Publications

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gene targeting in mouse embryonic stem cells with an adenoviral vector.we examined the ability of an e1, e3-defective adenoviral vector to act as a substrate for homologous recombination with chromosomal dna by including host chromosomal sequence from the mouse fgr locus that also contained a selectable marker. after infection of mouse embryonic stem cells, stable integration was selected for neomycin resistance and the efficiency of homologous recombination was evaluated. the adenoviral vector was capable of infecting mouse embryonic stem cells efficiently. betwee ...19958525428
human papillomavirus type 11 e1--e4 and l1 proteins colocalize in the mouse xenograft system at multiple time points.the most abundant viral mrna species in hpv 11-infected tissue consists of two exons, joining a segment of open reading frame (orf) e1 to orf e4, potentially encoding the e1--e4 protein. the l1 orf encodes the major capsid protein of hpv. our previous studies demonstrated colocalization of the hpv 11 e1--e4 and l1 proteins within the same cells of hpv 11-infected human foreskin implants grown in athymic mice (the mouse xenograft system) and removed 12 weeks after implantation. prior studies have ...19958525625
adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene inhibits vascular smooth muscle cell proliferation and neointima formation following balloon angioplasty of the rat carotid artery.vascular smooth muscle cell (vsmc) proliferation following arterial injury plays a critical role in a variety of vascular proliferative disorders, including atherosclerosis and restenosis after balloon angioplasty. in this study, we tested the hypothesis that localized arterial infection at the time of balloon angioplasty with an adenovirus (adv-tk) encoding the herpes simplex virus thymidine kinase gene (hsv-tk), followed by systemic ganciclovir administration, can inhibit vsmc proliferation an ...19958529096
evaluation of rubella virus e2 and c proteins in protection against rubella virus in a mouse model.an animal model is described that can provide further information for evaluating novel vaccines against rubella virus (rv). a group of mice was immunized with the lysate of insect cells infected by a recombinant baculovirus expressing e2 and c proteins of rv. another group of mice was immunized with the ra27/3 rubella vaccine. after 2 weeks, both groups of mice were challenged intramuscularly with live rv and the blood was drawn after 8, 24, 48 and 72 h. the presence of rubella challenge virus i ...19958533457
molecular modeling studies of hiv-1 reverse transcriptase nonnucleoside inhibitors: total energy of complexation as a predictor of drug placement and activity.computer modeling studies have been carried out on three nonnucleoside inhibitors complexed with human immunodeficiency virus type 1 (hiv-1) reverse transcriptase (rt), using crystal coordinate data from a subset of the protein surrounding the binding pocket region. results from the minimizations of solvated complexes of 2-cyclopropyl-4-methyl-5,11-dihydro-5h-dipyrido[3,2-b :2',3'-e][1,4] diazepin-6-one (nevirapine), alpha-anilino-2, 6-dibromophenylacetamide (alpha-apa), and 8-chloro-tetrahydro- ...19958535257
inhibition of the enteroviruses that cause acute hemorrhagic conjunctivitis (ahc) by benzimidazoles; enviroxime (ly 122772) and enviradone (ly 127123).enviradone (evird, (e)-1-[(1-methylethyl) sulfonyl]-6-(1-phenyl-1-propenyl)-1 h- benzimidazole-2-amine) and enviroxime (evirx, 2-amino-1-(isopropyl-sulfonyl)-6-benzimidazole phenyl ketone oxime) inhibited enterovirus 70 (ev70) and coxsackievirus a24 variant (ca24v) infection of conjunctival and laryngeal cells. on average, the continuous presence of 1-3 micrograms of evird or evirx/ml in cell cultures acutely infected with ev70 or ca24v inhibited virus production (> 2 log10 reduction) and 100% o ...19958540755
correlation of detectability of hepatitis c virus genome in saliva of elderly japanese symptomatic hcv carriers with their hepatic function.the hepatitis c virus (hcv) genome was sought in the saliva of 76 chronic hcv carriers (mean age nearly 60 years) in a rural japanese town, who had high serum titers of c-100 and anti-core second generation antibodies. in 27 samples (27 cases, 36%), the hcv-rna genome was detected by the reverse transcriptase - polymerase chain reaction with either of two sets of primers covering two regions of the hcv genome: the 5'noncoding region and the region encompassing the putative envelope (e1). transam ...19958557381
long-term humoral and cellular immunity induced by a single immunization with replication-defective adenovirus recombinant vector.this study examines the suitability of replication-defective adenovirus vectors for engineering recombinant vaccines. the immunological abilities and limitations of e1-deleted adenoviruses containing the lacz gene (ad-beta-gal) were investigated by examining the humoral and cellular immune responses to the beta-galactosidase protein. balb/c mice (h-2d) were given in a single injection of recombinant adenovirus. the cytotoxic t lymphocyte (ctl) response of spleen cells was evaluated. recognized t ...19958566039
a rapid method for the screening and typing of high risk hpvs using molecular biology techniques.to date over 60 different human papilloma virus (hpv) types have been described and novel hpv genomes are still being identified. the identification and taxonomy of papilloma viruses has become increasingly complex however, some types, especially hpv-16, -18 and to a lesser extent hpv-31 and -33, which are found in a high proportion of invasive cervical cancers and their metastases, are classified as high risk types for preventive reasons it is important to identify and classify the different hp ...19958572600
host dependent variation of hepatitis c virus: phylogenetic analyses.hepatitis c virus quasispecies in six patients from three families were separated by single strand conformation polymorphism analysis and by determination of nucleotide sequences of envelope regions containing the e1 gene segment and hypervariable region-1 of each quasispecies. four of the six patients had multiple quasispecies. phylogenetic analyses indicated that all quasispecies from one individual were highly homologous to each other. the homology was higher in the e1 gene segment than in hy ...19958572936
phylogenetic analysis of open reading frame 5 of field isolates of equine arteritis virus and identification of conserved and nonconserved regions in the gl envelope glycoprotein.the variation and phylogenetic relationship of open reading frame 5 (orf5) of 3 different laboratory strains of the original prototype bucyrus strain of equine arteritis virus (eav), the modified live virus vaccine (arvac, fort dodge laboratories), and 18 field isolates of eav from north america and europe were determined by comparison of their gene sequences. the viruses differed from the published sequence by between 3 (99.6% homology) and 94 (87.8%) nucleotides and by between 3 (98.8%) and 24 ...19958553578
[treatment of viral hepatitis].chronic forms of viral b,c and d hepatitis and fulminant hepatitis represent a serious healthcare problem. the study deals with the changes in the strategy in treating these diseases. during the chronic active hepatitis caused by the b hepatitis virus, the main aim of treatment is to cease multiplication of viruses, eliminate the clinical symptoms, prevent the development of cirrhosis, or the origin of hepatocellular carcinoma. the authors analyze the possibilities of the application of corticos ...19958556359
dna sequence of the deletion/insertion in early region 3 of ad5 dl309.dl309 is an adenovirus type 5 (ad5) mutant that has been extensively utilized for construction of ad5 mutants in early region 1 (e1), in developing vectors for use as viral vaccines, and in development of gene transfer vectors for gene therapy. ad5 dl309 has been useful for vector construction because of its altered xbai restriction pattern and lends itself to a variety of strategies for rescuing inserts or mutations into e1. it contains only one xbai site at 3.7 map units (m.u.) as compared to ...19958607286
hepatitis c virus genotyping by means of 5'-ur/core line probe assays and molecular analysis of untypeable samples.to test the theoretical possibility of 5'-ur mistyping between hepatitis c virus subtypes 1a and 1b, we combined a 5'-ur/core line probe assay (lipa) with a nested pcr system and retested 183 sera, previously genotyped as type 1a or 1b and originating mainly from western europe. eight percent of these were found to be wrongly subtyped. based on this method, 3 additional subtypes of type 1 were discovered (1d-1f). randomly selected european type 2 sera (n = 18) were tested with a similar type 2 5 ...19958578855
characterization of an adenovirus gene transfer vector containing an e4 deletion.we describe the construction and characterization of an adenovirus type 2 vector, ad2e4orf6, which has been modified in the e4 region to contain only open reading frame 6. when assayed in cultured cells, ad2e4orf6 virus replication is slightly delayed but viral dna synthesis, host-cell protein synthesis shut-off, and virus yield are indistinguishable from wild type. late protein synthesis is normal with the exception of fiber synthesis, which is reduced approximately 10-fold. despite the reduced ...19958590739
molecular genetic analysis of a female patient with pyruvate dehydrogenase deficiency: detection of a new mutation and differential expression of mutant gene product in cultured cells.a new 18 bp insertion mutation in the gene for the alpha subunit of pyruvate dehydrogenase (e1 alpha) was found in a female patient with congenital lactic acidaemia. cultured skin fibroblasts and epstein-barr virus-transformed lymphoblastoid cells from this patient showed decreased and normal pyruvate dehydrogenase complex (pdhc) activity, respectively. this 18 bp insertion was a de novo mutation, because it was not present in her parents. although this female patient was heterozygous for the no ...19958598635
late-onset hepatic failure: clinical features, serology and outcome following transplantation.a further series of 41 adult patients with late-onset hepatic failure was investigates with respect to aetiological factors, particularly hepatitis c and e, which have been identified since our earlier report of this condition. the increased use of transplantation and its impact on survival overall is assessed. comparison is made with 64 patients admitted over the same period with fulminant hepatic failure of non-a, non-b aetiology. screening for the hepatitis viruses revealed three cases of hep ...19958655952
vertical transmission of hiv-1 in mid-trimester gestation.between july, 1994 and march, 1995, 23 heart blood samples from fresh abortuses of hiv-1 seropositive pregnant women after elective termination of pregnancy between 18 and 25 weeks of gestation by prostaglandin e1 analogue vaginal administration were examined for polymerase chain reaction (pcr) of hiv-1 genome and p24 antigen to investigate the transplacental transfer of hiv-1 infection. all samples of fetal heart blood were positive for hiv-1 antibody (elisa), but negative for pcr and hiv-1 p24 ...19958717570
humoral immune response and natural killer activity in patients with mixed cryoglobulinemia.based on serological and molecular evidence of hepatitis c virus (hcv) infection in a significant proportion of patients with mixed cryoglobulinemia (mc), a direct association between hcv and mc has been suggested. the goal of the present study was to investigate the role played by hcv and by the immune response to the virus in the pathogenesis of mixed cryoglobulinemia.19958730486
development of cell lines capable of complementing e1, e4, and protein ix defective adenovirus type 5 mutants.the cloning capacity of currently available e1- and e3-deleted adenovirus (ad) vectors does not exceed 8 kb. to increase capacity and improve vector safety further, we have explored the possibility that early region 4 (e4) and the gene encoding protein ix (pix) might also be deleted. to generate cell lines expressing sufficient levels of e4 and pix proteins in trans in addition to e1-encoded proteins to complement mutations in these genes, we transformed 293 cells with constructs containing the ...19958664382
experimental quantification of transmission of genetically engineered pseudorabies virus.there is concern that live pseudorabies virus (prv) vaccine or prv vector vaccine strains may spread from vaccinated to unvaccinated pigs. moreover, it is feared that recombining prv vaccine strains with related vaccine or wild-type strains may lead to spread and survival of recombinant prv. to learn more about to what extent different prv vaccine strains could spread we used a previously described experimental model to study the transmission of intranasally inoculated prv mutant strains under e ...19958701591
expression of hepatitis c virus envelope proteins in transgenic mice.in an attempt to establish a model for hepatitis c virus (hcv) infection, we produced mice transgenic for the hcv envelope genes, e1 and e2, under the control of a regulatory element from hepatitis b virus. f1-generation mice from the established founders demonstrated expression of both e1 and e2 proteins as glycosylated forms in their organs including the liver. immunostaining revealed the localization of envelope proteins principally in the cytoplasm of hepatocytes around the hepatic central v ...19958847508
expression of hcv envelope proteins and the serological utility of the anti-e2 immune response.the 5' end of the hepatitis c virus (hcv) genome encodes structural proteins of the virion. the first gene encodes a highly basic core protein. immediately downstream of the core gene are regions which encode the envelope proteins (e1 and e2) of the virus. artificial expression and secretion of immunologically active envelope proteins have proven to be a substantial challenge due to the high degree of glycosylation and the existence of certain hydrophobic domains contained within these sequences ...19958875617
replication and expression of a recombinant sindbis virus in mosquitoes.a recombinant sindbis virus, te/3'2j/anti-s, containing lacrosse virus small segment cdna in antisense orientation, was inoculated into aedes triseriatus mosquitoes. virus replication and lac-anti-s rna expression were analysed temporally and spatially. te/3'2j/anti-s virus titre peaked at 5.0 log10 tcid50 in heads 6-9 days post infection (p.i.) and decreased to 3.4 log10 tcid50 by 37 days p.i. salivary glands contained 4.4 log10 tcid50 of virus 6 days p.i.; titres were lower in other organs. la ...19958825762
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 ...19959049331
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 ...19959049395
classifying hepatitis c virus genotypes.hepatitis c virus (hcv) is the major aetiological agent for blood-borne non-a, non-b hepatitis worldwide. since its discovery in 1989, at least 28 hcv genotypes have been reported, which differ by > 20% in the nucleotide sequence of the entire genome (approximately 9500 nucleotides) or the sequence of the e1 gene (576 nucleotides). different hcv genotypes have distinct geographical distributions, and may be associated with variations in viral replication and disease-inducing activity, as well as ...19959415133
a high capacity assay for inhibitors of human papillomavirus dna replication.the discovery of antiviral compounds against human papillomaviruses (hpv) has been hindered by the difficulties in culturing virus in vitro or assaying stable hpv dna replication. however, plasmids containing the hpv replication origin replicate transiently upon co-transfection with hpv e1 and e2 expression vectors. we have adapted this assay using secreted alkaline phosphatase (sap) as a reporter for rapid analysis of dna copy number. use of the sv40 early promoter in controlling sap expression ...19959636294
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 ...19969044743
use of bacterially expressed gst/ebna-1 fusion proteins for detection of antibodies in sera from patients with nasopharyngeal carcinoma and healthy donors.epstein-barr virus nuclear antigen-1 (ebna-1) is a protein expressed consistently in ebv infected cells and in ebv related malignant tissues. antibodies against ebna-1 may therefore possibly be used as a marker for disease screening. western blot analysis of serum antibodies was performed using gst (glutathione-s-transferase) fusion proteins containing different regions of ebna-1 as antigens. serum samples were collected from 38 patients with nasopharyngeal carcinoma (npc) and 38 healthy individ ...199610592788
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 ...19969099921
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 ...19969156798
genotype determination of hepatitis c virus from northern india: identification of a new subtype.hepatitis c virus (hcv) shows substantial nucleotide sequence diversity distributed throughout the viral genome, with many variants showing only 68-79% overall sequence homology. this has led to problems in diagnosis of hcv using commercial immunoassays. based on clustering of homologous sequences, various genotypes and subtypes of hcv have been described from different geographical regions. in the present study, 11 isolates from india were genotyped using sequence comparison for part of the non ...19968835354
processing pathways of the hepatitis c virus proteins.hepatitis c virus (hcv) is the major etiological agent of posttransfusion and community-acquired non-a, non-b hepatitis. it is an enveloped virus, grouped as a separate genus in the flaviviridae family. the plus-stranded rna genome encodes a polyprotein of about 3000 amino acids with the structural proteins core, e1 and e2 residing in the amino terminal quarter of the polyprotein and the nonstructural proteins ns2, ns3, ns4a, ns4b, ns5a and ns5b in the remainder. maturation of the structural pro ...19968836884
production of interleukin-2 in response to synthetic peptides from hepatitis c virus e1 protein in patients with chronic hepatitis c: relationship with the response to interferon treatment.the role of cellular immunity in the clearance of hepatitis c virus after interferon therapy has not yet been elucidated. here, we analyzed the t cell response to peptides from hepatitis c virus e1 protein in untreated and interferon-treated patients with chronic hepatitis c virus infection.19968836894
e7 protein of human papilloma virus-16 induces degradation of retinoblastoma protein through the ubiquitin-proteasome pathway.rb protein is a critical regulator of entry into the cell cycle, and loss of rb function by deletions, mutations, or interaction with dna viral oncoproteins leads to oncogenic transformation. we have shown that the human papilloma virus (hpv)-16 e7 gene is sufficient to induce the immortalization of mammary epithelial cells (mecs). surprisingly, the steady-state level of rb protein in these immortal cells was drastically decreased. here, we used pulse-chase analysis to show that the in vivo loss ...19968840974
suicide gene expression induced in tumour cells transduced with recombinant adenoviral, retroviral and plasmid vectors containing the erbb2 promoter.in order to exploit the tumour-specific nature of erbb2 expression for genetic prodrug-activation therapy, we have generated recombinant adenoviral, retroviral and plasmid vectors containing an expression cassette consisting of the erbb2 promoter and herpes simplex virus thymidine kinase coding sequence. in the case of the adenoviral vectors, the expression cassette was introduced into the e1 or e3 region of the genome. all of the vectors were capable of sensitizing erbb2-positive cells to the a ...19968986436
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 ...19968987625
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 ...19968991639
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 ...19969000092
nosocomial transmission of hepatitis c virus in haemodialysis patients.a systematic virological follow-up of 114 haemodialysis patients treated in the same unit showed that 37, including 17 pcr positive patients, were seropositive for hepatitis c virus (hcv). type 1b hcv was detected in 10 patients and was much more frequent in this population than in the whole population of patients treated in the hepatogastroenterology departments in southeastern france. the e1/e2 genomic region of seven type 1b hcv strains was sequenced. in four patients, a similar strain was de ...19968877762
humoral response after administration of e1-deleted adenoviruses: immune privilege of the subretinal space.an important limitation of e1-deleted recombinant adenoviruses in gene therapy is the immune response that they engender and that rapidly destroys transduced cells. transduced cells of the outer retina appear to be an exception. to determine whether differences in immune sequestration of the outer retina contribute to the increased stability of transgene expression in this tissue, we examined the systemic humoral response to an e1-deleted adenovirus injected into the subretinal space. subretinal ...19968886847
nucleotide sequence analysis of a major antigenic domain of the e1 glycoprotein of 22 rubella virus isolates.we have determined the nucleotide sequence of the region of the rubella virus genome which encodes amino acids 195-296 of the e1 glycoprotein (e1-195-296) from a panel of 22 rubella viruses obtained from europe, usa and asia between 1963-1995. e1-195-296 contains neutralizing and haemagglutinating determinants, and may represent a major antigenic domain. the nucleotide sequence divergence of the 22 rubella viruses compared to the therien strain sequence ranged from 0.65-7.14%. the greatest seque ...19968887486
interactions between pe2, e1, and 6k required for assembly of alphaviruses studied with chimeric viruses.during the assembly of alphaviruses, a preassembled nucleocapsid buds through the cell plasma membrane to acquire an envelope containing two virally encoded glycoproteins, e2 and e1. using two chimeric viruses, we have studied interactions between e1, e2, and a viral peptide called 6k, which are required for budding. a chimeric sindbis virus (sin) in which the 6k gene had been replaced with that from ross river virus (rr) produced wild-type levels of nucleocapsids and abundant pe2/e1 heterodimer ...19968892914
bovine viral diarrhea virus: characterization of a cytopathogenic defective interfering particle with two internal deletions.molecular characterization of bovine viral diarrhea virus pair 13 revealed that isolate cp13 is composed of a cytopathogenic (cp) defective interfering particle (di13) and a noncytopathogenic (noncp) helper virus. the di13 genome possesses two internal deletions of 1,611 and 3,102 nucleotides. except for a small fragment of the gene coding for glycoprotein e1, all structural protein genes are deleted together with most of the npro gene, the region coding for nonstructural proteins p7 and ns2. wh ...19968892949
sequence heterogeneity within the 5'-terminal region of the hepatitis gb virus c genome and evidence for genotypes.gb virus c is a positive-strand rna virus that is associated with hepatitis in humans. gb virus c bears some resemblance to hepatitis c virus in its genomic sequence and organization. however, unlike hepatitis c virus, an open reading frame possessing a complete core protein was not identified in the original isolate.19968895018
purification and in vitro-phospholabeling of secretory envelope proteins e1 and e2 of hepatitis c virus expressed in insect cells.the putative envelope glycoproteins of hepatitis c virus (hcv), e1 and e2, were expressed as recombinant, secretory proteins in sf9 insect cells through infection with recombinant baculoviruses. the influenza virus hemagglutinin signal sequence (hass) was inserted upstream of the hcv-cdnas in order to effect secretion. furthermore, a hexa-histidine tag for purification on a ni(2+)-nitrilotriacetic acid (ni(2+)-nta) column and a protein kinase a (pka) recognition sequence for in vitro-phospholabe ...19968896240
hepatitis c virus (hcv) subtype prevalence in chiang mai, thailand, and identification of novel subtypes of hcv major type 6.subtype analysis of hepatitis c viruses (hcvs) obtained from patients with chronic liver disease in chiang mai, thailand, was performed. of 46 hcv isolates, 13 (28%) were shown to belong to hcv subtype 3a (hcv-3a), 10 (22%) to belong to hcv-1a, 7 (15%) to belong to hcv-1b, 1 (2%) to belong to hcv-3b, and 1 (2%) to belong to a variant group, as determined from partial nucleotide sequences of the ns5b region of the viral genome. analysis of 5' untranslated region sequences identified five other is ...19968904416
antibody responses to hepatitis c envelope proteins in patients with acute or chronic hepatitis c.antibody responses to the hepatitis c virus (hcv) envelope proteins e1 and e2 were analyzed using two original assays in sera from 86 patients in different stages of disease. a western blot assay and an immunofluorescence assay (ifa) were developed using envelope proteins produced, respectively, in escherichia coli and in cv1 cells infected with a recombinant sv40. as a third method, the inno-lia hcv ab iii assay including e2 synthetic peptides was used. of 38 chronically infected patients posit ...19968915882
structure, genomic organization, replication and variability of hepatitis c virus.hepatitis c virus (hcv) is an enveloped, single-stranded rna virus that has been classified in the flaviviridae family. the genome of 9400 nucleotides comprises two non-coding regions in 5' and 3' flanking a large reading frame which codes for a polyprotein of 3000 amino acids; this polyprotein is further cleaved into structural (c, e1, e2) and non-structural (ns1, ns2, ns3, ns4, ns5) proteins. the positive rna acts as a cap-independent messenger; the transcription is mediated by the ns5 rna pol ...19968918741
characterization of the hla-restrictive elements of a rubella virus-specific cytotoxic t cell clone: influence of hla-dr4 beta chain residue 74 polymorphism on antigenic peptide-t cell interaction.the influence of glutamic acid (e)-alanine (a) dimorphism at position 74 of the dr4 beta chain on cytotoxic t cell recognition of an antigenic rubella virus peptide, e1(273-284), was studied using a panel of b cell lines and b cell transfectants expressing different hla-drb1 alleles as antigen-presenting cells and targets in 51cr-release assays. only b cell lines expressing the drb1*0403, drb1*0406 or drb1*0407 subtypes which shared a residue, e, at position 74 in the dr4 beta chain when sensiti ...19968921437
classical swine fever virus (csfv) envelope glycoprotein e2 containing one structural antigenic unit protects pigs from lethal csfv challenge.envelope glycoprotein e2, formerly called e1 or gp51-54, of classical swine fever virus (csfv) expressed in insect cells protects swine from classical swine fever. monoclonal antibodies directed against epitopes of domains b and c and subdomain a1 are neutralizing. the domains are located on two structural antigenic units in a proposed model of the antigenic structure of e2. one unit consists of nonconserved antigenic domains b and c and the other contains highly conserved antigenic domain a. we ...19968922467
[sindbis viruses of various geographic origin and differentiation of them from western equine encephalomyelitis viruses using the polymerase chain reaction].comparison of sindbis virus strains isolated in different regions of the world (in africa, australia, and europe, including russia and its nearest neighbors) in the polymerase chain reaction (pcr) by the primary gene structure of proteins nsp1 and e1 and in the neutralization test showed the greatest similarity between geographically close strains isolated in northern europe (kfl, karelia, 1381 and 1388, estonia). sindbis strains ar339 and babanki isolated in africa were similar to each other an ...19968928504
a helper-dependent adenovirus vector system: removal of helper virus by cre-mediated excision of the viral packaging signal.adenoviruses are attractive vectors for the delivery of foreign genes into mammalian cells for gene therapy. however, current vectors retain many viral genes that, when expressed at low levels, contribute to the induction of a host immune response against transduced cells. we have developed a helper-dependent packaging system for production of vectors that have large regions of the genome deleted. helper viruses were constructed with packaging signals flanked by loxp sites so that in 293 cells t ...19968942974
hepatitis a vaccination by intracutaneous low dose administration: a less expensive alternative.we investigated the immune response to three different intracutaneous (i.c.) doses of inactivated hepatitis a vaccine: 72, 144, and 216 elisa units (eu). the response was measured using a quotient score derived from a commercial enzyme-linked immunosorbent assay (havab abbott) and translated to iu per liter using a world health organization standard serum for hepatitis a virus antibody. the results were compared with the results obtained after an intramuscular (i.m.) full dose, i.e. 1,440 eu, at ...19968953668
[similarities and differences between western equine encephalomyelitis viruses with respect to genes for nonstructural protein nsp2 and structural proteins c and e2].genetic relationships of geographical isolates of the members of wee virus serocomplex (mcmillan, fort morgan, highlands j, and y62-33) were assessed by the polymerase chain reaction (pcr) and restriction analysis of the pcr products. oligonucleotide primers (21 nucleotides in length) were chosen for nsp2, nucleocapsid c, and e2-e1 protein genes based on the known primary structure of the mcmillan 16310-5614 genome (l. uryvayev et al., 1994, 1995). these primers were shown to differentiate well ...19968967065
biology of adenovirus vectors with e1 and e4 deletions for liver-directed gene therapy.recombinant adenoviruses with e1 sequences deleted efficiently transfer genes into a wide variety of target cells. antigen- and nonantigen-specific responses to the therapy lead to toxicity, loss of transgene expression, and difficulties with vector readministration. we have created new cell lines that allowed the isolation of more disabled adenovirus vectors that have both e1 and e4 deletions. studies with murine models of liver-directed gene therapy indicated that the e1- and e4-deleted vector ...19968971023
recombinant adenoviruses with large deletions generated by cre-mediated excision exhibit different biological properties compared with first-generation vectors in vitro and in vivo.in vivo gene transfer of recombinant e1-deficient adenoviruses results in early and late viral gene expression that elicits a host immune response, limiting the duration of transgene expression and the use of adenoviruses for gene therapy. the prokaryotic cre-lox p recombination system was adapted to generate recombinant adenoviruses with extended deletions in the viral genome (referred to here as deleted viruses) in order to minimize expression of immunogenic and/or cytotoxic viral proteins. as ...19968971024
transcriptional targeting of recombinant adenoviruses to human and murine melanoma cells.one potential avenue for future cancer therapy involves the specific targeting of effector genes to cancer cells throughout the body, including distant metastatic sites. as a first step toward this goal, we tested the ability of the transcriptional regulatory elements of the human and mouse tyrosinase genes to promote high levels of pigment cell-specific transcription. a construct consisting of 209 bp of the human tyrosinase promoter linked to two enhancer elements was demonstrated to drive high ...19968971169
glycoproteins e2 of the venezuelan and eastern equine encephalomyelitis viruses contain multiple cross-reactive epitopes.enzyme immunoassay (eia) with sixty types of monoclonal antibodies (mabs) was used to study cross-reactive epitopes on the attenuated and virulent strains of the eastern equine encephalomyelitis (eee) and venezuelan equine encephalomyelitis (vee) viruses. all three structural proteins of the eee and vee viruses were demonstrated to have both cross-reactive and specific antigenic determinants. the glycoprotein e1 of eee and vee viruses possesses three cross-reactive epitopes for binding to mabs. ...19968973533
quantitative and in vivo activity of adenoviral-producing cells made by cotransduction of a replication-defective adenovirus and a replication-enabling plasmid.achieving limited recombinant viral replication may provide a means of amplifying viral-mediated gene transfer in vivo. we have previously shown that cotransduction of an e1-defective adenovirus with a plasmid containing the deleted e1 functions into prostate carcinoma cells resulted in e1-defective virus production by those cells. the studies described here have extended these findings to more firmly establish the capacity of the trans complementation approach to achieve amplification of recomb ...19968853547
t- and b-cell responses to different hepatitis c virus antigens in patients with chronic hepatitis c infection and in healthy anti-hepatitis c virus--positive blood donors without viremia.as the host's immune response may determine the course of hepatitis c virus (hcv) infection, we studied the humoral and cellular immune responses to hcv-related antigens in subjects with different outcomes of hcv infection. lymphoproliferative responses and circulating antibodies to a panel of hcv core- and e1-related 25-mer peptides were examined in 10 healthy anti-hcv-seropositive blood donors (group a) and in 29 patients with chronic hepatitis c (group b). in addition, cellular recognition of ...19968855177
hepatitis c virus core protein: carboxy-terminal boundaries of two processed species suggest cleavage by a signal peptide peptidase.the expression and processing of hepatitis c virus core protein was analyzed. two protein bands, 21 kda (p21), corresponding to the full-length core, and 19 kda (p19), were detected as major products when core protein was expressed in the standard rabbit reticulocyte lysate system or in sf9 insect cells. core proteins with amino-terminal hexa-histidine tags were expressed which allowed the purification of the hexa-histidine p19 core with ni(2+)-nta columns. with the help of mass spectrometry, th ...19968862403
rubella reimmunization: comparative analysis of the immunoglobulin g response to rubella virus vaccine in previously seronegative and seropositive individuals.rubella virus (rv)-specific immunoglobulin g (igg) antibodies were studied in military recruits undergoing unselected immunization with live attenuated measles, mumps, and rubella virus (mmr) vaccine. three different whole-rv enzyme immunoassays (eias) and an epitope-specific eia with a synthetic peptide (bch-178c) representing a heutralization domain on the rv e1 envelope protein were used. before vaccination, 84.2, 87.7, and 84.5% of the subjects tested (n = 399) were found to be seropositive ...19968862587
adenoviral gene transfer of macrophage inflammatory protein-2 in rat lung.replication-defective adenoviral vectors are capable of localized transfer and expression of incorporated gene product in lung tissue. we have constructed an adenoviral vector that expresses rat macrophage inflammatory protein (mip)-2, a c-x-c chemokine specifically chemotactic for neutrophils, supernatants from 293 cells, infected with the adenoviral mip-2 (admip-2) construct, showed potent chemotactic activity and the ability of the admip-2 vector to transcribe and make functional protein was ...19968863686
murine humoral immune response against recombinant structural proteins of hepatitis c virus distinct from those of patients.we examined the humoral immune response to recombinant structural proteins of hepatitis c virus (hcv) such as c, e1 and e2 in immunized mice. mice showed high induction of antibodies against these three structural proteins. conformational and/or linear epitopes of these regions showed high responses in mice. comparison with patients revealed higher anti-e1 and anti-e2 responses in mice and 15 immunoreactive peptides which are unique to mice, especially 11 peptides from the e2 region. the hydroph ...19968867614
lymphocyte proliferative responses to recombinant hepatitis c virus antigens in patients with chronic hepatitis c.the purpose of the present study was to analyse lymphocyte proliferative responses to recombinant hepatitis c virus (hcv) antigens in chronic hepatitis c. four recombinant peptides derived from the ns3, core, e1 and e2/ns1 regions of the hcv genome were used as antigens in lymphocyte proliferative responses. forty-two patients, classified into various sub-groups, and 17 healthy control subjects were tested and the specific response was expressed as a stimulation index. responses were analysed wi ...19968872764
abortive infection of the baculovirus autographa californica nuclear polyhedrosis virus in sf-9 cells after mutation of the putative dna helicase gene.homologous recombination between the autographa californica nuclear polyhedrosis virus (acnpv) genome and a 0.6-kbp-long dna fragment derived from the putative dna helicase gene of bombyx mori nuclear polyhedrosis virus generates eh2-acnpv, an expanded-host-range acnpv mutant (s. maeda, s.g. kamita, and a. kondo, j. virol. 67:6234-6238, 1993). after inoculation at a high multiplicity of infection (moi), eh2-acnpv replicates efficiently in both the sf-9 (acnpv-permissive) and bmn (non-acnpv-permi ...19968709251
a gene transfer vector-cell line system for complete functional complementation of adenovirus early regions e1 and e4.the improvements to adenovirus necessary for an optimal gene transfer vector include the removal of virus gene expression in transduced cells, increased transgene capacity, complete replication incompetence, and elimination of replication-competent virus that can be produced during the growth of first-generation adenovirus vectors. to achieve these aims, we have developed a vector-cell line system for complete functional complementation of both adenovirus early region 1 (e1) and e4. a library of ...19968709289
usefulness and limitation of phylogenetic analysis for hepatitis c virus core region: application to isolates from egyptian and yemeni patients.we report here the nucleotide sequences of the core region of hcv isolates from egyptian and yemeni patients and the method for classifying these hcv isolates by phylogenetic analysis. sequence comparison suggested that the genotypes of these isolates were the same. preliminary phylogenetic analysis of the hcv core region indicated that the genotypes of both isolates were 1c. however, an additional phylogenetic tree of the hcv core region constructed using a greater number of hcv isolates than t ...19968712927
agonist-stimulated free calcium in subcellular compartments. delivery of recombinant aequorin to organelles using a replication deficient adenovirus vector.changes in the concentration of calcium ions ([ca2+]) within cellular organelles play a central role in controlling cellular function. we have engineered the ca2+ sensitive photoprotein aequorin to monitor selectively [ca2+] within defined subcellular compartments, namely the cytosol, nucleus and endoplasmic reticulum. dna encoding the engineered aequorins have been inserted into a replication deficient adenovirus (ad) type 5 e1-vector, under control of the cytomegalovirus (cmv) major immediate ...19968689671
different clinical behaviors of acute hepatitis c virus infection are associated with different vigor of the anti-viral cell-mediated immune response.the anti-viral t cell response is believed to play a central role in the pathogenesis of hepatitis c virus infection. since chronic evolution occurs in > 50% of hcv infections, the sequential analysis of the t cell response from the early clinical stages of disease may contribute to define the features of the t cell response associated with recovery or chronic viral persistence. for this purpose, 21 subjects with acute hepatitis c virus infection were sequentially followed for an average time of ...19968698862
investigation of murine cytomegalovirus latency and reactivation in mice using viral mutants and the polymerase chain reaction.studies with 6 ts mutants of mouse cytomegalovirus indicated that mutants tsm1, tsm2, tsm3, and tsm6, like wild-type (wt) virus, produced acute infection in mice, became latent, and were reactivated as infectious virus immunosuppression. using pcr, all five viruses expressed immediate-early (ie)-1, early (e)-1, and late (l, gb) genes during acute infection in all tissues examined (salivary glands, lung, spleen, liver, kidney, and heart). dna was present in most tissues during latent infection wi ...19968699162
visualization of hepatitis c virions and putative defective interfering particles isolated from low-density lipoproteins.hepatitis c virus (hcv) in highly infectious sera has been shown to be predominantly associated with low-density lipoproteins. to determine whether the association is specific to low-density lipoproteins (ldl) or very low-density lipoproteins (vldl), we fractionated hcv-containing plasma by a column chromatographic procedure known to separate these classes. hepatitis c virus rna detected by polymerase chain reaction (pcr) was associated primarily with the very low-density (vldl) fraction. howeve ...19968736235
molecular cloning and nucleotide sequence determination of three envelope genes of classical swine fever virus taiwan isolate p97.a strain of classical swine fever virus (csfv) has been isolated in taiwan. the cdna coding for three envelope glycoproteins e1, e2 and e3 were molecularly cloned from purified viral particles using the reverse transcription-polymerase chain reaction (rt-pcr) method and sequence-specific primers. the resulting pcr products (1113 bp for e1. 699 bp for e2 and 567 bp for e3) were cloned into the smai site of puc19 and then subjected to dna sequence analysis. data showed that nucleotide sequence of ...19968738176
characterization of the helicase and atpase activity of human papillomavirus type 6b e1 protein.human papillomavirus type 6b (hpv-6b) is one of the most common causes of human genital warts, an important sexually transmitted disease. discovery of antiviral therapies for this condition has been hampered by the inability to propagate the virus using standard tissue culture techniques and through difficulties in expressing sufficient recombinant viral proteins in vitro. replication of papillomavirus dna requires two viral proteins, e1 and e2. in an effort to establish assays to discover compo ...19968760430
cytoplasmic tail length influences fatty acid selection for acylation of viral glycoproteins.we report remarkable differences in the fatty acid content of thioester-type acylated glycoproteins of enveloped viruses from mammalian cells. the e2 glycoprotein of semliki forest virus contains mainly palmitic acid like most other palmitoylated proteins analysed so far. however, the other glycoprotein (e1) of the same virus, as well as the hef (haemagglutinin esterase fusion) glycoprotein of influenza c virus, are unique in this respect because they are acylated primarily with stearic acid. co ...19968761467
interaction between hepatitis c virus core protein and e1 envelope protein.hepatitis c virus has three structural genes named c, e1, and e2. the c gene encodes the core (capsid) protein and the e1 and e2 genes encode the envelope proteins. in an immunoprecipitation experiment, the e1 protein was found to be precipitated by an anti-core antibody in the presence but not in the absence of the core protein, indicating that the e1 protein can interact with the core protein. this interaction is independent of whether the e1 and the c genes are linked in cis or separated in d ...19968764026
antisense oligonucleotide inhibition of hepatitis c virus gene expression in transformed hepatocytes.genetic and biochemical studies have provided convincing evidence that the 5' noncoding region (5' ncr) of hepatitis c virus (hcv) is highly conserved among viral isolates worldwide and that translation of hcv is directed by an internal ribosome entry site (ires) located within the 5' ncr. we have investigated inhibition of hcv gene expression using antisense oligonucleotides complementary to the 5' ncr, translation initiation codon, and core protein coding sequences. oligonucleotides were evalu ...19968764029
disulfide bridge-mediated folding of sindbis virus glycoproteins.the sindbis virus envelope is composed of 80 e1-e2 (envelope glycoprotein) heterotrimers organized into an icosahedral protein lattice with t=4 symmetry. the structural integrity of the envelope protein lattice is maintained by e1-e1 interactions which are stabilized by intramolecular disulfide bonds. structural domains of the envelope proteins sustain the envelope's icosahedral lattice, while functional domains are responsible for virus attachment and membrane fusion. we have previously shown t ...19968764067
hepatitis c virus infection. biological and immunological features.the recent cloning and genomic identification of hepatitis c virus (hcv) by sensitive and specific immune techniques has allowed a better definition of both histopathological and clinical features of the previously not well defined non-a, non-b hepatitis. in this regard, antibodies to different hcv antigens are usually found during infection, even if some of them such as anti-e1 and anti-e2/ns1 have been shown to be associated with significant viraemic levels. acute hepatitis c is self-limiting ...19968766958
mechanisms of mutations inhibiting fusion and infection by semliki forest virus.semliki forest virus (sfv) infects cells by an acid-dependent membrane fusion reaction catalyzed by the virus spike protein, a complex containing e1 and e2 transmembrane subunits. e1 carries the putative virus fusion peptide, and mutations in this domain of the spike protein were previously shown to shift the ph threshold of cell-cell fusion (g91a), or block cell-cell fusion (g91d). we have used an sfv infectious clone to characterize virus particles containing these mutations. in keeping with t ...19968769412
analysis of hepatitis c virus isolates by serotyping and genotyping.hepatitis c virus (hcv)-positive sera from 106 chronically infected patients which had previously been genotyped were characterized by serotyping. genotypes were determined by a first-generation line probe assay (inno-lipa hcv) and by sequence analyses of the core, core-e1, and ns5b regions. hcv serotypes were determined by measuring type-specific antibodies to ns4-derived peptide antigens (murex hcv serotyping 1-6 assay). of 106 serum samples, serotype-specific antibodies were detected in 88 (s ...19968784590
dystrophin expression in muscles of mdx mice after adenovirus-mediated in vivo gene transfer.we have generated high-titer adenoviral recombinants (avr) expressing a 6.3-kb partial dystrophin cdna insert under the control of either the rous sarcoma virus (rsv) or cytomegalovirus (cmv) promoter. these avr preparations were free of both e1-containing avr and avr with a nonfunctional dystrophin expression cassette. with these optimal avr preparations, we have obtained a high degree of short-term (10 days) expression of a truncated (approximately 200 kd) dystrophin in dystrophin-deficient md ...19968788164
immunoreactive core peptides of hepatitis c virus produced in escherichia coli and in vitro dna amplification-restricted transcription-translation system.three kinds of hepatitis c virus (hcv) core peptides were produced directly and efficiently in e. coli: 1-120 aa of the c region as ncc, 1-157 aa as ncct and 1-190 aa as nccl. these peptides were estimated to be 16, 22 and 24 kda, respectively, by sds-polyacrylamide gel electrophoresis. the processing to produce p22 core protein observed in insect cells and mammalian systems did not occur in e. coli. these peptides were similarly reactive with serum antibody from patients with hepatitis c. a mut ...19968793834
development of a complementing cell line and a system for construction of adenovirus vectors with e1 and e2a deleted.although adenovirus vectors offer many advantages, it would be desirable to develop vectors with improved expression and decreased toxicity. toward this objective, an adenovirus vector system with deletion of both the el and e2a regions was developed. a 5.9-kb fragment of the adenovirus type 5 (ad5) genome containing the e2a gene and its early and late promoters was transfected into 293 cells. a complementing cell line, designated 293-c2, expressed the e2a mrna and protein and was found to compl ...19968794347
role of viral antigens in destructive cellular immune responses to adenovirus vector-transduced cells in mouse lungs.adenoviruses missing e1 have been used as gene delivery vectors to the lungs for the treatment of cystic fibrosis. transient expression of the recombinant gene and the development of inflammation have been two major limitations to the application of first-generation recombinant adenoviruses for gene therapy. studies with mouse models of liver- and lung-directed gene therapy suggested that cd8+ cytotoxic t lymphocytes (ctls) are effectors that contribute to extinction of transgene expression. the ...19968794368
cell surface expression of a functional rubella virus e1 glycoprotein by addition of a gpi anchor.rubella virus (rv) envelope glycoproteins e1 and e2 are targeted to the golgi as heterodimers. while e2 contains a transmembrane golgi retention signal, e1 is arrested in a pre-golgi compartment in the absence of e2, and appears to require heterodimerization in order to reach the golgi. various forms of e1 with deletions in the ectodomain or lacking the cytoplasmic (ct) and transmembrane (tm) domains, as well as the 29 c-terminal amino acid residues of the ectodomain were also retained intracell ...19968799810
structural requirements for initiation of translation by internal ribosome entry within genome-length hepatitis c virus rna.cap-independent translation of hepatitis c virus (hcv) rna is mediated by an internal ribosomal entry segment (ires) located within the 5' nontranslated rna (5'ntr), but previous studies provide conflicting views of the viral sequences which are required for translation initiation. these discrepancies could have resulted from the inclusion of less than full-length 5'ntr in constructs studied for translation or destabilization of rna secondary structure due to fusion of the 5'ntr to heterologous ...19968806485
virus-like particles and e1-e4 protein expressed from the human papillomavirus type 11 bicistronic e1-e4-l1 transcript.detection of e1-e4 protein in human papillomavirus (hpv 11)-infected tissue is tightly linked to detection of l1 major capsid protein. the only l1-containing transcript identified in hpv 11-infected tissue is the bicistronic e1-e4-l1 mrna, potentially encoding both the e1-e4 and the l1 proteins. it has not been established that these proteins can be expressed from the e1-e4-l1 transcript. the hpv 11 e1-e4-l1 sequence was cloned by reverse transcriptase polymerase chain reaction into the p1393 ve ...19968806486
characterization of cell lines allowing tightly regulated expression of hepatitis c virus core protein.a tetracycline-regulated system was used to generate cell lines allowing tightly controlled expression of a hepatitis c virus (hcv) cdna comprising the 5' noncoding, the core, and part of the e1 regions. production of 21-kda processed nucleocapsid protein could be regulated over a broad range by the concentration of tetracycline present in the culture medium. induction ratios of over 1,000-fold were found using an hcv core-luciferase fusion construct. core protein had an intracellular half-life ...19968806487
induction of systemic and mucosal immune responses in cotton rats immunized with human adenovirus type 5 recombinants expressing the full and truncated forms of bovine herpesvirus type 1 glycoprotein gd.we generated both replication-incompetent (had5-gd-e1 and had5-tgd-e1) and replication-competent (had5-gd-e3 and had5-tgd-e3) human adenovirus type 5 (had5) recombinants expressing the full (gd) or truncated form (tgd) of the glycoprotein gd gene of bovine herpevirus type 1 (bhv-1). recombinant gd and tgd expressed by had5-gd-e1 and had5-gd-e3 and by had5-tgd-e1 and had5-tgd-e3, respectively, were recognized by gd-specific monoclonal antibodies (mabs) directed against linear and conformational e ...19968806514
genotypes and multiple infections with hepatitis c virus in patients with haemophilia a in japan.hepatitis c virus (hcv) rna was tested for, and hcv genotypes determined, in 96 patients with haemophilia a in japan. of 88 patients aged > or = 10 years, 74 (84%) were positive for hcv rna at a frequency higher than that in patients aged less than 10 years (one of eight, 13%, p < 0.001). genotype i/1a was detected in 30(40%), ii/1b in 12 (16%), iii/2a in eight (11%), iv/2b in five (7%) and v/3a in 12 (16%); mixed infection with hcv of two different genotypes was identified in the remaining nine ...19968811642
preferential retention of the e6 and e7 regions of the human papilloma virus type 18 genome by human sperm cells.to determine if sperm cells differentially take up or retain different regions of human papilloma virus (hpv) type 18 genome.19968816629
encapsidated adenovirus minichromosomes allow delivery and expression of a 14 kb dystrophin cdna to muscle cells.adenovirus-mediated gene transfer to muscle is a promising technology for gene therapy of duchenne muscular dystrophy (dmd). however, currently available recombinant adenovirus vectors have several limitations, including a limited cloning capacity of approximately 8.5 kb, and the induction of a host immune response that leads to transient gene expression of 3-4 weeks in immunocompetent animals. gene therapy for dmd could benefit from the development of adenoviral vectors with an increased clonin ...19968817325
comparison of the protective efficacy of recombinant pseudorabies viruses against pseudorabies and classical swine fever in pigs; influence of different promoters on gene expression and on protection.the glycoprotein e (ge) locus in the genome of pseudorabies virus (prv) was used as an insertion site for the expression of glycoprotein e1 of classical swine fever virus (csfv). transcription of e1 in the recombinants m401, m402 or m403 was regulated by the gd promoter of prv, the immediate early gene promoter of human cytomegalovirus, or the ge promoter of prv, respectively. groups of four pigs were vaccinated once intramuscularly with 10(6) plaque forming units (p.f.u.) of the recombinant vir ...19968821642
induction of antibodies against structural proteins of hepatitis c virus in mice using recombinant adenovirus.replication-deficient recombinant adenoviruses expressing structural proteins of hepatitis c virus (hcv) were constructed. each recombinant lacks adenoviral e1a and e3 genes and bears expression units for hcv structural proteins. the expression units contain hcv cdnas coding for either the protein or core, one of two envelopes (e1 and e2) or all of these structural proteins (core, e1 and e2) under the control of the sr alpha promoter. in hela or hepg2 cells, the recombinants can express efficien ...19968821646
induction of apoptosis in murine clonal osteoblasts expressed by human t-cell leukemia virus type i tax by nf-kappa b and tnf-alpha.we investigated the effects of various cytokines in the presence of human t-cell leukemia virus type i (htlv-i) tax protein in murine clonal osteoblasts, mc3t3-e1 cells. skeletal remodeling by osteoclasts and osteoblasts is coordinated by cytokines, which are activated by htlv-i tax protein via nuclear factor-kappa b (nf-kappa b). mc3t3-e1 cells were cocultured with an irradiated htlv-i-producing lymphocyte cell line, mt-2. after coculture, the tumor necrosis factor-alpha (tnf-alpha) level in th ...19968822344
coupled pcr-restriction enzyme analysis for rapid identification of structural gene relationships among strains of eastern equine encephalitis virus.we have used restriction endonuclease digestion analysis of polymerase chain reaction (pcr)-amplified gene regions to rapidly examine individual structural gene relationships among field isolates of eastern equine encephalitis (eee) virus. the e1+ (e1 gene plus 292 nucleotides 3' of the coding region), e2, and c gene regions from north american (na) variety viruses and the e1 and c gene regions of south american (sa) variety viruses were successfully amplified by rt-pcr using a single primer set ...19968822635
detection and comparison of viral antigens in measles and rubella rashes.measles and rubella skin lesions were immunocytochemically compared by the avidin-biotin-peroxidase complex method for detecting viral antigens. cryostat sections of biopsied specimens of the skin were stained with mouse monoclonal antibodies to p protein of measles virus and to e1 protein of rubella virus. the measles virus antigen was concentrated in the corneal layer and the keratinocytes of the epidermis and in the surface part of the dermis in the biopsy secimens taken within 6 days after t ...19968824963
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