereby avoiding contamination of BCAF-containing supernatant by PMA and PHA. Supernatants produced under these conditions do not contain either IL-4 or IFN but contain traces of lymphotoxin and 2 to 10 ng/ml of IL-2. The T687 cell line will allow us to obtain a large volume of supernatant for biochemical study and purification of the molecule(s) responsible for BCAF activity. PMID:2497279

  17. Both apoB-48 and apoB-100 are synthesized by human enterocytes and secreted in hepatic bile

    SciTech Connect

    Rochette, C.; Bendayan, M.; Roy, C.C.; Milne, R.; Marcel, Y.; Levy, E. (Univ. of Montreal, Quebec (Canada))

    1990-02-26

    Using high resolution immunogold technique with polyclonal and monoclonal antibodies, the authors were able to show the presence of both forms of apoB (B-48 and B-100) in human enterocytes. Labeling for both isoproteins was present not only over the rough endoplasmic reticulum, but also on the apical vesicles, in multivesicular bodies and on microvilli indicated an internalization of apoB from the gut lumen. To examine the synthesis of apoB-100, a pulse of ({sup 3}H)-leucine was administered to human segments of intestine in explant culture. Newly synthesized apoB-100, confirmed by immunoprecipitation and immunoblot, represented 28% of total apoB production. The hypothesis that apoB-100 might be secreted in bile and internalized by the intestine, was tested by measuring apoB in human hepatic bile. The expression and immunoreactivity of both forms of apoB were obtained by using monoclonal antibodies which identify both B-48 and B-100 (1D1 and 2D8) or B-100 alone (3A10, 4G3, 5E11 and 22). While no epitopes were detected by 2D8 and 4G3, the distribution pattern for apoB was found by 1D1 (7.3%), 3A10 (31.2%), 5E11 (46.2%) and 22 (14.0%), suggesting that apoB fragments are secreted in bile. These findings provide evidence that apoB-100 is synthesized by the human gut and show that both isoproteins are consistent with the possibility that biliary apoB may be internalized by the enterocyte.

  18. Characterization of Proline-Serine-Rich Carboxyl Terminus in Human Sulfotransferase 2B1b: Immunogenicity, Subcellular Localization, Kinetic Properties, and Phosphorylation

    Microsoft Academic Search

    Dongning He; Charles N. Falany

    2006-01-01

    The human sulfotransferase (SULT) 2B1 gene is a member of the SULT2 gene family and encodes two isoforms, SULT2B1a and SULT2B1b. Although messages for both SULT2B1a and SULT2B1b are detectable in human tissues, only SULT2B1b has been identi- fied immunologically. Compared with other human SULTs, SULT2B1b has an extension at the proline- and serine-rich car- boxyl (PSC) end of about

  19. Organometallic B12-DNA conjugate: synthesis, structure analysis, and studies of binding to human B12-transporter proteins.

    PubMed

    Hunger, Miriam; Mutti, Elena; Rieder, Alexander; Enders, Barbara; Nexo, Ebba; Kräutler, Bernhard

    2014-10-01

    Design, synthesis, and structural characterization of a B12-octadecanucleotide are presented herein, a new organometallic B12-DNA conjugate. In such covalent conjugates, the natural B12 moiety may be a versatile vector for controlled in vivo delivery of oligonucleotides to cellular targets in humans and animals, through the endogenous B12 transport systems. Binding of the organometallic B12 octadecanucleotide to the three important human proteins of B12 transport was studied, to examine its structural suitability for the task of eventual in vivo oligonucleotide delivery. Binding was efficient with transcobalamin (TC), but not so efficient with the homologous glycoproteins intrinsic factor and haptocorrin. Binding of the B12 octadecanucleotide to TC suggests the capacity of the B12 moiety to serve as a natural vector for specific transport of single stranded, organometallic oligonucleotide loads from the blood stream into cells. PMID:25168390

  20. The Human Leukocyte Antigen–presented Ligandome of B Lymphocytes*

    PubMed Central

    Hassan, Chopie; Kester, Michel G. D.; de Ru, Arnoud H.; Hombrink, Pleun; Drijfhout, Jan Wouter; Nijveen, Harm; Leunissen, Jack A. M.; Heemskerk, Mirjam H. M.; Falkenburg, J. H. Frederik; van Veelen, Peter A.

    2013-01-01

    Peptides presented by human leukocyte antigen (HLA) molecules on the cell surface play a crucial role in adaptive immunology, mediating the communication between T cells and antigen presenting cells. Knowledge of these peptides is of pivotal importance in fundamental studies of T cell action and in cellular immunotherapy and transplantation. In this paper we present the in-depth identification and relative quantification of 14,500 peptide ligands constituting the HLA ligandome of B cells. This large number of identified ligands provides general insight into the presented peptide repertoire and antigen presentation. Our uniquely large set of HLA ligands allowed us to characterize in detail the peptides constituting the ligandome in terms of relative abundance, peptide length distribution, physicochemical properties, binding affinity to the HLA molecule, and presence of post-translational modifications. The presented B-lymphocyte ligandome is shown to be a rich source of information by the presence of minor histocompatibility antigens, virus-derived epitopes, and post-translationally modified HLA ligands, and it can be a good starting point for solving a wealth of specific immunological questions. These HLA ligands can form the basis for reversed immunology approaches to identify T cell epitopes based not on in silico predictions but on the bona fide eluted HLA ligandome. PMID:23481700

  1. Open-Channel Blockers at the Human a4b2 Neuronal Nicotinic Acetylcholine Receptor

    Microsoft Academic Search

    BRUNO BUISSON; DANIEL BERTRAND

    To extend our knowledge of the pharmacological profile of human a4b2 neuronal nicotinic receptors, we investigated the action of hexamethonium on the major brain human nicotinic acetylcholine receptor (nAChR) stably expressed in human em- bryonic kidney 293 cells. This compound displays all of the characteristics of an open-channel blocker at the human a4b2 nAChR: a voltage-dependent inhibition (more pronounced at

  2. Polyclonal human T lymphocyte activation results in the secondary functional activation of the human B lymphocyte.

    PubMed Central

    Bird, A G; Hammarström, L; Smith, C I; Britton, S

    1981-01-01

    The polyclonal T lymphocyte activators Con A and PHA were demonstrated to induce secretion of IgM and IgG as well as IgA antibodies in cultures of human peripheral blood lymphocytes. A similar response was seen in one-way mixed lymphocyte cultures and the kinetics, dose-response characteristics and optimal culture conditions are presented. The presence of functional T lymphocytes is a prerequisite for in vitro B lymphocyte activation in response to T lymphocyte mitogens. A narrow dose-response profile is a characteristic for both Con A and PHA and polyclonal B cell activation occurred at what have been previously regarded as suboptimal concentrations of these agents. The use of higher doses of these activators failed to generate immunoglobulin-secreting cells despite the presence of early and optimal levels of DNA synthesis in the cultures. PMID:6454516

  3. OATP1B1/1B3 activity in plated primary human hepatocytes over time in culture.

    PubMed

    Ulvestad, Maria; Björquist, Petter; Molden, Espen; Asberg, Anders; Andersson, Tommy B

    2011-11-01

    Primary human hepatocytes are widely used as an in vitro model for evaluation of drug metabolism and transport. However, it has been shown that the gene expression of many drug-metabolizing enzymes and transporters change in culture. The aim of the present study was to evaluate the activity of organic anion-transporting polypeptide 1B1 (OATP1B1) and 1B3 (OATP1B3) in plated primary human hepatocytes over time in culture. The uptake kinetics of the OATP1B1/1B3 substrate [(3)H]-estradiol-17?-d-glucuronide was determined in cells from five donors. An extensive and variable decrease in OATP1B1/1B3 activity and/or increase in passive diffusion was observed over time. Already after 6h in culture, the OATP1B1/1B3 activity was not possible to determine in liver cells from one donor, while after 24h, the uptake activity was not measurable in one additional donor. In the other three, the decrease in CL(int) (V(max)/K(m)) values ranged from 15% to 86% after 24h in culture compared to the values measured at 2h. Visual examination of OATP1B1 protein expression by confocal microscopy showed localization to the plasma membrane as expected, and an extensive decrease in OATP1B1 expression over time in culture supported the decline in activity. The significant reduction in SLCO1B1 and SLCO1B3 gene expression over time determined by RT-PCR also supported the loss of OATP1B1/1B3 activity. In conclusion, plated primary human hepatocytes are useful as an in vitro model for OATP1B1/1B3-mediated uptake studies, but the culture time may substantially change the uptake kinetics. PMID:21787759

  4. Human cytosolic sulfotransferase 2B1: isoform expression, tissue specificity and subcellular localization.

    PubMed

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

    2006-12-01

    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 3beta-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. PMID:17055258

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

    PubMed Central

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

    2007-01-01

    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. PMID:17055258

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

    PubMed

    Laffin, Brian; Petrash, J Mark

    2012-01-01

    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

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

    PubMed Central

    Laffin, Brian; Petrash, J. Mark

    2012-01-01

    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

  8. Molecular Surveillance of Enterovirus and Norwalk-Like Virus in Oysters Relocated to a Municipal-Sewage-Impacted Gulf Estuary

    Microsoft Academic Search

    Y. Carol Shieh; Ralph S. Baric; Jacquelina W. Woods; Kevin R. Calci

    2003-01-01

    An 18-month survey was conducted to examine the prevalence of enteric viruses and their relationship to indicators in environmentally polluted shellfish. Groups of oysters, one group per 4 weeks, were relocated to a coastal water area in the Gulf of Mexico that is impacted by municipal sewage and were analyzed for enteroviruses, Norwalk-like viruses (NLV), and indicator microorganisms (fecal coliform,

  9. Autocrine Transforming Growth Factor b Suppresses Telomerase Activity and Transcription of Human Telomerase Reverse Transcriptase in Human Cancer Cells1

    Microsoft Academic Search

    Hua Yang; Satoru Kyo; Masahiro Takatura; LuZhe Sun

    2001-01-01

    Because autocrine transforming growth factor b (TGF-b) can suppress carcinogenesis, which is often associated with telomerase activation, we studied whether autocrine TGF-b inhibits telomerase activity. Restoration of autocrine TGF-b activity in human colon carcinoma HCT116 cells after reexpression of its type II receptor (RII) led to a significant reduction of telomerase activity and the mRNA level of telomerase reverse transcriptase

  10. Cleavage of Interferon Regulatory Factor 7 by Enterovirus 71 3C Suppresses Cellular Responses

    PubMed Central

    Lei, Xiaobo; Xiao, Xia; Xue, Qinghua; Jin, Qi

    2013-01-01

    Enterovirus 71 (EV71) is a positive-stranded RNA virus which is capable of inhibiting innate immunity. Among virus-encoded proteins, the 3C protein compromises the type I interferon (IFN-I) response mediated by retinoid acid-inducible gene-I (RIG-I) or Toll-like receptor 3 that activates interferon regulatory 3 (IRF3) and IRF7. In the present study, we report that enterovirus 71 downregulates IRF7 through the 3C protein, which inhibits the function of IRF7. When expressed in mammalian cells, the 3C protein mediates cleavage of IRF7 rather than that of IRF3. This process is insensitive to inhibitors of caspase, proteasome, lysosome, and autophagy. H40D substitution in the 3C active site abolishes its activity, whereas R84Q or V154S substitution in the RNA binding motif has no effect. Furthermore, 3C-mediated cleavage occurs at the Q189-S190 junction within the constitutive activation domain of IRF7, resulting in two cleaved IRF7 fragments that are incapable of activating IFN expression. Ectopic expression of wild-type IRF7 limits EV71 replication. On the other hand, expression of the amino-terminal domain of IRF7 enhances EV71 infection, which correlates with its ability to interact with and inhibit IRF3. These results suggest that control of IRF7 by the 3C protein may represent a viral mechanism to escape cellular responses. PMID:23175366

  11. Characterization of an Enterovirus species E isolated from naturally infected bovine in China.

    PubMed

    Zhang, Haili; Liu, Hongtao; Bao, Jun; Guo, Yongli; Peng, Tongquan; Zhou, Pingping; Zhang, Wenlong; Ma, Bo; Wang, Junwei; Gao, Mingchun

    2014-10-13

    Bovine enteroviruses, which belong to the Picornaviridae family, can cause clinical symptoms in cattle and are excreted in feces. In this study, a cytolytic virus was isolated from Madin-Darby bovine kidney (MDBK) cells from fecal samples of bovine with severe diarrhea and hemorrhagic intestinal mucosa that had been originally diagnosed with bovine viral diarrhea (BVD) by a bovine viral diarrhea virus Ag point-of-care test (IDEXX, American). Random priming PCR was used to amplify underlying viral sequences and identify the isolated virus. Phylogenetic analysis indicated that the isolated virus closely matches the EV-E2 species, which is different from other Chinese strains previously isolated. The newly identified virus was named HLJ-3531/2013. We infected the sulking mice with the isolated virus. Reverse-transcription PCR, hematoxylin and eosin (HE) staining, serum neutralization (SN) test, and virus isolation from various tissues revealed that HLJ-3531/2013 can infect the intestine, liver, and lung of suckling mice. The present work is the first to report the reproduction of clinical symptoms by an isolated virus in an experimental infection model of animals and lays a solid foundation for the development of the pathogenesis of bovine enteroviruses. PMID:25102330

  12. Expression and immunogenicity of novel subunit enterovirus 71 VP1 antigens

    SciTech Connect

    Xu, Juan [China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China) [China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China); Department of Microbiology and Immunology, Nanjing Medical University (China); Wang, Shixia [China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China) [China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China); Department of Medicine, University of Massachusetts Medical School (United States); Gan, Weihua [Department of Pediatrics, The Second Affiliated Hospital, Nanjing Medical University (China)] [Department of Pediatrics, The Second Affiliated Hospital, Nanjing Medical University (China); Zhang, Wenhong [Department of Infectious Diseases, Huashan Hospital, Fudan University (China)] [Department of Infectious Diseases, Huashan Hospital, Fudan University (China); Ju, Liwen [School of Public Health, Fudan University (China)] [School of Public Health, Fudan University (China); Huang, Zuhu [Department of Infectious Diseases, The First Affiliated Hospital, Nanjing Medical University (China) [Department of Infectious Diseases, The First Affiliated Hospital, Nanjing Medical University (China); China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China); Lu, Shan, E-mail: shan.lu@umassmed.edu [Department of Infectious Diseases, The First Affiliated Hospital, Nanjing Medical University (China) [Department of Infectious Diseases, The First Affiliated Hospital, Nanjing Medical University (China); China-US Vaccine Research Center, The First Affiliated Hospital, Nanjing Medical University (China); Department of Medicine, University of Massachusetts Medical School (United States)

    2012-04-20

    Highlights: Black-Right-Pointing-Pointer EV71 is a major emerging infectious disease in many Asian countries. Black-Right-Pointing-Pointer Inactivated EV71 vaccines are in clinical studies but their safety and efficacy are unknown. Black-Right-Pointing-Pointer Developing subunit based EV71 vaccines is significant and novel antigen design is needed. Black-Right-Pointing-Pointer DNA immunization is an efficient tool to test the immunogenicity of VP1 based EV71 vaccines. Black-Right-Pointing-Pointer Multiple VP1 antigens are developed showing immunogenic potential. -- Abstract: Hand, foot, and mouth disease (HFMD) is a common viral illness in young children. HFMD is caused by viruses belonging to the enterovirus genus of the picornavirus family. Recently, enterovirus 71 (EV71) has emerged as a virulent agent for HFMD with severe clinical outcomes. In the current report, we conducted a pilot antigen engineering study to optimize the expression and immunogenicity of subunit VP1 antigen for the design of EV71 vaccines. DNA immunization was adopted as a simple technical approach to test different designs of VP1 antigens without the need to express VP1 protein in vitro first. Our studies indicated that the expression and immunogenicity of VP1 protein can be improved with alternated VP1 antigen designs. Data presented in the current report revealed novel pathways to optimize the design of VP1 antigen-based EV71 vaccines.

  13. Evaluation of a new automated microneutralization assay for the quantitative detection of neutralizing antibodies against enteroviruses.

    PubMed

    Rabenau, H; Weber, B

    1994-03-01

    An automated microneutralization assay for the quantitative detection of neutralizing antibodies (NA) against polioviruses and non-polio enteroviruses (NPEV) using a pipetting roboter (Tecan RSP 5072) was established and compared to the conventional manually performed test procedure. The qualitative neutralizing antibody detection was not significantly influenced by the assay system (manual or automated assay). Concerning the quantitative antibody detection, two-fold titre differences between the two test systems were observed in only 2.3% of the 260 serum samples investigated. The intra-assay and inter-assay variability of the quantitative detection of neutralizing antibodies using the automated assay proved to be very low. The quantitative detection of neutralizing antibodies using an automated pipetting robot permitted the testing of large numbers of samples within a shorter period and with less labour intensity as compared to the manually performed assay. Therefore it represents a valuable alternative to the conventional microneutralization test, especially for the serodiagnosis of non-polio enterovirus infections in large sample collectives, assessment of immunity to polioviruses and for seroepidemiological surveys. PMID:8061415

  14. Protection of salvianolic acid B for human endothelial cells against hydrogen peroxide-induced oxidative damage

    Microsoft Academic Search

    Jungang Zhang; Guangrong Zhao; Jinling Liu; Xiangwu Ji

    2009-01-01

    Salvianolic acid B(Sal B) is an active component of traditional Chinese medicine Salvia miltiorrhiza and is used to treat vascular diseases. To better understand its mechanism, the antioxidant capacities of Sal B was evaluated\\u000a with human endothelial cells under oxidative stress. Human endothelial cells were pretreated with Sal B for 12 h followed\\u000a by hydrogen peroxide for another 12 h.

  15. Human B cells secrete granzyme B when recognizing viral antigens in the context of the acute phase cytokine IL-21.

    PubMed

    Hagn, Magdalena; Schwesinger, Elisabeth; Ebel, Verena; Sontheimer, Kai; Maier, Julia; Beyer, Thamara; Syrovets, Tatiana; Laumonnier, Yves; Fabricius, Dorit; Simmet, Thomas; Jahrsdörfer, Bernd

    2009-08-01

    Human B cells are currently not known to produce the proapoptotic protease granzyme B (GrB) in physiological settings. We have discovered that BCR stimulation with either viral Ags or activating Abs in the context of the acute phase cytokine IL-21 can induce the secretion of substantial amounts of GrB by human B cells. Importantly, GrB response to viral Ags was significantly stronger in B cells from subjects recently vaccinated against the corresponding viruses as compared with unvaccinated subjects. GrB-secreting B cells featured a homogeneous CD19(+)CD20(+)CD27(-)CD38(-)IgD(-) phenotype, improved survival, and enhanced expression of costimulatory, Ag-presenting and cell-adhesion molecules. B cell-derived GrB was enzymatically active and its induction required the activation of similar signaling pathways as those in CTLs. Our findings suggest that GrB-secreting B cells support the early antiviral immune response against viruses with endosomal entry pathways, thereby counteracting overwhelming viral replication at the beginning of an infection until virus-specific T cells from draining lymph nodes arrive at the site of infection. Our data may also explain the elevated serum GrB levels found in the early phase of various viral diseases. PMID:19592644

  16. Identification of DNA of human origin based on amplification of human-specific mitochondrial cytochrome b region

    Microsoft Academic Search

    Hirokazu Matsuda; Yasuhisa Seo; Eiji Kakizaki; Shuji Kozawa; Eri Muraoka; Nobuhiro Yukawa

    2005-01-01

    Species-specific differences in a non-polymorphic region of the mitochondrial cytochrome b gene appear to be large enough to allow human-specific amplification of forensic DNA samples. We therefore developed a PCR-based method using newly designed primers to amplify a 157-bp portion of the human mitochondrial cytochrome b gene. The forward and reverse primers were designed to hybridize to regions of the

  17. Structural Basis for the Recognition of Human Cytomegalovirus Glycoprotein B by a Neutralizing Human Antibody

    PubMed Central

    Stump, Joachim D.; Wiegers, Anna-Katharina; Winkler, Thomas H.; Sticht, Heinrich; Mach, Michael; Muller, Yves A.

    2014-01-01

    Human cytomegalovirus (HCMV) infections are life-threating to people with a compromised or immature immune system. Upon adhesion, fusion of the virus envelope with the host cell is initiated. In this step, the viral glycoprotein gB is considered to represent the major fusogen. Here, we present for the first time structural data on the binding of an anti-herpes virus antibody and describe the atomic interactions between the antigenic domain Dom-II of HCMV gB and the Fab fragment of the human antibody SM5-1. The crystal structure shows that SM5-1 binds Dom-II almost exclusively via only two CDRs, namely light chain CDR L1 and a 22-residue-long heavy chain CDR H3. Two contiguous segments of Dom-II are targeted by SM5-1, and the combining site includes a hydrophobic pocket on the Dom-II surface that is only partially filled by CDR H3 residues. SM5-1 belongs to a series of sequence-homologous anti-HCMV gB monoclonal antibodies that were isolated from the same donor at a single time point and that represent different maturation states. Analysis of amino acid substitutions in these antibodies in combination with molecular dynamics simulations show that key contributors to the picomolar affinity of SM5-1 do not directly interact with the antigen but significantly reduce the flexibility of CDR H3 in the bound and unbound state of SM5-1 through intramolecular side chain interactions. Thus, these residues most likely alleviate unfavorable binding entropies associated with extra-long CDR H3s, and this might represent a common strategy during antibody maturation. Models of entire HCMV gB in different conformational states hint that SM5-1 neutralizes HCMV either by blocking the pre- to postfusion transition of gB or by precluding the interaction with additional effectors such as the gH/gL complex. PMID:25299639

  18. The (ProB27, ThrB28) human insulin analogue is more potent and more rapidly absorbed from subcutis than human insulin

    Microsoft Academic Search

    R Clausen; T G Jřrgensen; K H Jřrgensen; A H Johnsen; J J Led; K Josefsen

    2002-01-01

    Objective: To test the physiological properties of human insulin in which the amino acids Thr (B27) and Pro (B28) are interchanged (PT insulin). This was hypothesised to prevent dimerisation and accelerate the absorption from s.c. tissue without altering the affinity for the insulin receptor. Design: PT insulin was expressed in Pichia pastoris and processed in vitro. The purified compound was

  19. Structural analysis of human CCR2b and primate CCR2b by molecular modeling and molecular dynamics simulation

    Microsoft Academic Search

    Xiu-Fan Shi; Shixi Liu; Jinggong Xiangyu; Yaping Zhang; Jingfei Huang; Shuqiu Liu; Ci-Quan Liu

    2002-01-01

    CCR2b, a chemokine receptor for MCP-1, -2, -3, -4, plays an important role in a variety of diseases involving infection, inflammation, and\\/or injury, as well as being a coreceptor for HIV-1 infection. Two models of human CCR2b (hCCR2b) were generated by homology modeling and 1 ns restrained molecular dynamics (MD) simulation. In one only C113-C190 forms a disulfide bond (SS

  20. 1,25-dihydroxyvitamin D{sub 3} impairs NF-{kappa}B activation in human naive B cells

    SciTech Connect

    Geldmeyer-Hilt, Kerstin, E-mail: kerstin.hilt@charite.de [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany); Heine, Guido, E-mail: guido.heine@charite.de [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany) [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany); Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany); Hartmann, Bjoern, E-mail: bjoern.hartmann@charite.de [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany); Baumgrass, Ria, E-mail: baumgrass@drfz.de [Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany); Radbruch, Andreas, E-mail: radbruch@drfz.de [Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Deutsches Rheuma-Forschungszentrum Berlin, Chariteplatz 1, 10117 Berlin (Germany); Worm, Margitta, E-mail: margitta.worm@charite.de [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany)] [Allergie-Centrum-Charite, CCM, Klinik fuer Dermatologie und Allergologie, Charite - Universitaetsmedizin Berlin, Chariteplatz 1, 10117 Berlin (Germany)

    2011-04-22

    Highlights: {yields} In naive B cells, VDR activation by calcitriol results in reduced NF-{kappa}B p105 and p50 protein expression. {yields} Ligating the VDR with calcitriol causes reduced nuclear translocation of NF-{kappa}B p65. {yields} Reduced nuclear amount of p65 after calcitriol incubation results in reduced binding of p65 on the p105 promoter. {yields} Thus, vitamin D receptor signaling may reduce or prevent activation of B cells and unwanted immune responses, e.g. in IgE dependent diseases such as allergic asthma. -- Abstract: 1{alpha},25-dihydroxyvitamin D{sub 3} (calcitriol), the bioactive metabolite of vitamin D, modulates the activation and inhibits IgE production of anti-CD40 and IL-4 stimulated human peripheral B cells. Engagement of CD40 results in NF-{kappa}B p50 activation, which is essential for the class switch to IgE. Herein, we investigated by which mechanism calcitriol modulates NF-{kappa}B mediated activation of human naive B cells. Naive B cells were predominantly targeted by calcitriol in comparison with memory B cells as shown by pronounced induction of the VDR target gene cyp24a1. Vitamin D receptor activation resulted in a strongly reduced p105/p50 protein and mRNA expression in human naive B cells. This effect is mediated by impaired nuclear translocation of p65 and consequently reduced binding of p65 to its binding site in the p105 promoter. Our data indicate that the vitamin D receptor reduces NF-{kappa}B activation by interference with NF-{kappa}B p65 and p105. Thus, the vitamin D receptor inhibits costimulatory signal transduction in naive B cells, namely by reducing CD40 signaling.

  1. human-infective trypanosome T. b. rhodesi-ense has evolved a different way to escape

    E-print Network

    McGehee, Michael

    human-infective trypanosome T. b. rhodesi- ense has evolved a different way to escape killing that seem capable of killing human-infective trypanosomes13,14 but that do not cause kidney disease resistance to the TLFs present in most humans. SRA binds to APOL1 with high affinity and prevents either

  2. A Taxonomy of Distributed Human Computation Alexander J. Quinn, Benjamin B. Bederson

    E-print Network

    Golbeck, Jennifer

    of Maryland Department of Computer Science Human-Computer Interaction Lab College Park, MD 20742 USA {aq human- computer interaction and computer science, educators will require a common vocabulary to teachA Taxonomy of Distributed Human Computation Alexander J. Quinn, Benjamin B. Bederson University

  3. Posture Recognition with a 3D Human Model B. Boulay F. Bremond M. Thonnat

    E-print Network

    Paris-Sud XI, Université de

    of the large variety of postures due to the high degree of freedom in the human body. Moreover, for a samePosture Recognition with a 3D Human Model B. Boulay F. Bremond M. Thonnat INRIA - France INRIA.Thonnat@sophia.inria.fr ABSTRACT This paper proposes an approach to recognise human postures in video sequences, which combines a 2

  4. Improved crystallization of the coxsackievirus B3 RNA-dependent RNA polymerase

    SciTech Connect

    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, E-mail: arnaud.gruez@maem.uhp-nancy.fr [Centre National de la Recherche Scientifique and Universités d’Aix-Marseille I et II, UMR 6098, Architecture et Fonction des Macromolécules Biologiques, Ecole Supérieure d’Ingénieurs de Luminy-Case 925, 163 Avenue de Luminy, 13288 Marseille CEDEX 9 (France)

    2007-06-01

    The first crystal of a coxsackievirus RNA-dependent RNA polymerase is reported. 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 Ĺ resolution and that were suitable for structure determination.

  5. Tracking human antigen-specific memory B cells: a sensitive and generalized ELISPOT system

    Microsoft Academic Search

    Shane Crotty; Rachael D. Aubert; John Glidewell; Rafi Ahmed

    2004-01-01

    In the interest of better understanding the role of human memory B cells in protection against disease, we developed an assay to quantitate antigen-specific memory B cells in human blood. This assay utilizes a 6-day polyclonal stimulation of PBMC followed by an antigen-specific ELISPOT for the detection of memory B cells that have differentiated into antibody secreting cells (ASC) in

  6. Human Genome Diversity Project. Summary of planning workshop 3(B): Ethical and human-rights implications

    SciTech Connect

    NONE

    1993-12-31

    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.

  7. Epstein-Barr virus transformation induces B lymphocytes to produce human interleukin 10

    PubMed Central

    1993-01-01

    Interleukin 10 (IL-10) is a pleiotropic factor that enhances proliferation of activated human B lymphocytes and induces them to secrete high amounts of immunoglobulins. Here we show that several human B cell lines were able to constitutively secrete human (h)IL-10. Whereas none of the pre-B nor the plasmocytic cell lines tested produced hIL-10, 25 of the 36 tested mature B cell lines (lymphoblastoid and Burkitt lymphoma cell lines) secreted hIL-10. Moreover, 24 of these 25 hIL-10-producing B cell lines contained the Epstein-Barr virus (EBV) genome, suggesting a relationship between hIL- 10 production by human B cell lines and EBV expression. Accordingly, whereas polyclonal activation via triggering of surface immunoglobulins or CD40 antigen induced highly purified normal human B lymphocytes to produce only low (0.3-0.4 ng/ml) but significant amounts of hIL-10, EBV infection induced them to secrete high amounts of hIL-10 (4-9 ng/ml). Furthermore, addition of exogenous hIL-10, simultaneously to EBV infection, potentiated cell proliferation, whereas a blocking anti-IL- 10 antiserum inhibited it. Thus, hIL-10 produced by infected human B lymphocytes appears to be involved in the mechanisms of EBV-induced B cell proliferation. PMID:8381152

  8. Cytochrome P450 1B1 mRNA in the human central nervous system.

    PubMed Central

    Rieder, C R; Ramsden, D B; Williams, A C

    1998-01-01

    AIMS: To study the expression of CYP1B1 in a variety of human and rat cell lines as a means of identifying a new tool for the investigation of gene regulation. In addition, to identify the expression of cytochrome P450 1B1 (CYP1B1) in different regions of the central nervous system (CNS). METHODS: Reverse transcription-polymerase chain reaction followed by cloning and sequencing were used to detect the expression of CYP1B1 in human cell lines. Poly A+ mRNA from the human spinal cord and from different brain regions was analysed using a CYP1B1 probe labelled with 32PdCTP. RESULTS: Expression of CYP1B1 was shown in a human astrocytoma cell line (MOG-G-CCM). CYP1B1 mRNA was expressed in a variety of regions of the CNS but with a distinct regional specificity. Expression was highest in the putamen. CONCLUSIONS: The expression of CYP1B1 in a human astrocytoma enables this cell line to be used in further studies of regulation and function of this gene. The demonstration that CYP1B1 mRNA is expressed in a variety of regions of the CNS suggests a role for this gene in brain and spinal cord metabolism. The regional specificity of expression might explain the focal damage of certain human neurodegenerative diseases. PMID:9850336

  9. Human vitamin B12 absorption measurement by accelerator mass spectrometry using specifically

    E-print Network

    California at Davis, University of

    Human vitamin B12 absorption measurement by accelerator mass spectrometry using specifically to assimilate dietary vitamin B12. Assaying and understanding absorption and uptake of B12 is important because and showed plasma appearance and clearance curves consistent with the predicted behavior of the pure vitamin

  10. Anti-Enterovirus 71 Effects of Chrysin and Its Phosphate Ester

    PubMed Central

    Du, Jiang; Cui, Sheng; Yang, Fan; Jin, Qi

    2014-01-01

    Enterovirus 71 (EV71) can cause severe disease and even lead to death in children, and an effective antiviral drug is currently unavailable. The anti-EV71 effect of chrysin (5,7-dihydroxyflavone), a natural flavonoid commonly found in many plants, was tested in this report. By using the predicting program Autodock 4.0 and an in vitro protease inhibition assay, we found that chrysin could suppress viral 3Cpro activity. Replication of viral RNA and production of viral capsid protein and the infectious virion were strongly inhibited by chrysin, without noticeable cytotoxicity. Cytopathic effects on cells were also prevented. Diisopropyl chrysin-7-yl phosphate (CPI), the phosphate ester for chrysin, was generated through a simplified Atheron-Todd reaction to achieve stronger anti-viral activity. CPI was also able to bind with and inhibit viral 3Cpro activity in vitro. As expected, CPI demonstrated more potent antiviral activity against EV71. PMID:24598537

  11. An enterovirus 71 strain causes skeletal muscle damage in infected mice

    PubMed Central

    Lin, Peixin; Gao, Lulu; Huang, Yeen; Chen, Qing; Shen, Hong

    2015-01-01

    Objective: To study the target organs for enterovirus 71 (EV71) in infected suckling mice. Methods: 5-day-old BALB/c suckling mice were infected with an EV71 strain. Tissues of the infected mice were processed for histopathological examination, including immunohistochemistry, in situ hybridization, ultrastructural observation. Results: Some mice developed limb paralysis, trouble walking and loss of balance. Results of the histopathological study showed that a large amount of EV71 existed in the skeletal muscle tissues, accounting for the damage of the skeletal muscles. Conclusion: The EV71 clinical isolate used in this study presented evident myotropism. Skeletal muscles are important target organs for EV71 in the infected suckling mice. To clarify the relationship between EV71 infection and muscle diseases may contribute to a better understanding of the pathogenesis of EV71.

  12. Viral infection of engrafted human islets leads to diabetes.

    PubMed

    Gallagher, Glen R; Brehm, Michael A; Finberg, Robert W; Barton, Bruce A; Shultz, Leonard D; Greiner, Dale L; Bortell, Rita; Wang, Jennifer P

    2015-04-01

    Type 1 diabetes (T1D) is characterized by the destruction of the insulin-producing ?-cells of pancreatic islets. Genetic and environmental factors both contribute to T1D development. Viral infection with enteroviruses is a suspected trigger for T1D, but a causal role remains unproven and controversial. Studies in animals are problematic because of species-specific differences in host cell susceptibility and immune responses to candidate viral pathogens such as coxsackievirus B (CVB). In order to resolve the controversial role of viruses in human T1D, we developed a viral infection model in immunodeficient mice bearing human islet grafts. Hyperglycemia was induced in mice by specific ablation of native ?-cells. Human islets, which are naturally susceptible to CVB infection, were transplanted to restore normoglycemia. Transplanted mice were infected with CVB4 and monitored for hyperglycemia. Forty-seven percent of CVB4-infected mice developed hyperglycemia. Human islet grafts from infected mice contained viral RNA, expressed viral protein, and had reduced insulin levels compared with grafts from uninfected mice. Human-specific gene expression profiles in grafts from infected mice revealed the induction of multiple interferon-stimulated genes. Thus, human islets can become severely dysfunctional with diminished insulin production after CVB infection of ?-cells, resulting in diabetes. PMID:25392246

  13. Efficacy of IFN-?1 to Protect Human Airway Epithelial Cells against Human Rhinovirus 1B Infection

    PubMed Central

    Gulraiz, Fahad; Bellinghausen, Carla; Dentener, Mieke A.; Reynaert, Niki L.; Gaajetaan, Giel R.; Beuken, Erik V.; Rohde, Gernot G.; Bruggeman, Cathrien A.; Stassen, Frank R.

    2014-01-01

    Impaired interferon (IFN) production has been observed in various obstructive respiratory diseases. This contributes to enhanced sensitivity towards viral infections triggering acute exacerbations. To compensate for this impaired host IFN response, there is need to explore new therapeutic strategies, like exogenous administration of IFNs as prophylactic treatment. In the present study, we examined the protective potential of IFN-?1 and compared it with the previously established protecting effect of IFN-?. A549 cells and human primary bronchial epithelial cells were first treated with either IFN-? (500 IU/ml) or IFN-?1 (500 ng/ml) for 18 h. For infection, two approaches were adopted: i) Continuous scenario: after pre-treatment, cells were infected immediately for 24 h with human rhinovirus 1B (HRV1B) in IFN-containing medium, or were cultured for another 72 h in IFN-containing medium, and then infected for 24 h with HRV1B, ii) Pre-treatment scenario: IFN-containing medium was replaced after 18 h and cells were infected for 4 h either immediately after pre-treatment or after additional culturing for 72 h in IFN-free medium. The protective effect was evaluated in terms of reduction in the number of viral copies/infectious progeny, and enhanced expression of IFN-stimulated genes (ISGs). In both cell types and in both approaches, IFN-?1 and IFN-? treatment resulted in pronounced and long-lasting antiviral effects exemplified by significantly reduced viral copy numbers and diminished infectious progeny. This was associated with strong up-regulation of multiple ISGs. However, in contrast to the IFN-? induced expression of ISGs, which decreased over time, expression of ISGs induced by IFN-?1 was sustained or even increased over time. Here we demonstrate that the protective potential of IFN-?1 is comparable to IFN-?. Yet, the long-lasting induction of ISGs by IFN-?1 and most likely less incitement of side effects due to more localized expression of its receptors could make it an even more promising candidate for prophylactic treatment than IFN-?. PMID:24751942

  14. Expression of Human DNAJ (Heat Shock Protein-40) B3 in Humanized UDP-glucuronosyltransferase 1 Mice.

    PubMed

    Mitsugi, Ryo; Itoh, Tomoo; Fujiwara, Ryoichi

    2015-01-01

    The human DNAJB3 gene encodes a DNAJ (Heat shock protein 40; Hsp40) homolog, subfamily B, member 3 chaperone protein (DNAJB3), which can be down-regulated in disease conditions, as observed in decreased expression of DNAJB3 mRNA in peripheral blood mononuclear cells (PBMC) of obese patients. Recently, humanized UDP-glucuronosyltransferase (UGT) 1 mice (hUGT1 mice) were developed, in which the introduced human UGT1 gene contained a gene encoding human DNAJB3. In the present study, we analyzed the expression of human DNAJB3 mRNA in hUGT1 mice. Among the examined tissues, the testis had the highest expression of human DNAJB3 mRNA, while the lowest expression was observed in the liver. We found that the pattern of tissue-specific expression of mouse Dnajb3 in hUGT1 mice was very similar to that of human DNAJB3. We further demonstrated that the expression of human DNAJB3 in the liver was significantly reduced in high-fat-diet-fed hUGT1 mice compared to the expression level in the control mice, indicating that the expression of human DNAJB3 in hUGT1 mice could be similarly regulated in disease conditions such as obesity. Humanized UGT1 mice might therefore be useful to investigate the physiological role of human DNAJB3 in vivo. PMID:26147428

  15. Inferring nonneutral evolution from contrasting patterns of polymorphisms and divergences in different protein coding regions of enterovirus 71 circulating in Taiwan during 1998-2003

    PubMed Central

    2010-01-01

    Background Enterovirus (EV) 71 is one of the common causative agents for hand, foot, and, mouth disease (HFMD). In recent years, the virus caused several outbreaks with high numbers of deaths and severe neurological complications. Despite the importance of these epidemics, several aspects of the evolutionary and epidemiological dynamics, including viral nucleotide variations within and between different outbreaks, rates of change in immune-related structural regions vs. non-structural regions, and forces driving the evolution of EV71, are still not clear. Results We sequenced four genomic segments, i.e., the 5' untranslated region (UTR), VP1, 2A, and 3C, of 395 EV71 viral strains collected from 1998 to 2003 in Taiwan. The phylogenies derived from different genomic segments revealed different relationships, indicating frequent sequence recombinations as previously noted. In addition to simple recombinations, exchanges of the P1 domain between different species/genotypes of human enterovirus species (HEV)-A were repeatedly observed. Contrasting patterns of polymorphisms and divergences were found between structural (VP1) and non-structural segments (2A and 3C), i.e., the former was less polymorphic within an outbreak but more divergent between different HEV-A species than the latter two. Our computer simulation demonstrated a significant excess of amino acid replacements in the VP1 region implying its possible role in adaptive evolution. Between different epidemic seasons, we observed high viral diversity in the epidemic peaks followed by severe reductions in diversity. Viruses sampled in successive epidemic seasons were not sister to each other, indicating that the annual outbreaks of EV71 were due to genetically distinct lineages. Conclusions Based on observations of accelerated amino acid changes and frequent exchanges of the P1 domain, we propose that positive selection and subsequent frequent domain shuffling are two important mechanisms for generating new genotypes of HEV-A. Our viral dynamics analysis suggested that the importation of EV71 from surrounding areas likely contributes to local EV71 outbreaks. PMID:20868512

  16. Rapid Detection of Enteroviruses in Small Volumes of Natural Waters by Real-Time Quantitative Reverse Transcriptase PCR

    Microsoft Academic Search

    Jed A. Fuhrman; Xiaolin Liang; Rachel T. Noble

    2005-01-01

    Despite viral contamination of recreational waters, only bacterial, not viral, indicators are monitored routinely, due to a lack of rapid and cost-effective assays. We used negatively charged filters to capture enteroviruses from seawater and freshwater. Viral RNA was extracted using a commercial kit, and the viruses were quantified by real-time quantitative reverse transcriptase PCR (qRT-PCR). Poliovirus (6.6 to 330,000 virus

  17. Seven Strains of Enterovirus D68 Detected in the United States during the 2014 Severe Respiratory Disease Outbreak

    PubMed Central

    Brown, B. A.; Nix, W. A.; Sheth, M.; Frace, M.

    2014-01-01

    Clusters of severe respiratory disease in the United States were reported to the CDC beginning in August 2014. Enterovirus D68 (EV-D68) was identified from 83% (30/36) of initial severe cases. Investigations in August and September found severe EV-D68 cases to be widespread across the United States. We report seven EV-D68 genomes from the outbreak. PMID:25414503

  18. Detection by PCR of Enteroviruses in Cerebrospinal Fluid during a Summer Outbreak of Aseptic Meningitis in Switzerland

    Microsoft Academic Search

    MERI GORGIEVSKI-HRISOHO; JEAN-DANIEL SCHUMACHER; NEVENKA VILIMONOVIC; DANIEL GERMANN; LUKAS MATTER

    1998-01-01

    Enteroviruses (EV) are among the most common causes of aseptic meningitis. Standard diagnostic tech- niques are often too slow and lack sensitivity to be of clinical relevance. EV RNA can be detected withi n5hb y a commercially available reverse transcription-PCR (RT-PCR) test kit. Cerebrospinal fluid (CSF) samples from 68 patients presenting with aseptic meningitis during a summer outbreak in Switzerland

  19. The complete consensus sequence of coxsackievirus B6 and generation of infectious clones by long RT-PCR.

    PubMed

    Martino, T A; Tellier, R; Petric, M; Irwin, D M; Afshar, A; Liu, P P

    1999-10-01

    The full length sequence for the human pathogen coxsackievirus B6 (CVB6, Schmitt strain) has been determined. We used long RT-PCR to generate full length DNA amplicon of CVB6, and then directly sequenced the amplicons. One-step cloning of the full length amplicon enabled us to obtain an infectious clone of CVB6. RNA generated from CVB6 amplicon DNA or CVB6 clones, by transcription with T7 RNA polymerase, was demonstrated to be infectious upon transfection into HeLa cells in vitro. The CVB6 genome is characteristic of enteroviruses, with a 5'-non-translated region (743 nucleotides) followed by an open reading frame (encoding a 2184 amino acid polyprotein) and a 3'-non-translated region (100 nucleotides) and polyadenylated tail. The predicted amino acid sequence of CVB6 clustered with the other CVB serotypes and swine vesicular disease virus (SVDV). PMID:10500285

  20. Hepatitis B virus X gene induces human telomerase reverse transcriptase mRNA expression in cultured normal human cholangiocytes

    Microsoft Academic Search

    Sheng-Quan Zou; Zhen-Liang Qu; Zhan-Fei Li; Xin Wang

    AIM: To study the transcriptional regulation of human telomerase reverse transcriptase (hTERT) mRNA in normal human cholangiocytes (HBECs) after hepatitis B virus X (HBx) gene transfection and to elucidate the possible mechanism of HBV infection underlying cholangiocarcinoma. METHODS: HBECs were cultured in vitro and co-transfected with a eukaryotic expression vector containing the HBx coding region and a cloning vector containing

  1. B lymphocyte reconstitution after human bone marrow transplantation. Leu-1 antigen defines a distinct population of B lymphocytes.

    PubMed Central

    Antin, J H; Ault, K A; Rappeport, J M; Smith, B R

    1987-01-01

    Differences in the expression of Leu-1 (CD5) define two populations of recovering B cells after human marrow transplantation, Leu-1+ and Leu-1- B cells. The Leu-1+ B cells were polyclonal, of donor origin, and did not express detectable interleukin 2 receptor. Leu-1+ B cells generally appeared 2-4 wk after marrow grafting and often preceded the recovery of Leu-1- B cells. Acute and chronic graft vs. host disease (GvHD) resulted in the recovery of significantly fewer Leu-1+ B cells, whereas Leu-1- B cells were only decreased in acute GvHD. Multivariate analysis showed no significant effect of age, disease, prednisone or azathioprine, or ex vivo treatment of the marrow with anti-Leu-1 and complement on recovery of Leu-1+ and Leu-1- B cells, independent of the effects of GvHD. Leu-1+ B cells are a major lymphocyte population posttransplant. They may reflect a stage of differentiation of normal B cells or a separate B cell lineage. PMID:3112184

  2. Enterovirus 71 and coxsackievirus A16 3C proteases: binding to rupintrivir and their substrates and anti-hand, foot, and mouth disease virus drug design.

    PubMed

    Lu, Guangwen; Qi, Jianxun; Chen, Zhujun; Xu, Xiang; Gao, Feng; Lin, Daizong; Qian, Wangke; Liu, Hong; Jiang, Hualiang; Yan, Jinghua; Gao, George F

    2011-10-01

    Enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) are the major causative agents of hand, foot, and mouth disease (HFMD), which is prevalent in Asia. Thus far, there are no prophylactic or therapeutic measures against HFMD. The 3C proteases from EV71 and CVA16 play important roles in viral replication and are therefore ideal drug targets. By using biochemical, mutational, and structural approaches, we broadly characterized both proteases. A series of high-resolution structures of the free or substrate-bound enzymes were solved. These structures, together with our cleavage specificity assay, well explain the marked substrate preferences of both proteases for particular P4, P1, and P1' residue types, as well as the relative malleability of the P2 amino acid. More importantly, the complex structures of EV71 and CVA16 3Cs with rupintrivir, a specific human rhinovirus (HRV) 3C protease inhibitor, were solved. These structures reveal a half-closed S2 subsite and a size-reduced S1' subsite that limit the access of the P1' group of rupintrivir to both enzymes, explaining the reported low inhibition activity of the compound toward EV71 and CVA16. In conclusion, the detailed characterization of both proteases in this study could direct us to a proposal for rational design of EV71/CVA16 3C inhibitors. PMID:21795339

  3. Enterovirus 71 and Coxsackievirus A16 3C Proteases: Binding to Rupintrivir and Their Substrates and Anti-Hand, Foot, and Mouth Disease Virus Drug Design ?

    PubMed Central

    Lu, Guangwen; Qi, Jianxun; Chen, Zhujun; Xu, Xiang; Gao, Feng; Lin, Daizong; Qian, Wangke; Liu, Hong; Jiang, Hualiang; Yan, Jinghua; Gao, George F.

    2011-01-01

    Enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) are the major causative agents of hand, foot, and mouth disease (HFMD), which is prevalent in Asia. Thus far, there are no prophylactic or therapeutic measures against HFMD. The 3C proteases from EV71 and CVA16 play important roles in viral replication and are therefore ideal drug targets. By using biochemical, mutational, and structural approaches, we broadly characterized both proteases. A series of high-resolution structures of the free or substrate-bound enzymes were solved. These structures, together with our cleavage specificity assay, well explain the marked substrate preferences of both proteases for particular P4, P1, and P1? residue types, as well as the relative malleability of the P2 amino acid. More importantly, the complex structures of EV71 and CVA16 3Cs with rupintrivir, a specific human rhinovirus (HRV) 3C protease inhibitor, were solved. These structures reveal a half-closed S2 subsite and a size-reduced S1? subsite that limit the access of the P1? group of rupintrivir to both enzymes, explaining the reported low inhibition activity of the compound toward EV71 and CVA16. In conclusion, the detailed characterization of both proteases in this study could direct us to a proposal for rational design of EV71/CVA16 3C inhibitors. PMID:21795339

  4. The use of MR B+1 imaging for validation of FDTD electromagnetic simulations of human anatomies

    NASA Astrophysics Data System (ADS)

    Van den Berg, Cornelis A. T.; Bartels, Lambertus W.; van den Bergen, Bob; Kroeze, Hugo; de Leeuw, Astrid A. C.; Van de Kamer, Jeroen B.; Lagendijk, Jan J. W.

    2006-10-01

    In this study, MR B+1 imaging is employed to experimentally verify the validity of FDTD simulations of electromagnetic field patterns in human anatomies. Measurements and FDTD simulations of the B+1 field induced by a 3 T MR body coil in a human corpse were performed. It was found that MR B+1 imaging is a sensitive method to measure the radiofrequency (RF) magnetic field inside a human anatomy with a precision of approximately 3.5%. A good correlation was found between the B+1 measurements and FDTD simulations. The measured B+1 pattern for a human pelvis consisted of a global, diagonal modulation pattern plus local B+1 heterogeneties. It is believed that these local B+1 field variations are the result of peaks in the induced electric currents, which could not be resolved by the FDTD simulations on a 5 mm3 simulation grid. The findings from this study demonstrate that B+1 imaging is a valuable experimental technique to gain more knowledge about the dielectric interaction of RF fields with the human anatomy.

  5. Human LilrB2 Is a ?-Amyloid Receptor and Its Murine Homolog PirB Regulates Synaptic Plasticity in an Alzheimer’s Model

    PubMed Central

    Kim, Taeho; Vidal, George S.; Djurisic, Maja; William, Christopher M.; Birnbaum, Michael E.; Garcia, K. Christopher; Hyman, Bradley T.; Shatz, Carla J.

    2013-01-01

    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

  6. B7-H5 costimulates human T cells via CD28H

    E-print Network

    Zhu, Yuwen

    The B7/CD28 family has profound modulatory effects in immune responses and constitutes an important target for the development of novel therapeutic drugs against human diseases. Here we describe a new CD28 homologue (CD28H) ...

  7. DNMT3B7 expression related to MENT expression and its promoter methylation in human lymphomas.

    PubMed

    Alkebsi, Lobna; Handa, Hiroshi; Sasaki, Yoshiko; Osaki, Yohei; Yanagisawa, Kunio; Ogawa, Yoshiaki; Yokohama, Akihiko; Hattori, Hikaru; Koiso, Hiromi; Saitoh, Takayuki; Mitsui, Takeki; Tsukamoto, Norifumi; Nojima, Yoshihisa; Murakami, Hirokazu

    2013-12-01

    DNA methyltransferase (DNMT) 3B7 is the most expressed DNMT3B splice variant. It was reported that the loss of DNMT3B function led to overexpression of the MEthylated in Normal Thymocyes (MENT) and accelerated mouse lymphomagenesis. We investigated the DNMT3B7 expression and its relationship to MENT expression and promoter methylation in human lymphomas. DNMT3B7 and MENT expression were significantly (p<0.0001, p<0.01) higher in lymphomas than in non-malignant. Expression of DNMT3B7 and MENT were associated with MENT promoter hypomethylation. DNMT3B7 overexpression might interfere with the normal DNA methylation mechanism required for silencing the MENT proto-oncogene, and may accelerate human lymphomagenesis. PMID:24094886

  8. Rap2B promotes migration and invasion of human suprarenal epithelioma.

    PubMed

    Di, Jie-Hui; Qu, De-Bao; Lu, Zheng; Li, Lian-Tao; Cheng, Qian; Xin, Yong; Zhang, Long-Zhen; Zhang, Yanping; Zheng, Jun-Nian

    2014-09-01

    The aim of our study was to elucidate the role of Rap2B in the development of human suprarenal epithelioma and to investigate the effect of Rap2B on suprarenal epithelioma cells migration and invasion. We use tissue microarray and immunohistochemistry to evaluate Rap2B staining in 75 suprarenal epithelioma tissues and 75 tumor-adjacent normal renal tissues. And the expression of Rap2B protein in human suprarenal epithelioma cells and tissues was detected by western blot simultaneously. The role of Rap2B in suprarenal epithelioma cells migration and invasion was detected by using wound healing assay, cell migration assay, and matrigel invasion assay. After that, we performed western blot analysis and gelatin zymography to detect MMP-2 protein expression and enzyme activity. Our research showed that Rap2B expression was increased in tumor tissues compared with tumor-adjacent normal renal tissues. But no correlation was found between Rap2B expression and clinicopathological parameters. In addition, we found that Rap2B promoted the cell migration and invasion abilities, and Rap2B increased MMP-2 expression and enzyme activity in suprarenal epithelioma cells. Our data indicated that Rap2B expression is significantly increased in human suprarenal epithelioma and Rap2B can promote the cell migration and invasion abilities, which may provide a new target for the treatment of suprarenal epithelioma. PMID:24951956

  9. Isolation and Characterization of a Human Orphan UDP-Glucuronosyltransferase, UGT2B11

    Microsoft Academic Search

    Martin Beaulieu; Éric Lévesque; Dean W. Hum; Alain Bélanger

    1998-01-01

    Glucuronidation is an important metabolic pathway for both endogenous and exogenous compounds. To isolate novel UGT2B cDNA clones, human prostate and LNCaP cell cDNA libraries were screened using a pool of steroid-specific UGT2B cDNA as probes. We have isolated a novel human cDNA of 1.7 kb in length containing an open reading frame of 1587 pb which encodes a deduced

  10. A and B forms of the androgen receptor are expressed in a variety of human tissues

    Microsoft Academic Search

    Carol M. Wilson; Michael J. McPhaul

    1996-01-01

    Human genital skin fibroblasts contain both the full-length 110 K androgen receptor protein (AR-B, apparent Mr ? 110 000) and an 87 K N-terminally truncated AR isoform (AR-A, apparent Mr ? 87 000). These two AR species are structurally analogous to the A- and B-isoforms of the progesterone receptor (PR). We examined the distribution pattern of human AR isoforms in

  11. Immunostimulatory DNA inhibits IL4–dependent IgE synthesis by human B cells

    Microsoft Academic Search

    Anthony A. Horner; George F. Widhopf; Jan A. Burger; Kenji Takabayashi; Nadya Cinman; Arash Ronaghy; Hans L. Spiegelberg; Eyal Raz

    2001-01-01

    Background: Immunostimulatory sequence oligodeoxynucleotide (ISS-ODN) is a potent antiallergic immunomodulating agent in mice. However, few studies have addressed its antiallergic potential in human subjects. Objective: We sought to determine whether a phosphoro-thioate ISS-ODN could inhibit IL-4–dependent IgE synthesis by human B cells. Methods: Initially, nonatopic- and atopic-donor PBMCs were incubated with ISS-ODN or mutated oligodeoxynucleotide, and cytokine production and B-cell

  12. Functional expression of novel human and murine AKR1B genes

    PubMed Central

    Salabei, Joshua K.; Li, Xiao-Ping; Petrash, J. Mark; Bhatnagar, Aruni; Barski, Oleg A.

    2011-01-01

    The Aldo Keto Reductases (AKRs) are a superfamily of enzymes that catalyze the reduction of biogenic and xenobiotic aldehydes and ketones. AKR1B family has 2 known members in humans and 3 in rodents. Two novel gene loci, hereafter referred to as AKR1B15 in human and Akr1b16 in mouse have been predicted to exist within the AKR1B clusters. AKR1B15 displays 91% and 67% sequence identity with human genes AKR1B10 and AKR1B1, respectively while Akr1b16 shares 82–84% identity with murine Akr1b8 and Akr1b7. We tested the hypothesis that AKR1B15 and Akr1b16 genes are expressed as functional proteins in human and murine tissues, respectively. Using whole tissue mRNA, we were able to clone the full-length open reading frames for AKR1B15 from human eye and testes, and Akr1b16 from murine spleen, demonstrating that these genes are transcriptionally active. The corresponding cDNAs were cloned into pET28a and pIRES-hrGFP-1? vectors for bacterial and mammalian expression respectively. Both genes were expressed as 36 kDA proteins found in the insoluble fraction of bacterial cell lysate. These proteins, expressed in bacteria showed no enzymatic activity. However, lysates from COS-7 cells transfected with AKR1B15 showed a 4.8-fold (with p-nitrobenzaldehyde) and 3.3-fold (with DL-glyceraldehyde) increase in enzyme activity compared with untransfected COS-7 cells. The Akr1b16 transcript was shown to be ubiquitously expressed in murine tissues. Highest levels of transcript were found in heart, spleen, and lung. From these observations we conclude that the predicted AKR1B15 and 1b16 genes are expressed in several murine and human tissues. Further studies are required to elucidate their physiological roles. PMID:21276782

  13. Functional expression of novel human and murine AKR1B genes.

    PubMed

    Salabei, Joshua K; Li, Xiao-Ping; Petrash, J Mark; Bhatnagar, Aruni; Barski, Oleg A

    2011-05-30

    The Aldo Keto Reductases (AKRs) are a superfamily of enzymes that catalyze the reduction of biogenic and xenobiotic aldehydes and ketones. AKR1B family has 2 known members in humans and 3 in rodents. Two novel gene loci, hereafter referred to as AKR1B15 in human and Akr1b16 in mouse have been predicted to exist within the AKR1B clusters. AKR1B15 displays 91% and 67% sequence identity with human genes AKR1B10 and AKR1B1, respectively while Akr1b16 shares 82-84% identity with murine Akr1b8 and Akr1b7. We tested the hypothesis that AKR1B15 and Akr1b16 genes are expressed as functional proteins in human and murine tissues, respectively. Using whole tissue mRNA, we were able to clone the full-length open reading frames for AKR1B15 from human eye and testes, and Akr1b16 from murine spleen, demonstrating that these genes are transcriptionally active. The corresponding cDNAs were cloned into pET28a and pIRES-hrGFP-1? vectors for bacterial and mammalian expression, respectively. Both genes were expressed as 36kDa proteins found in the insoluble fraction of bacterial cell lysate. These proteins, expressed in bacteria showed no enzymatic activity. However, lysates from COS-7 cells transfected with AKR1B15 showed a 4.8-fold (with p-nitrobenzaldehyde) and 3.3-fold (with dl-glyceraldehyde) increase in enzyme activity compared with untransfected COS-7 cells. The Akr1b16 transcript was shown to be ubiquitously expressed in murine tissues. Highest levels of transcript were found in heart, spleen, and lung. From these observations we conclude that the predicted AKR1B15 and 1b16 genes are expressed in several murine and human tissues. Further studies are required to elucidate their physiological roles. PMID:21276782

  14. Further evidence for a human B cell activating factor distinct from IL-4.

    PubMed

    Diu, A; Février, M; Mollier, P; Charron, D; Banchereau, J; Reinherz, E L; Thčze, J

    1990-01-01

    Supernatants from activated human T cell clones were previously shown to contain B cell-activating factor (BCAF), an activity which results in polyclonal resting B cell stimulation. In the present study, we investigate the relationship between this activity and human interleukin-4 which was also shown to act on resting B cells. The supernatant of the T cell clone TT9 contains IL-4 but anti-IL-4 antiserum does not affect the response of B cells as measured by thymidine uptake or cell volume increase. Furthermore, IL-4 induces Fc epsilon-receptor (CD23) expression on 30% of unstimulated human B cells, whereas BCAF-containing supernatants from clone P2, that do not contain detectable amounts of IL-4, promote B cell proliferation without inducing CD23 expression. Our results therefore establish that IL-4 and BCAF are distinct activities and suggest that they trigger different activation pathways in human B cells. In addition, culture of B cells with T cell supernatants for 72 hr induces a three- to fourfold increase in the expression of HLA-DR, -DP, and -DQ antigens in 50% of B cells. The addition of inhibiting concentrations of anti-IFN-gamma, LT, or IL-4 antisera to the cultures does not change these results. Finally, 30% of B cells cultured with T cell supernatants leave the G1 phase of the cell cycle and 20% reach mitosis. Taken together, our findings further support the existence of a B cell-activating factor responsible for the activation of resting human B cells. PMID:2136716

  15. BDNF and its TrkB receptor in human fracture healing.

    PubMed

    Kilian, Olaf; Hartmann, Sonja; Dongowski, Nicole; Karnati, Srikanth; Baumgart-Vogt, Eveline; Härtel, Frauke V; Noll, Thomas; Schnettler, Reinhard; Lips, Katrin Susanne

    2014-09-01

    Fracture healing is a physiological process of repair which proceeds in stages, each characterized by a different predominant tissue in the fracture gap. Matrix reorganization is regulated by cytokines and growth factors. Neurotrophins and their receptors might be of importance to osteoblasts and endothelial cells during fracture healing. The aim of this study was to examine the presence of brain-derived neurotrophic factor (BDNF) and its tropomyosin-related kinase B receptor (TrkB) during human fracture healing. BDNF and TrkB were investigated in samples from human fracture gaps and cultured cells using RT-PCR, Western blot, and immunohistochemistry. Endothelial cells and osteoblastic cell lines demonstrated a cytoplasmic staining pattern of BDNF and TrkB in vitro. At the mRNA level, BDNF and TrkB were expressed in the initial and osteoid formation phase of human fracture healing. In the granulation tissue of fracture gap, both proteins--BDNF and TrkB--are concentrated in endothelial and osteoblastic cells at the margins of woven bone suggesting their involvement in the formation of new vessels. There was no evidence of BDNF or TrkB during fracture healing in chondrocytes of human enchondral tissue. Furthermore, BDNF is absent in mature bone. Taken together, BDNF and TrkB are involved in vessel formation and osteogenic processes during human fracture healing. The detection of BDNF and its TrkB receptor during various stages of the bone formation process in human fracture gap tissue were shown for the first time. The current study reveals that both proteins are up-regulated in human osteoblasts and endothelial cells in fracture healing. PMID:24984919

  16. The Thiazolobenzimidazole TBZE-029 Inhibits Enterovirus Replication by Targeting a Short Region Immediately Downstream from Motif C in the Nonstructural Protein 2C?

    PubMed Central

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

    2008-01-01

    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 2C, clustered at amino acid positions 224, 227, and 229, immediately downstream of NTPase/helicase motif C. The mutations were reintroduced, either alone or combined, into an infectious full-length CVB3 clone. In particular the mutations at positions 227 and 229 proved essential for the altered sensitivity of CVB3 to TBZE-029. Resistant virus exhibited cross-resistance to the earlier-reported antienterovirus agents targeting 2C, namely, guanidine hydrochloride, HBB [2-(alpha-hydroxybenzyl)-benzimidazole], and MRL-1237 {1-(4-fluorophenyl)-2-[(4-imino-1,4-dihydropyridin-1-yl)methyl]benzimidazole hydrochloride}. The ATPase activity of 2C, however, remained unaltered in the presence of TBZE-029. PMID:18337578

  17. Inhibition of human acid-sensing ion channel 1b by zinc

    PubMed Central

    Jiang, Qian; Zha, Xiang-Ming; Chu, Xiang-Ping

    2012-01-01

    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. PMID:22837807

  18. METABOLISM AND CYTOTOXICITY OF AFLATOXIN B 1 IN CYTOCHROME P-450EXPRESSING HUMAN LUNG CELLS

    Microsoft Academic Search

    Terry R. Van Vleet; Patrick J. Klein; Roger A. Coulombe Jr

    2002-01-01

    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.

  19. PANORAMIC CAPTURING AND RECOGNITION OF HUMAN ACTIVITY Xinding Sun, B. S. Manjunath

    E-print Network

    California at Santa Barbara, University of

    PANORAMIC CAPTURING AND RECOGNITION OF HUMAN ACTIVITY Xinding Sun, B. S. Manjunath Department for human movement recognition. Their method requires less computation, but is sensitive to variances in the movement. Little and Boyd [4] use the moments of moving points to represent the optic flow for the purpose

  20. Dynamic Control of Human Eye on Head System Bijoy K. Ghosh1, Indika B. Wijayasinghe1

    E-print Network

    Ghosh, Bijoy K.

    Dynamic Control of Human Eye on Head System Bijoy K. Ghosh1, Indika B. Wijayasinghe1 1. Laboratory.ghosh, indika.wijayasinghe}@ttu.edu Abstract: In this paper we study the human eye movement, when the eye shifts is restrained to remain fixed, eye movements obey Listing's constraint, which states that the allowed

  1. PostTranscriptional Regulation of Interleukin6 Gene Expression in Human Keratinocytes by Ultraviolet B Radiation

    Microsoft Academic Search

    Sven de Vos; Marion Brach; Anne Budnik; Markus Grewe; Friedhelm Herrmann; Jean Krutmann

    1994-01-01

    Exposure to increasing doses (290-315 nm) of ultraviolet (UV) B radiation is thought to profoundly affect human health. Studies on the biologic and molecular effects of UVB radiation on human skin are therefore of particular interest. There is experimental and clinical evidence to assume that UVB radiation-induced local and systemic inflammatory reactions might be mediated at least in part by

  2. UNIT 1B.3Phenotypic Analysis of Human Embryonic Stem Cells

    E-print Network

    Zandstra, Peter W.

    UNIT 1B.3Phenotypic Analysis of Human Embryonic Stem Cells Mark Ungrin,1 Michael O'Connor,2 Laboratory, BC Cancer Agency, Vancouver, British Columbia 3 StemCell Technologies, Inc, Vancouver, British Columbia ABSTRACT Human embryonic stem cells (hESCs) are an important tool for the study of develop- mental

  3. Direct Phenotypical and Functional Dysregulation of Primary Human B Cells by Human Immunodeficiency Virus (HIV) Type 1 In Vitro

    PubMed Central

    Perisé-Barrios, Ana Judith; Muńoz-Fernandez, María Ángeles; Pion, Marjorie

    2012-01-01

    Background Human immunodeficiency virus type 1 (HIV-1) induces a general dysregulation of immune system. Dysregulation of B cell compartment is generally thought to be induced by HIV-related immune activation and lymphopenia. However, a direct influence of HIV-1 particles on B cells was recently proposed as the third pathway of B cells dysregulation. Methods/Principal Findings We evaluated the direct and specific consequences of HIV-1 contact on activation, survival, proliferation and phenotype of primary B cells in vitro. Moreover, we examined expression of activation-induced cytidine deaminase (AID) mRNA that is responsible for class switch recombination (CSR) and somatic hypermutation (SHM). Here, we report that changes observed in cellular proliferation, phenotypes and activation of B cells could be caused by direct contact between HIV-1 particles and primary B cells in vitro. Finally, direct HIV-1-derived B cells activation led to the increase of AID mRNA expression and its subsequent CSR function was detected in vitro. Conclusion/Significance We showed that HIV-1 could directly induce primary B cells dysregulation triggering phenotypical and functional abilities of B cells in vitro that could explain in some extent early B-cell abnormalities in HIV disease. PMID:22768302

  4. Coxsackievirus B detection in cases of myocarditis, myopericarditis, pericarditis and dilated cardiomyopathy in hospitalized patients.

    PubMed

    Gaaloul, Imed; Riabi, Samira; Harrath, Rafik; Hunter, Timothy; Hamda, Khaldoun B; Ghzala, Assia B; Huber, Sally; Aouni, Mahjoub

    2014-12-01

    Coxsackieviruses B (CV?B) are known as the most common viral cause of human heart infections. The aim of the present study was to assess the potential role of CV?B in the etiology of infectious heart disease in hospitalized patients. The present study is based on blood, pericardial fluid and heart biopsies from 102 patients and 100 control subjects. All of the samples were examined for the detection of specific enteroviral genome using the reverse transcription polymerase chain reaction (RT?PCR) and sequence analysis. Immunohistochemical investigations for the detection of the enteroviral capsid protein, VP1, from the biopsies were performed. The samples were cultured on confluent KB monolayer cell line for possible virus isolation. The epidemiological data were also collected. CV?B was detected in 28 of the 102 patients. The sequence analysis demonstrated that 27 strains were identical to CV?B3 and only one strain was identical to CV?B1. Furthermore, VP1 in the heart biopsies was detected in enterovirus?positive cases, as revealed by RT?PCR. Pericarditis infection was more frequent than myocarditis (P<0.05) or myopericarditis (P=0.05). The epidemiological data demonstrate that CV?B heart infections occur mainly during autumn and winter, and young male adults are more susceptible than adolescents or adults (P<0.5). The present findings demonstrate a higher prevalence of viral heart infections, suggesting that CV?B may significantly contribute to heart infections. PMID:25241846

  5. Induction of c-myc expression in human B lymphocytes by B-cell growth factor and anti-immunoglobulin

    SciTech Connect

    Lacy, J.; Sarkar, S.N.; Summers, W.C.

    1986-03-01

    Purified human B lymphocytes were examined for transcriptional expression of c-myc in response to mitogenic stimulation by the method of in situ hybridization using /sup 35/S-labeled DNA probes. The level of c-myc expression increased 10- to 20-fold within 2 hr after the addition of anti-..mu.., formalinized Staphylococcus aureus Cowan strain I, or B-cell growth factor, as compared to resting B cells. After 72-96 hr of mitogenic stimulation, c-myc expression remained elevated 5-fold, but expression among individual cells had become more heterogeneous than at early time points. To determine whether c-myc expression in human B lymphocytes is phase specific within the cell cycle, mitogen-stimulated cells were sorted by DNA content into populations of cells in G/sub 0//G/sub 1/, S, and G/sub 2//M phases of the cell cycle. Examination of c-myc expression in phase-specific cells revealed that c-myc expression was elevated in all phases of the cell cycle, but it appeared to be maximally expressed in S phase. These studies suggest that c-myc expression in normal human B lymphocytes is cell-cycle dependent and remains elevated in all phases of the cycling cell.

  6. Comparative Gene Expression Profiling Identifies Common Molecular Signatures of NF-?B Activation in Canine and Human Diffuse Large B Cell Lymphoma (DLBCL)

    PubMed Central

    Mudaliar, Manikhandan A. V.; Haggart, Ross D.; Miele, Gino; Sellar, Grant; Tan, Karen A. L.; Goodlad, John R.; Milne, Elspeth; Vail, David M.; Kurzman, Ilene

    2013-01-01

    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 literature-derived NF-?B target gene set mapped to orthologous canine array probesets and human array probesets clearly separated the healthy and cancer samples in both datasets. The analysis demonstrated that for both human and canine DLBCL there is activation of the NF-?B/p65 canonical pathway, indicating that canine lymphoma could be used as a model to study NF-?B-targeted therapeutics for human lymphoma. To validate this, tissue arrays were generated for canine and human NHL and immunohistochemistry was employed to assess NF-?B activation status. In addition, human and canine B-cell lymphoma lines were assessed for NF-?B activity and the effects of NF-?B inhibition. PMID:24023754

  7. Competitive chemiluminescent enzyme immunoassay for vitamin B12 analysis in human milk

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BACKGROUND Few accurate data exist on the concentration of vitamin B12 in human milk. Binding of the vitamin to haptocorrin (HC) can interfere with the assay if not removed by pretreatment, and very low values can occur in women with poor B12 status. This study evaluated two competitive enzyme bind...

  8. Genetic analysis of immortalizing functions of Epstein-Barr virus in human B lymphocytes

    Microsoft Academic Search

    Wolfgang Hammerschmidt; Bill Sugden

    1989-01-01

    EPSTEIN-BARR virus (EBV), a herpes virus, infects human B lymphocytes in vitro and efficiently immortalizes them1. About 10 of the approximately 100 genes of EBV are expressed in recently immortalized B cells and although there is circumstantial evidence that at least three of these may contribute to the process of immortalization, there is no direct evidence that any particular gene

  9. Functional characterization of novel mutations in the human cytochrome b gene

    Microsoft Academic Search

    Frédéric Legros; Evi Chatzoglou; Paule Frachon; Hélčne Ogier de Baulny; Pascal Laforęt; Claude Jardel; Catherine Godinot; Anne Lombčs

    2001-01-01

    The great variability of the human mitochondrial DNA (mtDNA) sequence induces many difficulties in the search for its deleterious mutations. We illustrate these pitfalls by the analysis of the cytochrome b gene of 21 patients affected with a mitochondrial disease. Eighteen different sequence variations were found, five of which were new mutations. Extensive analysis of the cytochrome b gene of

  10. Human immunodeficiency virus, hepatitis B, C and D in Bangladesh's trucking industry: prevalence and risk factors

    Microsoft Academic Search

    Laura Gibney; Parwez Choudhury

    2001-01-01

    Background Human immunodeficiency virus (HIV) and hepatitis B and C, viral infections with shared percutaneous, mucosal and perinatal routes of transmission, are responsible for serious morbidity and mortality globally. In Bangladesh there is a dearth of research on prevalence and risk factors for these diseases. This study examines the prevalence of HIV and hepatitis (B, C, D) and risk factors

  11. Production of Colony-Stimulating Activity by Normal and Neoplastic Human B Lymphocytes

    Microsoft Academic Search

    V. Pistoia; R. Ghio; S. Roncella; F. Cozzolino; S. Zupo; M. Ferrarini

    1987-01-01

    Normal human B cells were purified from peripheral blood or tonsils and tested for their ability to release colony- stimulating activity (CSA) in short-term cultures. The tar- get cells used in the CSA assays were from peripheral blood or bone marrow. Unstimulated B cells produced CSA in amounts similar to those present in the GCT-conditioned medium used as a positive

  12. Amyloid b Peptide of Alzheimer's Disease Downregulates Bcl2 and Upregulates Bax Expression in Human Neurons

    Microsoft Academic Search

    Eric Paradis; Helene Douillard; Maria Koutroumanis; Cynthia Goodyer; Andrea LeBlanc

    1996-01-01

    Neuronal apoptosis is a suspected cause of neurodegenera- tion in Alzheimer's disease (AD). Increased levels of amyloid b peptide (Ab) induce neuronal apoptosis in vitro and in vivo. The underlying molecular mechanism of Ab neurotoxicity is not clear. The normal concentration of Ab in cerebrospinal fluid is 4 nM. We treated human neuron primary cultures with 100 nM amyloid b

  13. Infection of Human B Lymphocytes with Lymphocryptoviruses Related to Epstein-Barr Virus

    Microsoft Academic Search

    AMIR MOGHADDAM; JOACHIM KOCH; BETHANY ANNIS; FRED WANG

    Lymphocryptoviruses (LCVs) naturally infecting Old World nonhuman primates are closely related to the human LCV, Epstein-Barr virus (EBV), and share similar genome organization and sequences, biologic prop- erties, epidemiology, and pathogenesis. LCVs can efficiently immortalize B lymphocytes from the autologous species, but the ability of a given LCV to immortalize B cells from other Old World primate species is variable.

  14. Comparative Immunohistochemistry of Placental Corticotropin-Releasing Hormone and the Transcription Factor RelB-NF?B2 Between Humans and Nonhuman Primates.

    PubMed

    Rosen, Todd; Schulkin, Jay; Power, Michael; Tadesse, Serkalem; Norwitz, Errol R; Wen, Zhaoqin; Wang, Bingbing

    2015-01-01

    The transcription factor RelB-NF?B2, activated by the noncanonical NF?B pathway, positively regulates corticotropin-releasing hormone (CRH) and prostaglandin production in the term human placenta and may play an important role in the timing of human parturition. Here we explored whether RelB-NF?B2 signaling plays a role in parturition in nonhuman anthropoid primates. We performed immunohistochemical staining to assess the correlation between CRH and nuclear activity of RelB-NF?B2 heterodimers in term placentas from humans, 3 catarrhine primate species, and a single platyrrhine primate species. Consistent with our previous studies, the human placenta showed cytoplasmic staining for CRH and nuclear staining for RelB-NF?B2. Similar staining patterns were noted in the 3 catarrhine primates (chimpanzee, baboon, and rhesus macaque). The platyrrhine (marmoset) placentas stained positively for CRH and RelB but not for NF?B2. Catarrhine (but not platyrrhine) nonhuman primate term placentas demonstrate the same CRH staining and nuclear localization patterns of RelB and NF?B2 as does human placenta. These results suggest that catarrhine primates, particularly rhesus macaques, may serve as useful animal models to study the biologic significance of the noncanonical NF?B pathway in human pregnancy. PMID:25926400

  15. Structure and pre-B lymphocyte restricted expression of the VpreB in humans and conservation of its structure in other mammalian species.

    PubMed Central

    Bauer, S R; Kudo, A; Melchers, F

    1988-01-01

    DNA from several mammals, including humans, was found to contain one or more restriction enzyme digested DNA fragments which hybridized to the mouse VpreB gene under stringencies demonstrating at least 70% nucleotide sequence homologies, indicating that the VpreB locus may be widespread and highly conserved among mammals. A human VpreB genomic clone was isolated and sequenced. Two exons and the intervening intron are spaced almost identically as in the mouse VpreB1 gene, and show 76% sequence homology to the mouse gene. As in the mouse VpreB1 gene, the 5' end of the human VpreB gene contains characteristic features of Ig domains, while the 3' end is Ig non-related. This 3' Ig non-related structure of the VpreB gene(s) may, therefore, have existed before the speciation of humans and mice over 65 million years ago. Sequences encoding the entire putative second framework region and a stretch in the third framework region are identical in human and mouse VpreB. the human VpreB gene appears to be selectively expressed in human pre-B cell lines as an 0.85 kb poly(A)+ RNA. Its expression promises to be a useful marker for the detection of normal and malignant human pre-B lymphocytes. Images PMID:3258819

  16. Glycosylated human oxyhaemoglobin activates nuclear factor-?B and activator protein-1 in cultured human aortic smooth muscle

    PubMed Central

    Peiró, Concepción; Matesanz, Nuria; Nevado, Julián; Lafuente, Nuria; Cercas, Elena; Azcutia, Verónica; Vallejo, Susana; Rodríguez-Mańas, Leocadio; Sánchez-Ferrer, Carlos F

    2003-01-01

    Diabetic vessels undergo structural changes that are linked to a high incidence of cardiovascular diseases. Reactive oxygen species (ROS) mediate cell signalling in the vasculature, where they can promote cell growth and activate redox-regulated transcription factors, like activator protein-1 (AP-1) or nuclear factor-?B (NF-?B), which are involved in remodelling and inflammation processes. Amadori adducts, formed through nonenzymatic glycosylation, can contribute to ROS formation in diabetes. In this study, we analysed whether Amadori-modified human oxyhaemoglobin, glycosylated at either normal (N-Hb) or elevated (E-Hb) levels, can induce cell growth and activate AP-1 and NF-?B in cultured human aortic smooth muscle cells (HASMC). E-Hb (1 nM–1 ?M), but not N-Hb, promoted a concentration-dependent increase in cell size from nanomolar concentrations, although it failed to stimulate HASMC proliferation. At 10 nM, E-Hb stimulated both AP-1 and NF-?B activity, as assessed by transient transfection, electromobility shift assays or immunofluorescence staining. The effects of E-Hb resembled those of the proinflammatory cytokine tumour necrosis factor-? (TNF-?). E-Hb enhanced intracellular superoxide anions content and its effects on HASMC were abolished by different ROS scavengers. In conclusion, E-Hb stimulates growth and activates AP-1 and NF-?B in human vascular smooth muscle by redox-sensitive pathways, thus suggesting a possible direct role for Amadori adducts in diabetic vasculopathy. PMID:14504138

  17. Bayesian modeling of human concept learning Joshua B. Tenenbaum

    E-print Network

    Tenenbaum, Josh

    behavioral data. I focus on a simple abstract task of interest to both cognitive science and machine learning routinely but which com- puters are rarely capable of. Bridging machine learning and cognitive science that the examples are a random sample from the concept to be learned. The model gives precise fits to human behavior

  18. Complete Genome Sequences of Human Respiratory Syncytial Virus Genotype A and B Isolates from South Korea

    PubMed Central

    Yun, Mi-ran; Kim, A-Reum; Lee, Han Saem; Kim, Dae-Won; Lee, Wan-Ji; Kim, Kisoon; Kim, Sung Soon

    2015-01-01

    There is a paucity of complete genome sequence information for human respiratory syncytial virus (HRSV). To this end, we sequenced the complete genome sequences of HRSV genotype A (HRSV-A/IC688/12) and genotype B (HRSV-B/GW0047/14 and HRSV-B/IC0027/14). This information will increase the understanding of HRSV genetic diversity, evolution, pathogenicity, antigenicity, and transmissibility. PMID:25908140

  19. Inhibition of apolipoprotein B and triglyceride secretion in human hepatoma cells (HepG2)

    Microsoft Academic Search

    Mehrdad Haghpassand; Don Wilder; James B. Moberlyl

    Apolipoprotein B (apoB), the major protein com- ponent of triglyceride-rich lipoproteins, is assembled into a lipoprotein particle via a complex, multistep process. Recent studies indicate that triglyceride-rich lipoprotein assembly re- quires the activity of the heterodimeric protein, microsomal triglyceride transfer protein (MTP). We identified a novel inhibitor of apolipoprotein B secretion using the human hepatoma cell line, HepC2. CP-10447, a

  20. Clarithromycin Inhibits NF B Activation in Human Peripheral Blood Mononuclear Cells and Pulmonary Epithelial Cells

    Microsoft Academic Search

    TAKASHI ICHIYAMA; MIKI NISHIKAWA; TOMOMI YOSHITOMI; SHUNJI HASEGAWA; TOMOYO MATSUBARA; TAKASHI HAYASHI; SUSUMU FURUKAWA

    2001-01-01

    Macrolide antibiotics modulate the production of proinflammatory cytokines in vivo and in vitro. Tran- scription of the genes for these proinflammatory cytokines is regulated by nuclear factor kB (NF-kB). We ex- amined whether or not clarithromycin inhibits the activation of NF-kB induced by tumor necrosis factor alpha (TNF-a) or staphylococcal enterotoxin A (SEA) in human monocytic U-937 cells, a T-cell

  1. Human Parvovirus B19 as a Causative Agent for Rheumatoid Arthritis

    Microsoft Academic Search

    Yuichi Takahashi; Chihiro Murai; Shinobu Shibata; Yasuhiko Munakata; Tomonori Ishii; Keiko Ishii; Takako Saitoh; Takashi Sawai; Kazuo Sugamura; Takeshi Sasaki

    1998-01-01

    Human parvovirus B19 (B19) DNA was detected in the synovial tissues in 30 of 39 patients with rheumatoid arthritis (RA), and infrequently in those with osteoarthritis and traumatic joints. On the other hand, the expression of the B19 antigen VP-1 was specific (27\\/27) in RA synovium with active synovial lesions, but not in osteoarthritis and controls. The target cells of

  2. Transient expression, purification and characterization of bioactive human fibroblast growth factor 8b in tobacco plants

    Microsoft Academic Search

    H. H. Surya Kumar Potula; Sonal Roy Kathuria; A. K. Ghosh; T. K. Maiti; S. Dey

    2008-01-01

    cDNA of human fibroblast growth factor 8 isoform b (FGF8b) was cloned for the first time into a plant expression vector with\\u000a or without endoplasmic reticulum retention signal (KDEL) and was transiently expressed as His tagged fusion protein in Nicotiana tabacum leaves through Agrobacterium mediated gene transfer by vacuum infiltration method. Expression of FGF8b was confirmed by ELISA and Western

  3. Pharmacological effects of recombinant human tissue kallikrein on bradykinin B2 receptors

    PubMed Central

    Charest-Morin, Xavier; Raghavan, Arvind; Charles, Matthew L; Kolodka, Tadeusz; Bouthillier, Johanne; Jean, Mélissa; Robbins, Mark S; Marceau, François

    2015-01-01

    Tissue kallikrein (KLK-1), a serine protease, initiates the release of bradykinin (BK)-related peptides from low-molecular weight kininogen. KLK-1 and the BK B2 receptor (B2R) mediate beneficial effects on the progression of type 2 diabetes and renal disease, but the precise role of KLK-1 independent of its kinin-forming activity remains unclear. We used DM199, a recombinant form of human KLK-1, along with the isolated human umbilical vein, a robust bioassay of the B2R, to address the previous claims that KLK-1 directly binds to and activates the human B2R, with possible receptor cleavage. DM199 (1–10 nmol/L) contracted the isolated vein via the B2R, but in a tachyphylactic, kinin-dependent manner, without desensitization of the tissue to exogenously added BK. In binding experiments with recombinant N-terminally tagged myc-B2Rs expressed in HEK 293a cells, DM199 displaced [3H]BK binding from the rabbit myc-B2R, but not from the human or rat myc-B2Rs. No evidence of myc-B2R degradation by immunoblot analysis was apparent following treatment of these 3 myc-B2R constructs with DM199 (30 min, ?10 nmol/L). In HEK 293 cells stably expressing rabbit B2R-GFP, DM199 (11–108 pmol/L) elicited signaling-dependent endocytosis and reexpression, while a higher concentration (1.1 nmol/L) induced a partially irreversible endocytosis of the construct (microscopy), paralleled by the appearance of free GFP in cells (immunoblotting, indicative of incomplete receptor down-regulation). The pharmacology of DM199 at relevant concentrations (<10 nmol/L) is essentially based on the activity of locally generated kinins. Binding to and mild down-regulation of the B2R is possibly a species-dependent idiosyncratic response to DM199.

  4. Molecular cloning of cDNA for CENP-B, the major human centromere autoantigen

    Microsoft Academic Search

    William C. Earnshaw; Kevin E Sullivan; Paula S. Machlin; Carol A. Cooke; Donald A. Kaiser; Thomas D. Pollard; Naomi E Rothfield; Don W. Cleveland

    1987-01-01

    We have isolated a series of overlapping cDNA clones for ~95 % of the mRNA that encodes CENP-B, the 80-kD human centromere autoantigen recognized by patients with anticentromere antibodies. The cloned sequences encode a polypeptide with an apparent molecular mass appropriate for CENP-B. This polypeptide and CENP-B share three non-over- lapping epitopes. The first two are defined by monoclonal antibodies

  5. NF-kappaB blockade and oncogenic Ras trigger invasive human epidermal neoplasia

    Microsoft Academic Search

    Maya Dajee; Mirella Lazarov; Jennifer Y. Zhang; Ti Cai; Cheryl L. Green; Alan J. Russell; M. Peter Marinkovich; Shiying Tao; Qun Lin; Yoshiaki Kubo; Paul A. Khavari

    2003-01-01

    The nuclear factor NF-kappaB and oncogenic Ras can alter proliferation in epidermis, the most common site of human cancer. These proteins are implicated in epidermal squamous cell carcinoma in mice, however, the potential effects of altering their function are uncertain. Whereas inhibition of NF-kappaB enhances apoptosis in certain tumours, blockade of NF-kappaB predisposes murine skin to squamous cell carcinoma. Because

  6. Activation of Epstein-Barr virus latent genes protects human B cells from death by apoptosis

    Microsoft Academic Search

    Christopher D. Gregory; Caroline Dive; Sheila Henderson; Christopher A. Smith; Gwyn T. Williams; John Gordon; Alan B. Rickinson

    1991-01-01

    EPSTEiN-Barr virus (EBV), a human herpesvirus, establishes a persistent asymptomatic infection of the circulating B-lymphocyte pool1-3. The mechanism of virus persistence is not understood but, given the limited lifespan of most B cells in vivo, it seems most likely that EBV-infected cells must gain access to the long-lived memory B-cell pool. Here we show in an in vitro system that

  7. Association of Human Herpesvirus6B with Mesial Temporal Lobe Epilepsy

    Microsoft Academic Search

    Julie Fotheringham; Donatella Donati; Nahid Akhyani; Anna Fogdell-Hahn; Alexander Vortmeyer; John D. Heiss; Elizabeth Williams; Steven Weinstein; Derek A. Bruce; William D. Gaillard; Susumu Sato; William H. Theodore; Steven Jacobson

    2007-01-01

    Background Human herpesvirus-6 (HHV-6) is a b-herpesvirus with 90% seroprevalence that infects and establishes latency in the central nervous system. Two HHV-6 variants are known: HHV-6A and HHV-6B. Active infection or reactivation of HHV-6 in the brain is associated with neurological disorders, including epilepsy, encephalitis, and multiple sclerosis. In a preliminary study, we found HHV-6B DNA in resected brain tissue

  8. Treatment with ephrin B2 positively impacts the abnormal metabolism of human osteoarthritic chondrocytes

    Microsoft Academic Search

    Steeve Kwan Tat; Jean-Pierre Pelletier; Nathalie Amiable; Christelle Boileau; Martin Lavigne; Johanne Martel-Pelletier

    2009-01-01

    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

  9. Maintenance of Serological Memory by Polyclonal Activation of Human Memory B Cells

    NASA Astrophysics Data System (ADS)

    Bernasconi, Nadia L.; Traggiai, Elisabetta; Lanzavecchia, Antonio

    2002-12-01

    Production of antibodies can last for a lifetime, through mechanisms that remain poorly understood. Here, we show that human memory B lymphocytes proliferate and differentiate into plasma cells in response to polyclonal stimuli, such as bystander T cell help and CpG DNA. Furthermore, plasma cells secreting antibodies to recall antigens are produced in vivo at levels proportional to the frequency of specific memory B cells, even several years after antigenic stimulation. Although antigen boosting leads to a transient increase in specific antibody levels, ongoing polyclonal activation of memory B cells offers a means to maintain serological memory for a human lifetime.

  10. Relationship between constitutive nuclear factor-kappaB (NF-?B) and inhibitor kappaB-alpha (I?B-?) in an interferon-?-sensitive human Burkitt lymphoma cell line

    Microsoft Academic Search

    Pramod C. Rath

    2005-01-01

    The human Burkitt lymphoma Daudi cell line expresses constitutively active nuclear factor kappaB (NF-?B) in the nucleus in spite of high levels of inhibitor kappaB-alpha (I?B-?) in the cytoplasm. The antiproliferative response of these cells to interferon-? (IFN-?) correlated with the inhibition of the constitutive NF-?B activity by the cytokine. The present study shows that IFN-? caused an increase in

  11. Innate Response Activator (IRA) B Cells Reside in Human Tonsils and Internalize Bacteria In Vitro

    PubMed Central

    Pancotto, Laura; Ruggiero, Paolo; Rosa, Domenico; Manetti, Andrea; Romano, Antonio; Montagnani, Francesca; Bertholet, Sylvie; Castellino, Flora; Del Giudice, Giuseppe

    2015-01-01

    Innate response activator (IRA) B cells have been described in mice as a subset of B-1a B cells that produce granulocyte/macrophage colony-stimulating factor (GM-CSF) and have been found in the spleen upon activation. In humans, identification, tissue localization and functionality of these lymphocytes are poorly understood. We hypothesized that IRA B cells could reside in human palatine tonsils, which are a first line of defense from infection of the upper respiratory tract. In the present work, we used flow cytometry and confocal microscopy to identify and characterize human IRA (hIRA) B cells in tonsils. We show that CD19+CD20+GM-CSF+ B cells are present in the tonsils of all the subjects studied at a frequency ranging between ~0.2% and ~0.4% of the conventional CD19+CD20+GM-CSF- B cells. These cells reside within the B cell follicles, are mostly IgM+IgD+, express CD5 and show phagocytic activity. Our results support a role for hIRA B cells in the effector immune response to infections in tonsils. PMID:26066485

  12. E3 ubiquitin ligase Cbl-b suppresses human ORMDL3 expression through STAT6 mediation.

    PubMed

    Yang, Wei-Xia; Jin, Rui; Jiang, Chun-Ming; Wang, Xiao-Hua; Shu, Jin; Li, Ling; Zhu, Liang-Hua; Zhuang, Li-Li; Gao, Chao; Zhou, Guo-Ping

    2015-07-01

    Orosomucoid 1-Like Protein 3 (ORMDL3) is an asthma candidate gene and Casitas B lineage lymphoma b (Cbl-b), an E3 ubiquitin ligase, is a critical factor in maintaining airway immune tolerance. However, the association of Cbl-b with ORMDL3 for asthma is unclear. Here, we show that expression of ORMDL3 is significantly increased and shows a strong linear correlation with decreased Cbl-b in the peripheral blood of recurrent wheeze patients. To elucidate the molecular mechanisms underlying this correlation, we identified that Cbl-b suppressed the transcriptional activity and mRNA expression of ORMDL3 in vivo. Further investigation showed that phosphorylation of signal transducer and activator of transcription 6 (STAT6) was induced by interleukin 4 bound to the ORMDL3 promoter, while Cbl-b reduced the phosphorylation of STAT6. Our results show that Cbl-b suppresses human ORMDL3 expression through STAT6. PMID:26112603

  13. EVALUATION OF MIXED CELL TYPES AND 5-IODO-2'-DEOXYURIDINE TREATMENT UPON PLAQUE ASSAY TITERS OF HUMAN ENTERIC VIRUSES (JOURNAL VERSION)

    EPA Science Inventory

    Four continuous cell lines BGM, L-132, HEL-299, and RD were compared both when cultured separately and as mixtures for use in plaque assay titrations of human Adenovirus 1 and six human enterovirus serotypes. The effect of incubating these cell cultures in media containing IDU (5...

  14. Position 156 influences the peptide repertoire and tapasin dependency of human leukocyte antigen B*44 allotypes

    PubMed Central

    Badrinath, Soumya; Saunders, Philippa; Huyton, Trevor; Aufderbeck, Susanne; Hiller, Oliver; Blasczyk, Rainer; Bade-Doeding, Christina

    2012-01-01

    Background Polymorphic differences between donor and recipient human leukocyte antigen class I molecules can result in graft-versus-host disease due to distinct peptide presentation. As part of the peptide-loading complex, tapasin plays an important role in selecting peptides from the pool of potential ligands. Class I polymorphisms can significantly alter the tapasin-mediated interaction with the peptide-loading complex and although most class I allotypes are highly dependent upon tapasin, some are able to load peptides independently of tapasin. Several human leukocyte antigen B*44 allotypes differ exclusively at position 156 (B*44:02156Asp, 44:03156Leu, 44:28156Arg, 44:35156Glu). From these alleles, only the high tapasin-dependency of human leukocyte antigen B*44:02 has been reported. Design and Methods We investigated the influence of position 156 polymorphisms on both the requirement of tapasin for efficient surface expression of each allotype and their peptide features. Genes encoding human leukocyte antigen B*44 variants bearing all possible substitutions at position 156 were lentivirally transduced into human leukocyte antigen class I-negative LCL 721.221 cells and the tapasin-deficient cell line LCL 721.220. Results Exclusively human leukocyte antigen B*44:28156Arg was expressed on the surface of tapasin-deficient cells, suggesting that the remaining B*44/156 variants are highly tapasin-dependent. Our computational analysis suggests that the tapasin-independence of human leukocyte antigen B*44:28156Arg is a result of stabilization of the peptide binding region and generation of a more peptide receptive state. Sequencing of peptides eluted from human leukocyte antigen B*44 molecules by liquid chromatography-electrospray ionization-mass spectrometry (LTQ-Orbitrap) demonstrated that both B*44:02 and B*44:28 share the same overall peptide motif and a certain percentage of their individual peptide repertoires in the presence and/or absence of tapasin. Conclusions Here we report for the first time the influence of position 156 on the human leukocyte antigen/tapasin association. Additionally, the results of peptide sequencing suggest that tapasin chaperoning is needed to acquire peptides of unusual length. PMID:21993680

  15. CS1 (CRACC, CD319) induces proliferation and autocrine cytokine expression on human B lymphocytes.

    PubMed

    Lee, Jae Kyung; Mathew, Stephen O; Vaidya, Swapnil V; Kumaresan, Pappanaicken R; Mathew, Porunelloor A

    2007-10-01

    CS1 (CRACC, CD319), a member of the CD2 family of cell surface receptors, is implicated in the activation of NK cell-mediated cytotoxicity. Previous studies showed that CS1 is also expressed on activated B cells. However, the functional role of CS1 in human B-lymphocytes is not known. Two isoforms of CS1, CS1-L and CS1-S, are expressed in human NK cells that differentially regulate NK cell function. CS1-L contains immunoreceptor tyrosine-based switch motifs in its cytoplasmic domain whereas CS1-S lacks immunoreceptor tyrosine-based switch motifs. In this study, we show that human B lymphocytes express only the CS1-L isoform, and its expression is up-regulated upon B cell activation with various stimulators. Moreover, anti-CS1 mAb strongly enhanced proliferation of both freshly isolated as well as activated B cells. The enhanced proliferation effects of CS1 were most prominent on B cells activated by anti-CD40 mAbs and/or hrIL-4. The effects of CS1 on B cell proliferation were shown on both naive and memory B cells. Human cytokine microarray and quantitative real-time PCR results indicated that CS1 activation enhanced mRNA transcripts of flt3 ligand, lymphotoxin A, TNF, and IL-14. Neutralizing Abs against lymphotoxin A, TNF-alpha, and/or flt3 ligand abolished the ability of CS1 on the B cell proliferation. These results suggest that activation of B lymphocytes, through surface CS1, may be mediated through secretion of autocrine cytokines and CS1 may play a role in the regulation of B lymphocyte proliferation during immune responses. PMID:17878365

  16. Influence of Ultraviolet B Exposure on Immune Responses Following Hepatitis B Vaccination in Human Volunteers

    Microsoft Academic Search

    Annemarie Sleijffers; Johan Garssen; Frank R. de Gruijl; Greet J. Boland; Jan van Hattum; Willem A. van Vloten; Henk van Loveren

    2001-01-01

    Exposure to ultraviolet radiation can modulate immune responses in animal and humans. Remarkably, the ultraviolet-induced immunosuppression is not restricted to the exposed skin but is also found at other body sites, i.e., systemic immunosuppression. Effects of ultraviolet radiation on infections cannot be determined by experimentation on humans, but the effects of ultraviolet on vaccination may serve as a model. Moreover,

  17. Evaluation of a Real-Time Reverse Transcription-PCR Assay for Detection of Enterovirus D68 in Clinical Samples from an Outbreak in New York State in 2014.

    PubMed

    Zhuge, Jian; Vail, Eric; Bush, Jeffrey L; Singelakis, Lauren; Huang, Weihua; Nolan, Sheila M; Haas, Janet P; Engel, Helen; Della Posta, Millicent; Yoon, Esther C; Fallon, John T; Wang, Guiqing

    2015-06-01

    An outbreak of severe respiratory illness associated with enterovirus D68 (EV-D68) infection was reported in mid-August 2014 in the United States. In this study, we evaluated the diagnostic utility of an EV-D68-specific real-time reverse transcription-PCR (rRT-PCR) that was recently developed by the Centers for Disease Control and Prevention in clinical samples. Nasopharyngeal (NP) swab specimens from patients in a recent outbreak of respiratory illness in the lower Hudson Valley, New York State, were collected and examined for the presence of human rhinovirus or enterovirus using the FilmArray Respiratory Panel (RP) assay. Samples positive by RP were assessed using EV-D68 rRT-PCR, and the data were compared to results from sequencing analysis of partial VP1 and 5' untranslated region (5'-UTR) sequences of the EV genome. A total of 285 RP-positive NP specimens (260 from the 2014 outbreak and 25 from 2013) were analyzed by rRT-PCR; EV-D68 was detected in 74 of 285 (26.0%) specimens examined. Data for comparisons between rRT-PCR and sequencing analysis were obtained from 194 NP specimens. EV-D68 detection was confirmed by sequencing analysis in 71 of 74 positive and in 1 of 120 randomly selected negative specimens by rRT-PCR. The EV-D68 rRT-PCR showed diagnostic sensitivity and specificity of 98.6% and 97.5%, respectively. Our data suggest that the EV-D68 rRT-PCR is a reliable assay for detection of EV-D68 in clinical samples and has a potential to be used as a tool for rapid diagnosis and outbreak investigation of EV-D68-associated infections in clinical and public health laboratories. PMID:25854481

  18. Enterovirus 71 Induces Mitochondrial Reactive Oxygen Species Generation That is Required for Efficient Replication

    PubMed Central

    Cheng, Mei-Ling; Weng, Shiue-Fen; Kuo, Chih-Hao; Ho, Hung-Yao

    2014-01-01

    Redox homeostasis is an important host factor determining the outcome of infectious disease. Enterovirus 71 (EV71) infection has become an important endemic disease in Southeast Asia and China. We have previously shown that oxidative stress promotes viral replication, and progeny virus induces oxidative stress in host cells. The detailed mechanism for reactive oxygen species (ROS) generation in infected cells remains elusive. In the current study, we demonstrate that mitochondria were a major ROS source in EV71-infected cells. Mitochondria in productively infected cells underwent morphologic changes and exhibited functional anomalies, such as a decrease in mitochondrial electrochemical potential ??m and an increase in oligomycin-insensitive oxygen consumption. Respiratory control ratio of mitochondria from infected cells was significantly lower than that of normal cells. The total adenine nucleotide pool and ATP content of EV71-infected cells significantly diminished. However, there appeared to be a compensatory increase in mitochondrial mass. Treatment with mito-TEMPO reduced eIF2? phosphorylation and viral replication, suggesting that mitochondrial ROS act to promote viral replication. It is plausible that EV71 infection induces mitochondrial ROS generation, which is essential to viral replication, at the sacrifice of efficient energy production, and that infected cells up-regulate biogenesis of mitochondria to compensate for their functional defect. PMID:25401329

  19. Inhibition of Enterovirus 71 Replication by 7-Hydroxyflavone and Diisopropyl-Flavon7-yl Phosphate

    PubMed Central

    Du, Jiang; Cui, Sheng; Yang, Fan; Jin, Qi

    2014-01-01

    Enterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease, which has been continuously prevalent in Asia in recent years. In children, severe cases can lead to death, and no prophylactic or therapeutic measures against EV71 infection are available. The 3C proteases of EV71 play an important role in viral replication and are an ideal drug target. In previous work, we resolved the crystal structure for EV71 3Cpro. In this report, we took advantage of the automated docking program AutoDock 4.0 to simulate EV71 3Cpro-ligand conformation. 7-hydroxyflavone (HF) and its phosphate ester(FIP) were predicted to bind with EV71 3Cpro.In an in vitro protease inhibition assay, FIP inhibited EV71 3Cpro protease activity. Both flavones were highly active against EV71, protecting cells from EV71 infection. Replication of viral RNA and formation of EV71 plaque were all strongly inhibited in cells. These results indicated that HF and FIP may serve as potential protective agents in the treatment of patients with chronic EV71 infection. PMID:24664133

  20. Effect of Meteorological Conditions and Geographical Factors in the Onset of Enterovirus 71

    NASA Astrophysics Data System (ADS)

    Chen, Yu-An; Yu, Hwa-Lung

    2015-04-01

    Since it was first recognized in California in 1969, enterovirus 71 (EV71) infection has been a significant cause of neurological disorder and death in children worldwide. In 1998 a historic epidemic of EV71 infection caused hand-foot-and-mouth disease and herpangina in thousands of people in Taiwan. The impact of EV71 infection is greatest during the summer months in Asia, and epidemics recur with a seasonal pattern. It was reported that seasonal patterns of EV71 differed by geographical localities. Previous studies have also showed significant relationships between meteorological variables, in particular, temperature and relative humidity, and the seasonal epidemic patterns of EV71. However, important issues that remain unclear include the spatiotemporal pattern of the EV71 outbreaks in Taiwan, and what role of favorable meteorological conditions in the transmission of the disease in the space-time domain. Thus, this study used a semiparametric generalized additive model (GAM) to understand the association between EV71 and meteorological factors across space and time. This study utilized a population-based database containing space-time data for clinic and hospital visits (i.e., hospital location and appointment times) of EV71 occurring in children less than 18 years old in Taipei from 1998 to 2008. Meteorological data (i.e., temperature, rainfall, and relative humidity) for the study period were provided by the Taiwan Central Weather Bureau. This study expect to find out an important meteorological factor and threshold.

  1. Detection of porcine teschoviruses and enteroviruses by LightCycler real-time PCR.

    PubMed

    Krumbholz, Andi; Wurm, Rüdiger; Scheck, Oliver; Birch-Hirschfeld, Eckard; Egerer, Renate; Henke, Andreas; Wutzler, Peter; Zell, Roland

    2003-10-01

    Porcine picornaviruses comprising at least 23 serotypes grouped into six species were described as causative agents of neurological disorders, reproductive failure, and aphthae-like dermal lesions of swine. Other viruses such as classical swine fever virus (CSFV), African swine fever virus, pseudorabies virus (PRV), vesicular stomatitis virus, vesicular exanthema virus, porcine respiratory and reproductive syndrome virus, and porcine parvovirus (PPV) may cause diseases with similar clinical symptoms. Therefore, rapid and reliable PCR detection of the most frequent porcine picornaviruses is of interest. A real-time RT-PCR protocol employing LightCycler technology to detect all known serotypes of the three porcine enterovirus (PEV) cytopathic effect (CPE) groups was established. It uses three sets of primer pairs and group-specific hybridisation probes. The primer pairs were designed to amplify highly conserved sequences of the 5'-non-translated region (5'-NTR) of the relevant virus species. The one-step real-time PCR based on the LightCycler technology is more rapid and less contamination-prone than the nested RT-PCR and allows the precise quantitation of the virus load in the tested specimens. All acknowledged serotypes of the three PEV CPE groups and all tested field strains isolated from clinical specimens were detectable. Viruses of the PEV CPE group III can be distinguished from the closely related swine vesicular disease virus (SVDV). PMID:14500127

  2. Association of IP-10 gene polymorphism with susceptibility to Enterovirus 71 infection

    PubMed Central

    YANG, JING; CHEN, ZHEN-ZHEN; LV, TIE-GANG; LIU, PEI-PEI; CHEN, ZONG-BO

    2013-01-01

    Enterovirus 71 (EV71) often causes large outbreaks of diseases among children worldwide and its pathogenesis remains unclear. The aim of the present study was to investigate the association between interferon-inducible protein 10 (IP-10) polymorphism in children with EV71 infection. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were performed to analyze the gene polymorphisms of IP-10 (?1596C/T) in 58 EV71-infected and 48 control patients. The results showed that in EV71-infected patients the frequency of carrying CT + TT genotype and T allele is 10.3 and 6.0%, respectively, which is significantly lower than that of the controls (29.2 and 15.6%, respectively). Individuals with T allele had a lower risk of EV71 infection [odds ratio (OR) = 0.35, 95% confidence interval (CI), 0.13–0.89]. The results of this study indicated that ?1596T allele for the IP-10 gene may be a beneficial factor for EV71 infection. PMID:24648959

  3. Massive pulmonary hemorrhage in enterovirus 71-infected hand, foot, and mouth disease

    PubMed Central

    Lee, Dong Seong; Lee, Young Il; Ahn, Jeong Bae; Kim, Mi Jin; Kim, Jae Hyun; Kim, Nam Hee; Hwang, Jong Hee; Kim, Dong Wook; Lee, Chong Guk

    2015-01-01

    Hand, foot, and mouth disease (HFMD) is an acute, mostly self-limiting infection. Patients usually recover without any sequelae. However, a few cases are life threatening, especially those caused by enterovirus 71 (EV71). A 12-month-old boy was admitted to a primary hospital with high fever and vesicular lesions of the mouth, hands, and feet. After 3 days, he experienced 3 seizure episodes and was referred to our hospital. On admission, he was conscious and his chest radiograph was normal. However, 6 hours later, he suddenly lost consciousness and had developed a massive pulmonary hemorrhage that continued until his death. He experienced several more intermittent seizures, and diffuse infiltration of both lung fields was observed on chest radiography. Intravenous immunoglobulin, dexamethasone, cefotaxime, leukocyte-depleted red blood cells, fresh frozen plasma, inotropics, vitamin K, and endotracheal epinephrine were administered. The patient died 9 hours after intubation, within 3 days from fever onset. EV71 subgenotype C4a was isolated retrospectively from serum and nasopharyngeal swab by real-time reverse transcription-polymerase chain reaction. Here, we report a fatal case of EV71-associated HFMD with sudden-onset massive pulmonary hemorrhage and suspected encephalitis. PMID:25861335

  4. Rapid and highly sensitive detection of Enterovirus 71 by using nanogold-enhanced electrochemical impedance spectroscopy

    NASA Astrophysics Data System (ADS)

    Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Lu, Yu-Ning; Wang, Fang-Yu; Tsai, Li-Yun; Shieh, Juo-Yu; Yang, Jyh-Yuan; Juan, Chien-Chang; Tu, Lung-Chen; Chang, Chia-Ching

    2013-07-01

    Enterovirus 71 (EV71) infection is an emerging infectious disease causing neurological complications and/or death within two to three days after the development of fever and rash. A low viral titre in clinical specimens makes the detection of EV71 difficult. Conventional approaches for detecting EV71 are time consuming, poorly sensitive, or complicated, and cannot be used effectively for clinical diagnosis. Furthermore, EV71 and Coxsackie virus A16 (CA16) may cross react in conventional assays. Therefore, a rapid, highly sensitive, specific, and user-friendly test is needed. We developed an EV71-specific nanogold-modified working electrode for electrochemical impedance spectroscopy in the detection of EV71. Our results show that EV71 can be distinguished from CA16, Herpes simplex virus, and lysozyme, with the modified nanogold electrode being able to detect EV71 in concentrations as low as 1 copy number/50 ?l reaction volume, and the duration between sample preparation and detection being 11 min. This detection platform may have the potential for use in point-of-care diagnostics.

  5. Enterovirus infections in England and Wales, 2000-2011: the impact of increased molecular diagnostics.

    PubMed

    Kadambari, S; Bukasa, A; Okike, I O; Pebody, R; Brown, D; Gallimore, C; Xerry, J; Sharland, M; Ladhani, S N

    2014-12-01

    There have recently been significant changes in diagnostic practices for detecting enterovirus (EV) infections across England and Wales. Reports of laboratory-confirmed EV infections submitted by National Health Service (NHS) hospital laboratories to Public Health England (PHE) over a 12-year period (2000-2011) were analysed. Additionally, the PHE Virus Reference Department (VRD) electronic database containing molecular typing data from 2004 onwards was interrogated. Of the 13,901 reports, there was a decline from a peak of 2254 in 2001 to 589 in 2006, and then an increase year-on-year to 1634 in 2011. This increase coincided with increasing PCR-based laboratory diagnosis, which accounted for 36% of reported cases in 2000 and 92% in 2011. The estimated annual incidence in 2011 was 3.9/100,000 overall and 238/100,000 in those aged <3 months, who accounted for almost one-quarter of reported cases (n = 2993, 23%). During 2004-2011, 2770 strains were submitted for molecular typing to the VRD, who found no evidence for a predominance of any particular strain. Thus, the recent increase in reported cases closely reflects the increase in PCR testing by NHS hospitals, but is associated with a lower proportion of samples being submitted for molecular typing. The high EV rate in young infants merits further investigation to inform evidence-based management guidance. PMID:25039903

  6. Saururus chinensis (Lour.) Baill blocks enterovirus 71 infection by hijacking MEK1-ERK signaling pathway.

    PubMed

    Wang, Chunyang; Wang, Peng; Chen, Xiaoqing; Wang, Wei; Jin, Yu

    2015-07-01

    The aerial parts of Saururus chinensis (Lour.) Baill are a Chinese herbal medicine used for the treatment of edema and inflammatory diseases. However, the effect of this medicine on enterovirus 71 (EV71) infection has not been explored. Previous studies showed that MEK1-ERK signal pathway was required for efficient replication of EV71 infection and inhibition of this signal pathway has been shown to suppress virus infection. Here we show that the water extract of S. chinensis (Lour.) Baill (SCB) significantly blocks EV71 infection by inhibiting the activation of MEK1-ERK signal pathway with an IC50 of 8.9?g/mL. SCB at 30 and 60?g/mL blocked EV71-induced cytopathic effect (CPE) and production of infectious virion by 1.9 and 5.1logs, respectively. Virucidal assay suggested that SCB had no virucidal activity against EV71 and probably exerted its effect by targeting multiple steps in EV71 infection. Knockdown of MEK1 but not MEK2 blocked EV71 replication. And SCB treatment inhibited the activation of MEK1-ERK signal during EV71 infection. Furthermore, we found that rutin at 200?M, one of the major components of SCB, significantly suppressed EV71 induced CPE and inhibited viral replication in a dose dependent manner. Taken together, SCB inhibited EV71 infection by hijacking MEK1-ERK signal pathway and rutin was the responsible antiviral component of SCB. PMID:25912818

  7. Human immunodeficiency virus 1 Nef suppresses CD40-dependent immunoglobulin class switching in bystander B cells.

    PubMed

    Qiao, Xugang; He, Bing; Chiu, April; Knowles, Daniel M; Chadburn, Amy; Cerutti, Andrea

    2006-03-01

    Immunoglobulin class switching from immunoglobulin M (IgM) to IgG and IgA is central to immunity against viruses and requires the activation of B cells by T cells via CD154 (CD40 ligand) and cytokines. These molecules limit their signaling activity in immune cells by turning on negative feedback proteins, including IkappaB and SOCS. We show here that negative factor (Nef) protein, an immunosuppressive human immunodeficiency virus 1 protein expressed and released by infected cells, penetrates B cells both in vivo and in vitro. Nef suppressed immunoglobulin class-switch DNA recombination by inducing IkappaBalpha and SOCS proteins, which blocked CD154 and cytokine signaling via NF-kappaB and STAT transcription factors. Thus, human immunodeficiency virus 1 may evade protective T cell-dependent IgG and IgA responses by 'hijacking' physiological feedback inhibitors in B cells via Nef. PMID:16429138

  8. Infectious human papillomavirus type 31b: purification and infection of an immortalized human keratinocyte cell line

    Microsoft Academic Search

    Michelle A. Ozbun

    2002-01-01

    Human papillomaviruses (HPVs) are aetiological agents of human malignancies, most notably cervical cancers. The life-cycles of HPVs are dependent on epithelial differentiation, and this has impeded many basic studies of HPV biology. The organotypic (raft) culture system supports epithelial differentiation such that infectious virions are synthesized in raft tissues from epithelial cells that replicate extrachromosomal HPV genomes. The CIN-612 9E

  9. Interaction of human serum apolipoprotein B with sodium deoxycholate.

    PubMed

    Oeswein, J Q; Chun, P W

    1983-07-01

    Preparation of apolipoprotein B (Apo B)-deoxycholate (DOC) complexes by gel filtration chromatography in the presence of 20 mM DOC, pH 8.5, gave two populations of particles with 5% (peak I) and 13% (peak II) lipid remaining bound. These complexes were initially shown to be very large and elongated, with partition radii of approx. 131 +/- 0.5 A, weight average molecular weights of approx. 164 000 +/- 1 000, and an intrinsic viscosity of 80.19 +/- 2.21 ml/g. Additionally, they appeared very similar to native low-density lipoprotein on sodium dodecyl sulfate-polyacrylamide gels, giving one major band. Incubation of these samples for 10 days under nitrogen at 4 degrees C in the presence of antibiotics and protease inhibitor resulted in dissociation to many smaller subunits. Results of scanning molecular sieve chromatography and analytical ultracentrifugation showed that dissociation of these complexes was relatively slow and indicated the presence of at least two classes of components in fresh samples: one a very elongated complex with a radius directly correlated to the DOC/Apo B ratio and inversely correlated to sample aging; and another of much smaller radius which was independent of DOC/Apo B ratio but directly correlated to sample aging; indicating that these dissociated subunits interact with each other to an appreciable extent. Furthermore, these complexes were found to undergo a preferential hydration upon interaction with DOC, which may contribute to large changes in their effective specific volumes, as well as to dissociation of subunits. PMID:6882865

  10. Enzyme-linked immunosorbent assay for human plasma apolipoprotein B

    Microsoft Academic Search

    Jose M. Ordovas; Jeffity P. Peterson; Patricia Santaniello; Jeffrey S. Cohn; Peter W. F. Wilson; Ernst J. Schaefer

    A noncompetitive enzyme-linked immunosorbent assay (ELISA) has been developed for measuring total plasma apolipoprotein (apo) B using affinity purified polyclonal and monoclonal antibodies. Microtiter plates from different man- ufacturers were tested with regard to their IgG binding char- acteristics; only one plate yielded consistent coefficients of variation of less than 5%. The optimal plasma dilution in this assay was 1:3000.

  11. Development of Monoclonal Antibodies against Human CYP11B1 and CYP11B2

    PubMed Central

    Gomez-Sanchez, Celso E.; Qi, Xin; Velarde-Miranda, Carolina; Plonczynski, Maria W.; Parker, C. Richard; Rainey, William; Satoh, Fumitoshi; Maekawa, Takashi; Nakamura, Yasuhiro; Sasano, Hironobu; Gomez-Sanchez, Elise P.

    2014-01-01

    The final enzymes in the biosynthesis of aldosterone and cortisol are by the cytochrome P450 CYP11B2 and CYP11B1, respectively. The enzymes are 93% homologous at the amino acid level and specific antibodies have been difficult to generate. Mice and rats were immunized with multiple peptides conjugated to various immunogenic proteins and monoclonal antibodies were generated. The only peptide sequences that generated specific antibodies were amino acids 41-52 for the CYP11B2 and amino acids 80-90 for the CYP11B1 enzyme. The mouse monoclonal CYP11B2-41 was specific and sensitive for use in western blots and produced specific staining of the zona glomerulosa of normal adrenal glands. The rat monoclonal CYP11B1-80 also detected a single band by western blot and detected only the zona fasciculata. Triple immunofluorescence of the adrenal demonstrated that the CYP11B1 and the CYP11B2 did not co-localize, while as expected the CYP11B1 co-localized with the 17?-hydroxylase. PMID:24325867

  12. Seroepidemiology of Coxsackievirus A6, Coxsackievirus A16, and Enterovirus 71 Infections among Children and Adolescents in Singapore, 2008-2010

    PubMed Central

    Ang, Li Wei; Tay, Joanne; Phoon, Meng Chee; Hsu, Jung Pu; Cutter, Jeffery; James, Lyn; Goh, Kee Tai; Chow, Vincent Tak-Kwong

    2015-01-01

    Coxsackieviruses A6 (CV-A6) and A16 (CV-A16) and Enterovirus 71 (EV-A71) have caused periodic epidemics of hand, foot and mouth disease (HFMD) among children in Singapore. We conducted a cross-sectional study to estimate the seroprevalence of these enteroviruses among Singapore children and adolescents. The study was conducted between August 2008 and July 2010. It involved 700 Singapore residents aged 1–17 years whose residual sera were obtained following the completion of routine biochemical investigations in two public acute-care hospitals. The levels of neutralizing antibodies (NtAb) against CV-A6, CV-A16 and EV-A71 were analyzed by the microneutralization test. The age-specific geometric mean titer (GMT) of antibodies against each of the three enteroviruses and the 95% confidence intervals (CI) were calculated. The seroprevalence of CV-A6 and CV-A16 was high at 62.7% (95% CI: 59.1–66.2%) and 60.6% (95% CI: 56.9–64.1%), respectively. However, the seroprevalence of EV-A71 was significantly lower at 29.3% (95% CI: 26.0–32.8%). About 89.7% of the children and adolescents had been infected by at least one of the three enteroviruses by 13–17 years of age. About half (52.3%) were seropositive for two or all three enteroviruses, while only 16.1% had no NtAb against any of the three enteroviruses. High NtAb levels were observed in the younger age groups. CV-A6 and CV-A16 infections are very common among Singapore children and adolescents, while EV-A71 infections are less common. Infection is continually acquired from early childhood to adolescent age. PMID:26011735

  13. Selective enrichment in human megakaryocyte progenitors by the B203.13 surface differentiation antigen.

    PubMed

    Vitale, M; Zauli, G; Bassini, A; Visani, G; Rezzani, R; Croce, C M; Manzoli, F A

    1997-12-01

    The B203.13 monoclonal antibody specifically recognizes a subset (8-18.5%) of human CD34+ haemopoietic progenitors, which are significantly (P<0.05) enriched in megakaryocyte progenitors (CFU-meg). Moreover B203.13 expression was progressively lost as maturation proceeded along the megakaryocytic lineage. In fact: (i) CFU-meg derived from CD34+/B203.13+ showed a greater number of cells/colony with respect to CD34+/B203.13-; (ii) after 10 d of liquid cultures with 100 ng/ml of thrombopoietin, only a minority of CD34-derived cells positive for CD41 coexpressed B203.13; (iii) in situ immunocytochemical staining revealed that B203.13 was expressed exclusively on immature megakaryoblasts; (iv) circulating platelets did not express B203.13. PMID:9432019

  14. B7-H5 costimulates human T cells via CD28H

    PubMed Central

    Zhu, Yuwen; Yao, Sheng; Iliopoulou, Bettina P.; Han, Xue; Augustine, Mathew M.; Xu, Haiying; Phennicie, Ryan T.; Flies, Sarah J.; Broadwater, Megan; Ruff, William; Taube, Janis M.; Zheng, Linghua; Luo, Liqun; Zhu, Gefeng; Chen, Jianzhu; Chen, Lieping

    2013-01-01

    The B7/CD28 family has profound modulatory effects in immune responses and constitutes important targets for the development of novel therapeutic drugs against human diseases. Here we describe a new CD28 homolog (CD28H) that has unique functions in the regulation of the human immune response and is absent in mice. CD28H is constitutively expressed on all naive T cells. Repetitive antigenic exposure, however, induces a complete loss of CD28H on many T cells, and CD28H-negative T cells have a phenotype of terminal differentiation and senescence. After extensive screening in a receptor array, a B7-like molecule, B7 homolog 5 (B7-H5), was identified as a specific ligand for CD28H. B7-H5 is constitutively found in macrophages and could be induced on dendritic cells. The B7-H5/CD28H interaction co-stimulates human T cell growth and cytokine production, selectively via an AKT-dependent signaling cascade. Our study identifies a novel co-stimulatory pathway regulating human T cell responses. PMID:23784006

  15. Targeting Syk-activated B cells in murine and human chronic graft-versus-host disease.

    PubMed

    Flynn, Ryan; Allen, Jessica L; Luznik, Leo; MacDonald, Kelli P; Paz, Katelyn; Alexander, Kylie A; Vulic, Ante; Du, Jing; Panoskaltsis-Mortari, Angela; Taylor, Patricia A; Poe, Jonathan C; Serody, Jonathan S; Murphy, William J; Hill, Geoffrey R; Maillard, Ivan; Koreth, John; Cutler, Corey S; Soiffer, Robert J; Antin, Joseph H; Ritz, Jerome; Chao, Nelson J; Clynes, Raphael A; Sarantopoulos, Stefanie; Blazar, Bruce R

    2015-06-25

    Novel therapies for chronic graft-versus-host disease (cGVHD) are needed. Aberrant B-cell activation has been demonstrated in mice and humans with cGVHD. Having previously found that human cGVHD B cells are activated and primed for survival, we sought to further evaluate the role of the spleen tyrosine kinase (Syk) in cGVHD in multiple murine models and human peripheral blood cells. In a murine model of multiorgan system, nonsclerodermatous disease with bronchiolitis obliterans where cGVHD is dependent on antibody and germinal center (GC) B cells, we found that activation of Syk was necessary in donor B cells, but not T cells, for disease progression. Bone marrow-specific Syk deletion in vivo was effective in treating established cGVHD, as was a small-molecule inhibitor of Syk, fostamatinib, which normalized GC formation and decreased activated CD80/86(+) dendritic cells. In multiple distinct models of sclerodermatous cGVHD, clinical and pathological disease manifestations were not eliminated when mice were therapeutically treated with fostamatinib, though both clinical and immunologic effects could be observed in one of these scleroderma models. We further demonstrated that Syk inhibition was effective at inducing apoptosis of human cGVHD B cells. Together, these data demonstrate a therapeutic potential of targeting B-cell Syk signaling in cGVHD. PMID:25852057

  16. CENP-B box and pJa sequence distribution in human alpha satellite higher-order repeats (HOR)

    E-print Network

    Pavin, Nenad

    CENP-B box and pJa sequence distribution in human alpha satellite higher-order repeats (HOR) Marija Macgregor 3 June 2006 Key words: alphoid arrays, CENP-B box, higher-order repeat, human alpha satellite consensus alpha satellite higher-order repeats (HOR) and consensus distributions of CENP-B box and pJa motif

  17. PRODUCTION OF TUMOR NECROSIS FACTOR\\/CACHECTIN BY HUMAN B CELL LINES AND TONSILLAR B CELLS

    Microsoft Academic Search

    SUN-SANG J. SUNG; L. JUNG; JAY A. WALTERS; WEI CHEN; CHANG YI WANG; SHU MAN FU

    TNF\\/cachectin has diverse physiological functions and is produced by a wide va- riety ofcell types (reviewed in reference 1). Recently, evidence has accumulated that TNF may play regulatory roles in T and B lymphocyte activation, proliferation, anddifferentiation (2, 3) . ActivatedT cellshave been shown to produce aconsider- able amount of TNF (4-6) . However, the capacity of B cells to

  18. Purification and some properties of the small subunit of cytochrome b558 from human neutrophils.

    PubMed

    Yamaguchi, T; Hayakawa, T; Kaneda, M; Kakinuma, K; Yoshikawa, A

    1989-01-01

    We have attempted to purify the heme moiety of cytochrome b558 from human neutrophils. Cytochrome b558 was solubilized from the crude membrane fraction which was pretreated with both 1 M potassium phosphate buffer and 1% octyl glucoside at low ionic strength. The solubilization of cytochrome b558 was carried out efficiently with 1.6% octyl glucoside in the presence of 100 mM phosphate buffer. Solubilized cytochrome b558 was purified by hydroxylapatite, DEAE-Sephacel, and Mono Q fast protein liquid chromatography. The specific content of purified cytochrome b558 was 37 nmol/mg of protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of purified cytochrome b558 revealed a single band of 20,000 Da. The large subunit of cytochrome b558, which has been reported by others, could not be found in purified cytochrome b558 even with silver staining. The amino acid composition of the heme-containing moiety of cytochrome b558 was abundant in hydrophobic amino acids. The circular dichroism spectra of both oxidized and reduced b558-type cytochromes exhibited bilobed bands with wavelengths of crossover points closely corresponding to those of the maxima in the optical absorbance spectra at the Soret region. Furthermore, there were some differences in the shoulders and peak widths of CD spectra between oxidized and reduced b558-type cytochromes. These results indicate that this method provides the purification of the small subunit of human cytochrome b558 which is the heme-carrying subunit of cytochrome b558, and suggest that cytochrome b558 has heme-heme interaction and some conformational changes in the alternation of the redox state. PMID:2909509

  19. Human Immunoglobulin Specificity After Group B Arbovirus Infections

    PubMed Central

    McNair Scott, Robert; McCown, Jack M.; Russell, Philip K.

    1972-01-01

    Sera obtained during a dengue-2 epidemic from individuals with primary or secondary group B arbovirus infections contained cross-reactive antibody against several viruses in this group. Hemagglutination-inhibition antibody titers of the immunoglobulin M fractions from these sera revealed a rise in antibody specific for dengue-2. The acute and convalescent sera in over half of the secondary infection group also had low-level unchanging immunoglobulin M antibody titers to yellow fever which implicated yellow fever immunization as the primary infection. PMID:4629258

  20. Tumor necrosis factor alpha is an autocrine growth factor for normal human B cells.

    PubMed Central

    Boussiotis, V A; Nadler, L M; Strominger, J L; Goldfeld, A E

    1994-01-01

    Transcription of the human tumor necrosis factor alpha (TNF-alpha) gene is one of the earliest events that occurs after stimulation of B or T cells via their antigen receptors. Antibody directed at surface immunoglobulin (anti-Ig) on B cells has previously been shown to induce a rapid burst of TNF-alpha gene transcription, which can be blocked by the immunosuppressants cyclosporin A (CsA) and FK506. Here, TNF-alpha gene transcription is shown also to be highly and rapidly induced in human B cells after stimulation via the CD40 and interleukin 4 pathways, which similarly is inhibited by CsA and a panel of CsA or FK506 analogues that block calcineurin phosphatase activity. Endogenous TNF-alpha produced after stimulation was involved in B-cell proliferation since anti-TNF-alpha monoclonal antibody inhibited both anti-Ig- and anti-CD40-induced B-cell proliferative responses. Moreover, addition of TNF-alpha during stimulation resulted in augmentation of B-cell proliferation, which was also inhibited by anti-TNF-alpha monoclonal antibody. Although lymphotoxin alpha (LT-alpha) mRNA is induced by both pathways, it is not blocked by CsA, whereas LT-beta mRNA is constitutively expressed in B cells. Thus, TNF-alpha is a necessary autocrine growth factor for human B cells stimulated via two independent CsA-sensitive pathways and plays a role similar to that of interleukin 2 in T-cell proliferation. The autocrine nature of TNF-alpha in activated B cells implies a potential role for this cytokine in infection-related polyclonal B-cell expansion and in B-cell malignancies. Images PMID:7518925

  1. Plant stress and human health: Do human consumers benefit from UV-B acclimated crops?

    Microsoft Academic Search

    Marcel A. K. Jansen; Kathleen Hectors; Nora M. O’Brien; Yves Guisez; Geert Potters

    2008-01-01

    Plants are sessile organisms, and consequently cannot avoid exposure to stressful environmental conditions. Exposure to mild stress conditions can induce active acclimation responses, while more severe conditions cause metabolic disruptions. A common plant acclimation response to a variety of environmental stressors is the accumulation of antioxidants and secondary metabolites. For example, ultraviolet-B (UV-B) radiation impacts on the levels of a

  2. Human genes involved in hepatitis B virus infection

    PubMed Central

    Zeng, Zheng

    2014-01-01

    Persistent hepatitis B virus (HBV) infection is a significant public health problem because it is a major cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma (HCC). Roughly one-third of the world population has been infected with HBV and there are about 350 million (5%-6%) persistent carriers. HBV causes 80% of all liver cancer cases and is the second most important carcinogen, after smoking tobacco. There is an approximate 90% risk of becoming a persistent carrier following perinatal infection in infants born to e antigen positive carrier mothers and a 30% risk in pre-school children. Only 5%-10% of adults become persistent carriers following infection. Of individuals persistently infected with HBV, 10%-30% will develop liver cirrhosis and HCC. These highly variable outcomes in both clearance rates and disease outcomes in persistently infected individuals cannot be fully explained by differences in immunological, viral or environmental factors. Thus, differences in host genetic factors may affect the natural history of hepatitis B. PMID:24976707

  3. ADAM17 cleaves CD16b (Fc?RIIIb) in human neutrophils

    PubMed Central

    Wang, Yue; Wu, Jianming; Newton, Robert; Bahaie, Nooshin S.; Long, Chunmei; Walcheck, Bruce

    2012-01-01

    CD16b (Fc?RIIIb) is exclusively expressed by human neutrophils and binds IgG in immune complexes. Cell surface CD16b undergoes efficient ectodomain shedding upon neutrophil activation and apoptosis. Indeed, soluble CD16b is present at high levels in the plasma of healthy individuals, which appears to be maintained by the daily turnover of apoptotic neutrophils. At this time, the principal protease responsible for CD16b shedding is not known. We show that CD16b plasma levels were significantly decreased in patients administered a selective inhibitor targeting the metalloproteases ADAM10 and ADAM17. Additional analysis with inhibitors selective for ADAM10 or ADAM17 revealed that only inhibition of ADAM17 significantly blocked the cleavage of CD16b following neutrophil activation and apoptosis. CD16b shedding by ADAM17 was further demonstrated using a unique ADAM17 function-blocking mAb and a cell-based ADAM17 reconstitution assay. Unlike human CD16, however, mouse CD16 did not undergo efficient ectodomain shedding upon neutrophil stimulation or apoptosis, indicating that this mechanism cannot be modeled in normal mice. Taken together, our findings are the first to directly demonstrate that ADAM17 cleaves CD16 in human leukocytes. PMID:23228566

  4. Effect of increased HoxB4 on human megakaryocytic development

    SciTech Connect

    Zhong, Yiming [Department of Pathology, The Ohio State University, Columbus, OH (United States) [Department of Pathology, The Ohio State University, Columbus, OH (United States); Program in Molecular, Cellular, and Developmental Biology, The Ohio State University, Columbus, OH (United States); Sullenbarger, Brent [Department of Pathology, The Ohio State University, Columbus, OH (United States)] [Department of Pathology, The Ohio State University, Columbus, OH (United States); Lasky, Larry C., E-mail: Lasky.4@osu.edu [Department of Pathology, The Ohio State University, Columbus, OH (United States); Program in Molecular, Cellular, and Developmental Biology, The Ohio State University, Columbus, OH (United States)

    2010-07-30

    Research highlights: {yields} HoxB4 overexpression in human TF1 cells increased the expression of CD61 and CD41a. {yields} HoxB4 fusion protein enhanced megakaryocytic development of CD34{sup +} cord blood cells. {yields} Ectopic HoxB4 increased Tpo receptor expression and decreased c-Myb expression. {yields} HoxB4 RNA silencing increased c-Myb expression and decreased Fli-1 expression. -- Abstract: In order to produce clinically useful quantities of platelets ex vivo we may need to firstly enhance early self-renewal of hematopoietic stem cells (HSCs) and/or megakaryocyte (Mk) progenitors. The homeodomain transcription factor HoxB4 has been shown to be an important regulator of stem cell renewal and hematopoiesis; however, its effect on megakaryopoiesis is unclear. In this study, we investigated the effect of HoxB4 overexpression or RNA silencing on megakaryocytic development in the human TF1 progenitor cell line; we then used recombinant tPTD-HoxB4 fusion protein to study the effect of exogenous HoxB4 on megakaryocytic development of human CD34 positively-selected cord blood cells. We found that ectopic HoxB4 in TF1 cells increased the antigen expression of CD61and CD41a, increased the gene expression of thrombopoietin receptor (TpoR), Scl-1, Cyclin D1, Fog-1 and Fli-1 while it decreased c-Myb expression. HoxB4 RNA silencing in TF1 cells decreased the expression of CD61 and CD41a and decreased Fli-1 expression while it increased the expression of c-Myb. Recombinant tPTD-HoxB4 fusion protein increased the percentages and absolute numbers of CD41a and CD61 positive cells during megakaryocytic differentiation of CD34 positively-selected cord blood cells and increased the numbers of colony-forming unit-megakaryocyte (CFU-Mk). Adding tPTD-HoxB4 fusion protein increased the gene expression of TpoR, Cyclin D1, Fog-1 and Fli-1 while it inhibited c-Myb expression. Our data suggest that increased HoxB4 enhanced early megakaryocytic development in human TF1 cells and CD34 positively-selected cord blood cells primarily by upregulating TpoR and Fli-1 expression and downregulating c-Myb expression. Increasing HoxB4 expression or adding recombinant HoxB4 protein might be a way to expand Mks for the production of platelets for use in transfusion medicine.

  5. Murine and human b locus pigmentation genes encode a glycoprotein (gp75) with catalase activity

    SciTech Connect

    Halaban, R.; Moellmann, G. (Yale Univ. School of Medicine, New Haven, CT (USA))

    1990-06-01

    Melanogenesis is regulated in large part by tyrosinase, and defective tyrosinase leads to albinism. The mechanisms for other pigmentation determinants (e.g., those operative in tyrosinase-positive albinism and in murine coat-color mutants) are not yet known. One murine pigmentation gene, the brown (b) locus, when mutated leads to a brown (b/b) or hypopigmentated (B{sup lt}/B{sup lt}) coat versus the wild-type black (B/B). The authors show that the b locus codes for a glycoprotein with the activity of a catalase (catalase B). Only the c locus protein is a tyrosinase. Because peroxides may be by-products of melanogenic activity and hydrogen peroxide in particular is known to destroy melanin precursors and melanin, they conclude that pigmentation is controlled not only by tyrosinase but also by a hydroperoxidase. The studies indicate that catalase B is identical with gp75, a known human melanosomal glycoprotein; that the b mutation is in a heme-associated domain; and that the B{sup lt} mutation renders the protein susceptible to rapid proteolytic degradation.

  6. Heterologous expression, immunochemical and computational analysis of recombinant human interferon alpha 2b.

    PubMed

    Gull, Iram; Samra, Zahoor Qadir; Aslam, Muhammad Shahbaz; Athar, Muhammad Amin

    2013-12-01

    Interferon alpha 2b (IFN?-2b) is an important cytokine and used for antiviral and anticancer treatment. The low cost production of IFN?-2b with high biological activity is necessary to provide the interferon therapy to the hepatitis patients in Pakistan. In the present study, human interferon alpha 2b (hIFN?-2b) gene from a healthy person was cloned and overexpressed in E. coli BL21(DE3). The molecular weight of the expressed hIFN?-2b is 19 kDa. The over expressed recombinant hIFN?-2b was checked by ELISA using antibodies raised against commercially available hIFN?-2b. The biocomputational analysis of recombinant hIFN?-2b gene showed the 99.9% nucleotide sequence and 100% deduced amino acid sequence homology with reported sequences of IFN?-2b. The predicted 3D-structure showed mainly five ?-helices, one 310 helix and two disulfide bonds at Cys1-Cys98 and Cys129-Cys138. The amino acid sequence alignment indicated that the disulfide linkage position is conserved in all IFN? family members. On the basis of sequence homology among interferon alpha family, new potent variants of hIFN?-2b with enhance efficacy can be produced. Indigenous production of IFN?-2b from gene of local population will reduce the cost and increase tolerability of interferon therapy. PMID:23875128

  7. Imaging of nuclear factor ?B activation induced by ionizing radiation in human embryonic kidney (HEK) cells.

    PubMed

    Chishti, Arif Ali; Baumstark-Khan, Christa; Hellweg, Christine E; Reitz, Günther

    2014-08-01

    Ionizing radiation modulates several signaling pathways resulting in transcription factor activation. Nuclear factor kappa B (NF-?B) is one of the most important transcription factors that respond to changes in the environment of a mammalian cell. NF-?B plays a key role not only in inflammation and immune regulation but also in cellular radiation response. In response to DNA damage, NF-?B might inhibit apoptosis and promote carcinogenesis. Our previous studies showed that ionizing radiation is very effective in inducing biological damages. Therefore, it is important to understand the radiation-induced NF-?B signaling cascade. The current study aims to improve existing mammalian cell-based reporter assays for NF-?B activation by the use of DD-tdTomato which is a destabilized variant of red fluorescent protein tdTomato. It is demonstrated that exposure of recombinant human embryonic kidney cells (HEK/293 transfected with a reporter constructs containing NF-?B binding sites in its promoter) to ionizing radiation induces NF-?B-dependent DD-tdTomato expression. Using this reporter assays, NF-?B signaling in mammalian cells was monitored by flow cytometry and fluorescence microscopy. Activation of NF-?B by the canonical pathway was found to be quicker than by the genotoxin- and stress-induced pathway. X-rays activate NF-?B in HEK cells in a dose-dependent manner, and the extent of NF-?B activation is higher as compared to camptothecin. PMID:24880906

  8. Granzyme B produced by human plasmacytoid dendritic cells suppresses T-cell expansion.

    PubMed

    Jahrsdörfer, Bernd; Vollmer, Angelika; Blackwell, Sue E; Maier, Julia; Sontheimer, Kai; Beyer, Thamara; Mandel, Birgit; Lunov, Oleg; Tron, Kyrylo; Nienhaus, G Ulrich; Simmet, Thomas; Debatin, Klaus-Michael; Weiner, George J; Fabricius, Dorit

    2010-02-11

    Human plasmacytoid dendritic cells (pDCs) are crucially involved in the modulation of adaptive T-cell responses in the course of neoplastic, viral, and autoimmune disorders. In several of these diseases elevated extracellular levels of the serine protease granzyme B (GrB) are observed. Here we demonstrate that human pDCs can be an abundant source of GrB and that such GrB(+) pDCs potently suppress T-cell proliferation in a GrB-dependent, perforin-independent manner, a process reminiscent of regulatory T cells. Moreover, we show that GrB expression is strictly regulated on a transcriptional level involving Janus kinase 1 (JAK1), signal transducer and activator of transcription 3 (STAT3), and STAT5 and that interleukin-3 (IL-3), a cytokine secreted by activated T cells, plays a central role for GrB induction. Moreover, we find that the immunosuppressive cytokine IL-10 enhances, while Toll-like receptor agonists and CD40 ligand strongly inhibit, GrB secretion by pDCs. GrB-secreting pDCs may play a regulatory role for immune evasion of tumors, antiviral immune responses, and autoimmune processes. Our results provide novel information about the complex network of pDC-T-cell interactions and may contribute to an improvement of prophylactic and therapeutic vaccinations. PMID:19965634

  9. Nuclear envelope remodelling during human spermiogenesis involves somatic B-type lamins and a spermatid-specific B3 lamin isoform.

    PubMed

    Elkhatib, Razan; Longepied, Guy; Paci, Marine; Achard, Vincent; Grillo, Jean-Marie; Levy, Nicolas; Mitchell, Michael J; Metzler-Guillemain, Catherine

    2015-03-01

    The nuclear lamina (NL) is a filamentous protein meshwork, composed essentially of lamins, situated between the inner nuclear membrane and the chromatin. There is mounting evidence that the NL plays a role in spermatid differentiation during spermiogenesis. The mouse spermatid NL is composed of the ubiquitous lamin B1 and the spermatid-specific lamin B3, an N-terminally truncated isoform of lamin B2. However, nothing is known about the NL in human spermatids. We therefore investigated the expression pattern and localization of A-type lamins (A, C and C2) and B-type lamins (B1, B2 and B3) during human spermiogenesis. Here, we show that a lamin B3 transcript is present in human spermatids and that B-type lamins are the only lamins detectable in human spermatids. We determine that, as shown for their mouse counterparts, human lamin B3, but not lamin B2, induces strong nuclear deformation, when ectopically expressed in HeLa cells. Co-immunofluorescence revealed that, in human spermatids, B-type lamins are present at the nuclear periphery, except in the region covered by the acrosome, and that as the spermatid matures the B-type lamins recede towards the posterior pole. Only lamin B1 remains detectable on 33-47% of ejaculated spermatozoa. On spermatozoa selected for normal head density, however, this fell to <6%, suggesting that loss of the NL signal may be linked to complete sperm nucleus compaction. The similarities revealed between lamin expression during human and rodent spermiogenesis, strengthen evidence that the NL and lamin B3 have conserved functions during the intense remodelling of the mammalian spermatid nucleus. PMID:25477337

  10. Oltipraz-mediated Changes in Aflatoxin B, Biotransformation in Rat Liver: Implications for Human Chemointervention1

    Microsoft Academic Search

    Timo M. Buetler; Theo K. Bammler; John D. Hayes; David L. Eaton

    Oltipraz (OPZ) is currently being considered for human use to protect against aflatoxin B, (AFB)-induced hepatocarcinogenesis based on its proven protective effect in rats. The effectiveness of this treatment pre sumes that orthologous cytochrome P450 and glutathione S-transferase (GST) isozymes metabolize AFB in humans as they do in rats. In this study, alterations in the expression of multiple forms of

  11. Characterization of the cloned guinea pig leukotriene B 4 receptor: comparison to its human orthologue

    Microsoft Academic Search

    Yves Boie; Rino Stocco; Nicole Sawyer; Gillian M Greig; Stacia Kargman; Deborah M Slipetz; Gary P O'Neill; Takao Shimizu; Takehiko Yokomizo; Kathleen M Metters; Mark Abramovitz

    1999-01-01

    A cDNA clone coding for the guinea pig leukotriene B4 (BLT) receptor has been isolated from a lung cDNA library. The guinea pig BLT receptor has an open reading frame corresponding to 348 amino acids and shares 73% and 70% identity with human and mouse BLT receptors, respectively. Scatchard analysis of membranes prepared from guinea pig and human BLT receptor-transfected

  12. B and T Cells in the Lymphoid Tissues of Human Appendix

    Microsoft Academic Search

    Takashi Mizumoto

    1976-01-01

    Human appendix lymphoid cells (HAL) react very strongly to stimulation with concanavalin A, strongly to stimulation with phytohaemagglutinin, and weakly but definitely to stimulation with lipopolysaccharide. From the results of rosette formation assay, cytotoxicity tests with anti-T cell antiserum or anti-B cell antiserum, cell surface or intracellular immunoglobulin staining with fluorescein-conjugated rabbit anti-Fab of human immunoglobulin serum, and plaque-forming cell

  13. Production of Transgenic Tilapia with Brockmann Bodies Secreting [desThrB30] Human Insulin

    Microsoft Academic Search

    Bill Pohajdak; Marc Mansour; Olga Hrytsenko; J. Michael Conlon; L. Clayton Dymond; James R. Wright Jr

    2004-01-01

    Background. Tilapia are commercially important tropical fish which, like many teleosts, have anatomically discrete islet organs called Brockmann bodies. When transplanted into diabetic nude mice, tilapia islets provide long-term normoglycemia and mammalian-like glucose tolerance profiles. Methods. Using site-directed mutagenesis and linker ligation we have “humanized” the tilapia insulin gene so that it codes for [desThrB30] human insulin while maintaining the

  14. Sensitive method for the detection of fumonisin B 1 in human urine

    Microsoft Academic Search

    Prathapkumar H Shetty; Ramesh V Bhat

    1998-01-01

    Fumonisins are a group of carcinogenic mycotoxins that occur worldwide. This paper reports a sensitive method for the detection and quantitation of fumonisin B1 in human urine. Amberlite XAD-2 non-ionic polymeric adsorbent resin is used prior to strong anion-exchange (SAX) cartridge clean-up. As much as 100 ml of undiluted human urine can be loaded onto the column. Recoveries obtained using

  15. Alterations of nitric oxide synthase and xanthine oxidase activities of human keratinocytes by ultraviolet B radiation

    Microsoft Academic Search

    George Deliconstantinos; Vassiliki Villiotou; John C. Stavrides

    1996-01-01

    In the present study, we demonstrated that NO synthase (cNOS) and xanthine oxidase (XO) of human keratinocytes can be activated to release NO, Superoxide (O2?) and peroxynitrite (ONOO?) following exposure to ultraviolet B (UVB) radiation. We defined that this photo induced response may be involved in the pathogenesis of sunburn erythema and inflammation. Treatment of human keratinocytes with UVB (290–320

  16. Deposition of C3b and iC3b onto particulate activators of the human complement system. Quantitation with monoclonal antibodies to human C3

    PubMed Central

    1985-01-01

    Monoclonal antibodies were used to determine the number and molecular form of C3 bound to particulate activators of the complement (C) system by human serum. Sheep erythrocytes (E) coated with IgM (EIgM) and IgG (EIgG) were used to study activation of the classical pathway (CP). Yeast (Y), rabbit erythrocytes (ER), and five species of bacteria (Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae type 3, Streptococcus pyogenes, and Hemophilus influenzae type b) were used to study activation of the alternative pathway (AP). The deposition of C3b onto EIgM and EIgG incubated in C7-deficient human serum was dependent on the serum concentration. At all serum concentrations tested, there was complete conversion of C3b to iC3b. Kinetic analysis of C3b deposition and conversion to iC3b indicated that these events occurred almost simultaneously; the reaction was completed by 15 min. The deposition of C3 onto the AP activators ER and Y was also dependent on serum concentration, and ER, but not Y, required the presence of Mg- EGTA and thus the activation of only the AP. C3b deposition and conversion to iC3b on Y was complete in 15 min, with 82% of bound C3 converted to iC3b. For ER, maximum C3 deposition required 30 min in both the presence and absence of Mg-EGTA. However, after 1 h of incubation, 74% of bound C2 was iC3b in the absence of Mg-EGTA, compared with only 52% in the presence of Mg-EGTA. Thus, even on AP activators, a large portion of C3b may be converted to iC3b, and this conversion is probably controlled by elements on the particle's surface. Studies with the five species of bacteria yielded similar results. Approximately 3-5 X 10(4) molecules of C3 were bound per microorganism, with opsonization being completed in 30 min. Remarkably, only 16-28% of bound C3 was in the form of iC3b, even after 2 h of incubation. The presence or absence of Mg-EGTA, or the addition of purified CR1 to the reaction mixture, did not significantly effect the ratio of C3b to iC3b. Finally, SDS-PAGE and autoradiography of particle- bound 125I-C3 fragments confirmed that there was no conversion of iC3b to C3d,g or C3d. The data obtained about the opsonization of bacteria suggest that the predominant form of C3 that is encountered by inflammatory phagocytes may be C3b. PMID:2409200

  17. mRNA Decay Factor AUF1 Binds the Internal Ribosomal Entry Site of Enterovirus 71 and Inhibits Virus Replication

    PubMed Central

    Lin, Jing-Yi; Li, Mei-Ling; Brewer, Gary

    2014-01-01

    AU-rich element binding factor 1 (AUF1) has a role in the replication cycles of different viruses. Here we demonstrate that AUF1 binds the internal ribosome entry site (IRES) of enterovirus 71 (EV71) and negatively regulates IRES-dependent translation. During EV71 infection, AUF1 accumulates in the cytoplasm where viral replication occurs, whereas AUF1 localizes predominantly in the nucleus in mock-infected cells. AUF1 knockdown in infected cells increases IRES activity and synthesis of viral proteins. Taken together, the results suggest that AUF1 interacts with the EV71 IRES to negatively regulate viral translation and replication. PMID:25077793

  18. Study of enhanced immunogenecity of B7-1 gene transfected human hela cell line

    Microsoft Academic Search

    Xi He; Huilian Qin; Rong Xiang; Yuejian Zhang; Wenfei Ye; Qiuzao He

    1998-01-01

    Although cervical carcinoma cells may express the human papillomavirus protein E6 and E7, they fail to induce an effective\\u000a specific cytotoxic T lymphocyte response. Recent studies suggest that expression of CD-80 (B7-1) on tumor cells is effective\\u000a to induce antitumor immune responses.1,2 In our study, CD-80 gene was transfected into human Hela cell line with a CD-80 expression plasmid (B7-l*pcDNA3)

  19. Failure To Detect Functional Neutrophil B Helper Cells in the Human Spleen

    PubMed Central

    Nagelkerke, Sietse Quirijn; aan de Kerk, Daan Jacob; Jansen, Machiel Hugo; van den Berg, Timo Kars; Kuijpers, Taco Willem

    2014-01-01

    A novel role for human neutrophilic granulocytes was recently described, showing that these cells, upon entering the spleen, can be reprogrammed into a distinct B cell-helper neutrophil phenotype that is capable of eliciting B cell responses such as immunoglobulin secretion, class switch recombination and somatic hypermutation. Using similar protocols, we detected a homogeneous population of CD15highCD16high neutrophils in fresh human spleen samples, which did not differ in phenotype and function from blood neutrophils. No phenotypic characteristics of costimulatory nature were detected on splenic or circulating neutrophils, nor could we reproduce the immunoglobulin production of splenic B cells in the presence of splenic neutrophils, although B cell function and neutrophil activity were normal. Independent confirmation of a role for NBH cells is required. PMID:24523887

  20. Human tonsil intraepithelial B cells: a marginal zone-related subpopulation.

    PubMed Central

    Morente, M.; Piris, M. A.; Orradre, J. L.; Rivas, C.; Villuendas, R.

    1992-01-01

    AIMS: To determine if intraepithelial B cells in reactive human palatine tonsils were similar to the marginal zone cells of the spleen and Peyer's patches. METHODS: Reactive human palatine tonsils were studied using conventional methods of light microscopy, electron microscopy, and a panel of monoclonal antibodies for leucocyte common antigens. RESULTS: Clinically important numbers of marginal zone-related B cells around the mantle zone were absent in lymphoid follicles, but in the cryptal epithelium there were abundant lymphoid cells with centrocyte-like nuclei and clear cytoplasm, intermingled with macrophages and plasma cells. The immunophenotype of these intraepithelial B cells was distinctive and similar to that found in the splenic marginal zone cells (IgM+, IgD-, CD23-, CD10-, CD35+, CD21+, bc12+, KB61+). CONCLUSIONS: Intraepithelial B cells in human tonsil could represent the counterpart of the marginal zone described in Peyer's patches. Their presence within the epithelium could reflect the destination for the malignant B cells in the lymphoepithelial lesion of mucosa associated lymphoid tissue (MALT) lymphomas. Human palatine tonsil lymphoid tissue has morphological, immunophenotypic, and pathological features similar to those of MALT. Images PMID:1401174

  1. Genetic inactivation of TRAF3 in canine and human B-cell lymphoma.

    PubMed

    Bushell, Kevin R; Kim, Yukyoung; Chan, Fong Chun; Ben-Neriah, Susana; Jenks, Andrew; Alcaide, Miguel; Fornika, Daniel; Grande, Bruno M; Arthur, Sarah; Gascoyne, Randy D; Steidl, Christian; Morin, Ryan D

    2015-02-01

    Non-Hodgkin lymphomas (NHLs) are the most common cancer to affect pet dogs. In contrast to the many genes whose mutation contributes to lymphomagenesis in humans, relatively little is known about the acquired genetic alterations that lead to canine B-cell lymphomas (cBCLs). We performed a survey of 84 canine NHL tumors to identify genes affected by somatic point mutations. We found mutations affecting TRAF3, which encodes a negative regulator of nuclear factor (NF)-?B, to be a common feature of cBCLs, with mutations observed in 44% of tumors including a combination of somatic and rare germ-line variants. Overall, 30% of the tumors contained ?1 somatic TRAF3 mutation. The majority of mutations are predicted to cause loss of TRAF3 protein including those impacting reading frame and splicing. To determine whether TRAF3 loss might be relevant to human NHL, we also analyzed 148 human diffuse large B-cell lymphoma (DLBCL) tumors and identified loss of TRAF3 as a common event, affecting ?9% of DLBCLs, and reduced expression of TRAF3 among deleted cases. This study implicates mutations affecting NF-?B activity as a novel genetic commonality between human and canine NHLs and supports the potential utility of cBCLs with mutated TRAF3 as a model of the more aggressive activated B-cell subgroup of DLBCL. PMID:25468570

  2. Structural analysis of human CCR2b and primate CCR2b by molecular modeling and molecular dynamics simulation.

    PubMed

    Shi, Xiu-Fan; Liu, Shixi; Xiangyu, Jinggong; Zhang, Yaping; Huang, Jingfei; Liu, Shuqiu; Liu, Ci-Quan

    2002-07-01

    CCR2b, a chemokine receptor for MCP-1, -2, -3, -4, plays an important role in a variety of diseases involving infection, inflammation, and/or injury, as well as being a coreceptor for HIV-1 infection. Two models of human CCR2b (hCCR2b) were generated by homology modeling and 1 ns restrained molecular dynamics (MD) simulation. In one only C113-C190 forms a disulfide bond (SS model); in another the potential C32-C277 disulfide bond was formed (2SS model). Analysis of the structures and averaged displacements of Calpha atoms of the N-terminal residues shows that the main differences between the SS and 2SS models lie in a region D25YDYGAPCHKFD36; in the extracellular part of the 2SS model the accessible surfaces of N12, F23, Y26, Y28 and F35 are obviously raised and a more stable H-bond net is formed. The potential energy of the 2SS-water assembly finally fluctuated around -43,020 kJ x mol(-1), which is about 302 kJ x mol(-1) lower than that of the SS-water assembly. All these results suggest that the 2SS model is more favorable. The CCR2b genes of 17 primates were sequenced and four CCR2b models for primates Ateles paniscus (A. pan), Hylobates leucogyneus(H. leu), Papio cynocephalus (P. cyn) and Trachypithecus francoist ( T. fra) were generated based on the 2SS model. A comparison of hCCR2b with primate CCR2b also supports the importance of the region D25YDYGAPCHKFD36. Electrostatic potential maps of human and primate CCR2b all display the dipolar characteristics of CCR2b with the negative pole located in the extracellular part and a strong positive pole in the cytoplasmic part. Based on the CCR2b model, we suggest that the main functional residues fall in the D25YDYGAPCHKFD36 region, and the negative electrostatic feature is a non-specific, but necessary, factor for ligands or gp120/CD4 binding. PMID:12192431

  3. Expression of novel interleukin 2 binding molecules and their functional roles in human B cell differentiation.

    PubMed Central

    Tanaka, T; Saiki, O; Doi, S; Suemura, M; Negoro, S; Kishimoto, S

    1988-01-01

    Expressions and functional roles of novel IL-2 binding molecules (p70, 75) in the differentiation of B cells into Ig secreting cells were explored by using human several B cell lines and tonsillar B cells. Affinity-crosslinking studies revealed that five of nine B cell lines expressed p70 and p75 without detectable Tac antigen (p55) expression and the expression was associated with B cell maturation. In tonsillar B cells, small high-density B cells did not express p70 and p75, whereas large low-density B cells, which were thought to be activated in vivo, expressed them. Binding assays of radiolabeled IL-2 showed that the affinity of these molecules was intermediate (kD = 1-3 nM, 700-3,000 sites/cell). Furthermore, high concentrations of IL-2 (greater than 100 U/ml) induced Ig productions in large B cells and two of five cell lines. These results taken together suggest that B cells may express novel IL-2 binding molecules, associated with B cell differentiation and differentiate into Ig secreting cells by IL-2 through novel IL-2 binding molecules. Images PMID:2839549

  4. Differential Effects of Tacrolimus versus Sirolimus on the Proliferation, Activation and Differentiation of Human B Cells

    PubMed Central

    Xu, Luting; Mas, Valeria; Gallon, Lorenzo

    2015-01-01

    The direct effect of immunosuppressive drugs calcineurin inhibitor (Tacrolimus, TAC) and mTOR inhibitor (Sirolimus, SRL) on B cell activation, differentiation and proliferation is not well documented. Purified human B cells from healthy volunteers were stimulated through the B Cell Receptor with Anti-IgM + anti-CD40 + IL21 in the absence / presence of TAC or SRL. A variety of parameters of B cell activity including activation, differentiation, cytokine productions and proliferation were monitored by flow cytometry. SRL at clinically relevant concentrations (6 ng/ml) profoundly inhibited CD19+ B cell proliferation compared to controls whereas TAC at similar concentrations had a minimal effect. CD27+ memory B cells were affected more by SRL than naďve CD27- B cells. SRL effectively blocked B cell differentiation into plasma cells (CD19+CD138+ and Blimp1+/Pax5low cells) even at low dose (2 ng/ml), and totally eliminated them at 6 ng/ml. SRL decreased absolute B cell counts, but the residual responding cells acquired an activated phenotype (CD25+/CD69+) and increased the expression of HLA-DR. SRL-treated stimulated B cells on a per cell basis were able to enhance the proliferation of allogeneic CD4+CD25? T cells and induce a shift toward the Th1 phenotype. Thus, SRL and TAC have different effects on B lymphocytes. These data may provide insights into the clinical use of these two agents in recipients of solid organ transplants. PMID:26087255

  5. Neutrophils infiltrating ultraviolet B-irradiated normal human skin display high IL10 expression

    Microsoft Academic Search

    Gamze Piskin; Jan D. Bos; Marcel B. M. Teunissen

    2005-01-01

    Exposure to an erythemal dose of ultraviolet B (UVB) is known to induce interleukin (IL-10) expression in human skin. It is generally believed that this IL-10 is predominantly expressed by CD11b +HLA-DR + macrophages that infiltrate the UVB-exposed skin. This cytokine is presumed to contribute to the immunosuppressive effects of UVB by inhibiting cell-mediated immune responses. We recently demonstrated that

  6. Modulation of resistance to cisplatin by amphotericin B and aphidicolin in human larynx carcinoma cells

    Microsoft Academic Search

    L. Beketic-Oreskovic; Maja Osmak

    1995-01-01

    The aim of this study was to examine whether resistance to cisplatin [ cis-diamminedichloroplatinum (II)] (CDDP) could be overcome by amphotericin B, cyclosporin A and aphidicolin in two sublines of human larynx carcinoma HEp2 cells. The sensitivity of parental and cisplatin-resistant CA3 and CK2 cells to amphotericin B, cyclosporin A and aphidicolin, and also the effects of these drugs (given

  7. Modulation of resistance to cisplatin by amphotericin B and aphidicolin in human larynx carcinoma cells

    Microsoft Academic Search

    Lidija Beketic-Oreskovic; Maja Osmak

    1995-01-01

    The aim of this study was to examine whether resistance to cisplatin [cis-diamminedichloroplatinum (II)] (CDDP) could be overcome by amphotericin B, cyclosporin A and aphidicolin in two sublines of human larynx carcinoma HEp2 cells. The sensitivity of parental and cisplatin-resistant CA3 and CK2 cells to amphotericin B, cyclosporin A and aphidicolin, and also the effects of these drugs (given in

  8. Characterization of the human N?-terminal acetyltransferase B enzymatic complex

    Microsoft Academic Search

    Amagoia Ametzazurra; Cristina Gázquez; Marta Lasa; Esther Larrea; Jesús Prieto; Rafael Aldabe

    2009-01-01

    Background: Human N?-acetyltransferase complex B (hNatB) is integrated by hNaa20p (hNAT5\\/ hNAT3) and hNaa25p (hMDM20) proteins. Previous data have shown that this enzymatic complex is implicated in cell cycle progression and carcinogenesis. In yeast this enzyme acetylates peptides composed by methionine and aspartic acid or glutamic acid in their first two positions respectively and it has been shown the same

  9. Retrieval of human cytomegalovirus glycoprotein B from the infected cell surface for virus envelopment

    Microsoft Academic Search

    K. Radsak; M. Eickmann; T. Mockenhaupt; E. Bogner; H. Kern; A. Eis-Hiibinger; M. Reschke

    1996-01-01

    Summary Surface biotinylation of human cytomegalovirus (HCMV)-infected fibroblasts under pulse-chase conditions was used to define the cellular route of the dominant viral envelope glycoprotein gB into the cytoplasmic compartment of viral maturational envelopment. The results showed that a major fraction of gB was re-internalized from the infected cell surface prior to incorporation into the viral envelope. Viral particles carrying biotinylated

  10. Association of Human Herpesvirus6B with Mesial Temporal Lobe Epilepsy

    Microsoft Academic Search

    Julie Fotheringham; Donatella Donati; Nahid Akhyani; Anna Fogdell-Hahn; Alexander Vortmeyer; John D Heiss; Elizabeth Williams; Steven Weinstein; Derek A Bruce; William D Gaillard; Susumu Sato; William H Theodore; Steven Jacobson

    2007-01-01

    BackgroundHuman herpesvirus-6 (HHV-6) is a ?-herpesvirus with 90% seroprevalence that infects and establishes latency in the central nervous system. Two HHV-6 variants are known: HHV-6A and HHV-6B. Active infection or reactivation of HHV-6 in the brain is associated with neurological disorders, including epilepsy, encephalitis, and multiple sclerosis. In a preliminary study, we found HHV-6B DNA in resected brain tissue from

  11. Apolipoprotein B metabolism in humans: studies with stable isotope-labeled amino acid precursors

    Microsoft Academic Search

    Julian B. Marsh; Francine K. Welty; Alice H. Lichtenstein; Stefania Lamon-Fava; Ernst J. Schaefer

    2002-01-01

    This article reviews the literature from 1986 to early 2001 relating to apoB100 and apoB48 kinetics in humans using amino acid precursors labeled with stable isotopes. The following subjects are reviewed: (1) methodology; (2) normal individuals and the effects of aging; (3) diet; (4) hereditary dyslipidemias: familial hypercholesterolemia, familial combined hyperlipidemia, cholesteryl ester storage disease, cholesteryl ester transfer protein deficiency,

  12. Co-contamination of aflatoxin B1 and fumonisin B1 in food and human dietary exposure in three areas of China

    Microsoft Academic Search

    G. Sun; S. Wang; X. Hu; J. Su; Y. Zhang; Y. Xie; H. Zhang; L. Tang; J.-S. Wang

    2011-01-01

    Aflatoxins and fumonisins are ubiquitous foodborne toxicants and the co-occurrence of these mycotoxins in human foods represents a significant public health concern, which has been strongly associated with human aflatoxicosis, neural tube defects, as well as many types of primary cancers. In this study the co-contamination of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) in food and human dietary exposure

  13. HSPA12B inhibits lipopolysaccharide-induced inflammatory response in human umbilical vein endothelial cells

    PubMed Central

    Wu, Jun; Li, Xuehan; Huang, Lei; Jiang, Surong; Tu, Fei; Zhang, Xiaojin; Ma, He; Li, Rongrong; Li, Chuanfu; Li, Yuehua; Ding, Zhengnian; Liu, Li

    2015-01-01

    Heat shock protein A12B (HSPA12B) is a newly discovered member of the HSP70 protein family. This study investigated the effects of HSPA12B on lipopolysaccharide (LPS)-induced inflammatory responses in human umbilical vein endothelial cells (HUVECs) and the possible mechanisms involved. A HUVECs inflammatory model was induced by LPS. Overexpression of HSPA12B in HUVECs was achieved by infection with recombinant adenoviruses encoding green fluorescence protein-HSPA12B. Knockdown of HSPA12B was achieved by siRNA technique. Twenty four hours after virus infection or siRNA transfection, HUVECs were stimulated with 1 ?g/ml LPS for 4 hrs. Endothelial cell permeability ability was determined by transwell permeability assay. The binding rate of human neutrophilic polymorphonuclear leucocytes (PMN) with HUVECs was examined using myeloperoxidase assay. Cell migrating ability was determined by the wound-healing assay. The mRNA and protein expression levels of interested genes were analyzed by RT-qPCR and Western blot, respectively. The release of cytokines interleukin-6 and tumour necrosis factor-? was measured by ELISA. HSPA12B suppressed LPS-induced HUVEC permeability and reduced PMN adhesion to HUVECs. HSPA12B also inhibited LPS-induced up-regulation of adhesion molecules and inflammatory cytokine expression. By contrast, knockdown of HSPA12B enhanced LPS-induced increases in the expression of adhesion molecules and inflammatory cytokines. Moreover, HSPA12B activated PI3K/Akt signalling pathway and pharmacological inhibition of this pathway by Wortmannin completely abrogated the protection of HSPA12B against inflammatory response in HUVECs. Our results suggest that HSPA12B attenuates LPS-induced inflammatory responses in HUVECs via activation of PI3K/Akt signalling pathway. PMID:25545050

  14. Does the A-not-B error in adult pet dogs indicate sensitivity to human communication?

    PubMed

    Kis, Anna; Topál, József; Gácsi, Márta; Range, Friederike; Huber, Ludwig; Miklósi, Adám; Virányi, Zsófia

    2012-07-01

    Recent dog-infant comparisons have indicated that the experimenter's communicative signals in object hide-and-search tasks increase the probability of perseverative (A-not-B) errors in both species (Topál et al. 2009). These behaviourally similar results, however, might reflect different mechanisms in dogs and in children. Similar errors may occur if the motor response of retrieving the object during the A trials cannot be inhibited in the B trials or if the experimenter's movements and signals toward the A hiding place in the B trials ('sham-baiting') distract the dogs' attention. In order to test these hypotheses, we tested dogs similarly to Topál et al. (2009) but eliminated the motor search in the A trials and 'sham-baiting' in the B trials. We found that neither an inability to inhibit previously rewarded motor response nor insufficiencies in their working memory and/or attention skills can explain dogs' erroneous choices. Further, we replicated the finding that dogs have a strong tendency to commit the A-not-B error after ostensive-communicative hiding and demonstrated the crucial effect of socio-communicative cues as the A-not-B error diminishes when location B is ostensively enhanced. These findings further support the hypothesis that the dogs' A-not-B error may reflect a special sensitivity to human communicative cues. Such object-hiding and search tasks provide a typical case for how susceptibility to human social signals could (mis)lead domestic dogs. PMID:22460627

  15. Energetic and Molecular Water Permeation Mechanisms of the Human Red Blood Cell Urea Transporter B

    PubMed Central

    Azouzi, Slim; Gueroult, Marc; Ripoche, Pierre; Genetet, Sandrine; Colin Aronovicz, Yves; Le Van Kim, Caroline; Etchebest, Catherine; Mouro-Chanteloup, Isabelle

    2013-01-01

    Urea transporter B (UT-B) is a passive membrane channel that facilitates highly efficient permeation of urea. In red blood cells (RBC), while the major function of UT-B is to transport urea, it is assumed that this protein is able to conduct water. Here, we have revisited this last issue by studying RBCs and ghosts from human variants with defects of aquaporin 1 (AQP1) or UT-B. We found that UT-B's osmotic water unit permeability (pfunit) is similar to that of AQP1. The determination of diffusional permeability coefficient (Pd) allowed the calculation of the Pf/Pd ratio, which is consistent with a single-file water transport. Molecular dynamic simulations of water conduction through human UT-B confirmed the experimental finding. From these results, we propose an atomistic description of water–protein interactions involved in this permeation. Inside the UT-B pore, five water molecules were found to form a single-file and move rapidly along a channel by hydrogen bond exchange involving two critical threonines. We further show that the energy barrier for water located in the central region coincides with a water dipole reorientation, which can be related to the proton exclusion observed experimentally. In conclusion, our results indicate that UT-B should be considered as a new member of the water channel family. PMID:24376529

  16. Specificity of the coxsackievirus B4 VP4 capsid protein investigated in silico.

    PubMed

    Goffard, Anne; Sané, Famara; Soumillon, Magali; Hober, Didier

    2011-08-01

    The Enterovirus genus encompasses several species and various serotypes, like coxsackievirus-B1 (CV-B1) to CV-B6, and many variants. The role of these viruses, especially CV-B4, in the pathogenesis of type 1 diabetes is strongly suspected. It has been reported that antibodies directed towards the region of amino acids 11-30 of the VP4 capsid protein enhance the infection of human peripheral blood mononuclear cells with CV-B4. In order to predict the inter- and intra-serotype specificity of the region 11-30 of CV-B4 VP4, 362 available protein sequences of CV-B1 to -B6, CV-A9, and swine vesicular disease virus (SVDV) have been aligned and levels of homology have been calculated. Serine residue substitutions in this region of VP4 were observed without predictable subsequent modification of conformation or charge. The amino acids 16-24 region was the most variable. The sequence of amino acids 16-24 of the CV-B4E2 VP4 protein was highly homologous to those of other CV-B4 (64.4%) whereas there was no homology with CV-B3 and B5 and very low levels of homology with CV-B1 and B2 (3.3% and 9.9%, respectively). In conclusion, the bioinformatic analysis suggests that the region 16-24 of the VP4 capsid protein is the feature of the specificity of the target of infection-enhancing antibodies directed towards CV-B. PMID:21878192

  17. Vanillin protects human keratinocyte stem cells against ultraviolet B irradiation.

    PubMed

    Lee, Jienny; Cho, Jae Youl; Lee, Sang Yeol; Lee, Kyung-Woo; Lee, Jongsung; Song, Jae-Young

    2014-01-01

    Ultraviolet-B (UVB) irradiation is one of major factors which induce cellular damages in the epidermis. We investigated protective effects and mechanisms of vanillin, a main constituent of vanilla beans, against UVB-induced cellular damages in keratinocyte stem cells (KSC). Here, vanillin significantly attenuated UVB irradiation-induced cytotoxicity. The vanillin effects were also demonstrated by the results of the senescence-associated ?-galactosidase and alkaline comet assays. In addition, vanillin induced production of pro-inflammatory cytokines. Attempts to elucidate a possible mechanism underlying the vanillin-mediated effects revealed that vanillin significantly reduced UVB-induced phosphorylation of ataxia telangiectasia mutated (ATM), serine threonine kinase checkpoint kinase 2 (Chk2), tumor suppressor protein 53 (p53), p38/mitogen-activated protein kinase (p38), c-Jun N-terminal kinase/stress-activated protein kinase (JNK), S6 ribosomal protein (S6RP), and histone 2A family member X (H2A.X). UVB-induced activation of p53 luciferase reporter was also significantly inhibited by vanillin. In addition, while ATM inhibitor had no effect on the vanillin effects, mouse double minute 2 homolog (MDM2) inhibitor significantly attenuated suppressive effects of vanillin on UVB-induced activation of p53 reporter in KSC. Taken together, these findings suggest that vanillin protects KSC from UVB irradiation and its effects may occur through the suppression of downstream step of MDM2 in UVB irradiation-induced p53 activation. PMID:24184596

  18. Jumonji/Arid1b (Jarid1b) protein modulates human esophageal cancer cell growth.

    PubMed

    Kano, Yoshihiro; Konno, Masamitsu; Ohta, Katsuya; Haraguchi, Naotsugu; Nishikawa, Shimpei; Kagawa, Yoshinori; Hamabe, Atsushi; Hasegawa, Shinichiro; Ogawa, Hisataka; Fukusumi, Takahito; Noguchi, Yuko; Ozaki, Miyuki; Kudo, Toshihiro; Sakai, Daisuke; Satoh, Taroh; Ishii, Masaru; Mizohata, Eiichi; Inoue, Takeshi; Mori, Masaki; Doki, Yuichiro; Ishii, Hideshi

    2013-07-01

    Although esophageal cancer is highly heterogeneous and the involvement of epigenetic regulation of cancer stem cells is highly suspected, the biological significance of epigenetically modified molecules that regulate different subpopulations remains to be firmly established. Using esophageal cancer cells, we investigated the functional roles of the H3K4 demethylase Jumonji/Arid1b (Jarid1b) (Kdm5b/Plu-1/Rbp2-h1), an epigenetic factor that is required for continuous cell growth in melanoma. JARID1B knockdown resulted in the suppression of esophageal cancer cell growth, sphere formation and invasion ability and was associated with loss of epithelial marker expression. However, these inhibitory effects observed on tumor formation were reverted subsequent to subcutaneous inoculation of these cells into immune-deficient mice. These results indicated that JARID1B plays a role in maintaining cancer stem cells in the esophagus and justifies the rationale for studying the effects of continuous inhibition of this epigenetic factor in esophageal cancer. PMID:24649241

  19. Enterovirus 71 VPg Uridylation Uses a Two-Molecular Mechanism of 3D Polymerase

    PubMed Central

    Sun, Yuna; Wang, Yaxin; Shan, Chao; Chen, Cheng; Xu, Peng; Song, Mohan; Zhou, Honggang; Yang, Cheng; Xu, Wenbo; Shi, Pei-Yong

    2012-01-01

    VPg uridylylation is essential for picornavirus RNA replication. The VPg uridylylation reaction consists of the binding of VPg to 3D polymerase (3Dpol) and the transfer of UMP by 3Dpol to the hydroxyl group of the third amino acid Tyr of VPg. Previous studies suggested that different picornaviruses employ distinct mechanisms during VPg binding and uridylylation. Here, we report a novel site (Site-311, located at the base of the palm domain of EV71 3Dpol) that is essential for EV71 VPg uridylylation as well as viral replication. Ala substitution of amino acids (T313, F314, and I317) at Site-311 reduced the VPg uridylylation activity of 3Dpol by >90%. None of the Site-311 mutations affected the RNA elongation activity of 3Dpol, which indicates that Site-311 does not directly participate in RNA polymerization. However, mutations that abrogated VPg uridylylation significantly reduced the VPg binding ability of 3Dpol, which suggests that Site-311 is a potential VPg binding site on enterovirus 71 (EV71) 3Dpol. Mutation of a polymerase active site in 3Dpol and Site-311 in 3Dpol remarkably enables trans complementation to restore VPg uridylylation. In contrast, two distinct Site-311 mutants do not cause trans complementation in vitro. These results indicate that Site-311 is a VPg binding site that stabilizes the VPg molecule during the VPg uridylylation process and suggest a two-molecule model for 3Dpol during EV71 VPg uridylylation, such that one 3Dpol presents the hydroxyl group of Tyr3 of VPg to the polymerase active site of another 3Dpol, which in turn catalyzes VPg?VPg-pU conversion. For genome-length RNA, the Site-311 mutations that reduced VPg uridylylation were lethal for EV71 replication, which indicates that Site-311 is a potential antiviral target. PMID:23055549

  20. Evaluation of the stability of enterovirus 71 virus-like particle.

    PubMed

    Lin, Shih-Yeh; Chung, Yao-Chi; Chiu, Hsin-Yi; Chi, Wei-Kuang; Chiang, Bor-Luen; Hu, Yu-Chen

    2014-03-01

    Enterovirus 71 (EV71) is responsible for the outbreaks of hand-foot-and-mouth disease that caused significant mortality in children, but no vaccine is available yet. EV71 virus-like particle (VLP) is the empty capsid consisting of viral structural proteins but can elicit potent immune responses, rendering VLP a promising EV71 vaccine candidate. To evaluate whether VLP remains stable after long-term storage, which is crucial for advancing the VLP vaccine to the clinical setting, we evaluated the effects of NaCl concentration, buffers and temperatures on the VLP stability. We first validated the use of dynamic light scattering (DLS) for measuring the hydrodynamic diameter (?30-35 nm) of VLP, which was close to the VLP diameter (?25-27 nm) as measured by transmission electron microscopy (TEM). Using these techniques, we found that EV71 VLP remained stable for 5 months in sodium phosphate (NaPi) buffers with various NaCl concentrations. EV71 VLP also remained morphologically stable in NaPi, citrate and TE(+) buffers for 5 months, yet the enzyme-linked immunosorbent assay (ELISA) revealed that the VLP stored in citrate and TE(+) buffers partially lost the immunogenicity after 5 months. In contrast, the VLP stored in the NaPi buffer at 4°C remained stable macroscopically and microscopically for 5 months, as judged from the DLS, TEM and ELISA. The VLP stored at 25°C and 37°C also retained stability for 1 month, which would obviate the need of a cold chain during the shipping. These data altogether proved the stability of EV71 VLP and suggested that the VLP is amenable to bioprocessing and storage. PMID:24140131

  1. Chlorine dioxide inactivation of enterovirus 71 in water and its impact on genomic targets.

    PubMed

    Jin, Min; Shan, Jinyang; Chen, Zhaoli; Guo, Xuan; Shen, Zhiqiang; Qiu, Zhigang; Xue, Bin; Wang, Yongguang; Zhu, Dunwan; Wang, Xinwei; Li, Junwen

    2013-05-01

    To control the waterborne transmission of enterovirus 71(EV71), which is associated with hand foot and mouth disease (HFMD), it is essential to know the inactivation effectiveness of disinfectants on EV71 in water. In this article, we present a comparative analysis of the effects on EV71 following exposure to chlorine dioxide (ClO2) under different doses, pH, and temperature conditions. We show that the EV71 exhibited strong resistance to ClO2 (more than the MS2 standard) and that Ct value ranges required for a 4-log reduction of EV71 in buffered, disinfectant demand-free water at pH 7.2 and 20 °C by ClO2 were 4.24-6.62 mg/L·min according to the efficiency factor Hom model. ClO2 inactivation of the virus was temperature- and pH-dependent. The virucidal efficiency was higher at pH 8.2 than at pH 5.6 and pH 7.2 and higher at 36 °C than at 4 and 20 °C. In addition, we also observed the impact of ClO2 on the entire viral genome using RT-PCR, which indicated that the 5' noncoding region (5'-NCR) within the EV71 genome, specifically the 1-118 nt region, was the most easily damaged by ClO2 and correlated with viral infectivity. Our study has not only provided guidelines for EV71 disinfection strategies of waste and drinking water, but also confirmed the importance of the 5'-NCR for EV71 infectivity and may demonstrate a general inactivation by ClO2 of enteric virus by damaging the 5'-NCR. Furthermore, 5'-NCR can be used as a target region for PCR to investigate infectious virus contamination in environmental water and evaluate the inactivation effects of ClO2. PMID:23560857

  2. Enterovirus 71 Virion-Associated Galectin-1 Facilitates Viral Replication and Stability

    PubMed Central

    Lee, Pei-Huan; Liu, Chia-Ming; Ho, Tzong-Shiann; Tsai, Yi-Che; Lin, Chi-Cheng; Wang, Ya-Fang; Chen, Yuh-Ling; Yu, Chun-Keung; Wang, Shih-Min; Liu, Ching-Chuan; Shiau, Ai-Li; Lei, Huan-Yao; Chang, Chih-Peng

    2015-01-01

    Enterovirus 71 (EV71) infection causes a myriad of diseases from mild hand-foot-and-mouth disease or herpangina to fatal brain stem encephalitis complicated with pulmonary edema. Several severe EV71 endemics have occurred in Asia-Pacific region, including Taiwan, and have become a serious threat to children’s health. EV71 infection is initiated by the attachment of the virion to the target cell surface. Although this process relies primarily upon interaction between viruses and cell surface receptors, soluble factors may also influence the binding of EV71 to host cells.Galectin-1 has been reported to participate in several virus infections, but is not addressed in EV71. In this study, we found that the serum levels of galectin-1 in EV71-infected children were higher than those in non-infected people. In EV71 infected cells, galectin-1 was found to be associated with the EV71 VP1 and VP3 via carbohydrate residues and subsequently released and bound to another cell surface along with the virus. EV71 propagated from galectin-1 knockdown SK-N-SH cells exhibited lower infectivity in cultured cells and less pathogenicity in mice than the virus propagated from parental cells. In addition, this galectin-1-free EV71 virus was sensitive to high temperature and lost its viability after long-term storage, which could be restored following supplement of recombinant galectin-1. Taken together, our findings uncover a new role of galectin-1 in facilitating EV71 virus infection. PMID:25706563

  3. HspB1, HspB5 and HspB4 in Human Cancers: Potent Oncogenic Role of Some of Their Client Proteins

    PubMed Central

    Arrigo, André-Patrick; Gibert, Benjamin

    2014-01-01

    Human small heat shock proteins are molecular chaperones that regulate fundamental cellular processes in normal unstressed cells as well as in many cancer cells where they are over-expressed. These proteins are characterized by cell physiology dependent changes in their oligomerization and phosphorylation status. These structural changes allow them to interact with many different client proteins that subsequently display modified activity and/or half-life. Nowdays, the protein interactomes of small Hsps are under intense investigations and will represent, when completed, key parameters to elaborate therapeutic strategies aimed at modulating the functions of these chaperones. Here, we have analyzed the potential pro-cancerous roles of several client proteins that have been described so far to interact with HspB1 (Hsp27) and its close members HspB5 (?B-crystallin) and HspB4 (?A-crystallin). PMID:24514166

  4. Oxidation of PAH trans-Dihydrodiols by Human Aldo-Keto Reductase AKR1B10

    PubMed Central

    Quinn, Amy M.; Harvey, Ronald G.; Penning, Trevor M.

    2009-01-01

    AKR1B10 has been identified as a potential biomarker for human non-small cell lung carcinoma and as a tobacco exposure and response gene. AKR1B10 functions as an efficient retinal reductase in vitro, and may regulate retinoic acid homeostasis. However, the possibility that this enzyme is able to activate polycyclic aromatic hydrocarbon (PAH) trans-dihydrodiols to form reactive and redox-active o-quinones has not been investigated to date. AKR1B10 was found to oxidize a wide range of PAH trans-dihydrodiol substrates in vitro to yield PAH o-quinones. Reactions of AKR1B10 proceeded with improper stereochemistry, since it was specific for the minor (+)-benzo[a]pyrene-7S,8S-dihydrodiol diastereomer formed in vivo. However, AKR1B10 displayed reasonable activity in the oxidation of both the (?)-R,R and (+)-S,S stereoisomers of benzo[g]chrysene-11,12-dihydrodiol and oxidized the potentially relevant, albeit minor, (+)-benz[a]anthracene-3S,4S-dihydrodiol metabolite. We find that AKR1B10 is therefore likely to play a contributing role in the activation of PAH trans-dihydrodiols in human lung. AKR1B10 retinal reductase activity was confirmed in vitro and found to be 5- to 150-fold greater than the oxidation of PAH trans-dihydrodiols examined. AKR1B10 was highly expressed at the mRNA and protein levels in human lung adenocarcinoma A549 cells, and robust retinal reductase activity was measured in lysates of these cells. The much greater catalytic efficiency of retinal reduction compared to PAH trans-dihydrodiol metabolism suggests AKR1B10 may play a greater role in lung carcinogenesis through dysregulation of retinoic acid homeostasis than through oxidation of PAH trans-dihydrodiols. PMID:18788756

  5. Chromosomal mapping, gene structure and characterization of the human and murine RAB27B gene

    PubMed Central

    Ramalho, José S; Tolmachova, Tanya; Hume, Alistair N; McGuigan, Amanda; Gregory-Evans, Cheryl Y; Huxley, Clare; Seabra, Miguel C

    2001-01-01

    Background Rab GTPases are regulators of intracellular membrane traffic. The Rab27 subfamily consists of Rab27a and Rab27b. Rab27a has been recently implicated in Griscelli Disease, a disease combining partial albinism with severe immunodeficiency. Rab27a plays a key role in the function of lysosomal-like organelles such as melanosomes in melanocytes and lytic granules in cytotoxic T lymphocytes. Little is known about Rab27b. Results The human RAB27B gene is organised in six exons, spanning about 69 kb in the chromosome 18q21.1 region. Exon 1 is non-coding and is separated from the others by 49 kb of DNA and exon 6 contains a long 3' untranslated sequence (6.4 kb). The mouse Rab27b cDNA shows 95% identity with the human cDNA at the protein level and maps to mouse chromosome 18. The mouse mRNA was detected in stomach, large intestine, spleen and eye by RT-PCR, and in heart, brain, spleen and kidney by Northern blot. Transient over-expression of EGF-Rab27b fusion protein in cultured melanocytes revealed that Rab27b is associated with melanosomes, as observed for EGF-Rab27a. Conclusions Our results indicate that the Rab27 subfamily of Ras-like GTPases is highly conserved in mammals. There is high degree of conservation in sequence and gene structure between RAB27A and RAB27B genes. Exogenous expression of Rab27b in melanocytes results in melanosomal association as observed for Rab27a, suggesting the two Rab27 proteins are functional homologues. As with RAB27A in Griscelli Disease, RAB27B may be also associated with human disease mapping to chromosome 18. PMID:11178108

  6. Blockade of NF-?B activity in human prostate cancer cells is associated with suppression of angiogenesis, invasion, and metastasis

    Microsoft Academic Search

    Suyun Huang; Curtis A Pettaway; Hisanori Uehara; Corazon D Bucana; Isaiah J Fidler

    2001-01-01

    Since the NF-?B\\/relA transcription factor is constitutively activated in human prostate cancer cells, we determined whether blocking NF-?B\\/relA activity in human prostate cancer cells affected their angiogenesis, growth, and metastasis in an orthotopic nude mouse model. Highly metastatic PC-3M human prostate cancer cells were transfected with a mutated I?B? (I?B?M), which blocks NF-?B activity. Parental (PC-3M), control vector-transfected (PC-3M-Neo), and

  7. BIOS 5445 Human Ecology Dr. S.B. Malcolm Page -1 HUMAN ECOLOGY: BIOS 5445

    E-print Network

    Malcolm, Stephen

    Goal: To develop an appreciation for patterns of distribution and abundance of Homo sapiens the will to succeed. Our problem as humans is that much of what we do is governed by our genes acting selfishly purely selfish. So how do we resolve these issues? I don't pretend that this course will fully answer

  8. Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors.

    PubMed

    Kvell, Krisztian; Nguyen, Tuan H; Salmon, Patrick; Glauser, Frédéric; Werner-Favre, Christiane; Barnet, Marc; Schneider, Pascal; Trono, Didier; Zubler, Rudolf H

    2005-11-01

    Recently, using HIV-1-derived lentivectors, we obtained efficient transduction of primary human B lymphocytes cocultured with murine EL-4 B5 thymoma cells, but not of isolated B cells activated by CD40 ligation. Coculture with a cell line is problematic for gene therapy applications or study of gene functions. We have now found that transduction of B cells in a system using CpG DNA was comparable to that in the EL-4 B5 system. A monocistronic vector with a CMV promoter gave 32 +/- 4.7% green fluorescent protein (GFP)+ cells. A bicistronic vector, encoding IL-4 and GFP in the first and second cistrons, respectively, gave 14.2 +/- 2.1% GFP+ cells and IL-4 secretion of 1.3 +/- 0.2 ng/10(5) B cells/24 h. This was similar to results obtained in CD34+ cells using the elongation factor-1alpha promoter. Activated memory and naive B cells were transducible. After transduction with a bicistronic vector encoding a viral FLIP molecule, vFLIP was detectable by FACS or Western blot in GFP+, but not in GFP-, B cells, and 57% of sorted GFP+ B cells were protected against Fas ligand-induced cell death. This system should be useful for gene function research in primary B cells and development of gene therapies. PMID:16005685

  9. Characterization of the human N?-terminal acetyltransferase B enzymatic complex

    PubMed Central

    Ametzazurra, Amagoia; Gázquez, Cristina; Lasa, Marta; Larrea, Esther; Prieto, Jesús; Aldabe, Rafael

    2009-01-01

    Background Human N?-acetyltransferase complex B (hNatB) is integrated by hNaa20p (hNAT5/hNAT3) and hNaa25p (hMDM20) proteins. Previous data have shown that this enzymatic complex is implicated in cell cycle progression and carcinogenesis. In yeast this enzyme acetylates peptides composed by methionine and aspartic acid or glutamic acid in their first two positions respectively and it has been shown the same specificity in human cells. Methods We have silenced hNAA20 expression in hepatic cell lines using recombinant adenoviruses that express specific siRNAs against this gene and analyzed cell cycle progression and apoptosis induction after this treatment. Immunopurified hNatB enzymatic complexes from human cell lines were used for analyzing hNatB in vitro enzymatic activity using as substrate peptides predicted to be acetylated by NatB. Results hNAA20 silencing in hepatic cell lines reduces cell proliferation in a p53 dependent and independent manner. At the same time this treatment sensitizes the cells to a proapototic stimulus. We have observed that the hNatB complex isolated from human cell lines can acetylate in vitro peptides that present an aspartic or glutamic acid in their second position as has been described in yeast. Conclusion hNatB enzymatic complex is implicated in cell cycle progression but it exerts its effects through different mechanisms depending on the cellular characteristics. This is achievable because it can acetylate a great number of peptides composed by an aspartic or glutamic acid at their second residue and therefore it can regulate the activity of a great number of proteins. PMID:19660097

  10. UV-A-induced decrease in nuclear factor-kappaB activity in human keratinocytes.

    PubMed Central

    Djavaheri-Mergny, M; Gras, M P; Mergny, J L; Dubertret, L

    1999-01-01

    Previous reports have demonstrated an increase in nuclear factor-kappaB (NF-kappaB) activity in response to UV radiation. These studies have essentially focused on the DNA-damaging fraction of solar UV radiation (UV-B and UV-C). In contrast, the effects of UV-A radiation (320-400 nm) on NF-kappaB are not well known. In this study, we present evidence that UV-A radiation induces a marked decrease in NF-kappaB DNA-binding activity in NCTC 2544 human keratinocytes. In addition, NCTC 2544 keratinocytes pretreated with UV-A fail to respond to NF-kappaB inducers. Moreover, UV-A radiation induces a decrease in NF-kappaB-driven luciferase reporter gene expression in NCTC 2544 keratinocytes. The expression of the gene encoding IkappaBalpha (IkappaB is the NF-kappaB inhibitor), which is closely associated with NF-kappaB activity, is also reduced (3-fold) upon UV-A treatment. Our results indicate that the UV-A-induced decrease in NF-kappaB DNA-binding activity is associated with a decrease in the levels of the p50 and p65 protein subunits. This is the first evidence that an oxidative stress, such as UV-A radiation, may induce a specific decrease in NF-kappaB activity in mammalian cells, probably through degradation of NF-kappaB protein subunits. These findings suggest that UV-A could modulate the NF-kappaB-dependent gene expression. PMID:10051429

  11. Frequency and genotype of human parvovirus B19 among Iranian patients infected with HIV.

    PubMed

    Azadmanesh, Kayhan; Mohraz, Minoo; Kazemimanesh, Monireh; Aghakhani, Arezoo; Foroughi, Maryam; Banifazl, Mohammad; Eslamifar, Ali; Ramezani, Amitis

    2015-07-01

    The human parvovirus B19 (B19) usually causes a subclinical infection in immunocompetent individuals. Whereas immunocompromised individuals such as patients infected with HIV are at risk of persistent anemia due to B19 infection. Only few studies have been carried out on distribution and molecular epidemiology of B19 in Iran. We aimed to determine the frequency and genotype of B19 among Iranian patients infected with HIV. We conducted a survey on 99 HIV patients and 64 healthy controls. IgG and IgM antibodies against B19 were detected by ELISA and B19 DNA was assessed by nested PCR. PCR products were subjected to direct sequencing and classified after phylogenetic analysis. The prevalence of B19 immunoglobulin was 11.1% for IgG and 1% for IgM. B19 DNA was detected in 13.1% of cases. The prevalence of B19 IgG, IgM, and DNA in control group was 25%, 1.6%, and 9.4%, respectively. B19 IgG was significantly lower in HIV group than in normal controls. There was no significant difference regarding anemia between cases and controls. All sequenced B19 isolates belonged to genotype 1A with low genetic diversity. Our findings indicated that in the HAART era, the importance of B19 infections in HIV patients may be limited whereas persistent B19 viremia in the circulation of healthy controls raises a potential concern in blood donations. J. Med. Virol. 87:1124-1129, 2015. © 2015 Wiley Periodicals, Inc. PMID:25771750

  12. Identification of a germ line transcript from the unrearranged kappa gene in human B cells.

    PubMed Central

    Martin, D J; Van Ness, B G

    1989-01-01

    A novel kappa immunoglobulin-hybridizing mRNA in cell lines derived from human B cells arrested at several stages of development has been identified. Hybridization studies demonstrate that this 1.5-kilobase mRNA species is the spliced product of a precursor germ line transcript initiating upstream of the unrearranged JKappa locus. Images PMID:2573834

  13. Competitive chemiluminescent anzyme immunoassay for vitamin B12 analysis in human milk.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent discoveries of matrix interferences by haptocorrin (HC) in human milk and serum show that past analyses of vitamin B12 in samples with high HC content might have been inaccurate (Lildballe et al., 2009; Carmel & Agrawal, 2012). We evaluated two competitive enzyme-binding immunoassays for seru...

  14. Social closeness increases salivary progesterone in humans Stephanie L. Brown a,b,

    E-print Network

    Schultheiss, Oliver C.

    Social closeness increases salivary progesterone in humans Stephanie L. Brown a,b, , Barbara L Altruism Affiliation Social closeness We examined whether interpersonal closeness increases salivary, but not to differences in cortisol or other hormones such as testosterone (Zak et al., 2005). In the context of "cues

  15. Ingested human insulin inhibits the mosquito NF-żB-dependent immune response to Plasmodium falciparum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We showed previously that ingested human insulin activates the insulin/IGF-1 signaling pathway in Anopheles stephensi and increases the susceptibility of these mosquitoes to Plasmodium falciparum. In other organisms insulin can alter immune responsiveness through regulation of NF-kB transcription fa...

  16. B Cell Sensitization to Helminthic Infection Develops In Utero in Humans1

    Microsoft Academic Search

    Christopher L. King; Indu Malhotra; Peter Mungai; Alex Wamachi; John Kioko; John H. Ouma; James W. Kazura

    Human neonates are generally deficient in their ability to generate humoral immunity. This deficiency is thought to reflect physiologic immaturity of T and B cell function and lack of previous exposure to exogenous Ags. To determine whether neonatal humoral immunity can be modified by maternal helminth infection during pregnancy, we assessed Ig production by cord blood lymphocytes from healthy newborns

  17. Human endometrial stem cell neurogenesis in response to NGF and bFGF.

    PubMed

    Noureddini, Mahdi; Verdi, Javad; Mortazavi-Tabatabaei, Seyed Abdolreza; Sharif, Shiva; Azimi, Alireza; Keyhanvar, Peyman; Shoae-Hassani, Alireza

    2012-10-01

    The potential of cell therapy is promising in nerve regeneration, but is limited by ethical considerations about the proper and technically safe source of stem cells. We report the successful differentiation of human EnSCs (endometrial stem cells) as a rich source of renewable and safe progenitors into high-efficiency cholinergic neurons. The extracellular signals of NGF (nerve growth factor) and bFGF (basic fibroblast growth factor) could induce cholinergic neuron differentiation. ChAT (choline acetyltransferase), MAP2 (microtubule associated protein 2) and NF-l (neurofilament L) increased after administration of bFGF and NGF to the EnSC cultures. trkC and FGFR2 (fibroblast growth factor receptor 2), which belong to the NGF and bFGF receptors respectively, were determined in populations of EnSCs. NGF, bFGF and their combination differentially influenced human EnSCs high efficiency differentiation. By inducing cholinergic neurons from EnSCs in a chemically defined medium, we could produce human neural cells without resorting to primary culture of neurons. This in vitro method provides an unlimited source of human neural cells and facilitates clinical applications of EnSCs for neurological diseases. PMID:22804708

  18. The NADPH Metabolic Network Regulates Human aB-crystallin Cardiomyopathy and Reductive Stress in

    E-print Network

    The NADPH Metabolic Network Regulates Human aB- crystallin Cardiomyopathy and Reductive Stress is that the disease state can be linked to a disturbance in the oxidation/reduction environment of the cellAB in the Drosophila heart impaired cardiac function and increased heart tube dimensions, similar to the effects

  19. BDNF/TrkB signaling protects HT-29 human colon cancer cells from EGFR inhibition

    SciTech Connect

    Brunetto de Farias, Caroline [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil) [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); Children's Cancer Institute, 90420-140 Porto Alegre, RS (Brazil); Laboratory of Neuropharmacology and Neural Tumor Biology, Department of Pharmacology, Institute for Basic Health Sciences, Federal University of Rio Grande do Sul, 90050-170 Porto Alegre, RS (Brazil); National Institute for Translational Medicine (INCT-TM), 90035-003 Porto Alegre, RS (Brazil); Heinen, Tiago Elias; Pereira dos Santos, Rafael [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil) [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); Laboratory of Neuropharmacology and Neural Tumor Biology, Department of Pharmacology, Institute for Basic Health Sciences, Federal University of Rio Grande do Sul, 90050-170 Porto Alegre, RS (Brazil); National Institute for Translational Medicine (INCT-TM), 90035-003 Porto Alegre, RS (Brazil); Abujamra, Ana Lucia [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil) [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); Children's Cancer Institute, 90420-140 Porto Alegre, RS (Brazil); National Institute for Translational Medicine (INCT-TM), 90035-003 Porto Alegre, RS (Brazil); Schwartsmann, Gilberto [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil) [Cancer Research Laboratory, University Hospital Research Center (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); National Institute for Translational Medicine (INCT-TM), 90035-003 Porto Alegre, RS (Brazil); Department of Internal Medicine, School of Medicine, Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS (Brazil); and others

    2012-08-24

    Highlights: Black-Right-Pointing-Pointer BDNF protected HT-29 colorectal cancer cells from the antitumor effect of cetuximab. Black-Right-Pointing-Pointer TrkB inhibition potentiated the antitumor effect of cetuximab. Black-Right-Pointing-Pointer BDNF/TrkB signaling might be involved in resistance to anti-EGFR therapy. -- Abstract: The clinical success of targeted treatment of colorectal cancer (CRC) is often limited by resistance to anti-epidermal growth factor receptor (EGFR) therapy. The neurotrophin brain-derived neurotrophic factor (BDNF) and its receptor TrkB have recently emerged as anticancer targets, and we have previously shown increased BDNF levels in CRC tumor samples. Here we report the findings from in vitro experiments suggesting that BDNF/TrkB signaling can protect CRC cells from the antitumor effects of EGFR blockade. The anti-EGFR monoclonal antibody cetuximab reduced both cell proliferation and the mRNA expression of BDNF and TrkB in human HT-29 CRC cells. The inhibitory effect of cetuximab on cell proliferation and survival was counteracted by the addition of human recombinant BDNF. Finally, the Trk inhibitor K252a synergistically enhanced the effect of cetuximab on cell proliferation, and this effect was blocked by BDNF. These results provide the first evidence that increased BDNF/TrkB signaling might play a role in resistance to EGFR blockade. Moreover, it is possible that targeting TrkB could potentiate the anticancer effects of anti-EGFR therapy.

  20. MicroRNA-200b targets CREB1 and suppresses cell growth in human malignant glioma.

    PubMed

    Peng, Biao; Hu, Su; Jun, Qinming; Luo, Dongdong; Zhang, Xun; Zhao, Hailin; Li, Dan

    2013-07-01

    MicroRNAs can coordinately repress multiple target genes and interfere with the biological functions of the cell, such as proliferation and apoptosis. In the present study, we report that miR-200b was downregulated in malignant glioma cell lines and specimens. Overexpression of miR-200b suppressed the proliferation and colony formation of glioma cells. An oncogene encoding cAMP responsive element-binding protein 1 (CREB1), which has been shown to be an important transcription factor involved in the proliferation, survival, and metastasis of tumor cells, was here confirmed as a direct target gene of miR-200b. CREB1 was also found to be present at a high level in human glioma tissues. This was inversely correlated with miR-200b expression. Ectopic expression of CREB1 attenuated the growth suppressive phenotypes of glioma cells caused by miR-200b. These results indicate that miR-200b targets the CREB1 gene and suppresses glioma cell growth, suggesting that miR-200b shows tumor-suppressive activity in human malignant glioma. PMID:23543137

  1. In vivo dynamics of enterovirus protease revealed by fluorescence resonance emission transfer (FRET) based on a novel FRET pair

    SciTech Connect

    Hsu, Y.-Y. [Faculty of Biotechnology and Laboratory Science in Medicine, National Yang-Ming University, Taipei, Taiwan (China); Liu, Y.-N. [Faculty of Biotechnology and Laboratory Science in Medicine, National Yang-Ming University, Taipei, Taiwan (China); Wang Wenyen [Incubation Center, National Yang-Ming University, Taipei, Taiwan (China); Kao, Fu-Jen [Institute of Biophotonics, National Yang-Ming University, Taipei, Taiwan (China); Kung, S.-H. [Faculty of Biotechnology and Laboratory Science in Medicine, National Yang-Ming University, Taipei, Taiwan (China)]. E-mail: szkung@ym.edu.tw

    2007-02-23

    An in vivo protease assay suitable for analysis by fluorescence resonance energy transfer (FRET) was developed on the basis of a novel FRET pair. The specifically designed fusion substrate consists of green fluorescent protein 2 (GFP{sup 2})-peptide-red fluorescent protein 2 (DsRed2), with a cleavage motif for the enterovirus 2A protease (2A{sup pro}) embedded within the peptide region. FRET can be readily visualized in real-time from cells expressing the fusion substrate until a proteolytic cleavage by 2A{sup pro} from the input virus. The level of FRET decay is a function of the amount and infection duration of the inoculated virus as measured by a fluorometer assay. The FRET biosensor also responded well to other related enteroviruses but not to a phylogenetically distant virus. Western blot analysis confirmed the physical cleavage of the fusion substrate upon the infections. The study provides proof of principle for applying the FRET technology to diagnostics, screening procedures, and cell biological research.

  2. General primer-mediated polymerase chain reaction for detection of enteroviruses: application for diagnostic routine and persistent infections.

    PubMed Central

    Zoll, G J; Melchers, W J; Kopecka, H; Jambroes, G; van der Poel, H J; Galama, J M

    1992-01-01

    The aim of this study was to determine the applicability of the polymerase chain reaction (PCR) for routine diagnostic use and for the detection of persistent enteroviral infections. To this end, general primers were selected in the highly conserved part of the 5'-noncoding region of the enteroviral genome. They were tested on 66 different enterovirus serotypes. A specific fragment was amplified from 60 of 66 serotypes. An amplification product was not observed from coxsackievirus types A11, A17, and A24 and echovirus types 16, 22, and 23. Enteroviral RNA was detected by the PCR in routinely collected throat swabs and stool specimens that were found to be positive for enterovirus by isolation in tissue culture. Enteroviral RNA was detected in one of five myocardial biopsy specimens from patients with dilated cardiomyopathy, implicating virus persistence. No amplification product was obtained from eight control samples. Our results demonstrate the significance of the PCR for the detection of enteroviral RNA and, in particular, for the demonstration of persistent enteroviral infections. Images PMID:1370845

  3. Quercetin induces human colon cancer cells apoptosis by inhibiting the nuclear factor-kappa B Pathway

    PubMed Central

    Zhang, Xiang-An; Zhang, Shuangxi; Yin, Qing; Zhang, Jing

    2015-01-01

    Quercetin can inhibit the growth of cancer cells with the ability to act as chemopreventers. Its cancer-preventive effect has been attributed to various mechanisms, including the induction of cell-cycle arrest and/or apoptosis as well as the antioxidant functions. Nuclear factor kappa-B (NF-?B) is a signaling pathway that controls transcriptional activation of genes important for tight regulation of many cellular processes and is aberrantly expressed in many types of cancer. Inhibitors of NF-?B pathway have shown potential anti-tumor activities. However, it is not fully elucidated in colon cancer. In this study, we demonstrate that quercetin induces apoptosis in human colon cancer CACO-2 and SW-620 cells through inhibiting NF-?B pathway, as well as down-regulation of B-cell lymphoma 2 and up-regulation of Bax, thus providing basis for clinical application of quercetin in colon cancer cases. PMID:25829782

  4. Quercetin induces human colon cancer cells apoptosis by inhibiting the nuclear factor-kappa B Pathway.

    PubMed

    Zhang, Xiang-An; Zhang, Shuangxi; Yin, Qing; Zhang, Jing

    2015-01-01

    Quercetin can inhibit the growth of cancer cells with the ability to act as chemopreventers. Its cancer-preventive effect has been attributed to various mechanisms, including the induction of cell-cycle arrest and/or apoptosis as well as the antioxidant functions. Nuclear factor kappa-B (NF-?B) is a signaling pathway that controls transcriptional activation of genes important for tight regulation of many cellular processes and is aberrantly expressed in many types of cancer. Inhibitors of NF-?B pathway have shown potential anti-tumor activities. However, it is not fully elucidated in colon cancer. In this study, we demonstrate that quercetin induces apoptosis in human colon cancer CACO-2 and SW-620 cells through inhibiting NF-?B pathway, as well as down-regulation of B-cell lymphoma 2 and up-regulation of Bax, thus providing basis for clinical application of quercetin in colon cancer cases. PMID:25829782

  5. The Human Cytomegalovirus UL26 Protein Antagonizes NF-?B Activation

    PubMed Central

    Mathers, Chun; Schafer, Xenia; Martínez-Sobrido, Luis

    2014-01-01

    ABSTRACT Viral infection frequently triggers activation of host innate immune pathways that attempt to limit viral spread. The NF-?B pathway is a critical component that governs this response. We have found that the human cytomegalovirus (HCMV) UL26 protein antagonizes NF-?B activation. Upon infection, an HCMV strain lacking the UL26 gene (?UL26) induced the nuclear translocation of the NF-?B RelB subunit and activated expression and secretion of interleukin-6 (IL-6), an NF-?B target gene. The ?UL26 mutant was also more sensitive to challenge with tumor necrosis factor alpha (TNF-?), a canonical NF-?B inducer. Further, expression of UL26 in the absence of other viral proteins blocked NF-?B activation induced by either TNF-? treatment or infection with Sendai virus (SeV). Our results indicate that UL26 expression is sufficient to block TNF-?-induced NF-?B nuclear translocation and I?B degradation. Last, UL26 blocks TNF-?-induced I?B-kinase (IKK) phosphorylation, a key step in NF-?B activation. Combined, our results indicate that UL26 is part of a viral program to antagonize innate immunity through modulation of NF-?B signaling. IMPORTANCE The NF-?B signaling pathway regulates innate immunity, an integral host process that limits viral pathogenesis. Viruses have evolved mechanisms to modulate NF-?B signaling to ensure their replication. HCMV is a major cause of birth defects and disease in immunosuppressed populations. HCMV is known to actively target the NF-?B pathway, which is important for HCMV infection. Our results indicate that the HCMV UL26 gene is a key modulator of NF-?B pathway activity. We find the UL26 gene is both necessary and sufficient to block NF-?B activation upon challenge with antiviral cytokines. Further, UL26 attenuates the phosphorylation and activation of a key NF-?B activating kinase complex, IKK. Our study provides new insight into how HCMV targets the NF-?B pathway. Given its importance to viral infection, the mechanisms through which viruses target the NF-?B pathway highlight areas of vulnerability that could be therapeutically targeted to attenuate viral replication. PMID:25275128

  6. Human cytochrome P450 17A1 conformational selection: modulation by ligand and cytochrome b5.

    PubMed

    Estrada, D Fernando; Skinner, Andria L; Laurence, Jennifer S; Scott, Emily E

    2014-05-16

    Crystallographic studies of different membrane cytochrome P450 enzymes have provided examples of distinct structural conformations, suggesting protein flexibility. It has been speculated that conformational selection is an integral component of substrate recognition and access, but direct evidence of such substate interconversion has thus far remained elusive. In the current study, solution NMR revealed multiple and exchanging backbone conformations for certain structural features of the human steroidogenic cytochrome P450 17A1 (CYP17A1). This bifunctional enzyme is responsible for pregnenolone C17 hydroxylation, followed by a 17,20-lyase reaction to produce dehydroepiandrosterone, the key intermediate in human synthesis of androgen and estrogen sex steroids. The distribution of CYP17A1 conformational states was influenced by temperature, binding of these two substrates, and binding of the soluble domain of cytochrome b5 (b5). Notably, titration of b5 to CYP17A1·pregnenolone induced a set of conformational states closely resembling those of CYP17A1·17?-hydroxypregnenolone without b5, providing structural evidence consistent with the reported ability of b5 to selectively enhance 17,20-lyase activity. Solution NMR thus revealed a set of conformations likely to modulate human steroidogenesis by CYP17A1, demonstrating that this approach has the potential to make similar contributions to understanding the functions of other membrane P450 enzymes involved in drug metabolism and disease states. PMID:24671419

  7. Atrial fibrillation and sinus node dysfunction in human ankyrin-B syndrome: a computational analysis.

    PubMed

    Wolf, Roseanne M; Glynn, Patric; Hashemi, Seyed; Zarei, Keyan; Mitchell, Colleen C; Anderson, Mark E; Mohler, Peter J; Hund, Thomas J

    2013-05-01

    Ankyrin-B is a multifunctional adapter protein responsible for localization and stabilization of select ion channels, transporters, and signaling molecules in excitable cells including cardiomyocytes. Ankyrin-B dysfunction has been linked with highly penetrant sinoatrial node (SAN) dysfunction and increased susceptibility to atrial fibrillation. While previous studies have identified a role for abnormal ion homeostasis in ventricular arrhythmias, the molecular mechanisms responsible for atrial arrhythmias and SAN dysfunction in human patients with ankyrin-B syndrome are unclear. Here, we develop a computational model of ankyrin-B dysfunction in atrial and SAN cells and tissue to determine the mechanism for increased susceptibility to atrial fibrillation and SAN dysfunction in human patients with ankyrin-B syndrome. Our simulations predict that defective membrane targeting of the voltage-gated L-type Ca(2+) channel Cav1.3 leads to action potential shortening that reduces the critical atrial tissue mass needed to sustain reentrant activation. In parallel, increased fibrosis results in conduction slowing that further increases the susceptibility to sustained reentry in the setting of ankyrin-B dysfunction. In SAN cells, loss of Cav1.3 slows spontaneous pacemaking activity, whereas defects in Na(+)/Ca(2+) exchanger and Na(+)/K(+) ATPase increase variability in SAN cell firing. Finally, simulations of the intact SAN reveal a shift in primary pacemaker site, SAN exit block, and even SAN failure in ankyrin-B-deficient tissue. These studies identify the mechanism for increased susceptibility to atrial fibrillation and SAN dysfunction in human disease. Importantly, ankyrin-B dysfunction involves changes at both the cell and tissue levels that favor the common manifestation of atrial arrhythmias and SAN dysfunction. PMID:23436330

  8. TLR4 activates NF-{kappa}B in human ovarian granulosa tumor cells

    SciTech Connect

    Woods, Dori C., E-mail: dwoods2@partners.org [Vincent Center for Reproductive Biology, Vincent Obstetrics and Gynecology Service, Massachusetts General Hospital/Harvard Medical School, Boston, MA 02114 (United States); White, Yvonne A.R. [Vincent Center for Reproductive Biology, Vincent Obstetrics and Gynecology Service, Massachusetts General Hospital/Harvard Medical School, Boston, MA 02114 (United States)] [Vincent Center for Reproductive Biology, Vincent Obstetrics and Gynecology Service, Massachusetts General Hospital/Harvard Medical School, Boston, MA 02114 (United States); Dau, Caroline [University of California, San Francisco, School of Dentistry, San Francisco, CA 94143 (United States)] [University of California, San Francisco, School of Dentistry, San Francisco, CA 94143 (United States); Johnson, A.L. [Center for Reproductive Biology and Health, The Pennsylvania State University, University Park, PA 16802 (United States)] [Center for Reproductive Biology and Health, The Pennsylvania State University, University Park, PA 16802 (United States)

    2011-06-17

    Highlights: {yields} TLR4 is expressed in human ovarian granulosa tumor cells. {yields} Acting through TLR4, LPS and HSP60 induce a NF{kappa}B signaling cascade in human ovarian granulosa tumor cells. {yields} NF{kappa}B activation or inhibition did not alter chemosensitivity to TRAIL or cisplatin. -- Abstract: Previous studies have demonstrated expression of Toll-like receptors (TLRs) in the surface epithelium of normal ovaries (OSE) and in epithelial ovarian tumors. Most notably, OSE-derived cancers express TLR4, which activates the nuclear factor-kappa B (NF-{kappa}B) signaling cascade as a mediator of inflammatory response. Currently, there is considerable interest in elucidating the role of TLR-mediated signaling in cancers. Nevertheless, the expression of TLRs in granulosa cell tumors (GCTs) of the ovary, and the extent to which GCT expression of TLRs may influence cell-signaling pathways and/or modulate the efficacy of chemotherapeutics, has yet to be determined. In the present study, human GCT lines (COV434 and KGN) were utilized to evaluate expression of functional TLR4. TLR4 is expressed in GCT cell lines and ligation of TLR4 with bacterial lipopolysaccharide (LPS) led to I{kappa}B degradation and activation of NF-{kappa}B. NF-{kappa}B activation was confirmed by nuclear localization of NF-{kappa}B p65 following treatment with LPS and the naturally occurring ligand, HSP60. Notably, immunoneutralization of TLR4 blocked nuclear localization, and inhibition of NF-{kappa}B signaling attenuated LPS-induced TNF{alpha} plus increased doubling time in both cell lines. Contradictory to reports using human OSE cell lines, inhibition of NF-{kappa}B signaling failed to sensitize GCT lines to TRAIL or cisplatin. In summary, findings herein are the first to demonstrate a functional TLR-signaling pathway specifically in GCTs, and indicate that in contrast to OSE-derived cancers, inhibition of NF-{kappa}B does not sensitize GCTs to TRAIL or cisplatin.

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

    Microsoft Academic Search

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

    2005-01-01

    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

  10. Expression of MS4A and TMEM176 Genes in Human B Lymphocytes

    PubMed Central

    Zuccolo, Jonathan; Deng, Lili; Unruh, Tammy L.; Sanyal, Ratna; Bau, Jeremy A.; Storek, Jan; Demetrick, Douglas J.; Luider, Joanne M.; Auer-Grzesiak, Iwona A.; Mansoor, Adnan; Deans, Julie P.

    2013-01-01

    The MS4A gene family in humans includes CD20 and at least 15 other genes. CD20 exists as homo-oligomers in the plasma membrane, however different MS4A proteins expressed in the same cell may hetero-oligomerize. Given the importance of CD20 in B-cell function and as a therapeutic target, we sought to explore the potential for CD20 hetero-oligomerization with other MS4A proteins. We investigated expression in primary human B-cells of the four MS4A genes previously shown to be expressed in human B-cell lines (MS4A4A, MS4A6A, MS4A7, MS4A8B), as well as two genes comprising the closely related TMEM176 gene family, with a view to identifying candidates for future investigation at the protein level. TMEM176A and TMEM176B transcripts were either not detected, or were detected at relatively low levels in a minority of donor B-cell samples. MS4A4A and MS4A8B transcripts were not detected in any normal B-cell sample. MS4A6A and MS4A7 transcripts were detected at low levels in most samples, however the corresponding proteins were not at the plasma membrane when expressed as GFP conjugates in BJAB cells. We also examined expression of these genes in chronic lymphocytic leukemia (CLL), and found that it was similar to normal B-cells with two exceptions. First, whereas MS4A4A expression was undetected in normal B-cells, it was expressed in 1/14 CLL samples. Second, compared to expression levels in normal B-cells, MS4A6A transcripts were elevated in 4/14 CLL samples. In summary, none of the MS4A/TMEM176 genes tested was expressed at high levels in normal or in most CLL B-cells. MS4A6A and MS4A7 were expressed at low levels in most B-cell samples, however the corresponding proteins may not be positioned at the plasma membrane. Altogether, these data suggest that CD20 normally does not form hetero-oligomers with other MS4A proteins and that there are unlikely to be other MS4A proteins in CLL that might provide useful alternate therapeutic targets. PMID:23874341

  11. Expression and function of TRK-B and BDNF in human neuroblastomas.

    PubMed Central

    Nakagawara, A; Azar, C G; Scavarda, N J; Brodeur, G M

    1994-01-01

    There is considerable interest in the role of the TRK family of neuotrophin receptors in regulating growth and differentiation in normal and neoplastic nerve cells. A neuroblastoma is a common pediatric tumor derived from the neural crest, and the majority of favorable neuroblastomas express a high level of TRK-A mRNA. However, little is known about the expression or function of TRK-B in these tumors. TRK-B encodes a tyrosine kinase that binds to brain-derived neuotrophic factor (BDNF), as well as neurotrophin-3 (NT-3) and NT-4/5. We have studied the N-myc-amplified human neuroblastoma cell line, SMS-KCN, which expresses both TRK-B and BDNF. Exogenous BDNF induces tyrosine phosphorylation of TRK-B as well as phosphorylation of phospholipase C-gamma 1, the extracellular signal-regulated kinases 1 and 2, and phosphatidylinositol-3 kinase. BDNF also induces expression of the immediate-early genes c-FOS and NGFI-A but not NGFI-B or NGFI-C. In addition, BDNF appears to promote cell survival and neurite outgrowth. SMS-KCN cells also express TRK-A, which is phosphorylated in response to nerve growth factor. However, the downstream TRK-A signaling is apparently defective. Finally, we determined that in a series of 74 primary neuroblastomas, 36% express TRK-B mRNA, 68% express BDNF mRNA, and 31% express both. Truncated TRK-B appears to be preferentially expressed in more-differentiated tumors (ganglioneuromas and ganglioneuroblastomas), whereas full-length TRK-B is expressed almost exclusively in immature neuroblastomas with N-myc amplification. Our findings suggest that in TRK-B-expressing human neuroblastomas, BDNF promotes survival and induces neurite outgrowth in an autocrine or paracrine manner. The BDNF/TRK-B pathway may be particularly important for growth and differentiation of neuroblastomas with N-myc amplification. Images PMID:8264643

  12. Culture conditions profoundly impact phenotype in BEAS-2B, a human pulmonary epithelial model.

    PubMed

    Zhao, Fei; Klimecki, Walter T

    2015-08-01

    BEAS-2B, an immortalized, human lung epithelial cell line, has been used to model pulmonary epithelial function for over 30?years. The BEAS-2B phenotype can be modulated by culture conditions that include the presence or absence of fetal bovine serum (FBS). The popularity of BEAS-2B as a model of arsenic toxicology, and the common use of BEAS-2B cultured both with and without FBS, led us to investigate the impact of FBS on BEAS-2B in the context of arsenic toxicology. Comparison of genome-wide gene expression in BEAS-2B cultured with or without FBS revealed altered expression in several biological pathways, including those related to carcinogenesis and energy metabolism. Real-time measurements of oxygen consumption and glycolysis in BEAS-2B demonstrated that FBS culture conditions were associated with a 1.4-fold increase in total glycolytic capacity, a 1.9-fold increase in basal respiration, a 2.0-fold increase in oxygen consumed for ATP production and a 2.8-fold increase in maximal respiration, compared with BEAS-2B cultured without FBS. Comparisons of the transcriptome changes in BEAS-2B resulting from FBS exposure to the transcriptome changes resulting from exposure to 1??M sodium arsenite revealed that mRNA levels of 43% of the arsenite-modulated genes were also modulated by FBS. Cytotoxicity studies revealed that BEAS-2B cells exposed to 5% FBS for 8?weeks were almost 5 times more sensitive to arsenite cytotoxicity than non-FBS-exposed BEAS-2B cells. Phenotype changes induced in BEAS-2B by FBS suggest that culture conditions should be carefully considered when using BEAS-2B as an experimental model of arsenic toxicity. Copyright © 2014 John Wiley & Sons, Ltd. PMID:25524072

  13. CCDC22 deficiency in humans blunts activation of proinflammatory NF-?B signaling

    PubMed Central

    Starokadomskyy, Petro; Gluck, Nathan; Li, Haiying; Chen, Baozhi; Wallis, Mathew; Maine, Gabriel N.; Mao, Xicheng; Zaidi, Iram W.; Hein, Marco Y.; McDonald, Fiona J.; Lenzner, Steffen; Zecha, Agnes; Ropers, Hans-Hilger; Kuss, Andreas W.; McGaughran, Julie; Gecz, Jozef; Burstein, Ezra

    2013-01-01

    NF-?B is a master regulator of inflammation and has been implicated in the pathogenesis of immune disorders and cancer. Its regulation involves a variety of steps, including the controlled degradation of inhibitory I?B proteins. In addition, the inactivation of DNA-bound NF-?B is essential for its regulation. This step requires a factor known as copper metabolism Murr1 domain–containing 1 (COMMD1), the prototype member of a conserved gene family. While COMMD proteins have been linked to the ubiquitination pathway, little else is known about other family members. Here we demonstrate that all COMMD proteins bind to CCDC22, a factor recently implicated in X-linked intellectual disability (XLID). We showed that an XLID-associated CCDC22 mutation decreased CCDC22 protein expression and impaired its binding to COMMD proteins. Moreover, some affected individuals displayed ectodermal dysplasia, a congenital condition that can result from developmental NF-?B blockade. Indeed, patient-derived cells demonstrated impaired NF-?B activation due to decreased I?B ubiquitination and degradation. In addition, we found that COMMD8 acted in conjunction with CCDC22 to direct the degradation of I?B proteins. Taken together, our results indicate that CCDC22 participates in NF-?B activation and that its deficiency leads to decreased I?B turnover in humans, highlighting an important regulatory component of this pathway. PMID:23563313

  14. The TNF-family cytokine TL1A inhibits proliferation of human activated B cells.

    PubMed

    Cavallini, Chiara; Lovato, Ornella; Bertolaso, Anna; Pacelli, Luciano; Zoratti, Elisa; Zanolin, Elisabetta; Krampera, Mauro; Zamň, Alberto; Tecchio, Cristina; Cassatella, Marco A; Pizzolo, Giovanni; Scupoli, Maria T

    2013-01-01

    Death receptor (DR3) 3 is a member of the TNFR superfamily. Its ligand is TNF-like ligand 1A (TL1A), a member of the TNF superfamily. TL1A/DR3 interactions have been reported to modulate the functions of T cells, NK, and NKT cells and play a crucial role in driving inflammatory processes in several T-cell-dependent autoimmune diseases. However, TL1A expression and effects on B cells remain largely unknown. In this study, we described for the first time that B cells from human blood express significant amounts of DR3 in response to B cell receptor polyclonal stimulation. The relevance of these results has been confirmed by immunofluorescence analysis in tonsil and spleen tissue specimens, which showed the in situ expression of DR3 in antigen-stimulated B cells in vivo. Remarkably, we demonstrated that TL1A reduces B-cell proliferation induced by anti-IgM-antibodies and IL-2 but did not affect B-cell survival, suggesting that TL1A inhibits the signal(s) important for B-cell proliferation. These results revealed a novel function of TL1A in modulating B-cell proliferation in vitro and suggest that TL1A may contribute to homeostasis of effector B-cell functions in immune response and host defense, thus supporting the role of the TL1A/DR3 functional axis in modulating the adaptive immune response. PMID:23565196

  15. The TNF-Family Cytokine TL1A Inhibits Proliferation of Human Activated B Cells

    PubMed Central

    Cavallini, Chiara; Lovato, Ornella; Bertolaso, Anna; Pacelli, Luciano; Zoratti, Elisa; Zanolin, Elisabetta; Krampera, Mauro; Zamň, Alberto; Tecchio, Cristina; Cassatella, Marco A.; Pizzolo, Giovanni; Scupoli, Maria T.

    2013-01-01

    Death receptor (DR3) 3 is a member of the TNFR superfamily. Its ligand is TNF-like ligand 1A (TL1A), a member of the TNF superfamily. TL1A/DR3 interactions have been reported to modulate the functions of T cells, NK, and NKT cells and play a crucial role in driving inflammatory processes in several T-cell-dependent autoimmune diseases. However, TL1A expression and effects on B cells remain largely unknown. In this study, we described for the first time that B cells from human blood express significant amounts of DR3 in response to B cell receptor polyclonal stimulation. The relevance of these results has been confirmed by immunofluorescence analysis in tonsil and spleen tissue specimens, which showed the in situ expression of DR3 in antigen-stimulated B cells in vivo. Remarkably, we demonstrated that TL1A reduces B-cell proliferation induced by anti-IgM-antibodies and IL-2 but did not affect B-cell survival, suggesting that TL1A inhibits the signal(s) important for B-cell proliferation. These results revealed a novel function of TL1A in modulating B-cell proliferation in vitro and suggest that TL1A may contribute to homeostasis of effector B-cell functions in immune response and host defense, thus supporting the role of the TL1A/DR3 functional axis in modulating the adaptive immune response. PMID:23565196

  16. Murine and human b locus pigmentation genes encode a glycoprotein (gp75) with catalase activity.

    PubMed Central

    Halaban, R; Moellmann, G

    1990-01-01

    Melanogenesis is regulated in large part by tyrosinase (monophenol monooxygenase; monophenol, L-dopa:oxygen oxidoreductase, EC 1.14.18.1), and defective tyrosinase leads to albinism. The mechanisms for other pigmentation determinants (e.g., those operative in tyrosinase-positive albinism and in murine coat-color mutants) are not yet known. One murine pigmentation gene, the brown (b) locus, when mutated leads to a brown (b/b) or hypopigmented (Blt/Blt) coat versus the wild-type black (B/B). We show that the b locus codes for a glycoprotein with the activity of a catalase (hydrogen-peroxide:hydrogen-peroxide oxidoreductase, EC 1.11.1.6) (catalase B). Only the c locus protein is a tyrosinase. Because peroxides may be by-products of melanogenic activity and hydrogen peroxide in particular is known to destroy melanin precursors and melanin, we conclude that pigmentation is controlled not only by tyrosinase but also by a hydroperoxidase. Our studies indicate that catalase B is identical with gp75, a known human melanosomal glycoprotein; that the b mutation is in a heme-associated domain; and that the Blt mutation renders the protein susceptible to rapid proteolytic degradation. Images PMID:1693779

  17. B7-H3 is a potent inhibitor of human T cell activation: No evidence for B7-H3 and TREML2 interaction

    PubMed Central

    Leitner, Judith; Klauser, Christoph; Pickl, Winfried F.; Stöckl, Johannes; Majdic, Otto; Bardet, Anaďs F.; Kreil, David P.; Dong, Chen; Yamazaki, Tomohide; Zlabinger, Gerhard; Pfistershammer, Katharina; Steinberger, Peter

    2010-01-01

    B7-H3 belongs to the B7 superfamily, a group of molecules that costimulate or down-modulate T cell responses. Although it was shown that B7-H3 can inhibit T cell responses, several studies - most of them performed in murine systems - found B7-H3 to act in a costimulatory manner. In this study we have specifically addressed a potential functional dualism of human B7-H3 by assessing the effect of this molecule under varying experimental conditions as well as on different T cell subsets. We show that B7-H3 does not costimulate human T cells. In presence of strong activating signals, B7-H3 potently and consistently down-modulated human T cell responses. This inhibitory effect was evident when analyzing proliferation and cytokine production and affected naďve as well as pre-activated T cells. We furthermore demonstrate that B7-H3 - T cell interaction is characterized by an early suppression of IL-2 and that T cell inhibition can be reverted by exogenous IL-2. Since TREML2 has been recently described as costimulatory receptor of murine B7-H3 we have extensively analysed interaction of human B7-H3 with TREML2 (TLT2). In these experiments we found no evidence for such an interaction. Furthermore our data do not point to a role for murine TREML2 as a receptor for murine B7-H3. PMID:19544488

  18. Expression of the ammonia transporter family member, Rh B Glycoprotein, in the human kidney

    PubMed Central

    Han, Ki-Hwan; Lee, Hyun-Wook; Handlogten, Mary E.; Whitehill, Florence; Osis, Gunars; Croker, Byron P.; Clapp, William L.; Verlander, Jill W.

    2013-01-01

    The ammonia transporter family member, Rh B Glycoprotein (RhBG/Rhbg), is essential for ammonia transport by the rodent kidney, but in the human kidney mRNA but not protein expression has been reported. Because ammonia transport is fundamental for acid-base homeostasis, the current study addressed RhBG expression in the human kidney. Two distinct RhBG mRNA sequences have been reported, with different numbers of consecutive cytosines at nt1265 and thus encoding different carboxy-tails. Sequencing the region of difference in both human kidney and liver mRNA showed eight sequential cytosines, not seven as in some reports. Knowing the correct mRNA sequence for RhBG, we then assessed RhBG protein expression using antibodies against the correct amino acid sequence. Immunoblot analysis demonstrated RhBG protein expression in human kidney and immunohistochemistry identified basolateral RhBG in connecting segment (CNT) and the cortical and outer medullary collecting ducts. Colocalization of RhBG with multiple cell-specific markers demonstrated that that CNT cells and collecting duct type A intercalated cells express high levels of RhBG, and type B intercalated cells and principal cells do not express detectable RhBG. Thus, these studies identify the correct mRNA and thus protein sequence for human RhBG and show that the human kidney expresses basolateral RhBG protein in CNT, type A intercalated cells, and non-A, non-B cells. We conclude that RhBG can mediate an important role in human renal ammonia transport. PMID:23324176

  19. 17 beta-estradiol hydroxylation catalyzed by human cytochrome P450 1B1.

    PubMed Central

    Hayes, C L; Spink, D C; Spink, B C; Cao, J Q; Walker, N J; Sutter, T R

    1996-01-01

    The 4-hydroxy metabolite of 17 beta-estradiol (E2) has been implicated in the carcinogenicity of this hormone. Previous studies showed that aryl hydrocarbon-receptor agonists induced a cytochrome P450 that catalyzed the 4-hydroxylation of E2. This activity was associated with human P450 1B1. To determine the relationship of the human P450 1B1 gene product and E2 4-hydroxylation, the protein was expressed in Saccharomyces cerevisiae. Microsomes from the transformed yeast catalyzed the 4- and 2-hydroxylation of E2 with Km values of 0.71 and 0.78 microM and turnover numbers of 1.39 and 0.27 nmol product min-1.nmol P450-1, respectively. Treatment of MCF-7 human breast cancer cells with the aryl hydrocarbon-receptor ligand indolo[3,2-b]carbazole resulted in a concentration-dependent increase in P450 1B1 and P450 1A1 mRNA levels, and caused increased rates of 2-, 4-, 6 alpha-, and 15 alpha-hydroxylation of E2. At an E2 concentration of 10 nM, the increased rates of 2- and 4-hydroxylation were approximately equal, emphasizing the significance of the low Km P450 1B1-component of E2 metabolism. These studies demonstrate that human P450 1B1 is a catalytically efficient E2 4-hydroxylase that is likely to participate in endocrine regulation and the toxicity of estrogens. Images Fig. 1 PMID:8790407

  20. 17 beta-estradiol hydroxylation catalyzed by human cytochrome P450 1B1.

    PubMed

    Hayes, C L; Spink, D C; Spink, B C; Cao, J Q; Walker, N J; Sutter, T R

    1996-09-01

    The 4-hydroxy metabolite of 17 beta-estradiol (E2) has been implicated in the carcinogenicity of this hormone. Previous studies showed that aryl hydrocarbon-receptor agonists induced a cytochrome P450 that catalyzed the 4-hydroxylation of E2. This activity was associated with human P450 1B1. To determine the relationship of the human P450 1B1 gene product and E2 4-hydroxylation, the protein was expressed in Saccharomyces cerevisiae. Microsomes from the transformed yeast catalyzed the 4- and 2-hydroxylation of E2 with Km values of 0.71 and 0.78 microM and turnover numbers of 1.39 and 0.27 nmol product min-1.nmol P450-1, respectively. Treatment of MCF-7 human breast cancer cells with the aryl hydrocarbon-receptor ligand indolo[3,2-b]carbazole resulted in a concentration-dependent increase in P450 1B1 and P450 1A1 mRNA levels, and caused increased rates of 2-, 4-, 6 alpha-, and 15 alpha-hydroxylation of E2. At an E2 concentration of 10 nM, the increased rates of 2- and 4-hydroxylation were approximately equal, emphasizing the significance of the low Km P450 1B1-component of E2 metabolism. These studies demonstrate that human P450 1B1 is a catalytically efficient E2 4-hydroxylase that is likely to participate in endocrine regulation and the toxicity of estrogens. PMID:8790407

  1. IL-7 Dependence in Human B Lymphopoiesis Increases During Progression of Ontogeny from Cord Blood to Bone Marrow1

    PubMed Central

    Parrish, Yasmin Khan; Baez, Ineavely; Milford, Terry-Ann; Benitez, Abigail; Galloway, Nicholas; Rogerio, Jaqueline Willeman; Sahakian, Eva; Kagoda, Mercy; Huang, Grace; Hao, Qian-Lin; Sevilla, Yazmar; Barsky, Lora W.; Zielinska, Ewa; Price, Mary A.; Wall, Nathan R.; Dovat, Sinisa; Payne, Kimberly J.

    2009-01-01

    IL-7 is critical for B cell production in adult mice, however its role in human B lymphopoiesis is controversial. One challenge was the inability to differentiate human cord blood (CB) or adult bone marrow (BM) hematopoietic stem cells (HSCs) without murine stroma. Here, we examine the role of IL-7 in human B cell development using a novel, human-only model based on co-culturing human HSCs on primary human BM stroma. In this model, IL-7 increases human B cell production by >60-fold from both CB and adult BM HSCs. IL-7-induced increases are dose-dependent and specific to CD19+ cells. STAT5 phosphorylation and expression of the Ki-67 proliferation antigen, indicate that IL-7 acts directly on CD19+ cells to increase proliferation at the CD34+ and CD34? pro-B cell stages. Without IL-7, HSCS in CB, but not BM give rise to a small but consistent population of CD19LO B lineage cells that express EBF and PAX-5 and respond to subsequent IL-7 stimulation. Flt3 ligand, but not thymic stromal-derived lymhopoietin (TSLP), was required for the IL-7-independent production of human B lineage cells. As compared to CB, adult BM shows a reduction of in vitro generative capacity that is progressively more profound in developmentally sequential populations, resulting in a ~50-fold reduction in IL-7-dependent B lineage generative capacity. These data provide evidence that IL-7 is essential for human B cell production from adult BM and that IL-7-induced expansion of the pro-B compartment is increasingly critical for human B cell production during the progression of ontogeny. PMID:19299724

  2. Caveolin-1 Phosphorylation in Human Squamous and Epidermoid Carcinoma Cells: Dependence on ErbB1 Expression and Src Activation

    E-print Network

    Bertics, Paul J.

    Caveolin-1 Phosphorylation in Human Squamous and Epidermoid Carcinoma Cells: Dependence on ErbB1 cells and several squamous carcinoma cell lines. In all cases, EGF treatment induced the tyrosine human carcinoma cell lines that express various ErbB family members, including A431 epidermoid carcinoma

  3. Comparison of promoter activities for efficient expression into human B cells and haematopoietic progenitors with adenovirus Ad5\\/F35

    Microsoft Academic Search

    Marie-Pierre Cayer; Mathieu Drouin; Serey-Phorn Sea; Audrey Forest; Serge Côté; Carl Simard; Lucie Boyer; Annie Jacques; Nicolas Pineault; Daniel Jung

    2007-01-01

    Adenoviral gene transfer into human B lymphocytes and haematopoietic progenitors would allow the characterization of their function on cellular growth, differentiation and survival. Efficient gene expression is however strongly dependent on the promoter used. In this study, we investigated the relative strength of various promoters by following and measuring the expression of the reporter gene EYFP in human peripheral B

  4. 1-2B7B: Monoclonal antibody reacting to the 120 kDa polypeptide component of human epidermal hemidesmosomes

    Microsoft Academic Search

    X.-M. Zhang; Y. Horiguchi; M. Ueda; T. Yoshiki; S. Imamura

    1991-01-01

    Immunohistochemical and immunochemical analyses were performed on a monoclonal antibody designated 1-2B7B which was derived from immunizing mice with human prostate epithelial tissue. The 1-2B7B antigen was expressed not only along the acinous basement membrane zone (BMZ) of the prostate and testis, but also along the BMZ of the epithelia of several other organs including the skin, oesophagus, urinary bladder,

  5. Age and gender affect DNMT3a and DNMT3b expression in human liver.

    PubMed

    Xiao, Yongmei; Word, Beverly; Starlard-Davenport, Atena; Haefele, Aaron; Lyn-Cook, Beverly D; Hammons, George

    2008-06-01

    DNA methylation is catalyzed by a family of DNA methyltransferases (DNMTs) including the maintenance enzyme DNMT 1 and de novo methyltransferases DNMT 3a and DNMT 3b. Elevated levels of DNMTs have been found in cancer cells and in several types of human tumors. A polymorphism found in DNMT3b has been associated with increased risk for several cancers. The factors influencing DNMT expression in human tissues have not been clearly determined. he present study examined TDNMT3a and DNMT3b levels in human liver tissue samples and compared the effect of ageing, cigarette smoking, and gender. DNMT3a and DNMT3b expression levels in the samples from older individuals (56-78 years, n = 28) were both significantly higher than those of the younger group (16-48 years, n = 27) (73.2 +/- 3.4 vs 8.3 +/- 2.8 and 56.1 +/- 1.9 vs 17.5 +/- 5.7, respectively; p < 0.05). Levels of DNMT3b in females were significantly higher than those in males (75.4 +/- 2.2 vs 16.3 +/- 4.7; p < 0.05); however, DNMT3a levels were similar for females and males (52.7 +/- 2.7 vs 48.4 +/- 2.0). Expression levels of DNMT3a and DNMT3b were similar in smokers and nonsmokers (58.1 +/- 3.5 vs 60.8 +/- 3.1 and 54.5 +/- 2.3 vs 48.3 +/- 1.8, respectively). Genotyping for DNMT3b (C-->T) variant in this sample pool showed a frequency distribution of CC (41%), CT (50%), and TT (9%). The findings from this study suggest that ageing and gender may be important factors influencing DNA methylation status. PMID:17929180

  6. Roles of progesterone receptor a and B isoforms during human endometrial decidualization.

    PubMed

    Kaya, Hatice S; Hantak, Alison M; Stubbs, Lisa J; Taylor, Robert N; Bagchi, Indrani C; Bagchi, Milan K

    2015-06-01

    Progesterone, acting through the progesterone receptors (PGRs), is one of the most critical regulators of endometrial differentiation, known as decidualization, which is a key step toward the establishment of pregnancy. Yet a long-standing unresolved issue in uterine biology is the precise roles played by the major PGR isoforms, PGR-A and PGR-B, during decidualization in the human. Our approach, expressing PGR-A and PGR-B individually after silencing endogenous PGRs in human endometrial stromal cells (HESCs), enabled the analysis of the roles of these isoforms separately as well as jointly. Chromatin immunoprecipitation-sequencing in combination with gene expression profiling revealed that PGR-B controls a substantially larger cistrome and transcriptome than PGR-A during HESC differentiation. Interestingly, PGR-B directly regulates the expression of PGR-A. De novo motif analysis indicated that, although the 2 isoforms bind to the same DNA sequence motif, there are both common and unique neighboring motifs where other transcription factors, such as FOSL1/2, JUN, C/EBP?, and STAT3, bind and dictate the transcriptional activities of these isoforms. We found that PGR-A and PGR-B regulate overlapping as well as distinct sets of genes, many of which are known to be critical for decidualization and establishment of pregnancy. When PGR-A and PGR-B were coexpressed during HESC differentiation, PGR-B played a predominant role, although both isoforms influenced each other's transcriptional activity. This study revealed the gene networks that operate downstream of each PGR isoform to mediate critical functions, such as regulation of the cell cycle, angiogenesis, lysosomal activation, insulin receptor signaling, and apoptosis, during decidualization in the human. PMID:25875046

  7. Nickel induces oxidative burst, NF-?B activation and interleukin-8 production in human neutrophils.

    PubMed

    Freitas, Marisa; Gomes, Ana; Porto, Graça; Fernandes, Eduarda

    2010-11-01

    Many lines of evidence have suggested that oxidative stress and inflammation play a pivotal role in the toxicity of nickel salts. Considering that neutrophils are active participants in inflammatory processes, namely by producing high amounts of reactive oxygen species, the aim of the present study was to evaluate the putative activation of human neutrophils' oxidative burst by nickel. Subsequently, the influence of nickel in the pathways leading to NADPH oxidation in neutrophils was evaluated by measuring protein kinase C (PKC) activation. The effects of nickel on neutrophils' nuclear factor ?B (NF-?B) activation and on the production of the proinflammatory mediators interleukin (IL)-1?, IL-6, IL-8 and tumour necrosis factor ? were also evaluated. The results obtained showed that nickel, at concentrations that may be attained in vivo, stimulates the production of superoxide radical (O(2)(·-)), hydrogen peroxide (H(2)O(2)), and hypochlorous acid (HOCl) in human neutrophils in vitro, via activation of PKC. In addition, nickel was shown to activate NF-?B and to induce the production of IL-8 in these cells. These observations indicate that the sustained activation of human neutrophils by nickel may contribute for the long-term adverse effects on human health mediated by this metal. PMID:20632048

  8. Human\\/mouse radiation chimera generated from PBMC of B chronic lymphocytic leukemia patients with autoimmune hemolytic anemia produce anti-human red cell antibodies

    Microsoft Academic Search

    H Marcus; A Shimoni; D Ergas; A Canaan; B Dekel; D Ben-David; M David; E Sigler; Y Reisner; A Berrebi

    1997-01-01

    Previous studies performed in our laboratory have shown that B-CLL cells are involved in the production of anti-red cell auto-antibodies, providing a possible mechanism for the auto-immune hemolytic anemia occurring during the course of B-CLL. In order to confirm this hypothesis, we attempted to transfer human B-CLL with AIHA to immunodeficient mice. Peripheral blood mononuclear cells (PBMC) from 11 B-CLL

  9. Identification and Characterization of a Peptide That Specifically Binds the Human, Broadly Neutralizing Anti-Human Immunodeficiency Virus Type 1 Antibody b12

    PubMed Central

    Zwick, Michael B.; Bonnycastle, Lori L. C.; Menendez, Alfredo; Irving, Melita B.; Barbas, Carlos F.; Parren, Paul W. H. I.; Burton, Dennis R.; Scott, Jamie K.

    2001-01-01

    Human monoclonal antibody (MAb) b12 recognizes a conformational epitope that overlaps the CD-4-binding site of the human immunodeficiency virus type 1 (HIV-1) envelope. MAb b12 neutralizes a broad range of HIV-1 primary isolates and protects against primary virus challenge in animal models. We report here the discovery and characterization of B2.1, a peptide that binds specifically to MAb b12. B2.1 was selected from a phage-displayed peptide library by using immunoglobulin G1 b12 as the selecting agent. The peptide is a homodimer whose activity depends on an intact disulfide bridge joining its polypeptide chains. Competition studies with gp120 indicate that B2.1 occupies the b12 antigen-binding site. The affinity of b12 for B2.1 depends on the form in which the peptide is presented; b12 binds best to the homodimer as a recombinant polypeptide fused to the phage coat. Originally, b12 was isolated from a phage-displayed Fab library constructed from the bone marrow of an HIV-1-infected donor. The B2.1 peptide is highly specific for b12 since it selected only phage bearing b12 Fab from this large and diverse antibody library. PMID:11413337

  10. NF-?B Activation in Human Dental Pulp Stem Cells by TNF and LPS

    PubMed Central

    Chang, J.; Zhang, C.; Tani-Ishii, N.; Shi, S.; Wang, C.-Y.

    2006-01-01

    Post-natal human dental pulp stem cells (DPSCs) represent a unique precursor population in the dental pulp, which has multipotential and can regenerate a dentin/pulp-like structure. Because the dental pulp is frequently infected by oral bacteria due to dental decay, in this study, we examined whether lipopolysaccharide (LPS) and tumor necrosis factor (TNF) activated the immunologic transcription factor nuclear factor kappa B (NF-?B) in DPSCs. We found that both TNF and LPS activated the I-kappa B kinase complex (IKK) in DPSCs to induce the phosphorylation and degradation of I?B?, resulting in the nuclear translocation of NF-?B. Consistently, both TNF and LPS rapidly induced the expression of the NF-?B-dependent gene interleukin-8 (IL-8). However, unlike in monocytes, we found that LPS could not induce the phosphorylation of the NF-?B active subunit p65 in DPSCs. In summary, our studies suggest that DPSCs may be involved in immune responses during pulpal infection through activating NF-?B. PMID:16246929

  11. Effect of increased HoxB4 on human megakaryocytic development.

    PubMed

    Zhong, Yiming; Sullenbarger, Brent; Lasky, Larry C

    2010-07-30

    In order to produce clinically useful quantities of platelets ex vivo we may need to firstly enhance early self-renewal of hematopoietic stem cells (HSCs) and/or megakaryocyte (Mk) progenitors. The homeodomain transcription factor HoxB4 has been shown to be an important regulator of stem cell renewal and hematopoiesis; however, its effect on megakaryopoiesis is unclear. In this study, we investigated the effect of HoxB4 overexpression or RNA silencing on megakaryocytic development in the human TF1 progenitor cell line; we then used recombinant tPTD-HoxB4 fusion protein to study the effect of exogenous HoxB4 on megakaryocytic development of human CD34 positively-selected cord blood cells. We found that ectopic HoxB4 in TF1 cells increased the antigen expression of CD61and CD41a, increased the gene expression of thrombopoietin receptor (TpoR), Scl-1, Cyclin D1, Fog-1 and Fli-1 while it decreased c-Myb expression. HoxB4 RNA silencing in TF1 cells decreased the expression of CD61 and CD41a and decreased Fli-1 expression while it increased the expression of c-Myb. Recombinant tPTD-HoxB4 fusion protein increased the percentages and absolute numbers of CD41a and CD61 positive cells during megakaryocytic differentiation of CD34 positively-selected cord blood cells and increased the numbers of colony-forming unit-megakaryocyte (CFU-Mk). Adding tPTD-HoxB4 fusion protein increased the gene expression of TpoR, Cyclin D1, Fog-1 and Fli-1 while it inhibited c-Myb expression. Our data suggest that increased HoxB4 enhanced early megakaryocytic development in human TF1 cells and CD34 positively-selected cord blood cells primarily by upregulating TpoR and Fli-1 expression and downregulating c-Myb expression. Increasing HoxB4 expression or adding recombinant HoxB4 protein might be a way to expand Mks for the production of platelets for use in transfusion medicine. PMID:20599537

  12. Mapping of the human centromere protein B gene (CENPB) to chromosome 20p13 by fluorescence in situ hybridization

    SciTech Connect

    Naohiko, Seki [Kazusa DNA Research Institute, Chiba (Japan); Toshiyuki, Saito; Katsumi, Kitagawa [National Institute of Radiological Science, Chiba (Japan)] [and others

    1994-11-01

    Using a panel of rodent/human hybrid DNA, the two centromere protein genes (CENPB and CENPC) have been mapped to human chromosomes 20 and 12, respectively. We report here a regional assignment of the CENP-B-coding gene (CENPB) on human chromosomes using fluorescene in situ hybridization techniques.

  13. Inhibition of Interleukin-2 Gene Expression by Human Herpesvirus 6B U54 Tegument Protein

    PubMed Central

    Iampietro, Mathieu; Morissette, Guillaume; Gravel, Annie

    2014-01-01

    ABSTRACT Human herpesvirus 6B (HHV-6B) is a ubiquitous pathogen causing lifelong infections in approximately 95% of humans worldwide. To persist within its host, HHV-6B has developed several immune evasion mechanisms, such as latency, during which minimal proteins are expressed, and the ability to disturb innate and adaptive immune responses. The primary cellular targets of HHV-6B are CD4+ T cells. Previous studies by Flamand et al. (L. Flamand, J. Gosselin, I. Stefanescu, D. Ablashi, and J. Menezes, Blood 85:1263–1271, 1995) reported on the capacity of HHV-6A as well as UV-irradiated HHV-6A to inhibit interleukin-2 (IL-2) synthesis in CD4+ lymphocytes, suggesting that viral structural components could be responsible for this effect. In the present study, we identified the HHV-6B U54 tegument protein (U54) as being capable of inhibiting IL-2 expression. U54 binds the calcineurin (CaN) phosphatase enzyme, causing improper dephosphorylation and nuclear translocation of NFAT (nuclear factor of activated T cells) proteins, resulting in suboptimal IL-2 gene transcription. The U54 GISIT motif (amino acids 293 to 297), analogous to the NFAT PXIXIT motif, contributed to the inhibition of NFAT activation. IMPORTANCE Human herpesvirus 6A (HHV-6A) and HHV-6B are associated with an increasing number of pathologies. These viruses have developed strategies to avoid the immune response allowing them to persist in the host. Several studies have illustrated mechanisms by which HHV-6A and HHV-6B are able to disrupt host defenses (reviewed in L. Dagna, J. C. Pritchett, and P. Lusso, Future Virol. 8:273–287, 2013, doi:10.2217/fvl.13.7). Previous work informed us that HHV-6A is able to suppress synthesis of interleukin-2 (IL-2), a key immune growth factor essential for adequate T lymphocyte proliferation and expansion. We obtained evidence that HHV-6B also inhibits IL-2 gene expression and identified the mechanisms by which it does so. Our work led us to the identification of U54, a virion-associated tegument protein, as being responsible for suppression of IL-2. Consequently, we have identified HHV-6B U54 protein as playing a role in immune evasion. These results further contribute to our understanding of HHV-6 interactions with its human host and the efforts deployed to ensure its long-term persistence. PMID:25122797

  14. A Coxsackie B4 virus isolated in Yunnan in 2009 is a recombinant.

    PubMed

    Zhu, Yanju; Pan, Yue; Chen, Junying; Liu, Jiansheng; Chen, Wei; Ma, Shaohui

    2015-06-01

    Coxsackievirus B4 is a member of the species Enterovirus B in the Enterovirus genus of the Picornaviridae family. So far, there are only seven complete genome sequences of CVB4 published in GenBank database. In the study, the complete genome analysis of a Coxsackievirus B4 strain A155/YN/CHN/2009 isolated from a child with aseptic meningitis in Yunnan Province was performed. It had 85.1 and 83.3 % nucleotide similarity with prototype strain J.V.B Benschoten in the VP1 region and the complete genome, respectively. Phylogenetic analysis of VP1 region showed that A155/YN/CHN/2009 belongs to Genotype V circulating only in mainland of China. The results of Simplot and Bootscanning analysis implicated that A155 has recombined with other HEV-B viruses. PMID:25725901

  15. Conservation of Amino Acids in Human Rhinovirus 3C Protease Correlates with Broad-Spectrum Antiviral Activity of Rupintrivir, a Novel Human Rhinovirus 3C Protease Inhibitor

    Microsoft Academic Search

    S. L. Binford; F. Maldonado; M. A. Brothers; P. T. Weady; L. S. Zalman; J. W. Meador; D. A. Matthews; A. K. Patick

    2005-01-01

    The picornavirus 3C protease is required for the majority of proteolytic cleavages that occur during the viral life cycle. Comparisons of published amino acid sequences from 6 human rhinoviruses (HRV) and 20 human enteroviruses (HEV) show considerable variability in the 3C protease-coding region but strict conservation of the catalytic triad residues. Rupintrivir (formerly AG7088) is an irreversible inhibitor of HRV

  16. c-Src tyrosine kinase co-associates with and phosphorylates signal transducer and activator of transcription 5b which mediates the proliferation of normal human B lymphocytes.

    PubMed

    Cayer, M-P; Proulx, M; Ma, X-Z; Sakac, D; Gigučre, J-F; Drouin, M; Néron, S; Branch, D R; Jung, D

    2009-06-01

    c-Src is the normal human cellular protein homologue of the viral oncogene v-src. c-Src activity was reported recently to increase in CD40-activated human B lymphocytes, suggesting its involvement in proliferation. To elucidate the exact role of c-Src in this process, we investigated the effects of c-Src over-expression on normal B lymphocyte growth. B lymphocytes purified from human peripheral blood were infected with Ad5/F35 vector encoding either a constitutively active c-Src (c-Src/dominant-positive) or a dominant-negative c-Src (c-Src/DN). Little variation of B lymphocytes expansion could be observed between control enhanced yellow fluorescent protein and c-Src/dominant-positive-infected cells. In contrast, over-expression of c-Src/DN results in a 40% inhibition of B lymphocyte expansion. These results suggest that DN c-Src may compete with endogenous c-Src, resulting in partial inhibition of a transcriptional pathway involved in B lymphocyte proliferation. We demonstrate further that c-Src can phosphorylate signal transducer and activator of transcription 5b (STAT5b) on tyrosine 699 and that c-Src and STAT5b co-associate during B lymphocyte proliferation. These results confirm an important role for c-Src in the expansion of normal human B lymphocytes in vitro, in which c-Src may regulate STAT5b in the intracellular signalling pathway important for the proliferation of normal human B lymphocytes. PMID:19438593

  17. Enterovirus 71-induced autophagy increases viral replication and pathogenesis in a suckling mouse model

    PubMed Central

    2014-01-01

    Background We previously reported that Enterovirus 71 (EV71) infection activates autophagy, which promotes viral replication both in vitro and in vivo. In the present study we further investigated whether EV71 infection of neuronal SK-N-SH cells induces an autophagic flux. Furthermore, the effects of autophagy on EV71-related pathogenesis and viral load were evaluated after intracranial inoculation of mouse-adapted EV71 (MP4 strain) into 6-day-old ICR suckling mice. Results We demonstrated that in EV71-infected SK-N-SH cells, EV71 structural protein VP1 and nonstructural protein 2C co-localized with LC3 and mannose-6-phosphate receptor (MPR, endosome marker) proteins by immunofluorescence staining, indicating amphisome formation. Together with amphisome formation, EV71 induced an autophagic flux, which could be blocked by NH4Cl (inhibitor of acidification) and vinblastine (inhibitor of fusion), as demonstrated by Western blotting. Suckling mice intracranially inoculated with EV71 showed EV71 VP1 protein expression (representing EV71 infection) in the cerebellum, medulla, and pons by immunohistochemical staining. Accompanied with these infected brain tissues, increased expression of LC3-II protein as well as formation of LC3 aggregates, autophagosomes and amphisomes were detected. Amphisome formation, which was confirmed by colocalization of EV71-VP1 protein or LC3 puncta and the endosome marker protein MPR. Thus, EV71-infected suckling mice (similar to EV71-infected SK-N-SH cells) also show an autophagic flux. The physiopathological parameters of EV71-MP4 infected mice, including body weight loss, disease symptoms, and mortality were increased compared to those of the uninfected mice. We further blocked EV71-induced autophagy with the inhibitor 3-methyladenine (3-MA), which attenuated the disease symptoms and decreased the viral load in the brain tissues of the infected mice. Conclusions In this study, we reveal that EV71 infection of suckling mice induces an amphisome formation accompanied with the autophagic flux in the brain tissues. Autophagy induced by EV71 promotes viral replication and EV71-related pathogenesis. PMID:25139436

  18. Development of two potential diagnostic monoclonal antibodies against human cytomegalovirus glycoprotein B.

    PubMed

    Jun, Fan; Minhuan, Li; Yadan, Ma; Yaping, Huang; Hanying, Liang; Jianhua, Hu; Hangping, Yao; Weihang, Ma

    2012-07-01

    Human cytomegalovirus glycoprotein B (gB) represents a target for diagnosis and treatment in view of the role it plays in virus entry and spread. Nevertheless, to our knowledge, rare detection of a gB antigen has been reported in transplant patients and limited information is available about diagnostic gB monoclonal antibodies (mAbs). Our aim was to develop gB mAbs with diagnostic potential. Hydrophilic gB peptides (ST: amino acids 27-40, SH: amino acids 81-94) of favorable immunogenicity were synthesized and used to immunize BALB/c mice. Two mAbs, named ZJU-FH6 and ZJU-FE6, were generated by the hybridoma technique and limited serial dilution and then characterized by indirect ELISA, Western blotting, immunoprecipitation, and immunohistochemical staining. The mAbs displayed high titers of specific binding affinities for the ST and SH synthetic peptides at an mAb dilution of 1:60,000 and 1:240,000, respectively. Western blotting and immunoprecipitation indicated that these mAbs recognized both denatured and native gB of the Towne and AD169 strains. The mAbs, when used as the primary antibody, showed positive staining in cells infected with both Towne and AD169 strains. The mAbs were then tested on patients submitted to allogeneic hematopoietic stem cell transplantation. The gB antigen positivity rates of the patients tested using ZJU-FH6 and ZJU-FE6 were 62.0 and 63.0%, respectively. The gB antigen showed a significant correlation with the level of pp65 antigen in peripheral blood leukocytes. In conclusion, two potential diagnostic gB mAbs were developed and were shown to be capable of recognizing gB in peripheral blood leukocytes in a reliable manner. PMID:22618859

  19. Indole3-carbinol enhances ultraviolet B-induced apoptosis by sensitizing human melanoma cells

    Microsoft Academic Search

    D.-S. Kim; Y.-M. Jeong; S.-I. Moon; S.-Y. Kim; S.-B. Kwon; E.-S. Park; S.-W. Youn; K.-C. Park

    2006-01-01

    .  Indole-3-carbinol (I3C) has been found to act against several types of cancer, while ultraviolet B (UVB) is known to induce\\u000a the apoptosis of human melanoma cells. Here, we investigated whether I3C can sensitize G361 human melanoma cells to UVB-induced\\u000a apoptosis. We examined the effects of combined I3C and UVB (I3C\\/UVB) at various dosages. I3C (200 ?M)\\/UVB (50 mJ\\/cm2) synergistically reduced melanoma cell

  20. B Cell Differentiation and Immunoregulatory T Cell Function in Human Cord Blood Lymphocytes

    PubMed Central

    Tosato, Giovanna; Magrath, Ian T.; Koski, Irma R.; Dooley, Nancy J.; Blaese, R. Michael

    1980-01-01

    The functional maturity of T and B lymphocyte populations from human newborns was evaluated using a reverse hemolytic plaque assay to detect immunoglobulin-secreting cells generated in in vitro cultures stimulated with pokeweed mitogen (PWM), a T cell-dependent polyclonal activator, and the Epstein-Barr virus (EBV), a T cell-independent B cell activator. Cord blood lymphocytes failed to produce immunoglobulin in response to PWM, but did respond with immunoglobulin synthesis to stimulation with EBV. Co-culture experiments demonstrated that cord blood T cells would inhibit immunoglobulin production by adult cells stimulated with PWM, but not with EBV. Cord blood T cells did suppress immunoglobulin production by cord blood B cells when stimulated with a mixture of EBV and PWM, indicating that cord blood, in contrast to adult blood, contains a population of suppressor T cell precursors that are easily activated by PWM. Irradiation of the cord blood T cells with 2,000 rad eliminated the suppressor activity and revealed normal helper function for immunoglobulin (Ig) G, A, and M when these T cells were co-cultured with adult B cells. Cord blood B cells co-cultured with adult T cells or irradiated cord blood T cells did produce immunoglobulin in response to PWM, but the response was significantly lower than that of adult B cells, and only IgM was produced in these cultures. These studies demonstrate that both the T and B cells of the human newborn have significant functional differences compared with the functions of T and B lymphocyte populations in adults. PMID:6249850

  1. Nuclear factor kappa B is required for the production of infectious human herpesvirus 8 virions

    PubMed Central

    Blattman, Negin N.; Lagunoff, Michael; Blattman, Joseph N.; Corey, Lawrence

    2014-01-01

    Human herpesvirus 8 (HHV8) infection leads to potent activation of nuclear factor kappa B (NF?B) in primary and transformed cells. We used recombinant HHV8 (rKSHV.219) expressing green fluorescent protein under the constitutive cellular promoter elongation factor 2? and red fluorescent protein under an early HHV8 lytic gene promoter T1.1 to monitor replication during infection of human foreskin fibroblasts (HF), noting changes in NF?B activity. In primary HF, NF?B levels do not affect the ability of HHV8 to establish infection or maintain latency. Furthermore, there was no effect on the percent of cells undergoing reactivation from latency, and there were similar numbers of released and cell-associated HHV8 viral particles following reactivation in the presence of inhibitors. Reactivation of HHV8 in latently infected HF in the presence of NF?B inhibitors resulted in production of viral particles that did not efficiently establish infection, due to deficiencies in binding and/or entry into normally permissive cells. Exogenous expression of glycoprotein M, an envelope protein involved in viral binding and entry, was able to partially overcome the deficiency induced by NF?B inhibitors. Our data indicate that in primary cells, NF?B is not required for infection, establishment of latency, or entry into the lytic cycle, but is required for the expression of virion associated genes involved in the initial steps of virion infectivity. These studies suggest that strategies to inhibit NF?B may prevent HHV8 spread and should be considered as a potential therapeutic target for preventing HHV8 associated diseases. PMID:24795700

  2. Salvianolic acid B suppresses maturation of human monocyte-derived dendritic cells by activating PPAR?

    PubMed Central

    Sun, Aijun; Liu, Hongying; Wang, Shijun; Shi, Dazhuo; Xu, Lei; Cheng, Yong; Wang, Keqiang; Chen, Keji; Zou, Yunzeng; Ge, Junbo

    2011-01-01

    BACKGROUND AND PURPOSE Salvianolic acid B (Sal B), a water-soluble antioxidant derived from a Chinese medicinal herb, is known to be effective in the prevention of atherosclerosis. Here, we tested the hypothesis that the anti-atherosclerotic effect of Sal B might be mediated by suppressing maturation of human monocyte-derived dendritic cells (h-monDC). EXPERIMENTAL APPROACH h-monDC were derived by incubating purified human monocytes with GM-CSF and IL-4. h-monDC were pre-incubated with or without Sal B and stimulated by oxidized low-density lipoprotein (ox-LDL) in the presence or absence of PPAR? siRNA. Expression of h-monDC membrane molecules (CD40, CD86, CD1a, HLA-DR) were analysed by FACS, cytokines were measured by elisa and the TLR4-associated signalling pathway was determined by Western blotting. KEY RESULTS Ox-LDL promoted h-monDC maturation, stimulated CD40, CD86, CD1a, HLA-DR expression and IL-12, IL-10, TNF-? production; and up-regulated TLR4 signalling. These effects were inhibited by Sal B. Sal B also triggered PPAR? activation and promoted PPAR? nuclear translocation, attenuated ox-LDL-induced up-regulation of TLR4 and myeloid differentiation primary-response protein 88 and inhibited the downstream p38-MAPK signalling cascade. Knocking down PPAR? with the corresponding siRNA blocked these effects of Sal B. CONCLUSIONS AND IMPLICATIONS Our data suggested that Sal B effectively suppressed maturation of h-monDC induced by ox-LDL through PPAR? activation. PMID:21649636

  3. Evaluation of the threshold of stability for the human spine Martin L. Tanaka a,b,d,, Maury A. Nussbaum b,c

    E-print Network

    Ross, Shane

    Evaluation of the threshold of stability for the human spine Martin L. Tanaka a,b,d,Ă?, Maury A of the spine, paraspinal ligaments, core musculature, and neuro- muscular control system to maintain an upright

  4. Identification of a Neutralizing Epitope within Antigenic Domain 5 of Glycoprotein B of Human Cytomegalovirus

    PubMed Central

    Wiegers, Anna-Katharina; Sticht, Heinrich; Winkler, Thomas H.; Britt, William J.

    2014-01-01

    ABSTRACT Human cytomegalovirus (HCMV) is an important, ubiquitous pathogen that causes severe clinical disease in immunocompromised individuals, such as organ transplant recipients and infants infected in utero. The envelope glycoprotein B (gB) of HCMV is a major antigen for the induction of virus-neutralizing antibodies. We have begun to define target structures within gB that are recognized by virus-neutralizing antibodies. Antigenic domain 5 (AD-5) of gB has been identified as an important target for neutralizing antibodies in studies using human monoclonal antibodies (MAbs). Anti-AD-5 MAbs share a target site on gB, despite originating from different, healthy, HCMV-infected donors. Mutational analysis of AD-5 identified tyrosine 280 in combination with other surface-exposed residues (the YNND epitope) as critical for antibody binding. The YNND epitope is strictly conserved among different HCMV strains. Recombinant viruses carrying YNND mutations in AD-5 were resistant to virus-neutralizing MAbs. Competition enzyme-linked immunosorbent assays (ELISAs) with human HCMV-convalescent-phase sera from unselected donors confirmed the conserved antibody response for the YNND epitope in HCMV-infected individuals and, because a significant fraction of the gB AD-5 response was directed against the YNND epitope, further argued that this epitope is a major target of anti-AD-5 antibody responses. In addition, affinity-purified polyclonal anti-AD-5 antibodies prepared from individual sera showed reactivity to AD-5 and neutralization activity toward gB mutant viruses that were similar to those of AD-5-specific MAbs. Taken together, our data indicate that the YNND epitope represents an important target for anti-gB antibody responses as well as for anti-AD-5 virus-neutralizing antibodies. IMPORTANCE HCMV is a major global health concern, and a vaccine to prevent HCMV disease is a widely recognized medical need. Glycoprotein B of HCMV is an important target for neutralizing antibodies and hence an interesting molecule for intervention strategies, e.g., vaccination. Mapping the target structures of neutralizing antibodies induced by naturally occurring HCMV infection can aid in defining the properties required for a protective capacity of vaccine antigens. The data presented here extend our knowledge of neutralizing epitopes within gB to include AD-5. Collectively, our data will contribute to optimal vaccine design and development of antibody-based therapies. PMID:25320309

  5. Synthesis and Secretion of Procathepsin B and Cystatin C by Human Bronchial Epithelial Cells in Vitro: Modulation of Cathepsin B Activity by Neutrophil Elastase

    Microsoft Academic Search

    D. Burnett; M. Abrahamson; J. L. Devalia; R. J. Sapsford; R. J. Davies; D. J. Buttle

    1995-01-01

    Procathepsin B and cystatin C are found in human lung secretions. We investigated the capacity of human bronchial epithelial cells to synthesize and secrete these proteins. Immunoprecipitation of [35S]methionine-labeled proteins from cultured bronchial epithelial cell lysates, followed by denaturing gel electrophoresis and autoradiography, showed the presence of newly synthesized procathepsin B of Mr 42,000; no mature form was detected. Cathepsin

  6. Purification, crystallization and preliminary X-ray crystallographic analysis of human CCG1-interacting factor B.

    PubMed

    Padmanabhan, B; Kuzuhara, T; Mizuno, H; Horikoshi, M

    2000-11-01

    A novel human factor CIB (CCG1-interacting factor B) has been isolated using the yeast two-hybrid system. The 22 kDa CIB protein has been expressed in Escherichia coli, purified to homogeneity and crystallized in a form suitable for crystallographic studies. The protein was crystallized in the orthogonal space group P2(1)2(1)2(1), with unit-cell parameters a = 43.60 (2), b = 44.45 (1), c = 110.70 (5) A and one molecule in the asymmetric unit. The crystal diffracted beyond 2.2 A resolution using synchrotron radiation. PMID:11053859

  7. Human immunodeficiency virus type 1 Nef protein inhibits NF-kappa B induction in human T cells.

    PubMed Central

    Niederman, T M; Garcia, J V; Hastings, W R; Luria, S; Ratner, L

    1992-01-01

    Human immunodeficiency virus type 1 (HIV-1) can establish a persistent and latent infection in CD4+ T lymphocytes (W. C. Greene, N. Engl. J. Med. 324:308-317, 1991; S. M. Schnittman, M. C. Psallidopoulos, H. C. Lane, L. Thompson, M. Baseler, F. Massari, C. H. Fox, N. P. Salzman, and A. S. Fauci, Science 245:305-308, 1989). Production of HIV-1 from latently infected cells requires host cell activation by T-cell mitogens (T. Folks, D. M. Powell, M. M. Lightfoote, S. Benn, M. A. Martin, and A. S. Fauci, Science 231:600-602, 1986; D. Zagury, J. Bernard, R. Leonard, R. Cheynier, M. Feldman, P. S. Sarin, and R. C. Gallo, Science 231:850-853, 1986). This activation is mediated by the host transcription factor NF-kappa B [G. Nabel and D. Baltimore, Nature (London) 326:711-717, 1987]. We report here that the HIV-1-encoded Nef protein inhibits the induction of NF-kappa B DNA-binding activity by T-cell mitogens. However, Nef does not affect the DNA-binding activity of other transcription factors implicated in HIV-1 regulation, including SP-1, USF, URS, and NF-AT. Additionally, Nef inhibits the induction of HIV-1- and interleukin 2-directed gene expression, and the effect on HIV-1 transcription depends on an intact NF-kappa B-binding site. These results indicate that defective recruitment of NF-kappa B may underlie Nef's negative transcriptional effects on the HIV-1 and interleukin 2 promoters. Further evidence suggests that Nef inhibits NF-kappa B induction by interfering with a signal derived from the T-cell receptor complex. Images PMID:1527859

  8. Diversification of the ADH1B Gene during Expansion of Modern Humans

    PubMed Central

    Li, Hui; Gu, Sheng; Han, Yi; Xu, Zhi; Pakstis, Andrew J.; Jin, Li; Kidd, Judith R.; Kidd, Kenneth K.

    2013-01-01

    A variant allele, ADH1B*48His, also known as ADH1B*2, at the human Alcohol Dehydrogenase 1B gene (ADH1B) is strongly associated with alcoholism in some populations and has an unusual geographic distribution. Strong evidence implies selection has increased the frequency of this allele in some East Asian populations but does not fully explain its geographic pattern. We have studied haplotypes of ten single nucleotide polymorphisms (SNPs) and two short tandem repeat polymorphisms (STRPs) in the ADH1B region in 2,206 individuals from a worldwide set of populations. These SNPs and STRPs define nine common haplogroups most of which have distinct geographic patterns. The haplogroups H5 and H6, both with the derived ADH1B*48His allele, appear restricted to the Middle East and East Asia, respectively. The positively selected H7 is derived from H6 by a new regulatory region variant defining SNP rs3811801 restricted to East Asia. Age estimates of the haplogroups based on the STRPs also agree with the time of the migration events estimated by other studies. H7 is estimated to have expanded recently, around 2,800 years ago, and ancient DNA samples from North China confirm its presence about that time. The dating of the H7 expansion may help understand the selective force on the ADH1B gene. PMID:21592108

  9. Microcystins Induces Vascular Inflammation in Human Umbilical Vein Endothelial Cells via Activation of NF-?B

    PubMed Central

    Shi, Jun; Zhou, Jie; Zhang, Min

    2015-01-01

    Microcystins (MCs) produced by toxic cyanobacteria cause serious water pollution and public health hazard to humans and animals. However, direct molecular mechanisms of MC-LR in vascular endothelial cells (ECs) have not been understood yet. In this study, we investigated whether MC-LR induces vascular inflammatory process in cultured human umbilical vein endothelial cells (HUVECs). Our data demonstrated that MC-LR decreased HUVECs proliferation and tube formation and enhanced apoptosis. MC-LR also induced intracellular reactive oxygen species formation (ROS) in HUVECs. The MC-LR directly stimulated phosphorylation of NF-?B. Furthermore, MC-LR also increased cell adhesion molecules (ICAM-1 and VCAM-1) expression in HUVECs. Taken together, the present data suggested that MC-LR induced vascular inflammatory process, which may be closely related to the oxidative stress, NF-?B activation, and cell adhesion molecules expression in HUVECs. Our findings may highlight that MC-LR causes potential damage to blood vessels.

  10. Vitamin B12 and hepatitis C: Molecular biology and human pathology

    PubMed Central

    Lott, William B.; Takyar, Seyedtaghi S.; Tuppen, Joseph; Crawford, Darrell H. G.; Harrison, Michael; Sloots, Theo P.; Gowans, Eric J.

    2001-01-01

    Cobalamins are stored in high concentrations in the human liver and thus are available to participate in the regulation of hepatotropic virus functions. We show that cyanocobalamin (vitamin B12) inhibited the HCV internal ribosome entry site (IRES)-dependent translation of a reporter gene in vitro in a dose-dependent manner without significantly affecting the cap-dependent mechanism. Vitamin B12 failed to inhibit translation by IRES elements from encephalomyocarditis virus (EMCV) or classical swine fever virus (CSFV). We also demonstrate a relationship between the total cobalamin concentration in human sera and HCV viral load (a measure of viral replication in the host). The mean viral load was two orders of magnitude greater when the serum cobalamin concentration was above 200 pM (P < 0.003), suggesting that the total cobalamin concentration in an HCV-infected liver is biologically significant in HCV replication. PMID:11296247

  11. Transformed human bronchial epithelial cells (beas-2b) alter the growth and morphology of normal human bronchial epithelial cells in vitro

    Microsoft Academic Search

    Craig D. Albright; Raymond T. Jones; Eric A. Hudson; Joseph A. Fontana; Benjamin F. Trump; James H. Resau

    1990-01-01

    Normal human bronchial epithelial cells (BE) and adenovirus-12 SV40 hybrid virus transformed, non-tumorigenic human bronchial epithelial cells (BEAS-2B) were cultured for 7 days in a serum free hormone supplemented medium. BE cells after 3 days in culture were exposed to conditioned medium (CMt) from confluent BEAS-2B cells. By day 7, CMt-treated BE cells exhibited a lower colony forming efficiency (CFE),

  12. Molecular analysis of the prototype coxsackievirus B5 genome

    Microsoft Academic Search

    A. M. Lindberg; C. Polacek

    2000-01-01

    Summary.  ?To facilitate studies of the phylogenetic relationship between enteroviruses, in particular the prototype strain (Faulkner) of coxsackievirus B5 (CVB5F) and other CVB5 isolates and to facilitate studies of the interactions between CVB5F and the\\u000a target cell, the complete nucleotide sequence of the prototype has been determined. The complete sequence was collected from\\u000a three overlapping reverse transcription polymerase chain reaction (RT-PCR)

  13. Efficient gene transfer into normal human B lymphocytes with the chimeric adenoviral vector Ad5/F35.

    PubMed

    Jung, Daniel; Néron, Sonia; Drouin, Mathieu; Jacques, Annie

    2005-09-01

    The failure to efficiently introduce genes into normal cells such as human B lymphocytes limits the characterization of their function on cellular growth, differentiation and survival. Recent studies have shown that a new adenoviral vector Ad5/F35 can efficiently transduce human haematopoietic CD34+ progenitor cells. In this study, we compared the gene transfer efficiencies of the Ad5/F35 vector to that of the parental vector Ad5 in human B lymphocytes. Peripheral blood B cells obtained from healthy individuals were cultured in vitro using CD40-CD154 system. Normal B lymphocytes were infected with replication-defectives Ad5 and Ad5/F35, both containing the GFP reporter gene, and transduction efficiencies were monitored by flow cytometry. Ad5 was highly ineffective, infecting only about 5% of human B lymphocytes. In contrast, Ad5/F35 transduced up to 60% of human B lymphocytes and GFP expression could be detected for up to 5 days post infection. Importantly, physiology of B lymphocytes such as proliferation, viability and antibodies secretion were unaffected following Ad5/F35 transduction. Finally, we observed that memory B lymphocytes were more susceptible to Ad5/F35 infection than naďve B lymphocytes. Thus, our results demonstrate that the adenoviral vector Ad5/F35 is an efficient tool for the functional characterization of genes in B lymphopoiesis. PMID:16129448

  14. Genetic design of an optimized packaging cell line for gene vectors transducing human B cells

    Microsoft Academic Search

    E Hettich; A Janz; R Zeidler; D Pich; E Hellebrand; B Weissflog; A Moosmann; W Hammerschmidt

    2006-01-01

    Viral gene vectors often rely on packaging cell lines, which provide the necessary factors in trans for the formation of virus-like particles. Previously, we reported on a first-generation packaging cell line for gene vectors, which are based on the B-lymphotropic Epstein–Barr virus (EBV), a human ?-herpesvirus. This 293HEK-derived packaging cell line harbors a helper virus genome with a genetic modification

  15. Titration of human brain monoamine oxidase-A and-B by clorgyline andl-deprenil

    Microsoft Academic Search

    Christopher J. Fowler; Lars Oreland; Jan Marcusson; Bengt Winblad

    1980-01-01

    The interaction of clorgyline andl-deprenil with the-A and-B forms of human brain monoamine oxidase (MAO) has been studied. Both compounds inhibit cerebrocortical MAO in a manner consistent with a ‘suicide’ inactivation of the enzyme. The interaction of clorgyline with the-A form of the enzyme appears to take place almost entirely at specific binding sites, and the conditions required for this

  16. Enzyme plus light therapy to repair DNA damage in ultraviolet-B-irradiated human skin

    Microsoft Academic Search

    Helger Stege; Len Roza; Arie A. Vink; Markus Grewe; Thomas Ruzicka; Susanne Grether-Beck; Jean Krutmann

    2000-01-01

    Ultraviolet-B (UVB) (290-320 nm) radiation-induced cyclobutane pyrimidine dimers within the DNA of epidermal cells are detrimental to human health by causing mutations and immunosuppressive effects that presumably contribute to photocarcinogenesis. Conventional photoprotection by sunscreens is exclusively prophylactic in nature and of no value once DNA damage has occurred. In this paper, we have therefore assessed whether it is possible to

  17. Therapeutic Potential of Intracerebroventricular Replacement of Modified Human ?-Hexosaminidase B for GM2 Gangliosidosis

    Microsoft Academic Search

    Kazuhiko Matsuoka; Tomomi Tamura; Daisuke Tsuji; Yukie Dohzono; Keisuke Kitakaze; Kazuki Ohno; Seiji Saito; Hitoshi Sakuraba; Kohji Itoh

    2011-01-01

    To develop a novel enzyme replacement therapy for neurodegenerative Tay-Sachs disease (TSD) and Sandhoff disease (SD), which are caused by deficiency of ?-hexosaminidase (Hex) A, we designed a genetically engineered HEXB encoding the chimeric human ?-subunit containing partial amino acid sequence of the ?-subunit by structure-based homology modeling. We succeeded in producing the modified HexB by a Chinese hamster ovary

  18. Highly active antiretroviral therapy used to treat concurrent hepatitis B and human immunodeficiency virus infections

    Microsoft Academic Search

    Kazuki Nagai; Hiroo Hosaka; Shuichi Kubo; Noriko Nakamura; Masao Shinohara; Hiroko Nonaka

    1999-01-01

    :   We report a case of simultaneous infection with hepatitis B virus (HBV) and human immunodeficiency virus type 1 (HIV-1) in\\u000a a 26-year-old Japanese homosexual man. He was admitted to our hospital for acute hepatitis caused by HBV. At that time, HIV-1-antibody\\u000a (Ab) was not detected in his serum. After 6 months, he was readmitted to our hospital for further

  19. Generation and Characterization of High Affinity Humanized Fab Against Hepatitis B Surface Antigen

    Microsoft Academic Search

    Ashutosh Tiwari; Durgashree Dutta; Navin Khanna; Subrat K. Acharya; Subrata Sinha

    2009-01-01

    5S is a mouse monoclonal IgG1 that binds to the ‘a’ epitope of the Hepatitis B surface antigen (HBsAg) and tested positive\\u000a in an in vitro test for virus neutralization. We have earlier reported the generation of humanized single chain variable fragment\\u000a (scFv) from the same. In this article we report the generation of a recombinant Fab molecule by fusing

  20. Application of the Human Hepatitis B Virus Core Antigen from Transgenic Tobacco Plants for Serological Diagnosis

    Microsoft Academic Search

    Shinya Tsuda; Keiko Yoshioka; Takeshi Tanaka; Akiko Iwata; Akira Yoshikawa; Yuichiro Watanabe; Yoshimi Okada

    1998-01-01

    Background and objectives: The aim was to produce HBcAg from plants more cheaply than can be done by other currently available means, and to apply such antigen to immunoassay procedures for pretransfusion testing of donor blood. Materials and methods: Transgenic Nicotiana tabacum cv. SR-1 plants expressing the human hepatitis B virus (HBV) core antigen (HBcAg) gene were generated by Agrobacterium-mediated

  1. Overexpression of HSP70 prevents ultraviolet B-induced apoptosis of a human melanoma cell line

    Microsoft Academic Search

    Kyoung-Chan Park; Dong-Seok Kim; Hyun-Ok Choi; Kyu-Han Kim; Jin-Ho Chung; Hee-Chul Eun; Jae-Seon Lee; Jeong-Sun Seo

    2000-01-01

    \\u000a Abstract The heat shock response is a highly conserved reaction common to all cells and organisms. It has been reported that hyperthermic\\u000a treatment can induce the expression of the heat shock protein (HSP) and can protect cells from ultraviolet (UV) B radiation.\\u000a In this study, we evaluated the effects of induced HSP70 on resistance to UV radiation. G361 amelanotic human

  2. JARID1B expression in human melanoma and benign melanocytic skin lesions.

    PubMed

    Ku?bicki, Lukasz; Lange, Dariusz; Str?czy?ska-Niemiec, Anita; Chwirot, Barbara W

    2013-02-01

    It has been suggested that dynamically regulated expression of the JARID1B protein is required for the continuous growth of tumors and at the same time downregulated in melanoma. The majority of the data on a role of JARID1B in maintaining tumor growth has come from in-vitro and xenografting experiments, with only one immunohistochemical study involving human tissues. We compared JARID1B expression levels in human melanomas and benign nevi and analyzed patterns of spatial distributions of positive cells among different skin layers of the lesions. The expression of JARID1B was evaluated by immunohistochemistry in formalin-fixed paraffin-embedded samples of 30 nevi, 27 primary melanomas, four lymph node metastases, and one local recurrence of melanoma. Staining for JARID1B protein was stronger in melanomas compared with nevi. We also found a significant difference in the spatial distribution of positive cells in individual skin layers of nevi and melanomas. Staining of melanocytes located in granular and spinous layers of nevi was observed very rarely, whereas for melanomas, the mean percentage fractions of positive cells present in these layers exceeded the maximum values found for nevi. The spatial patterns and expression levels of JARID1B did not change significantly with melanoma progression and were similar for primary, metastatic, and recurrent melanomas. Contrary to earlier reports, this study shows enhanced expression of JARID1B by melanoma cells and indicates that such an enhancement may be an early event in the disease progression, is not correlated with melanoma invasiveness, and therefore may not be a suitable candidate as a prognostic marker. PMID:23262439

  3. B-Vitamin Levels in Human Milk among Different Lactation Stages and Areas in China

    PubMed Central

    Ren, Xiangnan; Yang, Zhenyu; Shao, Bing; Yin, Shi-an; Yang, Xiaoguang

    2015-01-01

    To determine the contents of B-vitamins in human milk in China, we analyzed 1778 human milk samples from the sample bank of the National High Technique R & D Program (863 Projects) which was a cross-sectional survey and covered 6419 human milk samples from healthy lactating mothers who were at different stages of lactation (0–330 days postpartum) in 11 provinces of China. The contents of free forms of six B-vitamins in these human milk samples were analyzed by using UPLC-MS/MS. The median concentrations of free form of 6 B-vitamins in colostrums, transitional milk, 15–180 d mature milk and 181-330 d mature milk were respectively as follows: thiamin 5.0 µg/L, 6.7 µg/L, 21.1 µg/L and 40.7 µg/L; riboflavin 29.3 µg/L, 40.6 µg/L, 33.6 µg/L and 29.6 µg/L; niacin 470.7 µg/L, 661.3 µg/L, 687.0 µg/L and 571.3 µg/L; vitamin B-6 4.6 µg/L, 16.1 µg/L, 62.7 µg/L and 80.7 µg/L; flavin adenine dinucleotide (FAD) 808.7 µg/L, 1162.8 µg/L, 1023.9 µg/L and 1057.2 µg/L; pantothenic acid 1770.9 µg/L, 2626.8 µg/L, 2213.0 µg/L and 1895.5 µg/L. The contents of 6 B-vitamins varied significantly among the different lactation stages and different areas (coastal area vs inland area, rural area vs urban area). The present study indicated that the concentrations of B-vitamins in colostrum were generally much lower than those in transitional milk and mature milk. Further studies are warranted for their roles and significance on B-vitamins in colostrum in nutrition and metabolism of neonates. PMID:26186707

  4. B cell responses to HIV and the development of human monoclonal antibodies.

    PubMed Central

    Boyd, J E; James, K

    1992-01-01

    In this review B cell responses in HIV-infected individuals are summarized together with the techniques used to date to produce human monoclonals to HIV and the properties of these antibodies. Profound disturbances in B cell responses are apparent both in vivo and in vitro. While there is evidence in vivo of marked polyclonal B cell activation, primary and secondary antibody responses are impaired. Similarly these cells exhibit spontaneous immunoglobulin secretion upon in vitro culture but do not readily respond to B cell mitogens and recall antigens including HIV. Furthermore, certain of these defects can be reproduced in normal B cells in vitro by incubation with HIV or HIV coded peptides. Individuals infected with HIV develop antibodies to HIV structural proteins (e.g. p17, p24, gp41 and gp120) and regulatory proteins (e.g. vif, nef, RT). Autoantibodies against a number of immunologically important molecules are also frequently observed. The anti-HIV antibodies are predominantly of the IgG1 isotype and exhibit a variety of effects on the virus in vitro. To date, using conventional immortalization strategies, an appreciable number of human monoclonals to HIV have been developed. These have been specific for gp41, gp120 and gag with antibodies of the former specificity predominating. The majority of these antibodies have been of the IgG1 isotype. Only a small number of the antibodies neutralize virus in vitro and most of these react with gp120. The neutralizing antibodies recognize conformational and carbohydrate epitopes or epitopes in amino acid positions 306-322. The predominant epitopes recognized by the anti-gp41 antibodies were in amino acid positions 579-620 and 644-662. A high percentage (congruent to 25%) of these antibodies enhance viral growth in vitro. The problems relating to the production of human monoclonals to HIV are discussed together with strategies that could be used in the future. PMID:1572084

  5. Genomic organization of the UGT2b gene cluster on human chromosome 4q13.

    PubMed

    Riedy, M; Wang, J Y; Miller, A P; Buckler, A; Hall, J; Guida, M

    2000-04-01

    The UDP-glucuronosyltransferases (UGTs) comprise a large family of proteins capable of detoxifying a wide variety of both endogenous and exogenous substrates. The primary function of this gene superfamily is to catalyze the glycosylation of substrates such as biogenic amines, steroids, bile acids, phenolic compounds and various other pharmacologically relevant compounds, including numerous carcinogens, toxic environmental pollutants and prescription drugs. This conjugation increases the solubility of these compounds, allowing them to be excreted more readily through hepatic or renal mechanisms. This paper describes the genomic characterization and chromosomal localization of three UGT2B genes which together comprise part of a large cluster of related sequences, including pseudogenes found on human chromosome 4q13. A genomic map spanning approximately 500-1000 kb of this region reveals the presence of three previously described UGT2B genes, at least two previously uncharacterized pseudogenes and a significant number of remnant gene fragments and places UGT2B4 between UGT2B7 and UGT2B15. Additionally, access to a large reference DNA bank allowed us to calculate allele frequencies for two UGT2B SNPs: D85R in UGT2B15 and Q458D in UGT2B4 amongst 803 unrelated individuals representing five ethnic populations. The data presented here suggest a recent evolutionary history of gene duplication, mutation and rearrangement. Furthermore, they suggest that a re-evaluation of the current description of the UGT2B gene family with respect to the number of specific genes, degree of allelic diversity and molecular evolution may be necessary. PMID:10803681

  6. BPR-3P0128 inhibits RNA-dependent RNA polymerase elongation and VPg uridylylation activities of Enterovirus 71.

    PubMed

    Velu, Arul Balaji; Chen, Guang-Wu; Hsieh, Po-Ting; Horng, Jim-Tong; Hsu, John Tsu-An; Hsieh, Hsing-Pang; Chen, Tzu-Chun; Weng, Kuo-Feng; Shih, Shin-Ru

    2014-12-01

    Enterovirus 71 (EV71) infections can cause hand, foot, and mouth disease with severe neurological complications. Because no clinical drug is available for treating EV71 infections, developing an efficient antiviral medication against EV71 infection is crucial. This study indicated that 6-bromo-2-[1-(2,5-dimethylphenyl)-5-methyl-1H-pyrazol-4-yl] quinoline-4-carboxylic acid (BPR-3P0128) exhibits excellent antiviral activity against EV71 (EC50 = 0.0029 ?M). BPR-3P0128 inhibits viral replication during the early post infection stage, targets EV71 RNA-dependent RNA polymerase and VPg uridylylation, and also reduces viral RNA accumulation levels and inhibits viral replication of EV71. PMID:25448086

  7. Negative modulation of Epstein-Barr virus episomes by a human B-cell growth factor.

    PubMed Central

    Morgan, J; Roome, A; Maizel, A; Sharma, S

    1989-01-01

    To study the effect of T-cell-derived BCGF-12kD on human B-cell autocrine growth-associated functions, we cultured Epstein Barr virus (EBV)-transformed normal B cells (LCL 72285) and Burkitt's lymphoma cells (Raji) in the presence or absence of BCGF-12kD. When cultured in media supplemented only with fetal calf serum, the LCL and Raji cell lines maintained relatively high levels of episomes. Although a similar level of proliferation could be maintained under defined culture conditions in media supplemented with BCGF-12kD, these conditions resulted in a time-dependent reduction of EBV sequences, as detected with EBV nuclear antigen (EBNA-1 and EBNA-2) gene probes. These results suggest that stimulation with T-cell-derived BCGF-12kD can alter a regulatory step which may be involved in the EBV transformation of B cells. Images PMID:2542626

  8. An Interaction between Glutathione and the Capsid Is Required for the Morphogenesis of C-Cluster Enteroviruses

    PubMed Central

    Ma, Hsin-Chieh; Liu, Ying; Wang, Chunling; Strauss, Michael; Rehage, Nina; Chen, Ying-Han; Altan-Bonnet, Nihal; Hogle, James; Wimmer, Eckard; Mueller, Steffen; Paul, Aniko V.; Jiang, Ping

    2014-01-01

    Glutathione (GSH) is the most abundant cellular thiol playing an essential role in preserving a reduced cellular environment. Cellular GSH levels can be efficiently reduced by the GSH biosynthesis inhibitor, L-buthionine sulfoximine (BSO). The aim of our study was to determine the role of GSH in the growth of two C-cluster enteroviruses, poliovirus type 1 (PV1) and coxsackievirus A20 (CAV20). Our results show that the growth of both PV1 and CAV20 is strongly inhibited by BSO and can be partially reversed by the addition of GSH. BSO has no effect on viral protein synthesis or RNA replication but it strikingly reduces the accumulation of 14S pentamers in infected cells. GSH-pull down assays show that GSH directly interacts with capsid precursors and mature virus made in the absence of BSO whereas capsid precursors produced under GSH-depletion do not bind to GSH. In particular, the loss of binding of GSH may debilitate the stability of 14S pentamers, resulting in their failure to assemble into mature virus. Immunofluorescence cell imaging demonstrated that GSH-depletion did not affect the localization of viral capsid proteins to the replication complex. PV1 BSO resistant (BSOr) mutants evolved readily during passaging of the virus in the presence of BSO. Structural analyses revealed that the BSOr mutations, mapping to VP1 and VP3 capsid proteins, are primarily located at protomer/protomer interfaces. BSOr mutations might, in place of GSH, aid the stability of 14S particles that is required for virion maturation. Our observation that BSOr mutants are more heat resistant and need less GSH than wt virus to be protected from heat inactivation suggests that they possess a more stable capsid. We propose that the role of GSH during enterovirus morphogenesis is to stabilize capsid structures by direct interaction with capsid proteins both during and after the formation of mature virus particles. PMID:24722315

  9. Activation of Ca2+-dependent K+ channels in human B lymphocytes by anti-immunoglobulin.

    PubMed Central

    MacDougall, S L; Grinstein, S; Gelfand, E W

    1988-01-01

    Many mammalian cell types exhibit Ca2+-dependent K+ channels, and activation of these channels by increasing intracellular calcium generally leads to a hyperpolarization of the plasma membrane. Their presence in B lymphocytes is as yet uncertain. Crosslinking Ig on the surface of B lymphocytes is known to increase the level of free cytoplasmic calcium ([Ca2+]i). However, rather than hyperpolarization, a depolarization has been reported to occur after treatment of B lymphocytes with anti-Ig. To determine if Ca2+-dependent K+ channels are present in B lymphocytes, and to examine the relationship between intracellular free calcium and membrane potential, we monitored [Ca2+]i by means of indo-1 and transmembrane potential using bis(1,3-diethylthiobarbituric)trimethine oxonol in human tonsillar B cells activated by anti-IgM. Treatment with anti-IgM induced a biphasic increase in [Ca2+]i and a simultaneous hyperpolarization. A similar hyperpolarization was induced by ionomycin, a Ca2+ ionophore. Delaying the development of the [Ca2+]i response by increasing the cytoplasmic Ca2+-buffering power delayed the hyperpolarization. Conversely, eliminating the sustained phase of the [Ca2+]i response by omission of external Ca2+ abolished the prolonged hyperpolarization. In fact, a sizable Na+-dependent depolarization was unmasked. This study demonstrates that in human B lymphocytes, Ca2+-dependent K+ channels can be activated by crosslinking of surface IgM. Moreover, it is likely that, by analogy with voltage-sensitive Ca2+ channels, Na+ can permeate through these ligand-gated Ca2+ "channels" in the absence of extracellular Ca2+. PMID:2448342

  10. Niacin Accelerates Intracellular ApoB Degradation by Inhibiting Triacylglycerol Synthesis in Human Hepatoblastoma (HepG2) Cells

    Microsoft Academic Search

    Fu-You Jin; Vaijinath S. Kamanna; Moti L. Kashyap

    2010-01-01

    The mechanism by which the potent drug niacin decreases apoB-containing atherogenic lipoproteins and prevents coronary disease is unclear. Utilizing human hepatoblastoma (HepG2) cells as an in vitro model, we have examined the effect of niacin on intracellular degradation of apoB and the regulatory mechanisms involved in apoB processing. Niacin significantly increased apoB degradation in a dose- and time-dependent manner. Treatment

  11. Dimeric Structure of a Human Apolipoprotein B mRNA Editing Protein and Cloning and Chromosomal Localization of Its Gene

    Microsoft Academic Search

    Paul P. Lau; Hui-Jia Zhu; Antonio Baldini; Chutaporn Charnsangavej; Lawrence Chan

    1994-01-01

    Apolipoprotein B (apoB) mRNA editing consists of a posttranscriptional C --> U conversion involving the first base of the codon CAA encoding glutamine-2153 to UAA, a stop codon, in apoB mRNA. Using a cloned rat cDNA as a probe, we cloned the cDNA and genomic sequences of the gene for a human apoB mRNA editing protein. Expression of the cDNA

  12. Lauroside B, a megastigmane glycoside from Laurus nobilis (bay laurel) leaves, induces apoptosis in human melanoma cell lines by inhibiting NF-?B activation.

    PubMed

    Panza, Elisabetta; Tersigni, Mariaroberta; Iorizzi, Maria; Zollo, Franco; De Marino, Simona; Festa, Carmen; Napolitano, Maria; Castello, Giuseppe; Ialenti, Armando; Ianaro, Angela

    2011-02-25

    Malignant melanoma is a highly aggressive tumor that frequently resists chemotherapy, so the search for new agents for its treatment is of great importance. In the present study, the antiproliferative propensity against human melanoma cell lines of lauroside B (1), a megastigmane glycoside isolated from Laurus nobilis (bay laurel) leaves, was investigated. This compound suppressed the proliferation of three human melanoma cell lines, namely, A375, WM115, and SK-Mel-28. The 1-induced inhibition of human melanoma cell proliferation was due to the induction of apoptosis, as demonstrated by FACS analysis with annexin V/PI staining and confirmed by activation of caspase-3 and by the cleavage of poly(ADP-ribose) polymerase (PARP). Growing evidence implicates NF-?B as an important contributor to metastasis and increased chemoresistance of melanoma. Thus, it was hypothesized that 1-induced apoptosis could be associated with suppression of NF-?B activation. The results showed that exposure of human melanoma cells to 1 inhibited I?B-? degradation and constitutive NF-?B DNA-binding activity as well as the expression, regulated by NF-?B, of two antiapoptotic genes, XIAP and c-FLIP. Induction of apoptosis by 1 in human aggressive melanoma cell lines has a potential high biological value. PMID:21188975

  13. Visfatin induces MUC8 and MUC5B expression via p38 MAPK/ROS/NF-?B in human airway epithelial cells

    PubMed Central

    2014-01-01

    Background Among a variety of inflammatory mediators, visfatin is a proinflammatory adipocytokine associated with inflammatory reactions in obesity, metabolic syndrome, chronic inflammatory disease, and autoimmune disease. However, the biological role of visfatin in secretion of major mucins in human airway epithelial cells has not been reported. Therefore, this study was conducted in order to investigate the effect and the brief signaling pathway of visfatin on MUC8 and MUC5B expression in human airway epithelial cells. Results Visfatin significantly induced MUC8 and MUC5B expression. Visfatin significantly activated phosphorylation of p38 MAPK. Treatment with SB203580 (p38 MAPK inhibitor) and knockdown of p38 MAPK by siRNA significantly blocked visfatin-induced MUC8 and MUC5B expression.Visfatin significantly increased ROS formation. Treatment with SB203580 significantly attenuated visfatin-induced ROS formation. Treatment with NAC (ROS scavenger) and DPI (NADPH oxidase inhibitor) significantly attenuated visfatin-induced MUC8 and MUC5B expression. However, treatment with NAC and DPI did not attenuate visfatin-activated phosphorylation of p38 MAPK. Visfatin significantly activated the phosphorylation of NF-?B. Treatment with PDTC (NF-?B inhibitor) significantly attenuated visfatin-induced MUC8 and MUC5B expression. Conclusions These results suggest that visfatin induces MUC8 and MUC5B expression through p38 MAPK/ROS/NF-?B signaling pathway in human airway epithelial cells. PMID:24885580

  14. The role of phosphodiesterase 4B in IL-8/LTB4-induced human neutrophil chemotaxis evaluated with a phosphodiesterase 4B inhibitor.

    PubMed

    Suzuki, Osamu; Goto, Ta?i; Yoshino, Toshiharu; Nakamura, Satoshi; Maeda, Hiroaki

    2015-06-01

    PDE4B was previously shown to be a dominant PDE4 subtype of neutrophils. However, its physiological role in the neutrophil function has not been evaluated. In this study, the inhibitory effects of a phosphodiesterase 4B (PDE4B)- selective inhibitor (compound A) and subtype non-selective PDE4 inhibitors (roflumilast and cilomilast) were evaluated in human peripheral blood cells. Compound A, roflumilast and cilomilast in a similar manner inhibited TNF-? production by LPS-stimulated human mononuclear cells. However, the inhibitory effect of compound A on IL-8 or LTB4-induced chemotactic response of neutrophils was modest even at the highest concentration (10 ?mol L-1), whereas roflumilast and cilomilast inhibited IL-8 or LTB4-induced neutrophil chemotaxis. Our results suggest that PDE4B does not play an important role during the chemotactic response of human neutrophils. PMID:26011935

  15. The next generation recombinant human cytomegalovirus vaccine candidates-beyond gB.

    PubMed

    Lilja, Anders E; Mason, Peter W

    2012-11-19

    Human cytomegalovirus (HCMV) infects the majority of the global population and persists within the infected host for life; infection of healthy adults rarely leads to severe acute clinical symptoms. In contrast, HCMV is a leading infectious cause of congenital disease and a common cause of complications in transplant recipients. A vaccine to prevent HCMV disease in these populations is a widely recognized medical need. We review recent advances in our understanding of the candidate vaccine antigens and published clinical trial data for the four most recent HCMV vaccine candidates: a gB subunit adjuvanted with MF59, a DNA vaccine expressing gB and pp65, alphavirus replicon particles (VRPs) expressing gB and a pp65-IE1 fusion protein, and a pp65 peptide vaccine. The candidates are safe, although some adverse events were reported for an adjuvanted variant of the pp65 peptide vaccine. The gB/MF59 vaccine elicited strong humoral responses with limited durability. The gB/pp65 DNA vaccine elicited cellular immunity, and the pp65 peptide vaccine elicited modest cellular immunity, but only when formulated with an adjuvant. Only the VRP vaccine expressing gB and pp65-IE1 elicited both humoral and cellular immunity. The gB/MF59 vaccine showed a short-term 50% efficacy at preventing infection of seronegative women and significantly reduced viremia and need for antivirals in solid organ transplant recipients, and the gB/pp65 DNA vaccine showed signs of clinical benefit in hematopoietic stem cell transplant recipients. Importantly, the partial efficacy of the subunit and DNA vaccines is new evidence that both humoral and cellular immunity contribute to controlling HCMV-related disease. These data show the clinical feasibility of a recombinant HCMV vaccine. We discuss areas for potential improvements in the next generation of vaccine candidates. PMID:23041121

  16. Substratum Compliance Regulates Human Trabecular Meshwork Cell Behaviors and Response to Latrunculin B

    PubMed Central

    Wood, Joshua A.; McKee, Clayton T.; Thomasy, Sara M.; Fischer, Marion E.; Shah, Nihar M.; Murphy, Christopher J.

    2011-01-01

    Purpose. To determine the impact of substratum compliance and latrunculin-B (Lat-B), both alone and together, on fundamental human trabecular meshwork (HTM) cell behavior. Lat-B is a reversible actin cytoskeleton disruptor that decreases resistance to aqueous humor outflow and decreases intraocular pressure. Methods. HTM cells were cultured on polyacrylamide hydrogels possessing values for compliance that mimic those reported for normal and glaucomatous HTM, or tissue culture plastic (TCP). Cells were treated with 0.2 ?M or 2.0 ?M Lat-B in dimethyl sulfoxide (DMSO) or DMSO alone. The impact of substratum compliance and/or Lat-B treatment on cell attachment, proliferation, surface area, aspect ratio, and migration were investigated. Results. HTM cells had profoundly decreased attachment and proliferation rates when cultured on hydrogels possessing compliance values that mimic those found for healthy HTM. The effect of Lat-B treatment on HTM cell surface area was less for cells cultured on more compliant hydrogels compared with TCP. HTM cell migration was increased on stiffer hydrogels that mimic the compliance of glaucomatous HTM and on TCP in comparison with more compliant hydrogels. Lat-B treatment decreased cellular migration on all surfaces for at least 7 hours after treatment. Conclusions. Substratum compliance profoundly influenced HTM cell behaviors and modulated the response of HTM cells to Lat-B. The inclusion of substratum compliance that reflects healthy or glaucomatous HTM results in cell behaviors and responses to therapeutic agents in vitro that may more accurately reflect in vivo conditions. PMID:22064990

  17. Development of Aflatoxin B1-Lysine Adduct Monoclonal Antibody for Human Exposure Studies

    PubMed Central

    Wang, Jia-Sheng; Abubaker, Salahaddin; He, Xia; Sun, Guiju; Strickland, Paul T.; Groopman, John D.

    2001-01-01

    Mouse monoclonal antibodies were developed against a synthetic aflatoxin B1 (AFB)-lysine–cationized bovine serum albumin conjugate. The isotype of one of these antibodies, IIA4B3, has been classified as immunoglobulin G1(?). The affinity and specificity of IIA4B3 were further characterized by a competitive radioimmunoassay. The affinities of IIA4B3 for AFB and its associated adducts and metabolites are ranked as follows: AFB-lysine > 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB > AFB = 8,9-dihydro-8-(N7-guanyl)-9-hydroxy-AFB > aflatoxin M1 > aflatoxin Q1. IIA4B3 had about a 10-fold higher affinity for binding to AFB-lysine adduct than to AFB when 3H-AFB–lysine was used as the tracer. The concentration for 50% inhibition for AFB-lysine was 0.610 pmol; that for AFB was 6.85 pmol. IIA4B3 had affinities at least sevenfold and twofold higher than those of 2B11, a previously developed antibody against parent AFB, for the major aflatoxin-DNA adducts 8,9-dihydro-8-(N7-guanyl)-9-hydroxy-AFB and 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB, respectively. An analytical method based on a competitive radioimmunoassay with IIA4B3 and 3H-AFB–lysine was validated with a limit of detection of 10 fmol of AFB-lysine adduct. The method has been applied to the measurement of AFB-albumin adduct levels in human serum samples collected from the residents of areas at high risk for liver cancer. PMID:11375185

  18. High Performance Liquid Chromatography-Mass Spectrometry (LC-MS) assay for polymyxin B1 and B2 in human plasma

    PubMed Central

    Thomas, Tiffany A; Broun, Emily C; Abildskov, Kirsten M; Kubin, Christine J; Horan, Jennifer; Yin, Michael T; Cremers, Serge

    2012-01-01

    Background Polymyxin B is an old antibiotic with increasing clinical relevance in the treatment of multi-drug resistant Gram-negative bacterial infections. However, current dosing regimens are largely empiric as clinical pharmacological characterization of the drug has been hindered by the lack of assays to measure polymyxin B in human plasma. Methods A high performance liquid chromatography-mass spectrometry (LC-MS) assay was developed to quantify polymyxin B1 and B2 in human plasma using pure calibrators. After purification with a solid-phase extraction column, polymyxin B1 and B2 were separated on a C18 column by gradient chromatography with an overall runtime of 12 minutes. Polymyxin B1 and B2 were ionized by positive electron spray ionization and the resulting ions specific to polymyxin B1 and B2 were monitored (selected ion recording). Results The dominant ions produced were [M + 2H]2+ at m/z 602.6 and 595.5 for polymyxin B1 and polymyxin B2, respectively. The assay was linear between concentrations of 100 and 2500 ng/ml, with inter-day precision of 5.9% and 3.4% at 100 ng/mL and 5.3% and 4.0% at 2000 ng/ml for polymyxin B1 and polymyxin B2, respectively. Accuracy was 80.2% and 82.2% at 100 ng/mL and 99.9% and 109.6% at 2000 ng/ml for polymyxin B1 and polymyxin B2, respectively. No interference from other drugs commonly administered with polymyxin B was detected. The performance of the assay is affected by gross hemolysis and hyperlipemia. The method was successfully applied to patient samples. Interestingly, in a single patient the ratio of B1 and B2 did not change over a period of 12 hours after administration of the drug. Conclusions A simple method for the simultaneous measurement of polymyxin B1 and polymyxin B2 in human plasma is described, which has the potential to optimize clinical use of this valuable antibiotic by permitting pharmacokinetic studies and therapeutic drug monitoring. PMID:22735673

  19. Effect of Cytochrome b5 Content on the Activity of Polymorphic CYP1A2, 2B6, and 2E1 in Human Liver Microsomes

    PubMed Central

    Zhang, Haifeng; Gao, Na; Liu, Tingting; Fang, Yan; Qi, Bing; Wen, Qiang; Zhou, Jun; Jia, Linjing; Qiao, Hailing

    2015-01-01

    Human cytochrome b5 (Cyt b5) plays important roles in cytochrome P450 (CYP)-mediated drug metabolism. However, the expression level of Cyt b5 in normal human liver remains largely unknown. The effect of Cyt b5 on overall CYP activity in human liver microsomes (HLM) has rarely been reported and the relationship between Cyt b5 and the activity of polymorphic CYP has not been systematically investigated. In this study, we found that the median value of Cyt b5 protein was 270.01 pmol/mg from 123 HLM samples, and 12- and 19-fold individual variation was observed in Cyt b5 mRNA and protein levels, respectively. Gender and smoking clearly influenced Cyt b5 content. In addition, we found that Cyt b5 protein levels significantly correlated with the overall activity of CYP1A2, 2B6, and 2E1 in HLM. However, when the CYP activities were sorted by single nucleotide polymorphisms (SNP), the effect of Cyt b5 protein on the kinetic parameters varied greatly. There were significant correlations between Cyt b5 content and Vmax and CLint of CYP1A2 wild-types (3860GG, 2159GG, and 5347CC) as well as homozygous mutants (163AA and 3113GG). In contrast to Vmax and CLint, the Km of CYP2B6 516GG and 785AA genotypes was inversely associated with Cyt b5 content. Correlations between Cyt b5 content and Vmax and CLint of CYP2E1 -1293GG, -1293GC, 7632TT, 7632TA, -333TT, and -352AA genotypes were also observed. In conclusion, Cyt b5 expression levels varied considerably in the Chinese cohort from this study. Cyt b5 had significant impact on the overall activity of CYP1A2, 2B6, and 2E1 in HLM and the effects of Cyt b5 protein on polymorphic CYP1A2, 2B6, and 2E1 activity were SNP-dependent. These findings suggest that Cyt b5 plays an important role in CYP-mediated activities in HLM and may possibly be a contributing factor for the individual variation observed in CYP enzyme activities. PMID:26046844

  20. Sanfilippo syndrome type B: cDNA and gene encoding human {alpha}-N-acetylglucosaminidase

    SciTech Connect

    Zhao, H.G.; Lopez, R.; Rennecker, J. [UCLA School of Medicine, Los Angeles, CA (United States)] [and others

    1994-09-01

    Deficiency of the lysosomal enzyme {alpha}-N-acetlyglucosaminidase underlies the type B Sanfilippo syndrome (MPS III B), a mucopolysaccharide storage disease with profound neurologic deterioration. We are acquiring tools to study the molecular basis of the disorder. The enzyme was purified from bovine testis; after ConA-, DEAE- and phenyl-Sepharose chromatography, it was subjected to SDS-PAGE without preheating. Of two bands of activity detected on the gel, 170 kDa and 87 kDa, the larger one, which coincided with a well-defined Coomassie blue band, was selected for sequence analysis. Degenerate 17-base oligonucleotides, corresponding to the ends of an internal 23 amino acid sequence, were used for RT-PCR of RNA from human fibroblasts. A 41-mer was synthesized from the sequence of the RT-PCR product and used to screen a human testis cDNA library. A number of cDNA inserts were isolated, all lacking the 5{prime} end and none longer than 1.7 kb. An additional 300 bp segment has been obtained by RACE. The cDNA sequence accounts for 9 of 11 peptides, allowing for species difference. Northern analysis of fibroblast RNA with a 1.5 kb cDNA probe showed the presence of a 3 kb mRNA; marked deficiency of this mRNA in two MPS III B fibroblast lines confirmed the authenticity of the cloned cDNA. While no homologous amino acid sequence has been found in a search of GenBank, the nucleotide sequence (interrupted by 4 introns) is present in a flanking region upstream of an unrelated gene on chromosome 17q11-21 (human 17{beta}-hydroxysteroid dehydrogenase). This must therefore be the chromosomal locus of the {alpha}-N-acetylglucosaminidase gene and of MPS III B.