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laboratory diagnosis of congenital human cytomegalovirus infection using polymerase chain reaction and shell vial culture.congenital hcmv infection was diagnosed at the 22th week of gestation. the infection was suspected because hcmv igm was detected in a serum sample obtained from the woman's husband. hcmv infection was detected in the amniotic fluid by polymerase chain reaction, shell vial culture (immunoperoxidase assay) and conventional virus isolation. serologic testing in paired sera of the woman and in umbilical cord blood for specific igm and iga remained negative. as serological data (preconceptional hcmv ...19921322865
a novel herpes simplex virus gene (ul49a) encodes a putative membrane protein with counterparts in other herpesviruses.comparative analysis of dna sequences located between the coding regions of genes ul49 and ul50 of herpes simplex virus types 1 and 2 (hsv-1 and -2) has revealed a small open reading frame (orf) of 91 and 87 codons respectively with the characteristics of a genuine protein-coding region. the predicted protein products are clearly related and exhibit features of membrane-inserted proteins, with potential n-proximal signal peptides and c-proximal membrane anchor regions. counterparts are present i ...19921322965
use of ganciclovir for cytomegalovirus infection.ganciclovir (9-(1,3-dihydroxy-2-propoxymethyl) guanine, dhpg) is an acyclovir analog with excellent antiviral activity against human cytomegalovirus (cmv). clinically, cmv infection occurs in from 60 to 90% of all renal transplant recipients and it is responsible for significant patient morbidity and graft loss. the likelihood of infection is closely related to the cmv status of both donor and recipient, with the greatest risk arising in the combination of a seronegative patient receiving a sero ...19921323340
a continuous sequence of more than 70 amino acids is essential for antibody binding to the dominant antigenic site of glycoprotein gp58 of human cytomegalovirus.antigenic domain 1 (ad-1) on glycoprotein gp58 of human cytomegalovirus was characterized in detail, using mouse and human monoclonal antibodies as well as human convalescent sera. series of procaryotically expressed fusion proteins and synthetic peptides of various lengths were used as sources of antigen. binding of antibodies was found to depend on a continuous sequence of more than 70 amino acids between residues 552 and 635 of gp58. the fine specificities for sequences involved in antibody b ...19921323695
failure to detect human cytomegalovirus dna in peripheral blood leukocytes of healthy blood donors by the polymerase chain reaction.the transmission of cytomegalovirus (cmv) by blood transfusion may have a major effect on certain immunocompromised patients. to protect susceptible blood recipients from infection, it is advisable to use blood components from cmv-seronegative donors. however, serologic tests are not capable of indicating which blood component actually harbors infectious virus and can transfer it to the recipient. therefore, a sensitive method is needed for the detection of the virus itself. there have been thre ...19921325693
the immediate early genes of human cytomegalovirus upregulate expression of the cellular genes myc and fos.human cytomegalovirus (hcmv) is an important pathogen of the lung. we determined whether the hcmv immediate early genes (ie1 and ie2) can alter the regulation of the cellular immediate early genes (c-fos and c-myc). plasmid constructs containing the promoter-regulatory regions c-myc or c-fos upstream of the reporter gene, chloramphemicol acetyl transferase, were co-transfected into t cells (jurkat cells), monocytes/macrophages (thp-1 cells), or human fibroblast cells with plasmid constructs cont ...19921325808
on the association of human beta 2 microglobulin with cell culture-grown human cytomegalovirus (hcmv).we studied the production of human beta 2 microglobulin (beta 2m) in mock-infected or human cytomegalovirus (hcmv) infected human embryonic lung fibroblasts (hel) and the association of human beta 2m with hcmv virions. titration of beta 2m by two-step sandwich enzyme immunoassay revealed that hel released considerable amounts of human beta 2m into the culture medium and that the production of beta 2m was significantly enhanced by hcmv infection. the concentration of human beta 2m in the culture ...19921326812
glycoprotein h of human cytomegalovirus (hcmv) forms a stable complex with the hcmv ul115 gene product.the human cytomegalovirus (hcmv) ul75 gene product is the homologue of herpes simplex virus type 1 (hsv-1) glycoprotein h (gh), a virion glycoprotein that is essential for infectivity and which is conserved among members of the alpha-, beta- and gamma-herpesviruses. it has previously been shown that hsv-1 gh forms a stable complex with hsv-1 gl, the product of the ul1 gene, and the formation of this complex facilitates the cell surface expression of gh. none of the open reading frames within the ...19921328481
the 72k ie1 and 80k ie2 proteins of human cytomegalovirus independently trans-activate the c-fos, c-myc and hsp70 promoters via basal promoter elements.growth-regulating cellular genes or genes encoding proteins involved in cell cycle control are likely to be major targets of viral gene products in the establishment of a cellular state favourable for a permissive infection. we have examined whether infection of permissive fibroblasts with human cytomegalovirus (hcmv) results in trans-regulation of such cellular genes. here we have shown that the proto-oncogenes c-fos and c-myc are specifically induced during immediate early (ie) and early times ...19921328493
e2f mediates dihydrofolate reductase promoter activation and multiprotein complex formation in human cytomegalovirus infection.the adenovirus immediate-early protein e1a activates the adenovirus e2 promoter and several cellular gene promoters through transcription factor e2f. the immediate-early proteins of human cytomegalovirus (hcmv) can complement an e1a-deficient adenovirus mutant and activate the adenovirus e2 promoter. hcmv also has been shown to activate the adenovirus e2 promoter. on the basis of these findings, we have investigated whether hcmv can activate the promoter of the cellular dihydrofolate reductase ( ...19921328853
point mutations in the dna polymerase gene of human cytomegalovirus that result in resistance to antiviral agents.three independently isolated mutants of human cytomegalovirus strain ad169 were found to be resistant to ganciclovir at a 50% effective dose of 200 microm. phosphorylation of ganciclovir was reduced 10-fold in mutant-infected cells compared with ad169-infected cells. all three mutants were also determined to be resistant to the nucleotide analogs (s)-1-[(3-hydroxy-2- phosphonylmethoxy)propyl]adenine (hpmpa) and (s)-1-[(3-hydroxy-2-phosphonylmethoxy)propyl]cytosine (hpmpc) and hypersensitive to t ...19921331515
strong trans activation of the human cytomegalovirus major immediate-early enhancer by p40tax of human t-cell leukemia virus type i via two repetitive tax-responsive sequence elements.the immediate-early 1 and 2 gene locus of human cytomegalovirus (hcmv) that encodes trans-activator proteins with effects on both homologous and heterologous promoters is expressed under control of a complex enhancer/promoter regulatory region. this enhancer contains four types of repetitive sequence elements with 17, 18, 19, and 21 bp that bind cellular transcription factors. although the hcmv enhancer acts as a powerful stimulator of transcription in most cell types examined, human t cells do ...19921331524
human herpesvirus 7 is a t-lymphotropic virus and is related to, but significantly different from, human herpesvirus 6 and human cytomegalovirus.an independent strain (ji) of human herpesvirus 7 (hhv-7) was isolated from a patient with chronic fatigue syndrome (cfs). no significant association could be established by seroepidemiology between hhv-7 and cfs. hhv-7 is a t-lymphotropic virus, infecting cd4+ and cd8+ primary lymphocytes. hhv-7 can also infect sup-t1, an immature t-cell line, with variable success. southern blot analysis with dna probes scanning 58.8% of the human herpesvirus 6 (hhv-6) genome and hybridizing to all hhv-6 strai ...19921332051
the procoagulant response of cytomegalovirus infected endothelial cells.the report describes the effect of an in vitro infection of human umbilical vein endothelial cells with human cytomegalovirus (cmv). the parameters studied are cellular procoagulant activity, secretion of plasminogen activator inhibitor (pai-1) and urokinase-type plasminogen activator (u-pa), activation and internalization of factor x and merocyanine 540 staining. the infection does not result in an increase in pai-1 and u-pa secretion, but it brings about a procoagulant response, which is relat ...19921332211
improved synthesis and biological evaluation of an acyclic thiosangivamycin active against human cytomegalovirus.we previously described the synthesis and in vitro antiviral activity of an acyclic thiosangivamycin analog (gupta et al., 1989a). in order to extend these initial studies, a new, multi-gram synthesis of 4-amino-7-[(2-hydroxy- ethoxy)methyl]pyrrolo]2,3-d]pyrimidine-5-thiocarboxamide (compound 229) was achieved in 5 steps from the known 5-amino-2-bromo-3,4-dicyanopyrrole in good overall yield. in plaque reduction assays with hcmv, compound 229 had an ic50 of 7 microm; in yield reduction assays th ...19921332597
monoclonal anti-idiotypes for the rapid detection of human cytomegalovirus.a novel enzyme-linked immunosorbent assay (elisa) was developed for human cytomegalovirus (hcmv) utilizing a monoclonal anti-idiotype specific for cmvb1, an antibody to hcmv. samples of hcmv were measured by their inhibition of the binding of cmvb1 to anti-idiotype. the elisa detected hcmv in a concentration-dependent manner from 20 to 0.6 x 10(3) pfu/ml, with 50% inhibition at approx. 3 x 10(3) pfu/ml. these data demonstrate the potential of anti-idiotype antibodies as the basis of simple and r ...19921333473
evaluation of neutralizing antibody titers against human cytomegalovirus in intravenous gamma globulin preparations. 19921334581
anti-viral activity of human recombinant heparin-binding proteins hbnf and mk.herpes simplex viruses bind to cell surface heparan sulfate proteoglycans, as a first step of viral infection. we report here that two recombinant heparin-binding proteins hbnf and mk inhibit infectivity of human herpes simplex viruses types 1 and 2 and human cytomegalovirus. carboxymethylated hbnf and mk, which retain affinity for heparin-sepharose, do not exhibit anti-viral activities. arguments are presented that anti-viral effects of hbnf and mk are due to the competition for the specific bi ...19921335242
discordant estimates of heterologous promoter activity as determined by reporter gene mrna levels and enzyme activity.in this study, the human cytomegalovirus (cmv) promoter fused to the lacz (beta-gal) reporter gene was transfected into neuroblastoma sk-n-be(2)-c cells, and phorbol ester-stimulated promoter activity assessed by both pcr quantitation of reporter gene mrna levels and enzyme activity. surprisingly, significant differences were observed in the induction profile of cmv promoter activity as judged by these two independent methods of analysis. for example, at 24 hrs post-transfection beta-gal activit ...19921335248
activity of different antiviral drug combinations against human cytomegalovirus replication in vitro.the effects of different antiviral drug combinations on the replication of various human cytomegalovirus (cmv) strains in human embryonic lung (hel) fibroblasts were evaluated. hpmpc combined with either ganciclovir, foscarnet or acyclovir showed additive to synergistic inhibition of cmv replication. combinations of zidovudine with hpmpc, ganciclovir, foscarnet or acyclovir also resulted in additive to synergistic inhibition of cmv replication. synergism tended to be higher for the clinical cmv ...19921337893
expression of a cytomegalovirus ie-1-factor viii cdna hybrid gene in transgenic mice.a construct containing the 5' end of the human cytomegalovirus major immediate early gene fused to the human coagulation factor viii cdna was used to produce transgenic mice. two out of five transgenic lines transcribed the construct. the expression was consistently seen in a limited number of tissues and was highest in muscle tissues. this is in contrast to the almost ubiquitous activity demonstrated in earlier studies with the ie-1 enhancer/promoter. human factor viii protein was detected immu ...19921338694
establishment and functional characterization of human herpesvirus 6-specific cd4+ human t-cell clones.in order to clarify the protective immune responses against a newly identified herpesvirus, human herpesvirus 6 (hhv-6), we established hhv-6-specific human t-cell clones and examined their functional properties. five cd3+cd4+cd8- t-cell clones, which proliferated in response to stimulation with two different strains of hhv-6 in the presence of autologous antigen-presenting cells but not with herpes simplex virus type 1 or human cytomegalovirus, were established from peripheral blood lymphocytes ...19921348547
human cytomegalovirus in rejected kidney grafts; detection by polymerase chain reaction.human cytomegalovirus (cmv) infections are frequently associated with graft rejection in the immunosuppressed patients following organ transplantation. thirty-four tissue samples from rejected kidneys and 18 samples from normal adult kidneys obtained from autopsies were investigated for the presence of cmv-dna by the polymerase chain reaction (pcr) and by immunohistochemistry. dna extracted from renal tissues after proteinase k digestion was specifically amplified in 32 cycles using primers whic ...19921355597
the dominant linear neutralizing antibody-binding site of glycoprotein gp86 of human cytomegalovirus is strain specific.bacterial fusion proteins, constructed from overlapping fragments of the open reading frame coding for gp86 of human cytomegalovirus (hcmv) strain ad169, were used to localize antigenic regions recognized by antibodies from human convalescent sera. a major domain for binding of conformation-independent antibodies was localized on fusion protein ap86, containing amino acids 15 to 142 of gp86. human antibodies, affinity purified on ap86, neutralized infectious virus in tissue culture. in addition, ...19921371164
infection of hematopoietic progenitor cells by human cytomegalovirus.the susceptibility of hematopoietic progenitor cells to infection by human cytomegalovirus (hcmv) was investigated using several strains of hcmv, including the recombinant strain rc256. rc256 is derived from the laboratory strain towne and contains the escherichia coli lacz gene coding for beta-galactosidase (beta-gal) regulated by an early hcmv promoter. expression of lacz allowed the detection of hcmv in individual hematopoietic cells. clonogeneic bone marrow (bm) progenitors, including cd34+ ...19921377049
the amino terminus of human cytomegalovirus glycoprotein b contains epitopes that vary among strains.we mapped three antigenic domains of continuous epitopes on human cytomegalovirus (cmv) glycoprotein b (gb) by reacting a panel of independently derived monoclonal antibodies with deletion mutants expressed transiently in cos-1 cells. one of these antigenic domains, dc2, maps in the last 75 amino acids of the carboxy terminus. these epitopes are conserved in strains towne and ad169, as well as in 19 clinical cmv isolates. elisas of dc2-reactive antibodies with a set of overlapping synthetic olig ...19921378884
monoclonal antibody e-13 (m-810) to human cytomegalovirus recognizes an epitope encoded by exon 2 of the major immediate early gene.monoclonal antibody (mab) e-13 to human cytomegalovirus is used widely for diagnostic and fundamental studies, and has been shown to be directed against an immediate early (ie) protein(s). to determine which viral antigen is detected by mab e-13, four subfragments from the open reading frame encoded by exons 2, 3 or 4 of ie-1 were cloned in the bacterial expression vector pros. the resulting fusion proteins contained amino acids 77 to 491 encoded by mainly exon 4, amino acids 25 to 78 encoded by ...19921383398
correction of alpha-l-fucosidase deficiency in fucosidosis fibroblasts by retroviral vector-mediated gene transfer.a full-length cdna clone encoding the lysosomal hydrolase alpha-l-fucosidase was cloned into two retroviral vectors, one using the human cytomegalovirus immediate-early promoter for expression, and the other, the retroviral long terminal repeat (ltr). high-titer amphotropic virus was produced for both constructs by infection of pa317 cells, and used to efficiently transduce the alpha-l-fucosidase gene into both human and canine fucosidosis fibroblasts. this resulted in correction of the alpha-l- ...19921525209
biochemical and immunological properties of rat recombinant interleukin-2 and interleukin-4.we have previously described the isolation and sequencing of cdna clones encoding rat interleukin-2 (il-2) and interleukin-4 (il-4). in the present study, we report the generation of stably transfected chinese hamster ovary (cho) cell lines which constitutively synthesize and secrete high levels of rat recombinant il-2 (ril-2) and il-4 (ril-4). the expression of the cytokine cdna sequences is driven by the human cytomegalovirus promoter/enhancer within the respective pee6. hcmv-gs vector constru ...19921551691
merocyanine-sensitized photoinactivation of enveloped viruses.a wide range of enveloped viruses, including human herpes simplex virus type 1, human cytomegalovirus, human t cell leukemia/lymphoma virus type i, human immunodeficiency virus type 1, sindbis virus, and friend erythroleukemia virus, are highly susceptible to merocyanine 540 (mc 540)-sensitized photoinactivation. by contrast, human pluripotent hematopoietic stem cells, red cells, factor viii, and von willebrand factor are much less sensitive. this suggests that mc 540 may be useful for the inact ...19921617187
antiherpesvirus activity and mode of action of sp-303, a novel plant flavonoid.sp-303, a natural plant flavonoid polymer of molecular weight 2,100 daltons, was found to have antiviral activity against two strains of type 1 herpes-type simplex virus, including a thymidine-kinase-deficient strain, and a strain of type 2 herpes simplex virus. the 50% effective concentrations (ec50s) were 1-2 microm. acyclovir, which was run in parallel, had values of 4-28 microm. surprisingly, the compound was inactive against human cytomegalovirus. sp-303 was also virucidal at 50 microm. int ...19937685261
the product of the ul31 gene of herpes simplex virus 1 is a nuclear phosphoprotein which partitions with the nuclear matrix.the nucleotide sequence of the ul31 open reading frame is predicted to encode a basic protein with a hydrophilic amino terminus and a nuclear localization signal. to identify its gene product, we constructed a viral genome in which the thymidine kinase gene was inserted between the ul31 and ul32 open reading frames. the thymidine kinase gene was then deleted, and in the process, the 5' terminus of the ul31 open reading frame was replaced with a 64-bp sequence in frame with the complete, authenti ...19937692079
anti-idiotypic mimicry of a neutralizing epitope on the glycoprotein b complex of human cytomegalovirus.experiments were carried out to investigate the ability of rabbit anti-idiotype antibodies (ab2), directed against an anti-human cytomegalovirus monoclonal antibody (ab1), to induce neutralizing antibodies specific for the immunodominant glycoprotein b viral complex. mice immunized with ab2 produced anti-ab2 (ab3) that was both antigen and idiotype specific with regard to ab1. we conclude that the ab2 antibodies mimicked a neutralizing epitope and acted as a network antigen for inducing a specif ...19937692085
cytofluorographic analysis of effects of interferons on expression of human cytomegalovirus proteins.the appearance of cytomegalovirus (cmv) proteins in infected fibroblasts was determined by flow cytometry. the sequential production of immediate early (ie), early (e), and late (l) proteins reacting with respective monoclonal antibodies (mabs) e13, 58/5, and 24/4 was determined in fibroblasts infected with the ad-169 strain of cmv. the percentage of cells expressing cmv proteins and the intensity of fluorescence within cells were determined from day 1 to day 7 post-infection. the effect of inte ...19937509340
detection of neutralizing antibodies against human cytomegalovirus: influence of strain variation.the influence of human cytomegalovirus (hcmv) strain variation on neutralizing antibody titers was investigated in sequential sera obtained from 12 organ transplant recipients (11 renal transplant recipients, 1 liver transplant patient) suffering from primary or secondary hcmv infection. cross-neutralization assays using either the international hcmv reference strain ad169 or individual clinical isolates from patients showed congruent results. restriction enzyme analysis of the hypervariable alp ...19938099944
cd13 (human aminopeptidase n) mediates human cytomegalovirus infection.human cytomegalovirus (hcmv) infects cells by a series of processes including attachment, penetration via fusion of the envelope with the plasma membrane, and transport of the viral dna to the nucleus. the details of the early events of hcmv infection are poorly understood. we have recently reported that cd13, human aminopeptidase n, a metalloprotease, is present on blood cells susceptible in vitro to hcmv infection (c. söderberg, s. larsson, s. bergstedt-lindqvist, and e. möller, j. virol. 67:3 ...19938105105
polymerase chain reaction detection of human cytomegalovirus in over 2000 blood specimens correlated with virus isolation and related to urinary virus excretion.the polymerase chain reaction (pcr) as applied to human cytomegalovirus (hcmv) detection should provide a valuable tool for rapid, reliable diagnosis of infection, thereby allowing prompt treatment. however, to date the high sensitivity of this technique and the lack of semi-quantitative interpretation have hindered establishing its validity for diagnosing systemic infection. we describe a rapid, simple, semi-quantitative pcr technique for hcmv detection. the validity of the technique was tested ...19938106600
detection of human cytomegalovirus dna in peripheral blood leukocytes by the polymerase chain reaction. 19938212125
antibodies to human cytomegalovirus 65-kilodalton fc binding protein in rheumatoid arthritis: idiotypic mimicry hypothesis of rheumatoid factor production.we previously reported that rheumatoid factors (rfs) might bear the internal image of fc gamma-binding proteins (fcbps) of herpes family viruses, suggesting the possibility that some rfs may be produced as antiidiotypic antibodies to anti-viral fcbp antibodies. since human cytomegalovirus (hcmv) has been implicated in the pathogenesis of ra, we made an attempt to detect antibodies to 65 kd major hcmv fcbp in sera and synovial fluid from patients with ra. western blotting was performed using hcmv ...19938218829
antibodies induced by a primary cytomegalovirus infection react with human herpesvirus 6 proteins.after a primary human cytomegalovirus (hcmv) infection, antibody titer to human herpes-virus 6 (hhv-6) rises. to determine if this occurs because of simultaneous infection with both viruses, serologic responses to these viruses were investigated among healthy women who received a live hcmv vaccine or acquired hcmv from an infected child. both vaccination and natural infection caused four-fold or greater titer rises to hhv-6. analysis of sera from 5 children with primary hhv-6 showed no serologic ...19938228344
murine cytotoxic t cell response specific for human cytomegalovirus glycoprotein b (gb) induced by adenovirus and vaccinia virus recombinants expressing gb.a murine model of the cytotoxic t lymphocyte (ctl) response to glycoprotein b (gb) of human cytomegalovirus (hcmv) was developed based on the use of adenovirus (ad) and vaccinia virus (vac) recombinants expressing gb. mice of different major histocompatibility haplotypes [cba (h-2k), balb/k (h-2k) and balb/c (h-2d)] infected with the ad-gb recombinant developed an ad-specific ctl response. however, only the h-2k mice developed a significant hcmv gb-specific ctl response, as indicated by the majo ...19938245869
the acyclovir legacy: its contribution to antiviral drug discovery.the discovery of acyclovir marked the beginning of an exciting era in antiviral research. early studies on the novel mode of action explained the selectivity of the compound and the remarkably narrow spectrum of activity against a subset of the herpesviruses. throughout the past decade many clinical trials have demonstrated the efficacy and safety of this drug. furthermore, the development of resistance does not appear to be a significant issue in normal individuals. acyclovir provided the stimu ...19938245880
[human cytomegalovirus, its significance in immune deficiency states, laboratory diagnosis, therapeutic possibilities].the authors report on the virological findings of 59 transplant recipients. the following procedures were used for the detection of human cytomegalovirus (hcmv) infection: detection of antiviral antibodies by elisa, the detection of virus-coded antigens in the patients' leucocytes (hcmv antigenemia test), "accelerated" virus isolation using immunofluorescence (if). serial examinations revealed the hcmv infection in 12 patients following organ transplantation. the antigenemia test proved to be po ...19938247512
human cytomegalovirus induces jc virus dna replication in human fibroblasts.jc virus, a human papovavirus, is the causative agent of the demyelinating brain disease progressive multifocal leucoencephalopathy (pml). pml is a rare but fatal disease which develops as a complication of severe immunosuppression. latent jc virus is harbored by many asymptomatic carriers and is transiently reactivated from the latent state upon immunosuppression. jc virus has a very restricted host range, with human glial cells being the only tissue in which it can replicate at reasonable effi ...19938248262
mutations of the human cytomegalovirus immediate-early 2 protein defines regions and amino acid motifs important in transactivation of transcription from the hiv-1 ltr promoter.the human cytomegalovirus (hcmv) immediate-early two (ie2) protein of 579 amino acids significantly activates expression from the human immunodeficiency virus (hiv) long terminal repeat (ltr) promoter. using a proviral hiv-1 genome with a mutated tat gene we demonstrate that the ie2 protein effects an increase in the steady-state level of viral rna similar to a level as from a wild-type proviral genome. the regions of the hcmv ie2 protein required for transactivation of the hiv-1 ltr promoter we ...19938337845
antisense suppression of transferrin receptor gene expression in a human hepatoma cell (huh-7) line.a recombinant plasmid carrying human transferrin receptor cdna in reverse orientation downstream from the human cytomegalovirus immediate early promoter/enhancer element was introduced into the huh-7 human hepatoma cell line by lipofection. cell surface transferrin binding and iron uptake from transferrin each decreased by about 50% in stable transfectants bearing integrated antisense dna expression vector. northern blot analysis indicated that the abundance of target transferrin receptor messag ...19938380064
polymorphonuclear cells are not sites of persistence of human cytomegalovirus in healthy individuals.polymorphonuclear leukocytes (pmnl) have been shown to harbour human cytomegalovirus (hcmv) in viraemic patients, but to date pmnl of asymptomatic healthy subjects have not been examined directly to determine whether this is a normal site of hcmv persistence. using the polymerase chain reaction (pcr), paired dna samples prepared from adherent peripheral blood mononuclear cells (pbmc), which are known to be a site of persistence of hcmv, and pmnl of 10 healthy adults were analysed. all of seven i ...19938381466
multiple mechanisms are implicated in the regulation of nf-kappa b activity during human cytomegalovirus infection.infection-induced activation of the human cytomegalovirus major immediate early enhancer/promoter has been shown to be regulated primarily by transcription factor nf-kappa b cis elements. however, the mechanism(s) by which human cytomegalovirus induces nf-kappa b activity is unknown. a study was therefore undertaken to determine how this virus would affect normal nf-kappa b regulation. viral infection of fibroblasts resulted in the specific stimulation of promoters containing major histocompatib ...19938381532
detection of human cytomegalovirus dna in paraffin sections of human brain by polymerase chain reaction and the occurrence of false negative results.paraffin-embedded necropsy material from 6 patients with human cytomegalovirus encephalitis (hcmve) corroborated by immunocytochemistry and 11 control cases were examined for the presence of human cytomegalovirus (hcmv) dna by a nested polymerase chain reaction (npcr). a characteristic 183 base pair (bp) fragment of the hcmv genome could readily be amplified in 4 cases of hcmve. in 2 cases of hcmve, viral dna could be demonstrated only sporadically by pcr, due most likely to inefficient dna extr ...19938382271
detection of human cytomegalovirus in ocular tissue by polymerase chain reaction and in situ dna hybridization.rapid and sensitive techniques with a high degree of accuracy are necessary for the diagnosis and management of cytomegalovirus (cmv) retinitis presenting with atypical clinical manifestations. light microscopy and immunohistochemical studies have limitations in the identification of this virus, but in situ dna hybridization offers a rapid, highly specific, and easily interpretable means of identifying cmv. a new procedure of enzymatic amplification of dna in vitro, called the polymerase chain r ...19938383072
a microneutralization enzyme immunoassay for antibody to human cytomegalovirus.we have developed a relatively rapid, sensitive and quantitative microneutralization assay for antibody to human cytomegalovirus (hcmv). cell monolayers in 96-well microtiter plates inoculated with pre-incubated virus-antibody mixtures were fixed after 3 days. infectious foci were stained with peroxidase-labeled human monoclonal antibody to a 64 kda immediate early antigen of hcmv, and the plates were read at od450. the 50% neutralization titer of the antibody was calculated. a study with 20 hum ...19938383161
2-acetylpyridine 5-[(dimethylamino)thiocarbonyl]-thiocarbonohydrazone (1110u81) potently inhibits human cytomegalovirus replication and potentiates the antiviral effects of ganciclovir.we studied the effects of 2-acetylpyridine 5-[(dimethylamino)thiocarbonyl]-thiocarbonohydrazone (1110u81 or a1110u), a potent inhibitor of the ribonucleotide reductases encoded by herpes simplex virus types 1 and 2 and by varicella-zoster virus, against human cytomegalovirus (hcmv) replication in infected mrc-5 cells. we show that 1110u81 is a potent inhibitor of hcmv dna replication (50% inhibitory concentration [ic50], 3.6 microm; ic90, 5.6 microm) and also potentiates the effects of ganciclov ...19938384821
[cytomegalovirus].since human cytomegalovirus (cmv) was discovered in 1950's the clinical pictures of cmv infection have been emphasized. during recent 30 years, the diagnosis and therapy of cmv infection have significantly progressed. especially the diagnostic techniques developed from serum titers (complemental fixation) to pcr (polymerase chain reaction). concerning the relation between cmv and neurological disorders of childhood, congenital cmv infection, classically called as cytomegalic inclusion disease, i ...19938384864
increase of interleukin 2 receptor and cd45ro antigen on lymphocytes cultured with human cytomegalovirus.to determine the expression of interleukin 2 receptor (il-2r) and the change of cd45ra and cd45ro antigen on lymphocytes in response to human cytomegalovirus (hcmv), peripheral blood mononuclear cells (pbmc) were cultured with hcmv antigen, and the expression of il-2r and of cd45ra and cd45ro antigens on lymphocytes was analyzed by flow cytometry. the expression of il-2r alpha (p55) and -beta (p75) was significantly increased on t cells obtained from cmv-seropositive donors following culture wit ...19938384933
an elisa using recombinant proteins for the detection of neutralizing antibodies against human cytomegalovirus.two prokaryotically expressed fusion proteins encompassing amino acids 484-650 (ad-1) and 27-100 (ad-2) of glycoprotein gp58/116 of human cytomegalovirus (hcmv) were purified from e. coli lysates and used in elisa to determine antibody levels in human sera. the specificity of the test was established by comparison of 116 randomly selected sera with commercially available hcmv-elisa tests. the recombinant polypeptides were then used for the analysis of antibody titers in 112 human sera and were c ...19938385703
comparison of immunological reactivity to two major antigens of homologous and ad169 human cytomegalovirus strains during primary infection in renal transplant patients.the igg and igm immunoblotting patterns for the major hcmv antigens p65 and p55/52 were studied in 5 renal transplant recipients during primary hcmv infections. sequential sera from the 5 patients were tested in parallel with standardized antigens derived from the reference strain ad169 and from the patient'own isolates. the immunoblotting patterns differed from patient to patient as well as between strain ad169 and the patient's homologous isolates. the differences consisted in a better respons ...19938385731
altering central nervous system physiology with a defective herpes simplex virus vector expressing the glucose transporter gene.because of their postmitotic nature, neurons are difficult subjects for gene transfer. to circumvent this, we have used a defective herpes simplex virus vector to overexpress the rat brain glucose transporter (gt) gene under the control of the human cytomegalovirus ie1 promoter. this vector, designated vie1gt, was propagated using a herpes simplex virus type 1 temperature-sensitive mutant, ts756. gt expressed from vie1gt was readily immunoprecipitated from membrane fractions of vie1gt-infected v ...19938386379
effect of interferon-alpha on immediate early gene expression of murine cytomegalovirus.interferon-alpha (ifn-alpha) significantly reduced the replication of murine cytomegalovirus (mcmv) in mouse embryo fibroblasts derived from the susceptible mouse strain c3h/hej. when infectious virus production was measured, a strong decrease in virus titer was observed in ifn-treated cells at a multiplicity of infection (moi) of 1 and 0.5 pfu/cell. analysis of virus-specified mrnas by northern blot assay revealed that ifn-alpha had a significant effect on the expression of viral mrnas at 48h. ...19938389790
identification of a dna-binding protein of human herpesvirus 6, a putative dna polymerase stimulatory factor.a 41k early nuclear antigen (p41), expressed in human herpesvirus type 6 (hhv-6)-infected t cells, was cloned by screening a cdna expression library with the anti-p41 monoclonal antibody (mab) c5. when expressed in mammalian cells, the cloned p41 protein comigrated with the authentic p41 protein from hhv-6-infected cells and localized to the nucleus. hhv-6 p41 shares 44% sequence identity with the human cytomegalovirus (hcmv) dna-binding protein, icp36 (ul44 gene product); p41 binds to ssdna wit ...19938389796
characterization of the major locus of immediate-early genes of rat cytomegalovirus.a major locus of rat cytomegalovirus (rcmv) immediate-early (ie) rna transcription was identified. a cdna library from rat embryo fibroblasts infected with rcmv under ie conditions was constructed and screened by using appropriate rcmv dna probes, revealing at least two ie genes (ie1 and ie2) transcribed from this locus by differential splicing. the first three exons (the first is noncoding) are spliced to exon 4 to form ie1 and to exon 5 to form ie2. the structural organization of the rcmv majo ...19938389919
a rabbit model for human cytomegalovirus-induced chorioretinal disease.ad169, a well-characterized laboratory strain of human cytomegalovirus (hcmv), was used to establish an animal model of progressive hcmv chorioretinal disease by injection of 10(5) pfu into the rabbit vitreous. chorioretinal, vitreous, and pulmonary disease were monitored by hcmv recovery, clinical observation, antigen localization, and histopathology. vitritis and focal areas of immune cellular infiltrates were seen in inner retinal layers on days 2-4 after inoculation. disease progressed with ...19938393056
high rate of concurrent genital infections with human cytomegalovirus and human papillomaviruses in cervical cancer patients.a case-control study evaluated a possible association between infection with human papillomavirus (hpv) or cytomegalovirus (cmv) and cervical cancer. seventy-eight patients with cervical cancer (cases) were compared with 55 age-compatible patients with uterine leiomyoma (controls). genital cmv and hpv infections were diagnosed by polymerase chain reaction amplification of viral dna in cervical tissues. smoking, first coitus or pregnancy before age 20, multiple sex partners, history of multiple p ...19938393059
translational inhibition mediated by a short upstream open reading frame in the human cytomegalovirus gpul4 (gp48) transcript.the human cytomegalovirus (cmv) virion glycoprotein gpul4 (gp48) gene expresses a transcript that contains three aug codons upstream from the one used to initiate synthesis of the gp48 protein. previously we reported that the second of these aug codons, aug2, was necessary but insufficient for inhibition of downstream translation (m. schleiss, c. r. degnin, and a. p. geballe, j. virol. 65:6782-6789, 1991). we now demonstrate that the coding information of the upstream open reading frame initiate ...19938394459
direct interaction of the human cytomegalovirus ie86 protein with the cis repression signal does not preclude tbp from binding to the tata box.the human cytomegalovirus major immediate-early gene encodes several protein isoforms which autoregulate the major immediate-early promoter (miep). one of these isoforms, the ie86 protein, represses the miep through a dna sequence located between the tata box and the transcription initiation site, designated the cis repression signal (crs). through mutational analysis, amino acid domains within ie86 responsible for binding the crs element were located at the c terminus. mutation of the putative ...19938394462
mapping a putative pyruvoyl decarboxylase active site to human cytomegalovirus open reading frame ul77.human cytomegalovirus is present as an apparently innocuous infection among a large proportion of the adult population that causes serious disease when congenitally transferred (1). human cytomegalovirus disease may become life threatening when found as an infection of immunocompromised transplant and aids patients (1,2). we now map a putative pyruvoyl decarboxylase enzyme prosthetic group, known to be essential to the active site of this class of enzymes, to human cytomegalovirus open reading f ...19938394702
identification of the gene product encoded by orf ul56 of the human cytomegalovirus genome.experiments were undertaken to identify the product of open reading frame (orf) ul56 of human cytomegalovirus (hcmv), the putative homolog of infected cell protein (icp) 18.5 of herpes simplex virus type 1 (hsv-1) that is thought to be involved in viral nucleocapsid maturation. northern blotting using a unique fragment of orf ul56 revealed a specific transcript of about 3.0 kb in hcmv-infected fibroblasts early and late postinfection (p.i.). two overlapping fragments of ul56 were subcloned for p ...19938395117
generation and effective enrichment of selectable human cytomegalovirus mutants using site-directed insertion of the neo gene.studies on the biology and function of human cytomegalovirus (hcmv) genes have been hampered by the limited number of viral mutants available for genetic analyses. we have developed a simple procedure to generate and enrich for hcmv recombinants. by inserting the bacterial neo gene, encoding neomycin/kanamycin phosphotransferase, into the large hcmv dna genome using homologous recombination, selectable mutants of this complex herpesvirus were isolated for the first time. the synthesis of neo fro ...19938395449
inhibition of human cytomegalovirus major immediate early gene expression by antisense rna expression vectors.we have used antisense oligonucleotides and expression vectors to inhibit human cytomegalovirus (hcmv) major immediate early (ie) gene expression. we find that oligonucleotides complementary to the hcmv 72k ie protein (ie1) coding region do inhibit hcmv infection, but this is non-specific. however, the use of certain antisense expression vectors, which express short oligonucleotides complementary to ie1, specifically inhibits ie1 expression at the protein level after introduction of ie expressio ...19938397287
diagnostics of persistent viruses: human cytomegalovirus as an example.infections with persistent viruses such as herpesviruses have become of significant clinical importance with the increasing number of immunocompromised patients at risk to suffer from severe disease. as antiviral chemotherapy is available for herpesvirus infections, the diagnostic methods for rapid and sensitive detection of symptomatic infection have been developed and recently refined. in human cytomegalovirus (hcmv), the use of recombinant viral antigens provides a rationale to improve serolo ...19938407251
analysis of acquired human cytomegalovirus infections by polymerase chain reaction.we used the polymerase chain reaction and primers corresponding to three regions of the human cytomegalovirus (hcmv) genome to study hcmvs isolated from 16 children attending a single day-care center and the father of two children in the same center. when we analyzed isolates with primers for the pp65 and major immediate-early genes, we observed nearly uniform amplification yielding products of predicted sizes. by contrast, primers for the a sequence demonstrated variability among hcmv strains, ...19938408568
effect of glycyrrhizin, cyclosporin a, and tumor necrosis factor alpha on infection of u-937 and mrc-5 cells by human cytomegalovirus.reactivation of latent or persistent human cytomegalovirus (hcmv) infection of monocytes or macrophages occurs under immunosuppressive conditions. we investigated the effect of glycyrrhizin (gl), cyclosporin a (csa), and tumor necrosis factor-alpha (tnf-alpha) on the viral dna synthesis and antigen expression of hcmv in u-937 and mrc-5 cells. although gl inhibited the viral antigen expression of hcmv in human monocytic cell line u-937 and human embryonic lung cell line mrc-5 in the study, as det ...19948283138
nucleotide sequence analysis of a 38.5-kilobase-pair region of the genome of human herpesvirus 6 encoding human cytomegalovirus immediate-early gene homologs and transactivating functions.human herpesvirus 6 (hhv-6) is prevalent in the human population, with primary infection occurring early in life. its predominant cd4+ t-lymphocyte tropism, its ability to activate human immunodeficiency virus type 1 (hiv-1) gene expression in vitro, and its upregulation of cd4 expression has led to speculation that hhv-6 may act as a positive cofactor in the progression of hiv infection to aids in individuals infected with both viruses. previous sequencing studies of restricted regions of the 1 ...19948289364
the effects of cytomegalovirus on human immunodeficiency virus replication in brain-derived cells correlate with permissiveness of the cells for each virus.human cytomegalovirus (hcmv) is commonly found in the brains of patients with aids and in some cases can be detected in the same cells as can human immunodeficiency virus type 1 (hiv-1). in this study, we analyzed the patterns of replication of hiv-1 and hcmv in singly infected cells and the effects of dual infection in human brain-derived cell lines of three different origins: neuroblastoma cell lines sk-n-mc and sy5y; astrocytoma/glioblastoma cell lines u373-mg and hs 683; and undifferentiated ...19948289398
human herpesvirus-7 (hhv-7).hhv-7 first isolated in 1990 from a healthy individual, is a ubiquitous agent. the second independent isolation of hhv-7 from a chronic fatigue syndrome patient was reported in 1992. the seroepidemiology of hhv-7 suggested that its prevalence rate in the u.s.a. population is > 85%; however, in japan a low prevalence rate has been reported. hhv-7 can be more readily isolated from the saliva than hhv-6. the primary infection of hhv-7 appears later in life than hhv-6. no disease has been reported t ...19947893982
enhancement human cytomegalovirus replication in a human lung fibroblast cell line by interleukin-8.we examined the effects of interleukin-8 (il-8) on cytomegalovirus (cmv) replication in human fibroblasts. exposure of fibroblasts to il-8 augmented both infectious virus production and replication of cmv, with concomitant increases in the levels of both the transcript of the cmv pp71 genome and the synthesis of the cmv late antigen. we also found that cmv selectively induced transcripts of the il-8 type 1 receptor in fibroblasts. these results suggest that il-8 also contributes to inflammatory ...19947933146
an efficient expression, purification and immunodetection system for recombinant gene products.we describe a modification of the mammalian expression vector prc/cmv, which drives expression of inserted genes from either the human cytomegalovirus (cmv) immediate-early promoter or the bacteriophage t7 rna polymerase promoter. the modification is designed to allow expression, simple purification and specific immunodetection of recombinant fusion proteins. the modified plasmid, termed ptag/cmv-neo, encodes a kozak consensus ribosome-binding site (rbs) and a 30-amino acid fusion tag peptide. t ...19947946324
elevated cytokine levels in synovial fluid of rheumatoid arthritis correlates with the presence of cytomegalovirus genome.synovial fluid aspirated from 34 patients with symptomatic rheumatoid arthritis (ra) was evaluated for the presence of human cytomegalovirus (cmv) genomic material using polymerase chain reaction (pcr), and for levels of interleukin 8 (il-8) and il-6 using enzyme-linked immunoadsorbence assay. il-8 and il-6 levels were significantly higher in cmv dna-positive ra patients than cmv dna-negative ra patients and at least 10-fold higher than in both corresponding control groups of patients with osteo ...19947948616
cytomegalovirus expression in minor salivary glands and chronic graft-versus-host disease.a systematic survey of human cytomegalovirus (cmv) was performed in 29 allogeneic bone marrow transplant (bmt) recipients. at day 100 a lip biopsy was performed and histological grading according to sale's score was compared with the immunohistochemical detection of the immediate early protein ie2 of hcmv. in 10 patients without chronic graft-versus-host disease (gvhd), 3 had lip biopsy grade 1, 7 had grade 0 sale's score and in 19 patients with chronic gvhd, 11 had grade 2, 1 had grade 1 and 7 ...19947951118
the extent of human cytomegalovirus replication in primary neurons is dependent on host cell differentiation.to study fetal brain infection with human cytomegalovirus (hcmv), an in vitro model was established using the human primary nontransformed neuronal cell line hcn-1a. on exposure to a mixture of factors promoting differentiation, hcn-1a cells differentiate into mature neurons. both undifferentiated and differentiated neurons were permissive to hcmv replication as assessed by immunohistochemistry and in situ dna hybridization. infectious center assays revealed that the ratio of virus-infected diff ...19947963724
cervical cytomegalovirus infection in prostitutes and in women attending a sexually transmitted disease clinic.this study aimed to determine the frequency of, and to define factors associated with, cervical shedding of cytomegalovirus (cmv) in highly sexually active women (licensed prostitutes) and in women attending a sexually transmitted disease (std) clinic. cervical specimens obtained from 195 licensed prostitutes and 187 std patients aged 17-50 years were compared for the presence of cervical cmv with specimens from 70 women of the same ages attending a gynecologic clinic. cervical cmv was identifie ...19947964646
rapid degradation of the heavy chain of class i major histocompatibility complex antigens in the endoplasmic reticulum of human cytomegalovirus-infected cells.human cytomegalovirus (hcmv) infection results in a marked reduction in the surface expression of class i major histocompatibility complex antigens on the host cells, which is thought to be one of the means for hcmv to evade the host immune system. to clarify the precise mechanism(s) of this phenomenon, we investigated the fate of the heavy chain of class i major histocompatibility complex antigens in hcmv-infected human embryonic lung fibroblasts (hel) by pulse-chase analysis and immunocytochem ...19947966584
detection of human cytomegalovirus genome in uterus tissue.to analyze persistent infection by human cytomegalovirus (hcmv) in vivo, specimens obtained from various sources and autopsied organs were examined for the presence of hcmv dna, mrna transcripts and antigens by polymerase chain reaction, in situ hybridization and immunostaining. the hcmv genome was detected in lung, liver, kidney, and blood vessels at an average positive rate of 15%. the highest pcr-positive rate was observed with cervical smears. subsequent examination of uterus tissues from pa ...19947979967
sequence variation of the amino-terminal antigenic domains of glycoprotein b of human cytomegalovirus strains isolated from chinese patients.to study the genetic variation of human cytomegalovirus (hcmv) in asian populations, the amino-terminal antigenic domains of glycoprotein b of hcmvs isolated from ethnic chinese transplant patients were cloned and sequenced. the nucleotide and encoded amino acid sequences were compared with published sequences of ad169 and towne laboratory strains. within the region sequenced, 9 out of 15 clinical isolates (60%) possessed a peptide configuration similar to that of strain ad169 while 6 isolates ( ...19947979986
structural analysis of the us-segment of a viable temperature sensitive human cytomegalovirus mutant.structural analysis of the us-segment of a viable temperature sensitive human cytomegalovirus mutant (ts9) by the use of restriction enzymes, specific amplifications by the polymerase chain reaction and dna sequencing, revealed deletions of open reading frames (orf) us14 and part of us15 in addition to that concerning us1 through us13 as reported previously [9]. it was further verified that the hind iii h-fragment and a major portion of the hind iii w-fragment were duplicated in inverted orienta ...19947979990
expression of protein kinase a and protein kinase c during ongoing human cytomegalovirus infection.during an hcmv infection, transcription of viral and cellular genes are mutually regulated. several cellular proteins have been implicated in the regulation of the hcmv major immediate early promoter (miep) which have been shown to respond to camp as well as activation of protein kinase c (pkc). we have examined the effect of an ongoing hcmv infection at the mrna level for the catalytic and regulatory subunits of protein kinase a (pka) and alpha and beta isoforms of pkc. there was a moderate ele ...19947980013
human cytomegalovirus latent infection of granulocyte-macrophage progenitors.we have investigated the interaction of human cytomegalovirus (cmv) with cultured primary granulocyte-macrophage progenitors, a suspected natural site of viral latency, and have established conditions for latent infection and reactivation in this cell population. progenitor cells from human fetal liver or bone marrow maintained a cd14+, cd15+, cd33+ cell surface phenotype during propagation in suspension culture. exposure to human cmv did not reduce growth or alter the phenotype of these cells d ...19947991550
replication of human cytomegalovirus in a rhabdomyosarcoma cell line depends on the state of differentiation of the cells.the replication of human cytomegalovirus (hcmv) was investigated in a new human rhabdomyosarcoma cell line (kfr) with morphological and biochemical characteristics of fetal striated muscle precursors (rhabdomyoblasts). kfr cells exhibited the unique property for spontaneous morphological transformation from a poorly-differentiated state into well-differentiated (myotube-like) rhabdomyoblasts. the poorly-differentiated rhabdomyoblasts promoted both complete viral gene expression and the productio ...19947998845
induction of endogenous human cytomegalovirus gene expression after differentiation of monocytes from healthy carriers.monocytes are one site of carriage of the human cytomegalovirus (hcmv) genome in healthy human carriers. however, as there are conflicting data detailing the level of hcmv gene expression during persistence in these cells, we have analyzed monocytes for evidence of viral immediate-early, early, and late transcription by using reverse transcription followed by pcr. we were unable to find evidence of hcmv lytic gene transcription in freshly isolated peripheral blood monocytes from hcmv-seropositiv ...19948107221
molecular evolution of herpesviruses: genomic and protein sequence comparisons.phylogenetic reconstruction of herpesvirus evolution is generally founded on amino acid sequence comparisons of specific proteins. these are relevant to the evolution of the specific gene (or set of genes), but the resulting phylogeny may vary depending on the particular sequence chosen for analysis (or comparison). in the first part of this report, we compare 13 herpesvirus genomes by using a new multidimensional methodology based on distance measures and partial orderings of dinucleotide relat ...19948107249
interleukin-11 expression in donor bone marrow cells improves hematological reconstitution in lethally irradiated recipient mice.lethally irradiated mice were transplanted with syngeneic bone marrow cells infected with a recombinant retrovirus vector containing the human interleukin-11 (hil-11) cdna under the control of the human cytomegalovirus (cmv) immediate early promoter. the hil-11 rna transcript from the vector was detected in the spleen and bone marrow of the recipient mice, and hil-11 protein accumulated in their serum. the hematological reconstitution of these mice was compared with recipient mice rescued with b ...19948112427
the immediate early genes of human cytomegalovirus upregulate tumor necrosis factor-alpha gene expression.cytomegalovirus (cmv) is an important cause of disease in the immunocompromised patient and cmv infection is associated with predominantly mononuclear inflammatory response. since products of the cmv immediate early (ie) gene region are potent trans-activators, we used the monocyte cell line thp-1 and a transient transfection assay to determine if these viral proteins upregulate expression of the tnf gene. the ie genes of cmv upregulated tnf gene activity as judged by increases in promoter activ ...19948113386
the equine herpesvirus type 1 glycoprotein homologous to herpes simplex virus type 1 glycoprotein m is a major constituent of the virus particle.glycoprotein 45 is a major envelope glycoprotein of equine herpesvirus type 1. the gene encoding this protein is located between map units 0.615 and 0.636 on the virus genome and evidence has suggested that it is encoded by gene 52, one of four genes within this region. using pcr we have amplified gene 52 and subsequently cloned it into a mammalian expression vector under the control of the human cytomegalovirus immediate early gene promoter. the gene was expressed in cos-7 cells and its product ...19948113768
involvement of shc in insulin- and epidermal growth factor-induced activation of p21ras.shc proteins are phosphorylated on tyrosine residues and associate with growth factor receptor-bound protein 2 (grb2) upon treatment of cells with epidermal growth factor (egf) or insulin. we have studied the role of shc in insulin- and egf-induced activation of p21ras in nih 3t3 cells overexpressing human insulin receptors (a14 cells). a14 cells are equally responsive to insulin and egf with respect to activation of p21ras. analysis of shc immunoprecipitates revealed that (i) both insulin and e ...19948114695
an endothelial cell-surface form of annexin ii binds human cytomegalovirus.human cytomegalovirus was shown to bind to human umbilical vein endothelial cells in a specific, saturable and calcium-dependent manner (kd = 7.9 pm (4 degrees c), 6469 virus binding sites/cell). affinity adsorption of detergent-prepared lysates of surface-radiolabeled endothelial cells to virions resulted in the identification of cell-derived proteins of approximate m(r) 36,000 and 32,000 that bound cytomegalovirus. protein sequencing of peptides obtained by cyanogen bromide cleavage demonstrat ...19948117306
tumor necrosis factor receptor p75 mediates cell-specific activation of nuclear factor kappa b and induction of human cytomegalovirus enhancer.the functional role of human tumor necrosis factor receptor (tnfr) p75 was studied by the use of tnfr p75-specific agonistic antibodies. human sw480t adenocarcinoma cells, stably transfected with a reporter construct containing beta-galactosidase under the control of human cytomegalovirus immediate early enhancer, were stimulated with anti-tnfr p75 polyclonal antiserum or monoclonal antibodies followed by measurement of beta-galactosidase activity and analysis by electrophoretic mobility shift a ...19948126005
application of pcr to multiple specimen types for diagnosis of cytomegalovirus infection: comparison with cell culture and shell vial assay.human cytomegalovirus (cmv) is a herpesvirus that is responsible for significant morbidity and mortality in congenitally infected infants and immunocompromised patients. antiviral therapies are available, thus making timely diagnosis of significant importance to at-risk patients. a pcr system was devised. the newly devised system, unlike previously described systems, can be applied to a wide variety of specimen types in a clinical microbiology laboratory setting. specimens from all sites routine ...19948126204
frequency distribution of cytomegalovirus envelope glycoprotein genotypes in bone marrow transplant recipients.using restriction analysis of polymerase chain reaction (pcr)-amplified dna, the cytomegalovirus (cmv) envelope glycoprotein (gb and gh) genotypes were determined for virus isolates from 128 bone marrow transplant recipients with fatal or nonfatal cmv. all isolates could be assigned to one of four gb and gh genotypes previously identified by dna sequencing studies. isolates of gb type 1 were more commonly found to be of gh type 2, whereas gb types 2-4 were more commonly linked to gh type 1. a sm ...19948133090
prokaryotic expression of a large fragment of the most antigenic cytomegalovirus dna-binding protein (ppul44) and its reactivity with human antibodies.we isolated and characterized from a lambda gt11 expression library clones expressing portions of human cytomegalovirus (hcmv)-p52. this nonstructural viral protein is encoded by ul44 and is known to be one of the best igm reactive antigens. the reactivity of these clones was studied with human antibody and the gene fragment coding for the most immune-reactive portion of p52 (aa 202-434) was cloned in a prokaryotic expression vector, pros, which overexpresses the antigen as a fusion protein to a ...19948175946
function of human cytomegalovirus glycoprotein b: syncytium formation in cells constitutively expressing gb is blocked by virus-neutralizing antibodies.we report that u373 glioblastoma cells constitutively producing human cytomegalovirus (hcmv) glycoprotein b (gb), the product of open reading frame ul55 of the hcmv genome, formed syncytia that contained 5 to 25 nuclei. flow cytometry with a panel of monoclonal antibodies (mabs) to the extracellular domain of hcmv gb showed that these cells expressed high densities of gb in the plasma membrane. we studied the properties of five clonal ub cell lines and the results are as follows. reactivity of a ...19948184537
helicase-primase complex of herpes simplex virus type 1: a mutation in the ul52 subunit abolishes primase activity.the ul52 gene product of herpes simplex virus type 1 (hsv-1) comprises one subunit of a 3-protein helicase-primase complex that is essential for replication of viral dna. the functions of the individual subunits of the complex are not known with certainty, although it is clear that the ul8 subunit is not required for either helicase or primase activity. examination of the predicted amino acid sequence of the ul5 gene reveals the existence of conserved helicase motifs; it seems likely, therefore, ...19948189507
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