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adeno-associated viral vector-mediated gene transfer of human blood coagulation factor ix into mouse liver.recombinant adeno-associated virus vectors (aav) were prepared in high titer (10(12) to 10(13) particles/ml) for the expression of human factor ix after in vivo transduction of murine hepatocytes. injection of aav-cmv-f.ix (expression from the human cytomegalovirus ie enhancer/promoter) into the portal vein of adult mice resulted in no detectable human factor ix in plasma, but in mice injected intravenously as newborns with the same vector, expression was initially 55 to 110 ng/ml. the expressio ...19989616156
influence of intravitreal injections of hpmpc and related nucleoside analogues on intraocular pressure in guinea pig eyes.cidofovir (hpmpc) is a potent long-acting anticytomegalovirus agent. in humans, its dose-limiting intravitreal toxicity results in the lowering of intraocular pressure (iop). the purpose of the present study was to determine the effects of hpmpc and various acyclic nucleoside phosphonate (anp) analogues when administered intravitreally in guinea pig eyes and to establish the structural and functional relation of these compounds in connection with their effects on the ciliary body and retina.19989620084
human cytomegalovirus persistently infects aortic endothelial cells.endothelial cells (ec) have been implicated as constituting an important cell type in the pathogenesis of human cytomegalovirus (hcmv). microvascular and macrovascular ec exhibit different biochemical and functional properties depending on the organ of origin. phenotypic differences between microvascular and macrovascular ec may alter the ability of these cells to support hcmv replication. in this study, we compared the replication of hcmv in primary macrovascular aortic ec (aec) with that in br ...19989621025
human herpesvirus 6 open reading frame u12 encodes a functional beta-chemokine receptor.human herpesvirus 6 (hhv- 6), which belongs to the betaherpesvirus subfamily and infects mainly t cells in vitro, causes acute and latent infections. hhv- 6 contains two genes (u12 and u51) that encode putative homologs of cellular g-protein-coupled receptors (gcr), while three other betaherpesviruses, human cytomegalovirus, murine cytomegalovirus, and human herpesvirus 7, have three, one, and two gcr-homologous genes, respectively. the u12 gene is expressed late in infection from a spliced mrna ...19989621074
in vitro generation of human cytomegalovirus pp65 antigenemia, viremia, and leukodnaemia.immunocompromised patients with disseminated human cytomegalovirus (hcmv) infection have circulating pmn carrying hcmv pp65 (antigenemia), infectious virus (viremia), and viral dna (leukodnaemia). because hcmv does not fully replicate in pmn, it is generally hypothesized that virions and viral materials are taken up by phagocytosis from fully permissive hcmv-infected endothelial cells. however, no experimental evidence has ever been provided for these pmn-endothelium interactions. pmn from 11 do ...19989637702
human cytomegalovirus: a viral complication in transplantation. 19989642503
the maastricht strain and england strain of rat cytomegalovirus represent different betaherpesvirus species rather than strains.the major immediate early (mie) locus of the maastricht strain of rat cytomegalovirus (rcmv) was found to comprise five exons of which the first is noncoding. the first three exons are spliced to either exon 4, generating ie1, or exon 5, generating ie2. an additional splicing event unique to rcmv (maastricht) was identified in exon 5, resulting in a 466-bp deletion. ie1 transcripts were detected exclusively during the ie phase of infection in vitro, whereas ie2 transcripts were detected during b ...19989657952
the acidic domain of pul37x1 and gpul37 plays a key role in transactivation of hcmv dna replication gene promoter constructions.transient complementation of human cytomegalovirus (hcmv) orilyt dna replication in permissive human diploid cells expressing replication genes under native promoters requires its ul36-38 gene products. two of the immediate early (ie) proteins encoded by this locus, pul37x1 and, to a lesser extent, gpul37, activated expression of hcmv early gene promoter constructions. the other ie protein encoded by the ul36-38 locus, pul36, and the early product, pul38, did not transactivate the hcmv early pro ...19989657958
antibody response to human cytomegalovirus (hcmv) glycoprotein b (gb) in aids patients with hcmv end-organ disease.human cytomegalovirus (hcmv)-specific antibody responses in hiv-1 infected individuals either with or without hcmv end-organ disease were examined to determine the whether development of hcmv disease was associated with a particular deficit in the antibody response. antiwhole hcmv, anti-glycoprotein b (gb), and neutralizing antibody levels were higher in hiv-1 infected individuals than in healthy immunocompetent subjects, particularly in patients with aids either with or without hcmv-associated ...19989661835
significance of reverse transcription polymerase chain reaction in the detection of human cytomegalovirus gene transcripts in thoracic organ transplant recipients.cytomegalovirus disease is a major cause of morbidity in transplant recipients. we have evaluated the clinical value of detecting viral mrna transcripts for the diagnosis of active infection leading to disease in recipients of thoracic organ transplants.19989662090
potential involvement of il-8 in the pathogenesis of human cytomegalovirus infection.the observations that several types of viruses induced interleukin (il)-8 production prompted us to investigate the interrelationship between il-8 and cytomegalovirus (cmv) infection. cmv infection caused il-8 production in a human monocytic cell line, thp-1, in dose- and time-dependent manners. moreover, cmv induced il-8 gene expression by concurrently activating transcription factors, nf-kappab and ap-1. furthermore, cmv infection of human fibroblast cell lines increased gene expression of a s ...19989665276
biological and immunogenic properties of rabies virus glycoprotein expressed by canine herpesvirus vector.in order to evaluate whether canine herpesvirus (chv) could be used as a live vector for the expression of heterologous immunogenes, we constructed a recombinant canine herpesvirus (chv) expressing glycoprotein (g protein) of rabies virus (rv). the gene of g protein was inserted within the thymidine kinase gene of chv yp11mu strain under the control of the human cytomegalovirus immediate early promoter. the g protein expressed by the recombinant chv was processed and transported to the cell surf ...19989682345
remission of iga nephropathy following treatment of cytomegalovirus infection with ganciclovir.following the detection of cytomegalovirus antigen in mesangial cells of some patients with iga nephropathy, an important role of human cytomegalovirus in the pathogenesis of iga nephropathy has been discussed. we studied a case of iga nephropathy with rapid deterioration of renal function associated with cytomegalovirus infection. following an infection of the upper respiratory tract, a 57-year-old woman developed with hematuria and acute renal failure. the histological diagnosis of iga nephrop ...19989696435
mutation in region iii of the dna polymerase gene conferring foscarnet resistance in cytomegalovirus isolates from 3 subjects receiving prolonged antiviral therapy.three human immunodeficiency virus-infected subjects with progressive cytomegalovirus (cmv) retinitis despite prolonged antiviral therapy had buffy coat cmv isolates that were resistant to both ganciclovir and foscarnet. genetic analysis of the resistant isolates showed that each contained a well-known ganciclovir resistance mutation in the viral ul97 phosphotransferase sequence, as well as a mutation (ala to val at codon 809, v809) in conserved region iii of the dna polymerase (pol) sequence. a ...19989697736
diagnostic significance and clinical impact of quantitative assays for diagnosis of human cytomegalovirus infection/disease in immunocompromised patients.in recent years several assays have been developed for quantitation of human cytomegalovirus (hcmv) in blood of immunocompromised (transplanted and aids) patients. it is currently agreed that the only reliable indication of the degree of dissemination of hcmv infection/disease is the measurement of hcmv in blood. diagnosis of hcmv end-organ disease (organ localizations) often does not benefit from quantitation of virus in blood, but requires detection and quantification of virus in samples taken ...19989699213
an antivirally active sulfated polysaccharide from sargassum horneri (turner) c. agardh.a sulfated polysaccharide was isolated from the hot water extract of a brown alga, sargassum horneri (turner) c. agardh. fucose was detected as the main component sugar of this polysaccharide. this compound showed potent antiviral activity against herpes simplex virus type 1, human cytomegalovirus and human immunodeficiency virus type 1. time-of-addition experiments suggested that it inhibited not only the initial stages of viral infection, such as attachment to and penetration into host cells, ...19989703258
the cyclin e promoter is activated by human cytomegalovirus 86-kda immediate early protein.human cytomegalovirus (hcmv) activates cyclin e/cdk2, which regulates cell cycle progression in g1 and s phase of the cell cycle. hcmv activation of cyclin e/cdk2 can be demonstrated in cells that are refractory to normal mitotic stimuli. this observation suggests that the virus has some means to overcome the stringent control on expression of cell cycle progression factors that is characteristic of cells in the g0 state. one of the mechanisms involved in activation of cyclin e/cdk2 is the induc ...19989705351
evaluation of the murex hybrid capture cytomegalovirus dna assay versus plasma pcr and shell vial assay for diagnosis of human cytomegalovirus viremia in immunocompromised patients.we evaluated a cytomegalovirus (cmv) 24-hour shell vial assay (sva), the murex hybrid capture cmv dna assay (hca), and a cmv plasma pcr for the detection of cmv viremia in renal and bone marrow transplant recipients and human immunodeficiency virus-infected patients. cmv viremia was detected by at least one method in 125 of 317 evaluable samples (39.4%) from 78 patients and was detected in 19.8% of samples by sva, 26.8% by hca, and 32.2% by plasma pcr. there was moderate to substantial agreement ...19989705391
factors influencing the effects of murine cytomegalovirus on the pancreas.background: as human cytomegalovirus (hcmv) infections are implicated in insulin-dependent diabetes mellitus (iddm), the effects of murine (m)cmv infection of inbred mice on the pancreas are of interest. results: inflammation and periacinar oedema peaked on day 3 and were replaced by a focal inflammation, but infected cells were rare. the islets were spared in c57bl mice. insulitis normally seen in non-obese diabetic (nod) mice was accelerated, but infected nod mice did not become glycosuric. is ...19989726035
evaluation of restriction fragment length polymorphism analysis of the ul10-ul13 genomic region for rapid identification of human cytomegalovirus strains.a sensitive semi-nested polymerase chain reaction (pcr) was established which allows rapid identification of human cytomegalovirus strains directly on clinical specimens, thereby permitting virus isolation and propagation on cell cultures to be avoided. the assay is based on restriction analysis of pcr products derived from the polymorphic ul10-ul13 region of the human cytomegalovirus genome. the method was evaluated using clinical samples from 23 subjects comprising 16 breast-feeding mothers an ...19989764560
extracellular signal-regulated kinase activity is sustained early during human cytomegalovirus infection.expression of many early viral genes during human cytomegalovirus (hcmv) infection is dependent on cellular transcription factors. several immediate-early and early viral promoters contain dna binding sites for cellular factors such as creb, ap-1, serum response factor, and elk-1, and these transcription factors can be activated by phosphorylation via the cellular mitogen-activated protein kinase (mapk) signal transduction cascade. to determine if the extracellular signal-regulated mapks, erk1 a ...19989765464
use of the human cytomegalovirus (hcmv) antigenemia assay for diagnosis and monitoring of hcmv infections and detection of antiviral drug resistance in the immunocompromised.quantification of viral load in blood has proven to be helpful in the follow-up of disseminated hcmv infections in immunocompromised patients.19989784143
discordant detection of human cytomegalovirus dna from peripheral blood mononuclear cells, granulocytes and plasma: correlation to viremia and hcmv infection.there exist only few data about the hcmv infection of single positive leukocyte subtypes in immunosuppressed patients. most reports describe hcmv coinfection of cells of the myelomonocytic line or even t- and b-cell populations. correlation of positive pcr findings from two major leukocyte fractions and plasma to viremia and hcmv infection in general should contribute to select suitable sources of hcmv dna for diagnostic purposes.19989785214
gene delivery to the neurulating embryo during culture.modulating expression of specific genes during embryogenesis will help elucidate their role in development. transient overexpression of specific genes can be accomplished by adding additional copies, or else antisense transcripts can be used to block expression. manipulation of gene expression requires an efficient, nontoxic gene delivery system. we compared a plasmid and a replication-defective adenovirus (ad5) as methods of delivering genes to the embryo during the neurulation stage of develop ...19989802190
na+-k+-cl- cotransport in human fibroblasts is inhibited by cytomegalovirus infection.we examined the effects of human cytomegalovirus (hcmv) infection on the na+-k+-cl- cotransporter (nkcc) in a human fibroblast cell line. using the cl--sensitive dye mqae, we showed that the mock-infected mrc-5 cells express a functional nkcc. 1) intracellular cl- concentration ([cl-]i) was significantly reduced from 53.4 +/- 3.4 mm to 35.1 +/- 3.6 mm following bumetanide treatment. 2) net cl- efflux caused by replacement of external cl- with gluconate was bumetanide sensitive. 3) in cl--deplete ...19989814982
cellular gene expression altered by human cytomegalovirus: global monitoring with oligonucleotide arrays.mechanistic insights to viral replication and pathogenesis generally have come from the analysis of viral gene products, either by studying their biochemical activities and interactions individually or by creating mutant viruses and analyzing their phenotype. now it is possible to identify and catalog the host cell genes whose mrna levels change in response to a pathogen. we have used dna array technology to monitor the level of approximately 6,600 human mrnas in uninfected as compared with huma ...19989826724
differentiation between acute primary and recurrent human cytomegalovirus infection in pregnancy, using a microneutralization assay.acute primary human cytomegalovirus (hcmv) infection in pregnancy, the major cause of congenital symptomatic infection, is often difficult to differentiate from recurrent infection, which presents a considerably smaller risk to the fetus. therefore, the diagnosis of primary infection in pregnancy is very important, especially if seroconversion is not documented and follow-up sera with declining igm-titers are not available. to investigate the value of the neutralizing antibody response against h ...19989829641
quanolirones i and ii, two new human cytomegalovirus protease inhibitors produced by streptomyces sp. wc76535.two new naphthacenequinone glycosides, quanolirones i (1) and ii (2) were isolated, together with the known compound galtamycin from the fermentation broth of streptomyces sp. wc76535. the structures 1 and 2 were established by analysis of their spectroscopic data and by comparison of their data to those of galtamycin. compounds 1, 2, and galtamycin showed inhibitory activity against hcmv protease with ic50 values of 14, 35, and 52 microm, respectively.19989834156
genetic analysis of a ganciclovir-resistant human cytomegalovirus mutant.we isolated a ganciclovir (gcv)-resistant human cytomegalovirus (hcmv) from a laboratory strain, ad169, and analysed the mutant. attempts were also made to identify directly the mutated gene. the 50% inhibitory concentration (ic50) of gcv for the mutant strain was five times higher than that of the wild-type strain. the mutant strain showed similar sensitivity to phosphonoacetic acid and cidofovir as the wild-type strain. these data suggest mutation in the ul97 gene encoding for the phosphotrans ...19989864050
thiazolo[4,5-d]pyrimidines. part ii. synthesis and anti-human cytomegalovirus activity in vitro of certain acyclonucleosides and acyclonucleotides derived from the guanine analogue 5-aminothiazolo[4,5-d]pyrimidine-2,7(3h,6h)-dione.the synthesis and in vitro antiviral activity of certain hydroxyalkoxymethyl, hydroxyalkyl, hydroxyalkenyl and phosphonoalkenyl derivatives of the guanine congener 5-aminothiazolo[4,5-d]pyrimidine-2,7(3h,6h)-dione are reported. the compounds of this study were selected for their structural similarity to acyclonucleosides with known anti-herpesvirus activity. 5-amino-3-[(z)-4-hydroxy-2-buten-1-yl]thiazolo[4,5-d]pyrimidine-2, 7(3h,6h)- dione was the only member of the series to display significant ...19989875377
3-hydroxyphthaloyl beta-lactoglobulin. iii. antiviral activity against herpesviruses.the spread of sexually transmitted diseases, including human immunodeficiency virus type 1 (hiv-1) and herpesvirus infections, has continued unabated despite educational efforts spearheaded as a response to the hiv-1 epidemic. this suggests the need for prophylactic measures, including the application of topical antiviral agents. chemical modification of bovine beta-lactoglobulin (beta-lg), the major protein of whey, by hydroxyphthalic anhydride (3hp) led to the generation of a potent hiv-1 inhi ...19989875389
investigation of the covalent modification of the catalytic triad of human cytomegalovirus protease by pseudo-reversible beta-lactam inhibitors and a peptide chloromethylketone.an investigation into the interaction between human cytomegalovirus (hcmv) protease and several beta-lactams, with characterization of the resulting acylenzymes using mass spectrometry, is reported. the time dependence of the inhibitors is highlighted by making comparisons of values obtained for inhibition and acylation. analysis of inactivated hcmv protease revealed a beta-lactam: protease stoichiometry of 1. subsequent enzymatic digestion with trypsin, peptide mapping using liquid chromatograp ...19989875526
strong conservation of the constitutive activity of the ie1/2 transcriptional control region in wild-type strains of human cytomegalovirus.the ie1/2 transcriptional control region of human cytomegalovirus (hcmv) drives the expression of the hcmv major immediate-early genes (ul123-122), which encode proteins crucial for initiation of the virus replicative cycle. nucleotide sequence polymorphism in this region of the viral genome could account for variations in the replication of hcmv wild-type strains. in order to test this hypothesis, the constitutive transcription-enhancing activity of the ie1/2 transcriptional control region deri ...19989880020
qualitative and quantitative detection of cytomegalovirus dna in sera by pcr as a clinical marker.the amount of human cytomegalovirus (cmv) dna in sera is considered to be a direct marker for cmv infection. we established conditions for nested pcr that detected one copy of cmv dna, and for competitive pcr, which detected five or more copies of cmv dna quantitatively. we tested 50 microl each of 16 freeze-stored and 5 fresh sera from patients, for cmv dna. in sera obtained from the same patient at different time points, small amounts of cmv dna were detected before the onset of cmv pneumonia. ...19989881658
polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein.human cytomegalovirus (hcmv) pp65 protein is the major constituent of viral dense bodies but is dispensable for viral growth in vitro. pp65 copurifies with a s/t kinase activity and has been implicated in phosphorylation of hcmv ie1 immediate-early protein and its escape from major histocompatibility complex 1 presentation. furthermore, the presence of pp65 correlates with a virion-associated kinase activity. to clarify the role of pp65, yeast two-hybrid system (ths) screening was performed to i ...19999882353
rna polymerase ii localizes at sites of human cytomegalovirus immediate-early rna synthesis and processing.pre-mrna synthesis in eukaryotic cells is preceded by the formation of a transcription initiation complex and binding of unphosphorylated rna polymerase ii (pol ii) at the promoter region of a gene. transcription initiation and elongation are accompanied by the hyperphosphorylation of the carboxy-terminal domain (ctd) of pol ii large subunit. recent biochemical studies provided evidence that rna processing factors, including those required for splicing, associate with hyperphosphorylated ctds fo ...19999889260
detection of serum antibodies against cytomegalovirus, varicella zoster virus and human herpesvirus 6 in patients with recurrent aphthous stomatitis.there has recently been renewed interest in the possible role of viruses in recurrent aphthous stomatitis (ras). in this study, sera from 22 patients with ras, 24 patients with oral lichen planus (olp) and 15 healthy controls were screened for igg and igm class antibodies to human cytomegalovirus (hcmv), varicella zoster virus (vzv) and human herpesvirus 6 (hhv-6). commercially available elisa and immunofluorescence kits were employed. there were no significant differences in the prevalence of i ...19999890451
transfusion of the hiv-seropositive patient: immunomodulation, viral reactivation, and limiting exposure to ebv (hhv-4), cmv (hhv-5), and hhv-6, 7, and 8. 19999924760
human cytomegalovirus inhibits transcription of the cc chemokine mcp-1 gene.in primary human diploid fibroblasts, infection with an unpurified stock of human cytomegalovirus induced accumulation of the cc chemokine mcp-1 in the cell culture medium. by 24 h postinfection, the level of mcp-1 returned to that in uninfected cultures. when cells were infected with uv-inactivated human cytomegalovirus, the induction of mcp-1 was still observed, but no reduction was seen by 24 h postinfection or later. this effect was the result of a decrease in the level of mcp-1 mrna present ...19999847345
focal transcriptional activity of murine cytomegalovirus during latency in the lungs.interstitial pneumonia is a frequent and critical manifestation of human cytomegalovirus (cmv) disease in immunocompromised patients, in particular in recipients of bone marrow transplantation. previous work in the murine cmv infection model has identified the lungs as a major organ site of cmv latency and recurrence. it was open to question whether the viral genome is transcriptionally silent or active during latency. transcription could be latency associated and thus be part of the latency phe ...19999847354
neoglycolipid conjugates of foscarnet with enhanced antiviral activity in cells infected with human cytomegalovirus and herpes simplex virus type 1.the synthesis of a series of neoglycolipid conjugates of foscarnet as potential drug targeting forms or lipophilic prodrugs of foscarnet is described. the compounds were obtained from suitably protected neoglycolipids, in which the lipid chain consisted of 12 to 20 carbon atoms, by ethoxycarbonylphosphonylation at the 6-hydroxyl or 4-hydroxyl group followed by deprotection. the in vitro antiviral activity of the compounds was determined in human foetal lung cells infected with human cytomegalovi ...199910628809
induction of protective immunity in chickens immunised with plasmid dna encoding infectious bursal disease virus antigens.direct dna inoculations were used to determine the efficacy of gene immunisation of chickens to elicit protective immune responses against infectious bursal disease virus (ibdv). the vp2 gene of ibdv strains gp40 and d78, and the vp2-vp4-vp3 encoding segment of strain d78 were cloned in an expression vector which consisted of human cytomegalovirus (hcmv) immediate early enhancer and promoter, adenovirus tripartite leader sequences and sv40 polyadenylation signal. for purification of vaccine-qual ...199910641338
protection of cattle against bovine leukemia virus (blv) infection could be attained by dna vaccination.the bovine leukemia virus (blv) envelope gene encoding extracellular glycoprotein gp51 and transmembrane glycoprotein gp30 was cloned into a vehicle expression vector under the human cytomegalovirus (cmv) intermediate early promoter. the intramuscular injection of this plasmid vector generated a cellular immune response. seven out of ten cows vaccinated with the dna construct resisted a drastic challenge (500 blv-infected lymphocytes as an infectious dose).199910824867
is there a risk of cytomegalovirus transmission during in vitro fertilization with donated oocytes?to define the risk of human cytomegalovirus (hcmv) transmission from donated oocytes.19999988402
epitope mapping of mouse monoclonal antibodies to the ppul83 lower matrix phosphoprotein of human cytomegalovirus.of nine mouse monoclonal antibodies (mabs) directed against the lower matrix protein (pp65; ppul83) of human cytomegalovirus (hcmv), all immunoprecipitated the 65-kda protein. only five were reactive by western blotting, however, and four of these mapped to linear antigenic epitopes located between amino acids 184-195 (mab c6), 343-357 (mab c11), 448-462 (mab c5), and 448-459 (mab c13). the epitope specificity of the fifth antibody (mab c3) and the four that recognised nonlinear sites could not ...199910022802
the leukocyte ig-like receptor (lir)-1 for the cytomegalovirus ul18 protein displays a broad specificity for different hla class i alleles: analysis of lir-1 + nk cell clones.leukocyte ig-like receptor (lir)-1 is a member of the ig superfamily which has been shown to bind the human cytomegalovirus mhc class i homologue ul-18 protein. in this study, we have analyzed the expression and function of lir-1 in human nk cells. we show that lir-1 is expressed by a subset of nk cells variable in size among different donors. when compared to the known hla class i-specific nk receptors, the expression of lir-1 was found to be partially overlapped with that of cd94-nkg2a or with ...199910050671
human cytomegalovirus infection in foci of langerhans cell histiocytosis.langerhans cell histiocytosis (lch) has been thought to be a disorder of immune regulation, and increasingly, evidence showing that the tissue damage in lch involves lymphokines and pro-inflammatory cytokines is reported. we detected human cytomegalovirus (hcmv)-dna in lch cells in the foci of lch lesions by immunohistochemistry, in situ hybridization and pcr. hcmv was detected in the nuclei and/or cytoplasm of lch cells in 9 of 27 lch cases by immunostaining. hcmv was probably an early antigen. ...199910071244
fractions of chemically oversulphated galactosaminoglycan sulphates inhibit three enveloped viruses: human immunodeficiency virus type 1, herpes simplex virus type 1 and human cytomegalovirus.a series of chemically oversulphated galactosaminoglycans (so3h:cooh ratio > or = 2) were tested in vitro as antiviral agents against human immunodeficiency virus type 1 (hiv-1), the aetiological agent of aids, and against herpes simplex virus type 1 and human cytomegalovirus, two agents responsible for opportunistic infections in hiv-infected people. the oversulphated derivatives displayed an increase in activity ranging from one to four orders of magnitude against the three viruses, as compare ...199910079877
the immediate early gene products of human cytomegalovirus increase vascular smooth muscle cell migration, proliferation, and expression of pdgf beta-receptor.evidence suggests that human cytomegalovirus (hcmv) infection contributes to the development of atherosclerosis and restenosis. because smooth muscle cell (smc) proliferation and migration are crucial events of both processes, and because pdgf beta-receptor modulates smc migration, we determined whether hcmv infection affects smc proliferation, migration, and pdgf beta-receptor expression. we employed a smc model in which hcmv infection leads to expression of only the immediate early (ie) hcmv g ...199910080946
frequent enrichment for cd8 t cells reactive against common herpes viruses in chronic inflammatory lesions: towards a reassessment of the physiopathological significance of t cell clonal expansions found in autoimmune inflammatory processes.we recently evidenced a dramatic enrichment for t cells reactive against epstein-barr virus (ebv) within inflamed joints of two rheumatoid arthritis patients. to assess the generality of this phenomenon and its relevance to autoimmunity, we studied the responses of cd8 t cells from patients with either acute or chronic inflammatory diseases (rheumatoid arthritis: n = 18, ankylosing spondylitis: n = 5, psoriatic arthritis: n = 4, reiter's syndrome: n = 3, arthrosis: n = 2, uveitis: n = 2, multipl ...199910092102
structure of adeno-associated virus vector dna following transduction of the skeletal muscle.the skeletal muscle provides a very permissive physiological environment for adeno-associated virus (aav) type 2-mediated gene transfer. we have studied the early steps leading to the establishment of permanent transgene expression, after injection of recombinant aav (raav) particles in the quadriceps muscle of mice. the animals received an raav encoding a secreted protein, murine erythropoietin (mepo), under the control of the human cytomegalovirus major immediate-early promoter and were sacrif ...19999971774
regulated expression of a sindbis virus replicon by herpesvirus promoters.we describe the use of herpesvirus promoters to regulate the expression of a sindbis virus replicon (sinrep/lacz). we isolated cell lines that contain the cdna of sinrep/lacz under the control of a promoter from a herpesvirus early gene which requires regulatory proteins encoded by immediate-early genes for expression. wild-type sindbis virus and replicons derived from this virus cause death of most vertebrate cells, but the cells discussed here grew normally and expressed the replicon and beta- ...19999971780
measurement of anti-human cytomegalovirus t cell reactivity in transplant recipients and its potential clinical use: a mini-review.by allowing direct determination of the frequencies of antigen-specific memory t cells in peripheral blood, novel techniques based on flow cytometry provide new diagnostic opportunities in various clinical settings, including organ transplantation. while the importance of the t cell compartment for the anti-human cytomegalovirus (hcmv) immune response is undisputed, efficient monitoring of this response was previously impossible because the conventional methods for measuring cd4+ and cd8+ t cell ...199910702713
human cytomegalovirus infection of immature dendritic cells and macrophages.a central aspect of human cytomegalovirus (hcmv) pathogenesis is the interaction of the virus with different antigen-presenting cell (apc) types of the host. in principle, a number of various cell types have the potential of antigen presentation when mhc ii expression is induced by appropriate stimuli. the most potent antigen presenters are monocytes/macrophages and dendritic cells (dcs), therefore called professional apcs. interestingly, these cells seem to be targets of productive hcmv infecti ...199910702719
human cytomegalovirus infection of human hematopoietic progenitor cells.for a number of years it has been well established that human cytomegalovirus (hcmv) can be transmitted by the cellular components of blood. hcmv is also associated with a number of hematologic disorders. although hcmv was thought to be present in blood cells in a latent or persistent form, it was not known how the virus was maintained and which cells were the carriers of hcmv. in addition to peripheral blood cells, there has been clinical evidence that hcmv may be associated with specific disor ...199910194116
inhibition of human immunodeficiency virus type 1 replication by the k10-k42 peptide of gap31 is due to induction of rapid but nonspecific precipitation of viral and nonviral proteins.the 33-amino acid peptide k10-k42 has previously been described as having potent anti-hiv-1 activity, and antiviral efficacy against hepatitis b and human cytomegalovirus in vitro. although the exact mechanism of antiviral activity was unknown, it was hypothesised that the k10-k42 peptide inhibited hiv-1 by interfering with one or more of the intracellular processes of reverse transcription, integration, and/or viral gene expression. we performed a series of experiments to identify and character ...199910195752
expression and characterization of a novel structural protein of human cytomegalovirus, pul25.human cytomegalovirus (hcmv) ul25 has recently been found to encode a new structural protein that is present in both virion and defective viral particles (c. j. baldick and t. shenk, j. virol. 70:6097-6105, 1996). in the present work a polyclonal antibody was raised against a prokaryotic pul25 fusion protein in order to investigate the biosynthesis and localization of the ul25 product (pul25) during hcmv replication in human fibroblasts. furthermore, pul25 was transiently expressed in its native ...199910196274
novel potential agents for human cytomegalovirus infection: synthesis and antiviral activity evaluation of benzothiadiazine dioxide acyclonucleosides.the first acyclonucleosides based on the benzothiadiazine dioxide system were synthesized following the silylation procedure. several acyclic moieties, including acetoxyethoxymethyl, benzyloxymethyl, and propargyloxymethyl groups, were introduced. two synthetic strategies were designed to selectively obtain the n-1 or n-3 derivatives. lipase-mediated deacylation was used for the deprotection of the acyclonucleosides. some of the benzothiadiazine dioxide acyclonucleosides, in particular 16, prove ...199910197958
human cytomegalovirus binding to human monocytes induces immunoregulatory gene expression.to continue our investigation of the cellular events that occur following human cmv (hcmv) infection, we focused on the regulation of cellular activation following viral binding to human monocytes. first, we showed that viral binding induced a number of immunoregulatory genes (il-1beta, a20, nf-kappab-p105/p50, and ikappabalpha) in unactivated monocytes and that neutralizing abs to the major hcmv glycoproteins, gb (ul55) and gh (ul75), inhibited the induction of these genes. next, we demonstrate ...199910202024
engagement of the cellular receptor for glycoprotein b of human cytomegalovirus activates the interferon-responsive pathway.cells respond to contact with human cytomegalovirus (hcmv) virions by initiating intracellular signaling and gene expression characteristic of the interferon (ifn)-responsive pathway. herein, we demonstrate that a principal mechanism of hcmv-induced signal transduction is via an interaction of the primary viral ligand, glycoprotein b (gb), with its cellular receptor. cells incubated with a purified, soluble form of gb resulted in the transcriptional upregulation of ifn-responsive genes oas and i ...199910207084
human cytomegalovirus replication and modulation of apoptosis in astrocytes.to characterize replication patterns and cytopathic effects during human cytomegalovirus (hcmv) infection of brain cells.199910225211
molecular characterization of the guinea-pig cytomegalovirus glycoprotein l gene.although the guinea pig cytomegalovirus (gpcmv) model is well suited to the study of vaccines for prevention of congenital cmv infection, there has been limited molecular characterization of gpcmv glycoproteins. since the in vivo co-expression of the human cytomegalovirus (hcmv) glycoprotein h (gh, gpul75) with glycoprotein l (gl, gpul115) may have relevance to cmv vaccine studies, these experiments were undertaken to test whether the gpcmv encodes a gl homolog. sequencing of the ecor i "g" frag ...199910226612
human cytomegalovirus inhibits ifn-alpha-stimulated antiviral and immunoregulatory responses by blocking multiple levels of ifn-alpha signal transduction.the type i ifns represent a primordial, tightly regulated defense system against acute viral infection. ifn-alpha confers resistance to viral infection by activating a conserved signal transduction pathway that up-regulates direct antiviral effectors and induces immunomodulatory activities. given the critical role of ifn-alpha in anti-human cytomegalovirus (hcmv) immunity and the profound ability of hcmv to escape the host immune response, we hypothesized that hcmv blocks ifn-alpha-stimulated re ...199910229853
quantitative analysis of latent human cytomegalovirus.cytomegalovirus latency depends on an interaction with hematopoietic cells in bone marrow and peripheral blood. the distribution of viral dna was investigated by pcr-driven in situ hybridization (pcr-ish), and the number of viral genomes per cell was estimated by quantitative competitive pcr during both experimental and natural latent infection. during experimental latent infection of cultured granulocyte-macrophage progenitors, the viral genome was detected in >90% of cells at a copy number of ...199910233941
synthesis and antiviral activity of 1-[1,5-dialkyl-1h-1,2,4-triazol-3-yl)methyl]thymines.cycloaddition of the intermediates 2 with 1-(cyanomethyl)-thymine 3 furnished the 1,2,4-triazolium salts 4, which rearranged spontaneously to the protonated salts 5. hydrolysis of 5, in situ, afforded the title compounds 6. compounds 6a-c were screened against hiv-1 (iiib), hiv-2 (rod), and human cytomegalovirus (hmcv) and showed poor or no activity, respectively.199910327888
stealth adaptation of an african green monkey simian cytomegalovirus.dna extracted from cultures of a cytopathic virus isolated from a patient with chronic fatigue syndrome was cloned into pbluescript plasmid. the nucleotide sequences of the plasmid inserts were analyzed using the blastn and blastx programs of the national center for biotechnology information. in confirmation of earlier studies, many of the sequences show partial homology to various regions within the genome of human cytomegalovirus (hcmv). the matching regions were unevenly distributed throughou ...199910331958
inhibition of influenza a virus reproduction by a ribozyme targeted against pb1 mrna.a ribozyme gene directed at a specific cleavage of mrna coding for pb1 protein, a component of rna-dependent rna-polymerase of influenza a virus, was constructed. the avian adenovirus celo virus-associated rna (va rna celo) promoter and human cytomegalovirus (cmv) promoter were used for the permanent expression of the ribozyme in cell lines. the cells were infected with influenza a virus strains a/singapore/1/57 and a/wsn/33, and the suppression of the virus reproduction and virus-specific prote ...199910333142
induction of neutralizing antibody against human cytomegalovirus (hcmv) with dna-mediated immunization of hcmv glycoprotein b in mice.immunization was accomplished by inoculating pcgb containing human cytomegalovirus (hcmv) glycoprotein b (gb) gene into balb/c mice intramuscularly. igm antibody was detected in all the immunized group. igg antibody was also found in all the tested mice with a mean peak antibody titer of 1:262 in three-times immunized groups. igg antibody appeared at 2 weeks postinoculation, raised peak levels at 7 weeks postinoculation and persisted over 6 months. neutralizing antibody was developed, and the pe ...199910338203
early detection of human cytomegalovirus infection after kidney transplantation by nucleic acid sequence-based amplification.the early detection of human cytomegalovirus infection after organ transplantation is a prerequisite for effective antiviral therapy. we evaluated the diagnostic value of monitoring the viral immediate-early (ie) 1 mrna expression in blood leukocytes by nucleic acid sequence-based amplification (nasba).199910342321
the transcriptional inhibitors, actinomycin d and alpha-amanitin, activate the hiv-1 promoter and favor phosphorylation of the rna polymerase ii c-terminal domain.actinomycin d and alpha-amanitin are commonly used to inhibit transcription. unexpectedly, however, the transcription of the human immunodeficiency virus (hiv-1) long terminal repeats (ltr) is shown to be activated at the level of elongation, in human and murine cells exposed to these drugs, whereas the rous sarcoma virus ltr, the human cytomegalovirus immediate early gene (cmv), and the hsp70 promoters are repressed. activation of the hiv ltr is independent of the nfkappab and tar sequences and ...199910347161
interstrain variation in the human cytomegalovirus dna polymerase sequence and its effect on genotypic diagnosis of antiviral drug resistance. adult aids clinical trials group cmv laboratories.the polymerase (pol) coding sequence was determined for 40 independent clinical cytomegalovirus isolates sensitive to ganciclovir and foscarnet. sequence alignments showed >98% interstrain homology and amino acid variation in only 4% of the 1, 237 codons. almost all variation occurred outside of conserved functional domains where resistance mutations have been identified.199910348781
domain mapping of the human cytomegalovirus ie1-72 and cellular p107 protein-protein interaction and the possible functional consequences.our previous work demonstrated that following human cytomegalovirus (hcmv) infection of fibroblasts, there was a protein-protein interaction between the hcmv ie1-72 immediate-early (ie) protein and the cellular p107 protein which resulted in the alleviation of p107-mediated transcriptional repression of e2f-responsive promoters. in a further characterization of this interaction, we now show that ie1-72 binds to the n-terminal portion of p107, not the c-terminal 'pocket' region that binds e2f-4, ...199910355776
stable production of a human growth hormone antagonist from cho cells adapted to serum-free suspension culture.human growth hormone (hgh) is a polypeptide with 191 amino acids and a molecular mass of 22 kda. an hgh analogue was created with a single amino acid substitution (glycine[g] 120 to arginine[r]) in the third alpha-helix of the hgh molecule. this hgh analogue, named hghg120r, was found to be an hgh antagonist. it is a parenteral drug candidate for treating conditions in which hgh levels are abnormally high, as found in type i diabetics. previously, a genetically engineered anchorage-dependent mou ...199910356250
dna microarrays of the complex human cytomegalovirus genome: profiling kinetic class with drug sensitivity of viral gene expression.we describe, for the first time, the generation of a viral dna chip for simultaneous expression measurements of nearly all known open reading frames (orfs) in the largest member of the herpesvirus family, human cytomegalovirus (hcmv). in this study, an hcmv chip was fabricated and used to characterize the temporal class of viral gene expression. the viral chip is composed of microarrays of viral dna prepared by robotic deposition of oligonucleotides on glass for orfs in the hcmv genome. viral ge ...199910364327
variation within the glycoprotein b gene of human cytomegalovirus is due to homologous recombination.human cytomegalovirus (hcmv) strains can be classified into different glycoprotein b (gb) genotypes. in a previous study, frequent intragenic variation of the gb gene was shown. the aim of this study was to analyse whether gb variation was due to homologous recombination. the gb gene of dna extracts derived from the peripheral blood leukocytes of 14 immunosuppressed patients was amplified by pcr and cloned. three variable sites of gb were analysed by restriction fragment analysis and dna sequenc ...199910374968
apoptosis induced by human cytomegalovirus infection can be enhanced by cytokines to limit the spread of virus.fas-mediated apoptosis is one of the immune effector pathways leading to the elimination of virus infected cells. in vivo, apoptotic signals are delivered to virus infected cells by fas-l and other cytokines secreted by specific t lymphocytes. cellular immune response appears to be essential in prevention of human cytomegalovirus (hcmv) disease. we have hypothesized that hcmv infection might directly or indirectly result in upregulation of fas receptor and in the presence of fas ligand, lead to ...199910390195
long-term persistence of immunoglobulin a (iga) and igm antibodies against human cytomegalovirus in solid-organ transplant recipients.the retrospective analysis of 494 solid-organ transplant recipients revealed that during the follow-up period (mean duration, 3.2 years) 184 (88%) of 209 anti-human cytomegalovirus (hcmv) immunoglobulin a (iga)-positive patients remained iga positive, as did 128 (74.85%) of 171 anti-hcmv igm-positive patients. we conclude that anti-hcmv iga and igm testing for management of clinically relevant hcmv infections in solid-organ transplant recipients is dispensable.199910391875
human cytomegalovirus stimulates cellular dihydrofolate reductase activity in quiescent cells.human cytomegalovirus (hcmv) productively infects quiescent fibroblasts in which the levels of deoxynucleotide triphosphates (dntps) and cell functions involved in dna metabolism are very low. since sufficient dntps levels are essential for human hcmv replication, host cell enzymes involved in the biosynthesis of dntps might be expected to be stimulated by viral infection in quiescent cells. we report that hcmv infection of quiescent fibroblasts stimulates the activity of cellular dihydrofolate ...199910393501
human cytomegalovirus infection decreases expression of thrombospondin-1 independent of the tumor suppressor protein p53.thrombospondin-1 (tsp-1) is a potent inhibitor of angiogenesis. it has been shown that promoter sequences of the tsp-1 gene can be transactivated by the wild-type tumor suppressor protein p53. as human cytomegalovirus (hcmv) infection inactivates wild-type p53 of various cell types, we investigated whether hcmv infection is associated with reduced tsp-1 production. we found, in conjunction with accumulated p53, that tsp-1 mrna and protein expression was significantly reduced in hcmv-infected cul ...199910393860
human cytomegalovirus and human herpesvirus 6 genes that transform and transactivate.this review is an update on the transforming genes of human cytomegalovirus (hcmv) and human herpesvirus 6 (hhv-6). both viruses have been implicated in the etiology of several human cancers. in particular, hcmv has been associated with cervical carcinoma and adenocarcinomas of the prostate and colon. in vitro transformation studies have established three hcmv morphologic transforming regions (mtr), i.e., mtri, mtrii, and mtriii. of these, only mtrii (ul111a) is retained and expressed in both tr ...199910398670
human cytomegalovirus, mhc class i and inhibitory signalling receptors: more questions than answers.the human cytomegalovirus ul18 protein, an mhc class i homologue, has been shown to bind to leucocyte immunoglobulin-like receptor (lir)-1, a member of a family of nine closely related immunoglobulin superfamily receptors expressed on leucocytes. the lirs are related to the natural killer (nk)-cell immunoglobulin-like receptors and to several other immunoreceptors. three groups of lir molecules have been defined: those containing cytoplasmic domain inhibitory signalling motifs, those with short ...199910399074
broadsheet. number 50: diagnosis of human cytomegalovirus infection and disease.human cytomegalovirus (cmv) remains an important cause of illness in immunocompromised individuals and the most common viral cause of congenital malformation. the tests available for diagnosis of cmv include serology, antigen detection, virus culture, tissue histopathology and nucleic acid detection. the diagnosis of cmv remains difficult because of the issues of virus latency, virus infection versus clinical disease and virus reactivation. the tests available and the use of these tests are unde ...199910399165
escape of human cytomegalovirus from hla-dr-restricted cd4(+) t-cell response is mediated by repression of gamma interferon-induced class ii transactivator expression.human cytomegalovirus (hcmv), a betaherpesvirus, is a pathogen which escapes immune recognition through various mechanisms. in this paper, we show that hcmv down regulates gamma interferon (ifn-gamma)-induced hla-dr expression in u373 mg astrocytoma cells due to a defect downstream of stat1 phosphorylation and nuclear translocation. repression of class ii transactivator (ciita) mrna expression is detected within the first hours of ifn-gamma-hcmv coincubation and results in the absence of hla-dr ...199910400755
three-dimensional visualization of tegument/capsid interactions in the intact human cytomegalovirus.the three-dimensional structure of the intact human cytomegalovirus (hcmv) was determined to 18-a resolution by electron cryomicroscopy and computer reconstruction. its capsid shell is composed of pentons, hexons, and triplexes arranged on a t = 16 icosahedral lattice and is identical to that of the b-capsid isolated from host cell nuclei. an icosahedrally ordered tegument layer formed by 960 copies of filamentous density is also visualized, which interacts with the pentons, hexons, and triplexe ...199910405351
immune responses and protection induced by dna vaccines encoding bovine parainfluenza virus type 3 glycoproteins.this study was designed to assess the parameters influencing the magnitude and type of immune responses generated to plasmids encoding the hemagglutinin/neuraminidase (hn) and fusion (f) proteins of bovine parainfluenzavirus type 3 (bpiv3). mice immunized with plasmids expressing hn or f under control of the rous sarcoma virus long terminal repeat promoter were primed, but they did not develop measurable immune responses. in contrast, strong humoral and cellular immune responses were induced wit ...199910405354
major product pp43 of human cytomegalovirus u(l)112-113 gene is a transcriptional coactivator with two functionally distinct domains.human cytomegalovirus u(l)112-113 encodes four phosphoproteins, pp84, pp50, pp43, and pp34, with common amino-termini. a previous report by kerry et al. (j. virol. 70, 373-382, 1996) demonstrated that u(l)112-113 products activate u(l)54 promoter in cooperation with immediate-early (ie) proteins. in this study, we identified a domain required for transcriptional activation in the pp43 protein, which consisted of two distinct regions: domain i (amino acids 272-296) and domain ii (amino acids 297- ...199910405360
rapid detection by reverse hybridization of mutations in the ul97 gene of human cytomegalovirus conferring resistance to ganciclovir.background of study: diseases due to human cytomegalovirus (hcmv) infection constitute a major threat in marrow and solid organ transplant recipients. ganciclovir (gcv) is widely used in prophylaxis and pre-emptive therapy of active hcmv infection. resistance to ganciclovir (gcv) may arise at variable frequency under gcv therapy and is conferred by mutations (i) in the ul97 gene (codons 460, 520, and 591-607) encoding a phosphotransferase which is essential for monophosphorylation of gcv and, to ...199910405892
evaluation of the amplisensor pcr and the sharp signal detection system for the early prediction of symptomatic cmv infection in solid transplant recipients.cytomegalovirus (cmv) is associated with high morbidity and mortality in transplant patients. specific antiviral treatment at an early stage of cmv infection may effectively ameliorate, but not eliminate cmv disease in these patients. presently, the pp65 antigenemia test on peripheral leukocytes is the method most widely used for predicting and monitoring transplant patients for active cmv infection. nucleic acid amplification methods are less well defined since they lack standardisation.199910405895
depletion of extracellular rantes during human cytomegalovirus infection of endothelial cells.human cytomegalovirus (cmv) infection results in pneumonitis in bone-marrow and lung-transplant recipients. the source of cmv infection contributing to the onset of pneumonitis is unclear, but may involve infection of the lung endothelium in the presence of infiltrating mononuclear cells. viral infection stimulates the host cell to express chemokines as signals to recruit specific immune cells to the site of injury. cmv encodes a chemokine receptor that may function to reduce host cell expressio ...199910423397
indolocarbazoles: potent, selective inhibitors of human cytomegalovirus replication.in our search for new, safer anti-hcmv agents, we discovered that the natural product arcyriaflavin a (la) was a potent inhibitor of hcmv replication in cell culture. a series of analogues (symmetrical indolocarbazoles) was synthesised to investigate structure activity relationships in this series against a range of herpes viruses (hcmv, vzv, hsv1, and 2). this identified a number of novel, selective and potent inhibitors of hcmv, 12,13-dihydro-2,10-difluoro-5h-indolo[2,3-a]pyrrolo[3,4-c]carbazo ...199910428375
minicircle: an improved dna molecule for in vitro and in vivo gene transfer.minicircles are a new form of supercoiled dna molecule for nonviral gene transfer which have neither bacterial origin of replication nor antibiotic resistance marker. they are thus smaller and potentially safer than the standard plasmids currently used in gene therapy. they were obtained in e. coli by att site-specific recombination mediated by the phage lambda integrase, which was used to excise the expression cassette from the unwanted plasmid sequences. we produced two minicircles containing ...199910435105
increased duration of transgene expression in the lung with plasmid dna vectors harboring adenovirus e4 open reading frame 3.for gene therapy to be effective in the treatment of chronic diseases, plasmid dna (pdna) vectors that provide persistent expression of therapeutic levels of the transgene product are desirable. studies in the lung with adenovirus vectors showed that products of the adenovirus e4 region can act both in cis and in trans to increase the duration of expression when transcription of the transgene was under the control of the human cytomegalovirus (cmv) promoter. to determine if these e4-encoded prot ...199910446923
discovery of 1,6-naphthyridines as a novel class of potent and selective human cytomegalovirus inhibitors. 199910447945
anti-repression of rna polymerase ii transcription by pyrrole-imidazole polyamides.pyrrole-imidazole polyamides are ligands that bind in the minor groove of dna with high affinity and sequence selectivity. molecules of this class have been shown to disrupt specific transcription factor-dna interactions and to inhibit basal and activated transcription from various rna polymerase ii and iii promoters. a set of eight-ring hairpin-motif pyrrole-imidazole polyamides has been designed to bind within the binding site for the human cytomegalovirus (cmv) ul122 immediate early protein 2 ...199910451376
development of biopharmaceuticals in plant expression systems: cloning, expression and immunological reactivity of human cytomegalovirus glycoprotein b (ul55) in seeds of transgenic tobacco.plant seeds offer unique opportunities for the production and delivery of oral subunit vaccines. we have used the immunodominant glycoprotein b complex of human cytomegalovirus (hcmv), introduced into tobacco plants, as a model system for studying the merit of this promising approach. given the advantages of expressing proteins in seeds, a novel expression vector was developed incorporating regulatory sequences of glutelin, the major rice seed storage protein, to direct synthesis of recombinant ...199910462237
target structures of the cd8(+)-t-cell response to human cytomegalovirus: the 72-kilodalton major immediate-early protein revisited.cell-mediated immunity plays an essential role in the control of infection with the human cytomegalovirus (hcmv). however, only a few cd8(+)-t-cell epitopes are known, with the majority being contained in the pp65 phosphoprotein, which is believed to dominate the cd8(+)-t-cell response to hcmv. here, we have readdressed the issue of cd8(+) t cells specific for the 72-kda major immediate-early protein (ie-1), which is nonstructural but is found very early and throughout the replicative cycle. usi ...199910482568
the development of an automated in situ assay for the detection of human cytomegalovirus in peripheral blood leukocytes.human cytomegalovirus (hcmv) infections are common in immunosuppressed patients, especially transplant recipients and patients with aids. the utility of an automated in situ hybridization (ish) assay for the rapid detection of hcmv immediate early mrna was evaluated using cytospin (shandon lipshaw, inc., pittsburgh, pa) prepared leukocytes from peripheral blood samples. in this study, the detection of hcmv immediate early protein by immunofluorescent antibody staining of the standard shell vial ...199910488758
a simple and rapid method for preparation of viral dna from cell associated cytomegalovirus.in the field of human cytomegalovirus pathogenesis there is growing interest in analyzing recent clinical isolates rather than cell culture adapted laboratory strains. however, true low passage isolates are strictly cell associated prior to cell culture adaptation and only a minor fraction of cells are infected at low passage number. both conditions hinder the preparation of pure viral dna. to date, genetic analyses had been carried out mostly with supernatant associated cytomegalovirus. a rapid ...199910488769
distribution of human cmv-specific memory t cells among the cd8pos. subsets defined by cd57, cd27, and cd45 isoforms.chronic antigenic stimulation has been associated with peripheral blood expansions of cd8pos. t cells characterized by cd57 expression, loss of cd27 expression, and reversal of the cd45ro(bright) /ra(dim) phenotype usually associated with immunological memory towards a cd45ro(dim) /ra(bright) phenotype. however, the relationship and functional significance of these subset(s) has remained controversial. here, this issue was addressed using a novel flow cytometric technique that allows simultaneou ...199910508265
site-specific recombination in mammalian cells expressing the int recombinase of bacteriophage hk022.the int gene of bacteriophage hk022, coding for the integrase protein, was cloned in a mammalian expression vector downstream of the human cytomegalovirus (cmv) promoter. green monkey kidney cells (cos-1) and mouse embryo fibroblast cells (nih3t3) transiently transfected with the recombinant plasmid express the integrase protein. co-transfection of this plasmid with reporter plasmids for site-specific recombination and pcr analyses show that the integrase promotes site-specific integration as we ...199910532317
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