| 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 ... | 1995 | 8572600 |
| 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 ... | 1995 | 8572936 |
| lack of correlation between sendai virus p/c mrna structure and its utilization of two aug start sites from alternate reading frames: implications for viral bicistronic mrnas. | the polycistronic p/c mrna of sendai virus encodes five proteins (c', p, c, y1, and y2) each of which initiates from a distinct start site. two major proteins, p and c, are expressed in approximately equimolar amounts from two consecutive augs in overlapping reading frames. to better understand the mechanism of expression of the c protein from a downstream aug, site-directed mutants of the p/c mrna were created and expressed in cos1 cells. the secondary structure of the mrna was examined to dete ... | 1996 | 8573577 |
| efficient generation of recombinant adenoviruses using adenovirus dna-terminal protein complex and a cosmid bearing the full-length virus genome. | an efficient method of constructing recombinant adenoviruses (ads) has been established. the expression unit to be introduced into recombinant ad was first inserted into the unique swa i site of the full-length ad genome cloned in a cassette cosmid. the cassette bearing the expression unit was then cotransfected into human embryonic kidney 293 cells together with the ad dna-terminal protein complex digested at several sites with eco t22i or ase i/ecori. the use of the parent ad dna-terminal prot ... | 1996 | 8577762 |
| 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 ... | 1995 | 8578855 |
| 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 ... | 1995 | 8590739 |
| cis and trans requirements for stable episomal maintenance of the bpv-1 replicator. | papillomavirus genomes are maintained as multicopy nuclear plasmids in transformed cells. to address the mechanisms by which the viral dna is stably propagated in the transformed cells, we have constructed a cell line ch04.15 expressing constitutively the viral proteins e1 and e2, that are required for initiation of viral dna replication. we show that these viral proteins are necessary and sufficient for stable extrachromosomal replication. using the cell line ch04.15, we have shown that the bov ... | 1996 | 8598191 |
| therapeutic antitumor response after immunization with a recombinant adenovirus encoding a model tumor-associated antigen. | recombinant adenovirus (rad), deleted of critical genes that enable viral replication and replaced with genes encoding heterologous proteins, has been shown to be a safe and effective vector in gene therapy studies. to evaluate a potential role for rad as an immunogen, we used two different replication-defective type 2 rads encoding the model ag, beta-galactosidase (beta-gal). to determine whether rad elicited the kind of immune responses therapeutic in an anti-tumor setting, the beta-gal-expres ... | 1996 | 8598466 |
| 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 ... | 1995 | 8598635 |
| recombinant adenovirus deleted of all viral genes for gene therapy of cystic fibrosis. | recombinant adenoviruses are being developed for gene therapy of inherited disorders such as cystic fibrosis because they efficiently transduce recombinant genes into nondividing cells in vivo. first generation recombinant adenoviruses, rendered defective by deletion of sequences spanning e1a and e1b, express low levels of early and late viral genes that activate destructive cellular immune responses. current strategies for improving recombinant adenoviruses attempt to inactivate other essential ... | 1996 | 8599194 |
| immune response to epitopes of hepatitis c virus (hcv) structural proteins in hcv-infected humans and chimpanzees. | hepatitis c virus (hcv)-infected humans and chimpanzees were studied for reactivity with linear epitopes in hcv h strain structural proteins. in 10 hcv-infected patients, epitopes were mostly mapped to the capsid and e1 proteins but not to e2. however, serum from 1 hcv-infected blood donor with a high anti-capsid titer reacted with multiple epitopes including e2. by contrast, antibody to capsid epitopes was seen in sera from hcv-rechallenged chimpanzees but not from chronically infected animals. ... | 1996 | 8603958 |
| 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 ... | 1995 | 8607286 |
| processing of the e1 glycoprotein of hepatitis c virus expressed in mammalian cells. | the structural part of the hepatitis c virus (hcv) genome encodes a capsid protein, c and two envelope glycoproteins, e1 and e2, released from the virus polyprotein precursor by signalase(s) cleavage(s). the processing of e1 was investigated by infecting simian cells with recombinant vaccinia viruses expressing parts of the hcv structural proteins. when the predicted e1 sequence was expressed alone (amino acid residues 174-370 of the polyprotein) or with the capsid protein gene (residues 1-370). ... | 1996 | 8609471 |
| genetic conservation of highlands j viruses. | we studied molecular evolution of the mosquito-borne alphavirus highlands j (hj) virus by sequencing pcr products generated from 19 strains isolated between 1952 and 1994. sequences of 1200 nucleotides including portions of the e1 gene and the 3' untranslated region revealed a relatively slow evolutionary rate estimated at 0.9-1.6 x 10(-4) substitutions per nucleotide per year. phylogenetic trees indicated that all hj viruses descended from a common ancestor and suggested the presence of one dom ... | 1996 | 8610461 |
| dimerization of rubella virus capsid protein is not required for virus particle formation. | rubella virus (rv) virions contain, in addition to the rna genome, a capsid protein (c) and two envelope glycoproteins (e1 and e2). the c protein in isolated virions has been reported to be a disulfide-linked dimer (c2). there are two cysteine residues (cys-152 and cys-196) within the c protein. to define the role of disulfide-linked c2 dimer in virion formation, site-directed mutagenesis was used to alter the cys-152 residue to serine. the association of mutant c protein with envelope glycoprot ... | 1996 | 8614992 |
| analysis of hpv1 e4 complexes and their association with keratins in vivo. | the hpv1 e4 gene encodes a family of abundant nonstructural late proteins whose role in the virus life cycle is unknown. their localization to keratin filaments when expressed in cultured epithelial cells has suggested a possible involvement in virus release by disturbing the integrity of the infected cell. here we show that in naturally occurring hpv1-induced tumors, the majority of the e4 protein (>95%) exists as complexes which do not contain keratins. the identification of discrete species o ... | 1996 | 8615013 |
| control of p53 expression by adenovirus 12 early region 1a and early region 1b 54k proteins. | the level of p53 is markedly increased in human cells in response to expression of the ad12 e1a proteins and, quite separately, to the ad12 e1b 54k protein. the behaviour of p53 in these two circumstances has been examined using a549 cells infected with ad12 dl620 (a mutant virus which does not express the larger e1b protein and is replication-defective) and human skin fibroblasts expressing the ad12 e1b 54k protein (hsf 54k). in normal and e1a-expressing a549 cells, p53 is located predominantly ... | 1996 | 8615026 |
| assembly of the sindbis virus spike protein complex. | the sindbis virus glycoproteins e1 and e2 are organized into 80 trimers of heterodimers within the virus envelope. using pulse-chase protocols and chemical crosslinkers, we have found that e1 and e2 precursor, pe2, rapidly assemble into heterodimers and then into trimers of heterodimers after translocation into the endoplasmic reticulum. e1 folds into its mature conformation within the endoplasmic reticulum via at least three intermediates differing in the configurations of their disulfide bonds ... | 1996 | 8623521 |
| a replication-defective human adenovirus recombinant serves as a highly efficacious vaccine carrier. | in this manuscript, an e1 and e3 deleted adenoviral recombinant expressing the rabies virus glycoprotein (g protein) under the control of the cytomegalovirus early promoter was tested for induction of a rabies virus-specific immune response in mice. the construct was found to induce neutralizing antibodies and cytolytic t cells to rabies virus. mice vaccinated with the adenoviral construct either by the systemic route or by application into the airways were protected against a subsequent infecti ... | 1996 | 8623532 |
| deduced consensus sequence of sindbis virus strain ar339: mutations contained in laboratory strains which affect cell culture and in vivo phenotypes. | the consensus sequence of the sindbis virus ar339 isolate, the prototype alphavirus, has been deduced. the results presented here suggest (i) that a substantial proportion of the sequence divergence evident between the consensus sequence and sequences of laboratory strains of ar339 has resulted from selection for efficient growth in cell culture, (ii) that many of these changes affect the virulence of the virus in animal models, and (iii) that such modified genetic backgrounds present in laborat ... | 1996 | 8627724 |
| the complete dna sequence and genomic organization of the avian adenovirus celo. | the complete dna sequence of the avian adenovirus chicken embryo lethal orphan (celo) virus (fav-1) is reported here. the genome was found to be 43,804 bp in length, approximately 8 kb longer than those of the human subgenus c adenoviruses (ad2 and ad5). this length is supported by pulsed-field gel electrophoresis analysis of genomes isolated from several related fav-1 isolates (indiana c and ote). the genes for major viral structural proteins (illa, penton base, hexon, pvi, and pviii), as well ... | 1996 | 8627769 |
| synergistic induction of osteocalcin gene expression: identification of a bipartite element conferring fibroblast growth factor 2 and cyclic amp responsiveness in the rat osteocalcin promoter. | fibroblast growth factors (fgfs) are important regulators of calvarial osteoblast growth and differentiation. we have studied the regulation of the osteoblast-specific gene osteocalcin (oc) by fgf2 in phenotypically immature mc3t3-e1 calvarial osteoblastic cells. fgf2 markedly induces oc mrna accumulation in mc3t3-e1 cells in the presence of forskolin (fsk). similarly, oc promoter activity (luciferase reporter) is up-regulated 6-10-fold by fgf2/fsk or by fgf2/8-bromo cyclic amp. half-maximal ind ... | 1996 | 8631781 |
| two species of rev proteins, with distinct n termini, are expressed by caprine arthritis encephalitis virus. | several cdna clones representing alternatively spliced rev-specific transcripts were isolated from a cdna library prepared from himalayan tahr cells infected with caprine arthritis encephalitis virus (caev). we previously characterized two rev-like cdna species, d1 and d2, and a tat e1 cdna containing the rev coding sequence downstream to the tat. in these cdnas, the rev coding domain derives its amino terminus from the n terminus of env, which is spliced to the 3' open reading frame encoding th ... | 1996 | 8642706 |
| chimeric hepatitis b virus core particles as probes for studying peptide-integrin interactions. | an rgd-containing epitope from the foot-and-mouth disease virus (fmdv) vp1 protein was inserted into the e1 loop of the hepatitis b virus core (hbc) protein. this chimeric protein was expressed at high levels in escherichia coli and spontaneously assembled into virus-like particles which could be readily purified. these fusion particles elicited high levels of both enzyme-linked immunosorbent assay- and fmdv-neutralizing antibodies in guinea pigs. the chimeric particles bound specifically to cul ... | 1996 | 8648742 |
| 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 ... | 1995 | 8655952 |
| the e1 protein is mandatory for pore formation by semliki forest virus spikes. | insect cells (aedes albopictus, clone c6/36) were infected with various variants of semliki forest virus including the wild type using the sfv replicon system. the variants included deletion mutants lacking one of the structural proteins and a mutant with a point mutation in p62 (sql). the latter mutation results in a failure to process p62 to e2 and e3. after infection of the cells with different variant viruses and subsequent expression of viral proteins in the host cell plasma membrane low ph ... | 1996 | 8659114 |
| 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 ... | 1995 | 8664382 |
| 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 ... | 1996 | 8689671 |
| 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 ... | 1996 | 8698862 |
| 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 ... | 1996 | 8699162 |
| 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 ... | 1995 | 8701591 |
| 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 ... | 1996 | 8709251 |
| 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 ... | 1996 | 8709289 |
| 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 ... | 1996 | 8712927 |
| 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 ... | 1995 | 8717570 |
| 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. | 1995 | 8730486 |
| 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 ... | 1996 | 8736235 |
| 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 ... | 1996 | 8738176 |
| 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 ... | 1996 | 8760430 |
| 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 ... | 1996 | 8761467 |
| 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 ... | 1996 | 8764026 |
| 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 ... | 1996 | 8764029 |
| 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 ... | 1996 | 8764067 |
| 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 ... | 1996 | 8766958 |
| 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 ... | 1996 | 8769412 |
| 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 ... | 1996 | 8784590 |
| 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 ... | 1996 | 8788164 |
| 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 ... | 1996 | 8793834 |
| 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 ... | 1996 | 8794347 |
| 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 ... | 1996 | 8794368 |
| 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 ... | 1996 | 8799810 |
| 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 ... | 1996 | 8806485 |
| 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 ... | 1996 | 8806486 |
| 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 ... | 1996 | 8806487 |
| 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 ... | 1996 | 8806514 |
| 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 ... | 1996 | 8811642 |
| 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. | 1996 | 8816629 |
| 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 ... | 1996 | 8817325 |
| 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 ... | 1996 | 8821642 |
| 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 ... | 1996 | 8821646 |
| 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 ... | 1996 | 8822344 |
| 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 ... | 1996 | 8822635 |
| 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 ... | 1996 | 8824963 |
| 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 ... | 1995 | 8825762 |
| 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 ... | 1996 | 8835354 |
| 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 ... | 1996 | 8836884 |
| 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. | 1996 | 8836894 |
| 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 ... | 1996 | 8840974 |
| 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 ... | 1995 | 8847508 |
| 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 ... | 1996 | 8853547 |
| 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 ... | 1996 | 8855177 |
| 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 ... | 1996 | 8862403 |
| 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 ... | 1996 | 8862587 |
| 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 ... | 1996 | 8863686 |
| 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 ... | 1996 | 8867614 |
| 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 ... | 1996 | 8872764 |
| 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 ... | 1995 | 8875617 |
| 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 ... | 1996 | 8877762 |
| 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 ... | 1996 | 8886847 |
| 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 ... | 1996 | 8887486 |
| 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 ... | 1996 | 8892914 |
| 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 ... | 1996 | 8892949 |
| 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. | 1996 | 8895018 |
| 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 ... | 1996 | 8896240 |
| 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 ... | 1996 | 8904416 |
| 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 ... | 1996 | 8915882 |
| 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 ... | 1996 | 8918741 |
| 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 ... | 1996 | 8921437 |
| 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 ... | 1996 | 8922467 |
| [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 ... | 1996 | 8928504 |
| 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 ... | 1996 | 8942974 |
| 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 ... | 1996 | 8953668 |
| [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 ... | 1996 | 8967065 |
| 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 ... | 1996 | 8971023 |
| 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 ... | 1996 | 8971024 |
| 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 ... | 1996 | 8971169 |
| 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. ... | 1996 | 8973533 |
| recombinational history and molecular evolution of western equine encephalomyelitis complex alphaviruses. | western equine encephalomyelitis (wee) virus (togaviridae: alphavirus) was shown previously to have arisen by recombination between eastern equine encephalomyelitis (eee)- and sindbis-like viruses (c. s. hahn, s. lustig, e. g. strauss, and j. h. strauss, proc. natl. acad. sci. usa 85:5997-6001, 1988). we have now examined the recombinational history and evolution of all viruses belonging to the wee antigenic complex, including the buggy creek, fort morgan, highlands j, sindbis, babanki, ockelbo, ... | 1997 | 8985391 |
| formation of native hepatitis c virus glycoprotein complexes. | the hepatitis c virus (hcv) glycoproteins (e1 and e2) interact to form a heterodimeric complex, which has been proposed as a functional subunit of the hcv virion envelope. as examined in cell culture transient-expression assays, the formation of properly folded, noncovalently associated e1e2 complexes is a slow and inefficient process. due to lack of appropriate immunological reagents, it has been difficult to distinguish between glycoprotein molecules that undergo productive folding and assembl ... | 1997 | 8985401 |
| 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 ... | 1996 | 8986436 |