| reactivity of synthetic peptides representing selected sections of hepatitis c virus core and envelope proteins with a panel of hepatitis c virus-seropositive human plasma. | a series of 54 synthetic peptides, 15-20 residues long, that represented selected parts of the structural proteins of hepatitis c virus (hcv) were tested for immunoreactivity with a panel of 45 plasma samples from potential blood donors who were known to be seropositive for anti-hcv. most of the ten peptides that represented the core protein showed reactivity with most of the panel samples. all except one of the 20 peptides that represented non-hypervariable regions of envelope proteins e1 and e ... | 1997 | 8986952 |
| synthesis of soluble rubella virus spike proteins in two lepidopteran insect cell lines: large scale production of the e1 protein. | the two envelope glycoproteins of rubella virus (rv), e1 of 58 kda and e2 of 42-47 kda, were individually expressed in lepidopteran spodoptera frugiperda as well as in trichoplusia ni insect cells using baculovirus vectors. the authentic signal sequences of e1 and e2 were replaced with the honeybee melittin signal sequence, allowing efficient entrance into the secretory pathway of the insect cell. in addition, the hydrophobic transmembrane anchors at the carboxyl termini of e1 and e2 proteins we ... | 1996 | 8987625 |
| detection of rubella virus-specific immunoglobulin m antibodies with a baculovirus-expressed e1 protein. | the structural proteins of rubella virus (rv) were expressed in insect cells by using the baculovirus expression vector system. the recombinant e1 envelope glycoprotein was purified by immunoaffinity chromatography and used to detect rv-specific immunoglobulin m antibodies in a time-resolved fluoroimmunoassay. correlation analysis between the reactivities of antibodies against this recombinant e1 and the reactivities against authentic rv antigen shows that purified e1 can detect rv antibodies of ... | 1996 | 8991639 |
| role of glycoprotein pe2 in formation and maturation of the sindbis virus spike. | sindbis virus envelope assembly is a multistep process resulting in the maturation of a rigid, highly ordered t=4 icosahedral protein lattice containing 80 spikes composed of trimers of e1-e2 heterodimers. intramolecular disulfide bonds within e1 stabilize e1-e1 associations required for envelope formation and maintenance of the envelope's structural integrity. the structural integrity of the envelope protein lattice is resistant to reduction by dithiothreitol (dtt), indicating that e1 disulfide ... | 1997 | 8995682 |
| evolutionary analysis of variants of hepatitis c virus found in south-east asia: comparison with classifications based upon sequence similarity. | variants of hepatitis c virus (hcv) have been classified by nucleotide sequence comparisons in different regions of the genome. many investigators have defined the ranges of sequence similarity values or evolutionary distances corresponding to divisions of hcv into types, subtypes and isolates. using these criteria, novel variants of hcv from vietnam, thailand and indonesia have been classified as types 7, 8, 9, 10 and 11, many of which can be further subdivided into between two to four subtypes ... | 1996 | 9000092 |
| classification of hepatitis c virus variants in six major types based on analysis of the envelope 1 and nonstructural 5b genome regions and complete polyprotein sequences. | the phylogenetic status of recently described isolates of hepatitis c virus (hcv) from vietnam, thailand and indonesia (previously classified as types 7, 8, 9, 10 and 11) was re-analysed by the neighbour-joining method instead of the unweighted pair-group method with arithmetic mean (upgma) that was first used by the discoverers of these strains. the analysis of complete amino acid sequences and of nucleotide sequences of the envelope 1 (672 nt) and nonstructural 5b (1092 nt) genomic regions per ... | 1997 | 9010284 |
| casein kinase ii phosphorylates bovine papillomavirus type 1 e1 in vitro at a conserved motif. | the e1 protein of bovine papillomavirus type 1 (bpv-1) is a phosphoprotein which specifically binds and unwinds the virus replication origin by atp-dependent helicase activity. the el protein has been shown to be multiply phosphorylated in vivo, although the sites of modification are incompletely mapped. examination of the predicted amino acid sequence of all available e1 proteins revealed strong conservation between amino acids 25 and 60 of a motif consisting of a serine residue followed by a s ... | 1997 | 9010301 |
| purification of rubella virus e1-e2 protein complexes by immunoaffinity chromatography. | a murine monoclonal antibody directed against the e1 membrane glycoprotein of rubella virus was immobilized on an n-hydroxysuccinimide-activated chromatographic support. the antibody was used to purify rubella virus e1-e2 protein complexes from tween-80/diethyl ether extracts of cell culture supernatants containing virus particles. the adsorption behaviour of immunosorbents with ligand densities of 2.9, 5.4 and 11.1 mg monoclonal antibody per millilitre of gel was investigated using batchwise co ... | 1997 | 9015274 |
| the origin of hepatitis c virus genotypes. | for many rna viruses, relatively recent times of origin of extant viruses are implied by the high rate of substitution observed in longitudinal studies. however, extrapolation of short-term rates of substitution can give misleading estimates of times of divergence. we show here that the common ancestor of different types of hepatitis c virus (hcv) is older than previously thought. the rate of hcv sequence change was measured amongst a cohort of individuals infected following administration of an ... | 1997 | 9018053 |
| nucleotide sequence of the barmah forest virus genome. | barmah forest virus (bfv) is an atypical alphavirus [dalgarno, l., short, n. j., hardy, c. m., bell, j. r., strauss, j. h., and marshall, i. d. (1984). virology 133, 416-426] and has been classified as the sole known member of a seventh alphavirus serocomplex. the complete nucleotide sequence of bfv genomic rna is 11,488 nucleotides in length excluding the poly(a) tail. two long open reading frames in the rna encode a nonstructural polyprotein of 2411 amino acids and a structural polyprotein of ... | 1997 | 9018152 |
| detection of type 2-like t-helper cells in hepatitis c virus infection: implications for hepatitis c virus chronicity. | one striking clinical feature of hepatitis c virus (hcv) infection is that more than 50% of patients with acute hepatitis c will develop chronic infection. to investigate its possible mechanisms, we examined the activation of type 2-like t-helper (th2-like) cells relating to the development of chronicity. peripheral blood cd4+ t-cell proliferation and cytokine secretion in response to a panel of recombinant hcv antigens including core (c22), envelope 1 (e1), e2, nonstructural (ns) protein 4 (c10 ... | 1997 | 9021963 |
| induction of cytotoxic t-cell response against hepatitis c virus structural antigens using a defective recombinant adenovirus. | a replication-defective recombinant adenovirus (rad), radcmv-ce1, containing core and e1 genes of hepatitis c virus (hcv) was constructed. radcmv-ce1 was able to express core and e1 proteins both in mice and human cells. immunization of balb/c mice with radcmv-ce1 induced a specific cytotoxic t-cell response against the two hcv proteins. this response was characterized using a panel of 60 synthetic 14- or 15-mer overlapping peptides (10 amino-acid overlap) spanning the entire sequence of these p ... | 1997 | 9021966 |
| construction of adenovirus vectors through cre-lox recombination. | two barriers prevent adenovirus-based vectors from having wide application. one is the difficulty of making new adenoviruses, and the second is the strong immunological reaction to viral proteins. here we describe uses of cre-lox recombination to overcome these problems. first, we demonstrate a simple method for constructing e1-substituted adenoviruses. second, we demonstrate a method to construct adenovirus vectors carrying recombinant genes in place of all of the viral genes, so-called gutless ... | 1997 | 9032314 |
| generation of cytotoxic t lymphocytes against immunorecessive epitopes after multiple immunizations with adenovirus vectors is dependent on haplotype. | currently, adenovirus (ad) is being considered as a vector for the treatment of cystic fibrosis as well as other diseases. however, the cytotoxic t lymphocyte (ctl) response to ad could limit the effectiveness of such approaches. since the ctl response to virus infection is often focused on one or a few immunodominant epitopes, one approach to circumvent this response is to create vectors that lack these immunodominant epitopes. the effectiveness of this approach was tested by immunizing mice wi ... | 1997 | 9032363 |
| genetic characterization of ruminant pestiviruses: sequence analysis of viral genotypes isolated from sheep. | historically, the genus pestivirus was believed to contain three species of viruses; bovine viral diarrhea virus (bvdv), border disease virus (bdv) and classical swine fever virus (csfv). however, based on limited sequence analysis of a small number of pestiviral isolates from domestic livestock, evidence has recently emerged indicating that at least four distinct genotypes exist. in an attempt to gain a better understanding of the degree of viral variation among ruminant pestiviruses, the entir ... | 1997 | 9037733 |
| e1a rna transcripts amplify adenovirus-mediated tumor reduction. | previous work by this group has established that e1-defective, recombinant adenoviruses can be replication-enabled by the codelivery of a plasmid encoding the deleted e1 functions, a strategy now designated conditional replication-enablement system for adenovirus (cresa). in the studies reported here, the original replication-enabling plasmid was replaced by two separate plasmids that encoded the necessary e1a and e1b functions, respectively. an rna transcript encoding the requisite e1a function ... | 1996 | 9044743 |
| brefeldin a and monensin arrest cell surface expression of membrane glycoproteins and release of rubella virus. | the maturation of rubella virus (rv) glycoproteins e2 and e1 was examined by using brefeldin a (bfa) and monensin. bfa, which induces the rapid redistribution of golgi enzymes residing in the golgi complex into the endoplasmic reticulum (er), was used to locate the intracellular site for the modification of carbohydrate side-chains on rv e1 and e2 proteins. the monovalent ionophore monensin, which inhibits intracellular transport of proteins through the er-golgi complex, was used to block the tr ... | 1995 | 9049331 |
| sequence analysis of hepatitis c virus genotypes 1 to 5 reveals multiple novel subtypes in the benelux countries. | hepatitis c virus (hcv) isolates from a cohort of 315 patients from the benelux countries (belgium, the netherlands, luxembourg) were genotyped by means of reverse hybridization inno-lipa (line probe assay). genotypes 1a, 1b, 2a, 2b, 3a, 4a and 5a were detected. from the cohort, isolates representing all types and those showing an aberrant lipa pattern were further analysed by sequencing parts of the 5' utr, core (nt 1 to 326; aa residues 1 to 108) and core/e1 (nt 477 to 924; aa residues 159 to ... | 1995 | 9049395 |
| derivatives of activated h-ras lacking c-terminal lipid modifications retain transforming ability if targeted to the correct subcellular location. | to examine the ability of ras to activate signal transduction pathways in the absence of lipid modifications, fusion proteins were constructed that target raswt or activated ras61l to cellular membranes as integral membrane proteins, using the first transmembrane domain of the e1 protein of avian infectious bronchitis virus (ibv), which contains a cis-golgi targeting signal. golgi-targeted derivatives of activated ras were completely inactive in transformation assays. however, when examined in f ... | 1997 | 9050994 |
| conditional repression of the e2 transcription unit in e1-e3-deleted adenovirus vectors is correlated with a strong reduction in viral dna replication and late gene expression in vitro. | an e1-e3-deleted recombinant adenovirus vector expressing the hybrid protein tetr-krab has been produced. in this virus, adtg9562, the e2 transcription is regulated by tetr-krab and teto sequences inserted in cis. in the absence of tetracycline, a strong reduction in e2a gene expression, viral dna replication, and late gene expression was observed in noncomplementing a549 cells, and a reduction in viral growth was seen in the e1-expressing 293 cells. in contrast, there was no repression in the p ... | 1997 | 9060700 |
| interleukin-10 immunoadhesin production by a replication-defective adenovirus. | the present study examined the use of replication-defective adenovirus for in vitro production of an immunoadhesin. a recombinant adenovirus, rendered replication defective by deletion of the e1 gene, was constructed to contain the murine interleukin-10 gene fused in frame with the hinge, ch2, and ch3 domains of the murine immunoglobulin gamma 1 heavy chain constant region gene under the control of the human cytomegalovirus promoter. the resultant recombinant virus, ad5.hcmv.mil-10:hfc, was used ... | 1997 | 9075770 |
| [comparison between hgv genome and hgbv-c genome]. | new hepatitis viruses, hepatitis g virus(hgv) and hepatitis gb virus c(hgbv-c), were reported from two groups of researchers. now these two are thought to be similar but hgv genome(u44402) and hgbv-c genome(u36380) do not have the same sequence. we compare these two sequences in both nucleotide and aminoacid analyses. homology of nucleotide between hgv and hgbv-c is 83.8% in 5'nc region, 88.8% in core, 85.1% in e1, 85.7% in e2, 85.1% in ns2-3, 84.5% in ns4a, 86.8% in ns4b-5a, 88.6% in ns5b and 1 ... | 1997 | 9086765 |
| a novel family of viral death effector domain-containing molecules that inhibit both cd-95- and tumor necrosis factor receptor-1-induced apoptosis. | molluscum contagiosum virus proteins mc159 and mc160 and the equine herpesvirus 2 protein e8 share substantial homology to the death effector domain present in the adaptor molecule fas-associated death domain protein (fadd) and the initiating death protease fadd-like interleukin-1beta-converting enzyme (flice) (caspase-8). fadd and flice participate in generating the death signal from both tumor necrosis factor receptor-1 (tnfr-1) and the cd-95 receptor. the flow of death signals from tnfr-1 occ ... | 1997 | 9092488 |
| the role of rubella-immunoblot and rubella-peptide-eia for the diagnosis of the congenital rubella syndrome during the prenatal and newborn periods. | rubella infection during the first trimester of pregnancy can cause the congenital rubella syndrome (crs). patients with crs were shown to have a decreased humoral and cellular immunity. it is not known whether asymptomatic newborns who had experienced intrauterine infection with rubella virus (rv) differ in their antibody response from newborns with crs. in this study we compared both groups for a difference which might be a useful diagnostic criterion for crs during the prenatal and newborn pe ... | 1997 | 9093941 |
| mutational analysis of the human papillomavirus type 16 e1--e4 protein shows that the c terminus is dispensable for keratin cytoskeleton association but is involved in inducing disruption of the keratin filaments. | the function of the human papillomavirus (hpv) e4 proteins is unknown. in cultured epithelial cells the proteins associate with the keratin intermediate filaments (ifs) and, for some e4 types, e.g., hpv type 16 (hpv-16), induce collapse of the keratin networks. an n-terminal leucine-rich motif (llxll) is a conserved feature of many e4 proteins. in a previous study we showed that deletion of this region from the hpv-1 and -16 e4 proteins abrogated the localization of the mutant proteins to the ke ... | 1997 | 9094627 |
| the use of an e1-deleted, replication-defective adenovirus recombinant expressing the rabies virus glycoprotein for early vaccination of mice against rabies virus. | an e1-deleted, replication-defective adenovirus recombinant of the human strain 5 expressing the rabies virus glycoprotein, termed adrab.gp, was tested in young mice. mice immunized at birth with the adrab.gp construct developed antibodies to rabies virus and cytokine-secreting lymphocytes and were protected against subsequent challenge. maternal immunity to rabies virus strongly interferes with vaccination of the offspring with a traditional inactivated rabies virus vaccine. the immune response ... | 1997 | 9094641 |
| echovirus 1 replication, not only virus binding to its receptor, vla-2, is required for the induction of cellular immediate-early genes. | induction of immediate-early genes c-jun, junb, and c-fos was demonstrated during echovirus 1 infection in a human osteogenic sarcoma (hos) cell line. tenfold induction was seen at 10 h postinfection, corresponding approximately to the end of the first replication cycle of the virus. echovirus 1 uses vla-2 integrin as its cellular receptor, and ligand binding by integrin is known to trigger signal transduction pathways ultimately activating immediate-early genes. in the present study, however, v ... | 1997 | 9094704 |
| unwinding of the box i element of a herpes simplex virus type 1 origin by a complex of the viral origin binding protein, single-strand dna binding protein, and single-stranded dna. | the herpes simplex virus type 1 (hsv-1) genome contains three origins of replication: oril and two copies of oris. these origins contain specific sequences, box i and box ii, linked by an at-rich segment, that are recognized by an hsv-1-encoded origin binding protein (ul9 protein) which also possesses dna helicase activity. despite its intrinsic helicase activity, the ul9 protein is unable to unwind oris or the box i element of oris, either in the presence or absence of the hsv-1-encoded single- ... | 1997 | 9096307 |
| transgenic expression of hepatitis c virus structural proteins in the mouse. | although hepatitis c virus (hcv) is a leading cause of morbidity and mortality worldwide, the role of viral cytopathic effects remains unclear. to study the biosynthesis of hcv structural proteins and their pathogenic role, we constructed transgenic mice, expressing type 1b hcv structural proteins (core, e1, and e2) in liver tissues. two liver-specific promoters were used. the mouse major urinary protein (mup) promoter has been shown to be developmentally regulated with little or no expression i ... | 1997 | 9096613 |
| investigation of the pattern of diversity of hepatitis c virus in relation to times of transmission. | the rate of sequence change of hcv in vivo was used to date the spread of hcv genotype 1b in european, usa and japanese populations. silent substitution rates of 0.0011 and 0.0017 substitutions per site per year were observed in the ns5 and e1 regions by sequence comparisons from a cohort of individuals infected from a common source of infection 17 years previously. mean silent substitution frequencies of 0.169 and 0.224 in ns5 and e1, respectively, were observed amongst type 1b variants infecti ... | 1997 | 9097281 |
| plasmid dna-based immunization for hepatitis c virus structural proteins: immune responses in mice. | plasmid dna-based immunization has been shown to be an effective means of vaccination in animal models. in this study, the immune responses to various hepatitis c virus structural protein antigens were evaluated using this technique. | 1997 | 9098018 |
| adenovirus-mediated in vivo gene transfer rapidly protects ornithine transcarbamylase-deficient mice from an ammonium challenge. | the purpose of this study was to determine the time of onset, duration, and the efficacy of in vivo gene transfer in protecting the ornithine transcarbamylase deficient spf/y mouse from an acute ammonium challenge. the animals were challenged with ammonia (10 mmol/kg nh4cl) 1, 2, 7, 14, or 28 d after the administration of a recombinant adenoviral construct deleted in e1 and with a temperature sensitive mutation in e2. although there was no protection with the control lacz virus, the ornithine tr ... | 1997 | 9098855 |
| definition of three minimal t helper cell epitopes of rubella virus e1 glycoprotein. | to characterize t cell-recognized epitopes on rubella virus (rv) e1 glycoprotein, il-2-dependent rv-specific t cell lines were established from 14 rubella-seropositive healthy donors. the responses of these lines were studied by using a panel of 94 partially overlapping synthetic peptides of 15 amino acids (aa) length covering the known nucleotide sequence of rve1 glycoprotein. two to seven peptide-defined epitopes were recognized by the t cell lines, but a large interindividual variation was fo ... | 1996 | 9099921 |
| development and characterization of sv40 immortalized rat submandibular acinar cell lines. | rat submandibular salivary gland acinar cells were transfected by capo4 precipitation using a plasmid containing a replication-defective simian virus (sv40) genome. out of 27 clonal cell lines, two were shown to have moderate to high levels of cytodifferentiation and salivary gland acinar cell function. functional studies with the two cell lines indicated that the beta-adrenergic agonist, isoproterenol, vasoactive intestinal peptide, and prostaglandin e1 were effective activators of intracellula ... | 1997 | 9112124 |
| in vivo and in vitro gene transfer and expression in rat intestinal epithelial cells by e1-deleted adenoviral vector. | the intestine is proposed to be an attractive target site for somatic gene therapy due to a large mass of proliferating tissue and stem cells in the crypts. previous studies using a retroviral vector have shown that a reporter gene, bacterial beta-galactosidase (beta-gal), can be transferred and expressed in the small intestinal epithelial cell. however, transduction efficiency is relatively low in rat and mice intestines. in the present study, we employed an e1-deleted adenoviral vector (which ... | 1997 | 9113515 |
| major differences between whv and hbv in the regulation of transcription. | studies were carried out to further characterize enhancer and promoter elements on the woodchuck hepatitis virus (whv) genome. we were able to confirm the existence of whv promoters analogous to the major promoters of the related human hepatitis b virus (hbv) and of an enhancer analogous to the recently described whv e2 element (ueda, k., wei, y., and ganem, d., virology 217, 413, 1996). however, we were unable to identity an enhancer analogous to the e1 element of (hbv), despite the fact that t ... | 1997 | 9123867 |
| transmission of the hepatitis c virus in an hemodialysis unit: evidence for nosocomial infection. | hepatitis c virus (hcv) infection is a frequent feature in hemodialysis (hd) patients. the way of viral transmission is difficult to establish, but in previous studies the role of blood transfusions and of hd treatment duration, and the possibility of nosocomial transmission of the virus have been suggested. we present here the results of a virological follow-up of hcv infection in our hd unit in 1993-1994, and a molecular study of viral strains that led to a possible reconstruction of viral spr ... | 1997 | 9128794 |
| the entire nucleotide sequences of two gb virus c/hepatitis g virus isolates of distinct genotypes from japan. | recently, putative viral agents responsible for human non-a to e hepatitis have been independently reported by two groups of investigators and designated gb virus c (gbv-c) and hepatitis g virus (hgv), respectively. the entire nucleotide sequences were determined for two viral genomes isolated from japanese blood donors with gbv-c rna. one of them (gt230) had a total genomic length of 9390 nucleotides (nt) with 5' and 3' untranslated regions of 551 and 313 nt, while the other (gt110) had genomic ... | 1997 | 9129645 |
| analysis of mother-to-infant transmission of hepatitis c virus: quasispecies nature and buoyant densities of maternal virus populations. | mother-to-infant transmission of hepatitis c virus (hcv) was analyzed by sequencing of viral rna and semiquantitative polymerase chain reaction following ultracentrifugation of maternal sera. in two mother-infant pairs, the hypervariable region 1 (hvr1) and carboxyl terminus of envelope 1 (e1) were sequenced. both viral sequences in the infants were less diverse than those of their mothers. although the e1 sequences were almost identical in each mother-infant pair, the hvr1 sequences of the infa ... | 1997 | 9139088 |
| analyses of disulfides present in the rubella virus e1 glycoprotein. | the surface of rubella virus contains the glycoproteins e1 and e2. the e1 protein induces neutralizing antibodies and has been implicated in the process of recognition of cellular receptors. to gain information on the structural organization of the e1 protein we have analyzed the disulfide bonds present within this molecule. the reactivity of the protein with radioactively labeled iodoacetic acid indicates that all 20 cysteine residues present in the ectodomain of the e1 protein are involved in ... | 1997 | 9143273 |
| delineation of regions important for heteromeric association of hepatitis c virus e1 and e2. | hepatitis c virus (hcv) is the major causative agent of blood-borne non-a non-b hepatitis. the persistence of hcv infection is believed to reflect escape from the host immunosurveillance system by mutations in hypervariable region 1 (hvr1) of the envelope protein 2 (e2). two envelope proteins of hcv, e1 and e2, have been reported to form a heteromeric complex but the exact organization of the viral envelope proteins remains uncertain. we examined the interaction of e1 and e2 by far- western blot ... | 1997 | 9143310 |
| increased cyclic amp response to forskolin in epstein-barr virus-transformed human b-lymphocytes derived from schizophrenics. | phorbol 12-myristate-13-acetate (pma), a protein kinase c (pkc) activator, elevated basal cyclic amp levels and enhanced isoproterenol-, prostaglandin e1- (pge1), forskolin- and cholera toxin-stimulated cyclic amp accumulation in epstein-barr virus (ebv)-transformed human b-lymphocytes. staurosporine, a pkc inhibitor, significantly antagonized the increase in cyclic amp accumulation produced by pma, whereas the inactive phorbol ester, 4 alpha-phorbol 12,13-didecanoate (4 alpha pdd), had no effec ... | 1997 | 9151357 |
| long-term gene delivery into the livers of immunocompetent mice with e1/e4-defective adenoviruses. | we have compared the in vitro and in vivo behaviors of a set of isogenic e1- and e1/e4-defective adenoviruses expressing the lacz gene of escherichia coli from the rous sarcoma virus long terminal repeat. infection of tumor-derived established cell lines of human origin with the doubly defective adenoviruses resulted in (i) a lower replication of the viral backbone that correlated with reduced levels of e2a-specific rna and protein, (ii) a significant shutoff of late gene and protein expression, ... | 1997 | 9151856 |
| role of the first and third extracellular domains of cxcr-4 in human immunodeficiency virus coreceptor activity. | the cxcr-4 chemokine receptor and cd4 behave as coreceptors for cell line-adapted human immunodeficiency virus types 1 and 2 (hiv-1 and hiv-2) and for dual-tropic hiv strains, which also use the ccr-5 coreceptor. the cell line-adapted hiv-1 strains lai and ndk and the dual-tropic hiv-2 strain rod were able to infect cd4+ cells expressing human cxcr-4, while only lai was able to infect cells expressing the rat homolog of cxcr-4. this strain selectivity was addressed by using human-rat cxcr-4 chim ... | 1997 | 9151868 |
| diminishing adenovirus gene expression and viral replication by promoter replacement. | the adenovirus e4 promoter was replaced by a synthetic promoter composed of a minimal tata box and five consensus 17-mer yeast gal4-binding-site elements. the viral vectors, which also contained human factor ix (hfix) cdna driven by rous sarcoma virus long terminal repeat in the e1 region, were then constructed and expanded in 293 cells permanently expressing gal4/vp16 fusion protein. viral replication and expression of adenovirus e4 genes and late genes (hexon and fiber) were evaluated in vitro ... | 1997 | 9151874 |
| complete correction of hyperbilirubinemia in the gunn rat model of crigler-najjar syndrome type i following transient in vivo adenovirus-mediated expression of human bilirubin udp-glucuronosyltransferase. | recombinant adenoviral vectors are useful for the in vivo expression of genes in hepatocytes. adenoviral vectors deleted in e1a, e1b, and e3b were constructed and used to study in vivo expression of the major human bilirubin udp-glucuronosyltransferase isoform (hug br1) under the transcriptional control of the cytomegalovirus (cmv) immediate-early promoter-enhancer (h5.010cmv hugbr1). as a control, a recombinant adenoviral vector containing the beta-galactosidase reporter gene driven by the cmv ... | 1996 | 9156798 |
| gene transfer into enteric neurons of the rat small intestine in organ culture using a replication defective recombinant herpes simplex virus type 1 (hsv1) vector, but not recombinant adenovirus vectors. | we have designed a system in which to test gene transfer into gut neurons consisting of an organ culture of neonatal rat small intestine. the tissue was exposed to herpes simplex- and adenovirus-derived vectors: (1) a temperature-sensitive herpes simplex virus-1 (hsv1) vector (tsk-beta gal) containing the lacz gene encoding beta-galactosidase (beta-gal), under the transcriptional control of the hsv1 immediate-early 3 (ie3) promoter; (2) rad35, an e1-/e3- replication-deficient adenovirus expressi ... | 1997 | 9176519 |
| recognition of contiguous allele-specific peptide elements in the rubella virus e1 envelope protein. | peptides which bind to human hla-drb1 class ii molecules in an allele-specific fashion were derived from the immunodominant e1 envelope protein of rubella virus. two nonoverlapping e1 peptide epitopes were recognized by rubella virus-specific t cells in the context of independent hla alleles when presented either separately or as a contiguous polypeptide containing both epitopes. direct binding analysis of potential peptide epitopes to distinct hla molecules provides a direct approach for select ... | 1997 | 9178465 |
| predicted secondary structure of the hepatitis g virus and gb virus-a 5'untranslated regions consistent with an internal ribosome entry site. | we describe comparative sequence analysis of 20 isolates of the recently discovered hepatitis g virus (hgv) and propose a model of the rna secondary structure at the 3' end of the 5' untranslated region (utr) of this virus. a single aug starting at nucleotide position 552, which was in-frame and continuous with the putative polyprotein, was conserved in all 20 isolates and appeared to be the most likely site for the initiation of polyprotein synthesis. this consensus aug was 14 amino acid residu ... | 1997 | 9181526 |
| recombinant adenovirus synthesizing cell surface-anchored beta hcg induces bioneutralizing antibodies in rats. | a recombinant adenovirus (re-ad) has been constructed that synthesizes a cell surface-anchored form of the beta-subunit of human chorionic gonadotropin (beta hcg). this was achieved by in-frame fusion of beta hcg cdna at its c terminus with the gene sequences coding for the vesicular stomatitis virus glycoprotein (vsvg) transmembrane domain. the fusion protein gene was placed under the control of human cytomegalovirus (hcmv) immediate early promoter and this expression cassette was inserted into ... | 1997 | 9185867 |
| high level of transgene expression in cell cultures and in the mouse by replication-incompetent adenoviruses harboring modified vai genes. | replication-incompetent adenoviruses are currently used in gene therapy trials. most of the work designed to increase the expression from these vectors concerns the modification of cis sequences of the foreign transcription unit, so as to improve the transcription level or the stability of the mrna. in this report, we show that an alternative strategy based on the coexpression of modified vai genes can efficiently increase gene expression both in cell cultures and in animals. the vai rna is synt ... | 1997 | 9188608 |
| analysis of hepatitis c virus core protein interaction domains. | hepatitis c virus (hcv) core protein forms the internal viral coat that encapsidates the genomic rna and is enveloped in a host cell-derived lipid membrane. as the single capsid protein, core should be capable of multimerization but attempts to produce virus-like particles following expression of hcv structural proteins have not been successful. in this study, we have analysed the interaction capacity of full-length and truncated hcv core using the yeast two-hybrid system. full-length core conta ... | 1997 | 9191926 |
| mutation rate of gb virus c/hepatitis g virus over the entire genome and in subgenomic regions. | a patient on maintenance hemodialysis was infected with a recently discovered putative non-a to -e hepatitis virus designated gb virus c (gbv-c) or hepatitis g virus (hgv) by transfusion. the viral isolate was recovered from the patient soon after she turned positive for gbv-c/hgv rna in serum (gs185) and 8.4 years thereafter (gs193), and the entire nucleotide sequences were determined. they both had a genomic length of 9391 nucleotides with a defective c gene made of only 42 nucleotides. betwee ... | 1997 | 9201215 |
| high prevalence of hepatitis c virus infection in patients with b-cell lymphoproliferative disorders in italy. | starting from the observation that a number of consecutive patients with non-hodgkin's lymphoma (nhl) resulted positive for hepatitis c virus (hcv) antibodies on routine testing, we set up a survey for hcv contact prevalence in all patients with lymphoproliferative disorders (lpd) followed in our institution. we searched for hcv antibodies by a third-generation elisa technique, followed by a confirmation test (riba iii); serum viral rna and hcv genotype were investigated by a rt-pcr technique. w ... | 1997 | 9209002 |
| different affinity of monoclonal antibodies for conserved neutralizing epitopes on two strains of rubella virus. | there is, apparently, only one serological type of rubella virus (rv) in the population, although several isolates exist with different characteristics. some authors failed to detect significant differences among rv strains by neutralization, hemagglutination inhibition, and enzyme immunoassay using polyclonal and monoclonal antibodies, but differences in growth, plaque morphology, and temperature sensitivity between vaccine and wild-type strains were shown by chantler et al. (3) with the purpos ... | 1997 | 9210284 |
| evaluation of hepatitis c virus envelope proteins expressed in e. coli and insect cells for use as tools for antibody screening. | the two envelope proteins of hepatitis c virus, e1 and e2, were expressed in e. coli and, as secretory proteins, in sf9 insect cells using recombinant baculoviruses. co-infection of insect cells with e1 and e2-recombinant baculoviruses was performed, which has been shown to result in formation of e1-e2 dimers. all envelope proteins were purified by ni2+-nta chromatography and used for screening of serum samples in a hcv eia assay. serum samples of normal blood donors, chronically hcv-infected pa ... | 1997 | 9210602 |
| fibroblast growth factor receptor signaling activates the human interstitial collagenase promoter via the bipartite ets-ap1 element. | interstitial collagenases participate in the remodeling of skeletal matrix and are regulated by fibroblast growth factor (fgf). a 0.2-kb fragment of the proximal human interstitial collagenase [matrix metalloproteinase (mmp1)] promoter conveys 4- to 8-fold induction of a luciferase reporter in response to fgf2 in mc3t3-e1 osteoblasts. by 5'-deletion, this response maps to nucleotides -100 to -50 relative to the transcription initiation site. the 63- bp mmp1 promoter fragment -123 to -61 confers ... | 1997 | 9212060 |
| sequence variation in the early genes e1e4, e6 and e7 of human papilloma virus type 6. | the majority of condylomata acuminata (anogenital warts) are caused by infection with human papilloma virus type 6 (hpv-6). we have sequenced the hpv-6 early genes, e1-e4, e6 and e7 from wart biopsy dna samples sourced from the uk and usa and derived a consensus sequence for these genes and the proteins they encode. when compared to the prototype hpv-6b sequence, published over 12 years ago, the e1-e4 consensus sequence showed 3/91 (3.3%) amino acid changes, the e6 consensus sequence showed 1/15 ... | 1997 | 9213393 |
| two new lignans with activity against influenza virus from the medicinal plant rhinacanthus nasutus. | two new lignans, rhinacanthin e (1) and rhinacanthin f (2), were isolated from the aerial parts of the plant rhinacanthus nasutus. their structures were established by detailed spectroscopic analysis. these compounds show significant antiviral activity against influenza virus type a. | 1997 | 9214738 |
| detection of expressed chloramphenicol acetyltransferase in the saliva of culex pipiens mosquitoes. | mosquito salivary glands play an important role in the transmission of arthropod-borne pathogens. the ability to express genes in mosquitoes would be a powerful approach to characterize salivary gland genes, and to reveal important vector determinants of pathogen transmission. here we report the use of a double subgenomic sindbis (dssin) virus, designated te/3'2j/cat, and a packaged sindbis replicon virus, designated rep5/cat/26s, to express chloramphenicol acetyltransferase (cat) protein in the ... | 1997 | 9219368 |
| presence of a neutralizing domain in isolates of rubella virus in cordoba, argentina. | we studied the presence of a neutralizing epitope of rubella virus (rv) in locally circulating strains in cordoba, argentina, using binding by the monoclonal antibody (mab) h3. this epitope is contained in a sequence of the e1 glycoprotein (e1208-239) represented by the synthetic peptide sp15. h3 mab showed specific binding to sp15 by enzyme-linked immunosorbent assay (elisa). one wild-type postnatal isolate, four clones derived from this isolate, and one congenital isolate were reactive with h3 ... | 1997 | 9220172 |
| hepatitis c virus envelope proteins bind lactoferrin. | hepatitis c virus (hcv) has two envelope proteins, e1 and e2, which form a heterooligomer. during dissection of interacting regions of hcv e1 and e2, we found the presence of an interfering compound or compounds in skim milk. here we report that human as well as bovine lactoferrin, a multifunctional immunomodulator, binds two hcv envelope proteins. as determined by far-western blotting, the bacterially expressed e1 and e2 could bind lactoferrin in human milk directly separated or immunopurified ... | 1997 | 9223490 |
| the nonstructural proteins of the hepatitis c virus: structure and functions. | the hepatitis c virus is the major causative agent of nona-nonb hepatitis worldwide. although this virus cannot be cultivated in cell culture, several of its features have been elucidated in the past few years. the viral genome is a single-stranded, 9.5kb long rna molecule of positive polarity. the viral genome is translated into a single polyprotein of about 3000 amino acids. the virally encoded polyprotein undergoes proteolytic processing by a combination of cellular and viral proteolytic enzy ... | 1997 | 9224925 |
| dna vaccination for the induction of immune responses against hepatitis c virus proteins. | recent analysis of clinical and experimental cases of hepatitis c virus (hcv) infection suggest the possible role of the viral nucléocapsid (c), the nonstructural protein 3 (ns3) and the envelope glycoproteins e1 and/or e2 in the mounting of immune responses capable to control infection (botarelli et al., gastroenterology, 1993, 104, 580-587; choo et al., proc. natl acad. sci. usa, 1994, 91, 1294-1298). we have used dna-based immunization to study the immune responses that can be induced by inje ... | 1997 | 9234532 |
| advances in adenoviral vectors: from genetic engineering to their biology. | ad2 and ad5 belong to a group of human cytolytic viruses that target the respiratory airways for reproduction, whereas latent infections establish within other tissues. signals therefore exist that control this dichotomic process in different cell types, perhaps including cis and/or trans elements of viral origin. since 1993, ad2- and ad5-based adenoviruses lacking all or part of the e1 regulatory region have been undergoing evaluation in phase i trials that target cancer and cystic fibrosis. th ... | 1997 | 9240963 |
| outbreak of hepatitis c virus infection in patients with hematologic disorders treated with intravenous immunoglobulins: different prognosis according to the immune status. | the influence of immunodeficiency on the course of hepatitis c virus (hcv) infection is still debated, although a worsening effect has been suggested. we compared the characteristics of hepatitis c in two groups of hematologic patients with different levels of immunocompetence who acquired the same virus strain after treatment with contaminated intravenous immunoglobulins (ivig). indications for ivig therapy were idiopathic thrombocytopenic purpura (itp) in six patients and hypogammaglobulinemia ... | 1997 | 9242566 |
| genomic characterization of brazilian hepatitis c virus genotypes 1a and 1b. | parts of 5' non-coding (5' nc) and of e1 envelope regions of the hepatitis c virus (hcv) genome were amplified from sera of 26 brazilian anti-hcv antibody-positive patients using the reverse transcription-polymerase chain reaction (rt-pcr). fourteen samples were pcr positive with primers from the 5' nc region and 8 of them were also positive with primers from the e1 region. a genomic segment of 176 bp from the e1 region of 7 isolates was directly sequenced from pcr products. the sequences were c ... | 1997 | 9246231 |
| human b cells secreting immunoglobulin g to glutamic acid decarboxylase-65 from a nondiabetic patient with multiple autoantibodies and graves' disease: a comparison with those present in type 1 diabetes. | antibodies to glutamic acid decarboxylase-65 (gad65) are present in a number of autoimmune disorders, such as insulin-dependent (type 1) diabetes mellitus (iddm), stiff man syndrome, and polyendocrine autoimmune disease. antibodies to gad in iddm patients usually recognize conformation-dependent regions on gad65 and rarely bind to the second isoform, glutamic acid decarboxylase-67 (gad67). in contrast, those present in stiff man syndrome and polyendocrine disease commonly target the second isofo ... | 1997 | 9253351 |
| development and characterization of a binary gene expression system based on bacteriophage t7 components in adenovirus vectors. | to explore the utility of the bacteriophage t7 binary system in adenovirus (ad) vectors we constructed three ad5-based vectors containing the t7 rna polymerase (t7pol) gene in either early region 1 (e1) or e3. the recombinant ad vectors were either deficient (adt7pol1, adt7pol2) or competent (adt7pol3) for replication in human cells other than ad5 transformed (293) cells. to test the ability of the t7 polymerase produced by these vectors to drive gene expression, a reporter vector was constructe ... | 1997 | 9256069 |
| review: molecular epidemiology of hepatitis c virus. | molecular techniques have been used to investigate the epidemiology of hepatitis c virus (hcv) at several different levels. at a global level, the time of divergence of the diverse hcv genotypes isolated from different geographical regions has been estimated from the rate of divergence observed among a cohort of individuals infected from a common source. estimates of more than 300 years for virus subtypes and more than 500-2000 years for virus types are consistent with their current geographical ... | 1997 | 9257244 |
| use of reverse-transcription polymerase chain reaction for detection of rubella virus rna in cell cultures inoculated with clinical samples. | a recently developed reverse transcription-nested polymerase chain reaction (rt-npcr) method for rubella virus (rv) rna detection was assessed in a series of african green monkey kidney (agmk) cell cultures inoculated with clinical samples from patients with suspected rv infection. results were compared with those of conventional virus isolation/identification. the assay included an internal control of amplification consisting of a synthetic rna molecule mimicking the rv e1 target sequence. a se ... | 1997 | 9258938 |
| serological and molecular analysis of hepatitis c virus envelope regions 1 and 2 during acute and chronic infections in chimpanzees. | acute and chronic hepatitis c virus infections were investigated retrospectively in chimpanzees that had been infected from a single source. anti-e1 and anti-e2 were detected in two of three chimpanzees with a chronic infection, but were first detected 1 to 2 years after inoculation. sequence evolution of the e1 region in three animals over a period of 9 to 11 years revealed a mutation rate of 1.02 to 2.23 x 10(-3) base substitutions per site per year. the acute phase viremia levels in acute inf ... | 1997 | 9260695 |
| the hepatitis c virus ns4a protein: interactions with the ns4b and ns5a proteins. | hepatitis c virus encodes a large polyprotein precursor that is proteolytically processed into at least 10 distinct products, in the order nh2-c-e1-e2-p7-ns2-ns3-ns4a-ns4b-ns5a-ns5b -cooh. a serine proteinase encoded in the n-terminal 181 residues of the ns3 nonstructural protein is responsible for cleavage at four sites (3/4a, 4a/4b, 4b/5a, and 5a/5b) in the nonstructural region. ns4a, a 54-residue nonstructural protein which forms a stable complex with the ns3 proteinase, is required as a cofa ... | 1997 | 9261364 |
| detection of serologic neutralizing antibodies against hpv-11 in patients with condyloma acuminata and cervical dysplasia using an in vitro assay. | this study was designed to investigate if neutralizing antibodies against hpv-11 are detectable in the serum of patients with condyloma acuminata (ca) or cervical intraepithelial neoplasia (cin) using an in vitro infectivity assay for hpv-11. purified hpv-11 virions were extracted from xenografted condyloma tissues implanted into athymic mice and used to infect cultured neonatal human foreskin keratinocytes (hfk) and an immortalized adult skin cell line (hacat). the presence of hpv-11-specific e ... | 1997 | 9264579 |
| herpes simplex viral vectors expressing bcl-2 are neuroprotective when delivered after a stroke. | considerable interest has focused on the possibility of using viral vectors to deliver genes to the central nervous system for the purpose of decreasing necrotic neuronal injury. to that end, we have previously shown that a herpes simplex virus (hsv) vector expressing bcl-2 could protect neurons from ischemia. in that study, vector was delivered before the ischemia. however, for such gene therapy to be of clinical use, vectors must be protective even if delivered after the onset of the insult. i ... | 1997 | 9270490 |
| persistent transgene expression in mouse liver following in vivo gene transfer with a delta e1/delta e4 adenovirus vector. | extensive in vivo gene transfer studies in animal models and human gene therapy clinical trials with e1-deleted adenovirus vectors have demonstrated transience of transgene expression due to direct cytopathic effects of the vectors and host immune response to virally expressed proteins. in order to overcome these difficulties, we have recently developed packaging cell lines which support the growth of adenovirus vectors containing lethal deletions in both e1 and e4 gene regions. here we demonstr ... | 1997 | 9274715 |
| adenovirus-mediated expression of green fluorescent protein. | a recombinant replication-deficient adenovirus has been generated that expresses a mutant of the aquorea victoria green fluorescent protein (gfp) under the control of the strong cmv promoter by insertion into the e1 region (adv-gfp). high expression of gfp was found in different cell types after infection with the recombinant virus that could be easily detected by fluorescence microscopy. in human umbilical vein endothelial cells (huvec), expression levels had already reached a maximum after 2 d ... | 1997 | 9274728 |
| characterization of an overlapping cd8+ and cd4+ t-cell epitope on rubella capsid protein. | a synthetic peptide corresponding to rubella virus capsid protein residues 263 to 275 which contains an epitope recognized by a cloned cd4+ cytotoxic t-lymphocyte (ctl) line was used to induce cd8+ t-cell lines specific to this peptide. a peptide-specific cd8+ ctl clone was derived and characterized. this peptide-specific cd8+ ctl clone exhibited cytotoxicity against target cells infected by a vaccinia recombinant virus expressing rubella virus capsid protein, but not by target cells infected by ... | 1997 | 9281508 |
| persistent echovirus infection of mouse cells expressing the viral receptor vla-2. | mouse cells are not susceptible to infection with echovirus 1 (ev-1) because they lack the viral receptor, human vla-2. two mouse fibroblast cell lines, l cells and 3t3 cells, were made susceptible to ev-1 infection after transformation with cdnas of human vla-2. after ev-1 infection, l cell transformants of human vla-2 (alpha2beta1 l cells) develop cytopathic effect (cpe) as expected, while 3t3 cell transformants of human vla-2 (alpha2beta1 3t3 cells) or the alpha2 subunit of human vla-2 (alpha ... | 1997 | 9281509 |
| molecular epidemiology of rubella by nucleotide sequences of the rubella virus e1 gene in three east asian countries. | twenty-six strains of rubella virus were compared with each other for a molecular epidemiologic study of the virus in three east asian countries, using the e1 gene of 1443 nucleotides and the following 41 nucleotides in a noncoding region. nucleotide substitution rates among strains were 0.0-9.4/100 nucleotides. a phylogenetic tree drawn indicated that 2 of 3 chinese strains were quite different from the other 24 strains; all isolates in the 1960s were classified into a single group independent ... | 1997 | 9291305 |
| characterization of truncated forms of hepatitis c virus glycoproteins. | hepatitis c virus (hcv) glycoproteins (e1 and e2) both contain a carboxy-terminal hydrophobic region, which presumably serves as a membrane anchor. when they are expressed in animal cell cultures, these glycoproteins, in both mature complexes and misfolded aggregates, are retained in the endoplasmic reticulum. the effect of carboxy-terminal deletions on hcv glycoprotein secretion and folding was examined in this study. sindbis and/or vaccinia virus recombinants expressing truncated forms of thes ... | 1997 | 9292018 |
| [a comparative phylogenetic analysis of the coding portion of the genes for structural protein e1 in the shi-min' strain and in other strains of the classic hog cholera virus]. | the primary nucleotide sequence of the fragment of e1 (gp51-55) gene (772 bp in length) of virulent strain shi-min' of classical swine fever virus (csfv) has been determined. multiple alignments of the e1 gene fragments of various strains and isolates of csfv, which are homologues to the cloned fragment, has been carried out; the phylogenetic tree has been plotted and the consensus sequence has been determined. the strain shi-min', which was used as a control csfv strain in former soviet republi ... | 1997 | 9297292 |
| characteristics of nucleotide substitution in the hepatitis c virus genome: constraints on sequence change in coding regions at both ends of the genome. | comparison of complete genome sequences for different variants of hepatitis c virus (hcv) reveals several different constraints on sequence change. synonymous changes are suppressed in coding regions at both 5' and 3' ends of the genome. no evidence was found for the existence of alternative reading frames or for a lower mutation frequency in these regions. instead, suppression may be due to constraints imposed by rna secondary structures identified within the core and ns5b genes. nonsynonymous ... | 1997 | 9302317 |
| sequence variation and phylogenetic analysis of the 5' terminus of hepatitis g virus. | we determined the nucleotide and deduced amino acid sequence of the 5' terminus of the hepatitis g virus (hgv) genome from isolates of varied geographical origins. our analysis showed that the putative 5' non-coding region (ncr) contains several blocks of highly conserved sequences that may be useful for the development of a reverse transcriptase-polymerase chain reaction (rt-pcr) assay for detection of hgv rna. overall, the degree of conservation within the 669-nucleotide (nt) 5'terminal sequen ... | 1997 | 9310927 |
| bovine papillomavirus type 1 e1 and simian virus 40 large t antigen share regions of sequence similarity required for multiple functions. | the full-length product of the bovine papillomavirus type 1 (bpv-1) e1 translational open reading frame is required for viral dna replication in vivo and in vitro. e1 is a multifunctional protein whose properties include atp binding, acting as an atpase-dependent dna helicase, dna binding to the bpv-1 origin of viral dna replication, and association with the e2 transcriptional transactivator, e2ta, a second viral protein involved in dna replication. all of these properties are thought to be impo ... | 1997 | 9311841 |
| expression of gp19k increases the persistence of transgene expression from an adenovirus vector in the mouse lung and liver. | activation of the cellular immune system and subsequent lysis of vector-transduced cells by adenovirus- or transgene-specific cytotoxic t lymphocytes have been shown to limit transgene expression in animal models. the adenovirus gp19k gene product associates with major histocompatibility complex class i proteins and prevents their maturation by sequestering them in the endoplasmic reticulum. gp19k has been shown to block the ability of adenovirus-specific cytotoxic t lymphocytes to recognize vir ... | 1997 | 9311844 |
| characterization of an endoplasmic reticulum retention signal in the rubella virus e1 glycoprotein. | rubella virus contains three structural proteins, capsid, e2, and e1. e2 and e1 are type i membrane glycoproteins that form a heterodimer in the endoplasmic reticulum (er) before they are transported to and retained in the golgi complex, where virus assembly occurs. the bulk of unassembled e2 and e1 subunits are not transported to the golgi complex. we have recently shown that e2 contains a golgi-targeting signal that mediates retention of the e2-e1 complex (t. c. hobman, l. woodward, and m. g. ... | 1997 | 9311850 |
| the formation of intramolecular disulfide bridges is required for induction of the sindbis virus mutant ts23 phenotype. | the sindbis virus envelope protein spike is a hetero-oligomeric complex composed of a trimer of glycoprotein e1-e2 heterodimers. spike assembly is a multistep process which occurs in the endoplasmic reticulum (er) and is required for the export of e1 from the er. pe2 (precursor to e2), however, can transit through the secretory pathway and be expressed at the cell surface in the absence of e1. although oligomer formation does not appear to be required for the export of pe2, there is evidence tha ... | 1997 | 9311853 |
| the nucleocapsid-binding spike subunit e2 of semliki forest virus requires complex formation with the e1 subunit for activity. | alphaviruses, such as semliki forest virus (sfv), mature by budding at the plasma membrane (pm) of infected cells and enter uninfected ones by a membrane fusion process in the endosomes. both processes are directed by the p62/e2-e1 membrane protein heterodimer of the virus. the p62 protein, or its mature form e2, provides a cytoplasmic protein domain for interaction with the nucleocapsid (nc) of the virus, and the e1 protein functions as a membrane fusogen. we have previously shown that the p62/ ... | 1997 | 9311874 |
| baculoviral display of the green fluorescent protein and rubella virus envelope proteins. | the ability to display heterologous proteins and peptides on the surface of different types of bacteriophage has proven extremely useful in protein structure/function studies. to display such proteins in a eucaryotic environment, we have produced a vector allowing for fusion of proteins to the amino-terminus of the autographa californica nuclear polyhedrosis virus (acnpv) major envelope glycoprotein, gp64. such fusion proteins incorporate into the baculoviral virion and display the flag epitope ... | 1997 | 9325155 |
| identification of strain-specific nucleotide sequences in e1 and ns4 genes of rubella virus vaccine strains in japan. | strain-specific nucleotide sequences of e1 and ns4 genes in five strains of a live rubella virus vaccine manufactured in japan were identified for comparison, using 2389 nucleotides (1443 nucleotides of the e1 gene, 41 of the 3' terminal region following the e1 gene and 905 of the ns4 gene). sequences of the e1 gene in three strains (matsuura, tcrb19 and to-336) were identified. takahashi and matsuba strains shared common sequences, but were discriminated by the sequence of the ns4 gene. these f ... | 1997 | 9330471 |
| constitutive expression of murine ctla4ig from a recombinant adenovirus vector results in prolonged transgene expression. | the administration of soluble muctla4ig around the time of adenovirus vector mediated gene transfer into murine hepatocytes has been shown to markedly prolong transgene expression, diminish the formation of adenovirus neutralizing antibody, decrease t cell proliferative response and infiltration into the liver without causing irreversible systemic immunosuppression. in this study, an e1/e3-deleted adenovirus vector constitutively expressing murine ctla4ig (ad.rsv-muctla4ig) was constructed in or ... | 1997 | 9338015 |
| a simian virus 40 large t-antigen segment containing amino acids 1 to 127 and expressed under the control of the rat elastase-1 promoter produces pancreatic acinar carcinomas in transgenic mice. | the simian virus 40 large t antigen induces tumors in a wide variety of tissues in transgenic mice, the precise tissues depending on the tissue specificity of the upstream region controlling t-antigen expression. expression of mutant t antigens that contain a subset of the protein's activities restricts the spectrum of tumors induced. others showed previously that expression of a mutant large t antigen containing the n-terminal 121 amino acids (t1-121) under control of the lymphotropic papovavir ... | 1997 | 9343166 |
| different susceptibilities of genetic variants of hepatitis c virus (hcv) to interferon (ifn). | genetic variants of hcv may have different degrees of resistance to ifn and may therefore influence the outcome of ifn therapy. however, selection of hcv variants by ifn has not been investigated in detail. in this paper, heteroduplex analysis was used to monitor major changes of hcv populations in 4 chronically infected patients under ifn therapy. we found that a major variant of the hcv 5' non-coding region (5' ncr) emerged in a responder. in other patients although no new variant of the 5' nc ... | 1997 | 9349304 |
| crystal structure of the i domain from integrin alpha2beta1. | we have determined the high resolution crystal structure of the i domain from the alpha-subunit of the integrin alpha2beta1, a cell surface adhesion receptor for collagen and the human pathogen echovirus-1. the domain, as expected, adopts the dinucleotide-binding fold, and contains a metal ion-dependent adhesion site motif with bound mg2+ at the top of the beta-sheet. comparison with the crystal structures of the leukocyte integrin i domains reveals a new helix (the c-helix) protruding from the ... | 1997 | 9353312 |
| echovirus 1 interaction with the human very late antigen-2 (integrin alpha2beta1) i domain. identification of two independent virus contact sites distinct from the metal ion-dependent adhesion site. | the human integrin very late antigen (vla)-2 (cd49b/cd29) mediates interactions with collagen and is the receptor for echovirus 1. binding sites for both collagen and echovirus 1 have been mapped to the i domain within the alpha2 subunit of the vla-2 alpha2beta1 heterodimer. although murine vla-2 interacts with collagen, it does not bind virus. we have used isolated human-murine chimeric i domains expressed as glutathione s-transferase fusion proteins in escherichia coli to identify two groups o ... | 1997 | 9353313 |
| sequence variation and phylogenetic analysis of envelope glycoprotein of hepatitis g virus. | a transfusion-transmissible agent provisionally designated hepatitis g virus (hgv) was recently identified. in this study, we examined the variability of the hgv genome by analysing sequences in the putative envelope region from 72 isolates obtained from diverse geographical sources. the 1561 nucleotide sequence of the e1/e2/ns2a region of hgv was determined from 12 isolates, and compared with three published sequences. the most variability was observed in 400 nucleotides at the n terminus of e2 ... | 1997 | 9367362 |
| oligomerization-dependent folding of the membrane fusion protein of semliki forest virus. | the spikes of alphaviruses are composed of three copies of an e2-e1 heterodimer. the e1 protein possesses membrane fusion activity, and the e2 protein, or its precursor form, p62 (sometimes called pe2), controls this function. both proteins are, together with the viral capsid protein, translated from a common c-p62-e1 coding unit. in an earlier study, we showed that the p62 protein of semliki forest virus (sfv) dimerizes rapidly and efficiently in the endoplasmic reticulum (er) with the e1 prote ... | 1997 | 9371630 |
| hepatitis c virus e2 protein purified from mammalian cells is frequently recognized by e2-specific antibodies in patient sera. | the envelope protein of hepatitis c virus (hcv) is composed of two membrane-associated glycoproteins, e1 and e2. to obtain hcv e2 protein as a secretory form at a high level, we constructed a recombinant chinese hamster ovary (cho) cell line expressing a c-terminal truncated e2 (e2t) fused to human growth hormone (hgh), cho/hghe2t. the hghe2t fusion protein was purified from the culture supernatant using anti-hgh mab affinity chromatography at approximately 80% purity. the purified hghe2t protei ... | 1997 | 9374479 |
| membrane fusion activity of semliki forest virus in a liposomal model system: specific inhibition by zn2+ ions. | semliki forest virus (sfv) has been shown previously to fuse efficiently with cholesterol- and sphingolipid-containing liposomal model membranes in a low-ph-dependent manner. several steps can be distinguished in this process, including low-ph-induced irreversible binding of the virus to the liposomes, facilitated by target membrane cholesterol, and subsequent fusion of the viral membrane with the liposomal bilayer, specifically catalyzed by target membrane sphingolipid. binding and fusion are m ... | 1997 | 9375004 |