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human cytomegalovirus early infection, acute rejection, and major histocompatibility class ii expression in transplanted lung. molecular, immunocytochemical, and histopathologic investigations.the present study aimed to investigate the relationship between acute rejection and human cytomegalovirus (hcmv) infection, as well as the coexpression of hla-dr and immediate-early (ie) viral antigens, in 143 transbronchial biopsies and bronchoalveolar lavage fluids of 32 lung transplant recipients. we investigated the occurrence of morphologically overt viral infection with conventional histopathology, the expression of ie antigens with single labeling immunohistochemistry, the coexpression of ...19968610354
definition of a human herpesvirus-6 betaherpesvirus-specific domain in glycoprotein gh that governs interaction with glycoprotein gl: substitution of human cytomegalovirus glycoproteins permits group-specific complex formation.formation of the glycoprotein gh/gl heterooligomer has important implications for understanding the pathology of human herpesvirus-6(hhv-6)-associated disease because this complex is essential for infectivity and fusogenic cell-to-cell spread. definition of the hhv-6 gh domain involved in protein-protein interactions was addressed by targeting regions defined by conserved cysteines identified by alignment of gh amino acid sequences representative of all herpesvirus subfamilies. studies using sit ...19968610443
dimerization of the human cytomegalovirus protease: kinetic and biochemical characterization of the catalytic homodimer.the single-chain 28 kda human cytomegalovirus (hcmv) protease catalytic domain containing the a143q mutation has been kinetically and conformationally characterized. the specific activity of the hcmv a143q protease (hcmvp) increases as the protease concentration increases, suggesting that this protease oligomerizes at high protein concentration to form a more active species. both cross-linking and light-scattering studies of hcmvp show the existence of a homodimer with an apparent molecular mass ...19968611517
defined large-scale alterations of the human cytomegalovirus genome constructed by cotransfection of overlapping cosmids.we have constructed defined human cytomegalovirus (cmv) mutants by cotransfecting overlapping cosmid clones spanning the 230-kbp genome. using this strategy, we have introduced a 13-kbp region of dna from a virulent strain of cmv into a defined position within the avirulent cmv(towne) genome. although more than 80% of the genome of these recombinant viruses was derived from towne dna, their plaque morphology more closely resembled that of toledo. to date, cmv is the largest virus and requires th ...19968627734
structure-activity relationships among 2-substituted 5,6-dichloro-, 4,6-dichloro-, and 4,5-dichloro-1-[(2-hydroxyethoxy) methyl]- and -1-[(1,3-dihydroxy-2-propoxy) methyl]benzimidazoles.the sodium salt of 2,5,6-trichlorobenzimidazole (8a) was condensed with [2-(benzyloxy)ethoxy]-methyl chloride (9) and [1,3-bis(benzyloxy)-2-propoxy]methyl chloride (18) to provide the corresponding protected acyclic nucleosides 10a and 19a, which on debenzylation afforded 2,5,6-trichloro-1-[(2-hydroxyethoxy)methyl]benzimidazole (11a) and 2,5,6-trichloro-1-[(1,3-dihydroxy-2-propoxy)methyl] benzimidazole (20a), respectively. a similar condensation of 2,4,6-trichlorobenzimidazole (2a) and 2,4,5-tri ...19968632412
polymerase chain reaction for prenatal diagnosis of congenital human cytomegalovirus infection.the reliability of the polymerase chain reaction (pcr) for prenatal diagnosis of human cytomegalovirus (hcmv) infection was determined by retrospective testing of 35 amniotic fluids identified previously as positive or negative for hcmv by virus isolation. amniocentesis was performed in 26 pregnant women with primary hcmv infection at 14-36 weeks gestation, 3-21 weeks after maternal infection. blood samples were obtained from 20 fetuses for igm determination and/or virus isolation. amniotic flui ...19958636720
comparison of benzimidazole nucleosides and ganciclovir on the in vitro proliferation and colony formation of human bone marrow progenitor cells.recently we have shown that certain benzimidazole ribonucleosides are potent and selective inhibitors of human cytomegalovirus (hcmv) replication. because antiviral drugs used to treat hcmv and human immunodeficiency virus (hiv) infections can suppress marrow progenitors, we have evaluated the most promising of the new benzimidazoles for their effects on human bone marrow cells in vitro. in an initial study of the bone marrow toxicity of one of the most active compounds, 100 microm 2-bromo-5,6-d ...19968639416
inhibition of human cytomegalovirus ul80 protease by specific intramolecular disulfide bond formation.a symmetrically substituted disulfide compound, cl13933, was identified as a potent inhibitor of human cytomegalovirus ul80 protease. two types of inhibited protease were observed, depending on inhibitor concentration. at high concentrations, cl13933 formed a covalent adduct with the protease on cys residues. at lower concentrations, this compound induced specific intramolecular disulfide formation between cys84 and cys87, and between cys138 and cys161. in contrast, cys202 did not form disulfide ...19968639545
differentiation of the immortalized adult neuronal progenitor cell line hc2s2 into neurons by regulatable suppression of the v-myc oncogene.a regulatable retroviral vector in which the v-myc oncogene is driven by a tetracycline-controlled transactivator and a human cytomegalovirus minimal promoter fused to a tet operator sequence was used for conditional immortalization of adult rat neuronal progenitor cells. a single clone, hc2s2, was isolated and characterized. two days after the addition of tetracycline, the hc2s2 cells stopped proliferating, began to extend neurites, and expressed the neuronal markers tau, neun, neurofilament 20 ...19968643664
retrieval of human cytomegalovirus glycoprotein b from the infected cell surface for virus envelopment.surface biotinylation of human cytomegalovirus (hcmv)-infected fibroblasts under pulse-chase conditions was used to define the cellular route of the dominant viral envelope glycoprotein gb into the cytoplasmic compartment of viral maturational envelopment. the results showed that a major fraction of gb was re-internalized from the infected cell surface prior to incorporation into the viral envelope. viral particles carrying biotinylated gb were subsequently released into the culture medium. vira ...19968645095
quantitation of cytomegalovirus dna and characterization of viral gene expression in bronchoalveolar cells of infected patients with and without pneumonitis.cytomegalovirus (cmv) is often present in bronchoalveolar lavage (bal) fluid of immunosuppressed patients without cmv pneumonitis. the amount of viral dna within bal cells of patients with definite cmv pneumonitis and of viral shedders was quantitated by polymerase chain reaction (pcr) and the extent of cmv gene expression within bal cells was defined by reverse transcription - pcr. no viral dna was detected in 6 viral shedders, and 12 had low copy numbers (mean, 72 copies/10(5) bal cells; media ...19968648201
cellular or viral protein binding to a cytomegalovirus promoter transcription initiation site: effects on transcription.we have previously shown that the ie2 protein of human cytomegalovirus (cmv) represses its own synthesis by binding to the major immediate-early promoter (m. p. macias and m. f. stinski, proc. natl. acad. sci. usa 90:707-711, 1993). the binding of a viral protein (ie2) and a cellular protein in the region of the transcription start site was investigated by site-specific mutational analysis and electrophoretic mobility shift assay. the viral protein and the cellular protein require different but ...19968648697
human cytomegalovirus immediate-early 2 (ie2) protein can transactivate the human hsp70 promoter by alleviation of dr1-mediated repression.the immediate-early 1 and 2 (ie1 and ie2, respectively) proteins of human cytomegalovirus are known transcription factors, which regulate the expression of viral and cellular genes. transcriptional activation by ie2 is dependent on the presence of a tata motif in target promoters, and ie2 can interact directly with the tata-binding protein (tbp) component of tfiid. tbp is known to be the target for transcriptional repression by the cellular dr1 protein, and this factor has been shown to repress ...19968648740
cloning, expression and characterization of the proteinase from human herpesvirus 6.after the u53 gene encoding the proteinase from human herpesvirus 6 (hhv-6) was sequenced, it was expressed in escherichia coli, and the activity of the purified, recombinant hhv-6 proteinase was characterized quantitatively by using synthetic peptide substrates mimicking the release and maturation cleavage sites in the polyprotein precursors of hhv-6, human cytomegalovirus (cmv), murine cmv, and epstein-barr virus. despite sharing 40% identity with other betaherpesvirus proteinases such as huma ...19968648756
repression by a differentiation-specific factor of the human cytomegalovirus enhancer.we detected a novel nuclear protein, mrf, that binds to multiple sites on the modulator which is located upstream of the human cytomegalovirus major immediate early gene enhancer. the expression of mrf is differentiation specific; the dna binding activity is present in nuclear extracts from undifferentiated tera-2 and thp-1 cells, but significantly reduced after these cells are induced to differentiate. in undifferentiated cells the enhancer activity is repressed by the modulator and upon differ ...19968649988
characterization of the human cytomegalovirus ul105 gene and identification of the putative helicase protein.we have characterized the transcription unit for the human cytomegalovirus ul105 gene and identified its putative protein product. the ul105 gene product is proposed to mediate helicase activity in the assembled helicase-primase complex. the two other putative proteins in this complex are the gene products of ul102 (primase-associated factor) and ul70 (primase). using northern blot analysis we have determined that the ul105 transcript is a 3.4-kb message that can be detected as early as 24 hr po ...19968659123
effects of desferrioxamine on human cytomegalovirus replication and expression of hla antigens and adhesion molecules in human vascular endothelial cells.desferrioxamine (dfo), commonly used in therapy as a chelator of ferric ion in disorders of iron overload, is a potent inhibitor of human cytomegalovirus (hcmv) replication in cultured fibroblast cells. moreover, dfo has immunomodulatory activity both in vitro and in vivo. we studied dfo effects on hcmv replication in cultured human endothelial cells and on the expression of several cell surface molecules, which mediate interactions of endothelial cells with other cell types in the immune system ...19958665150
detection of human cytomegalovirus dna: how, when and where?although several methods have been utilized for the detection and quantification of human cytomegalovirus (hcmv) dna, all of them can be divided into three groups: (i) detection of hcmv dna directly in tissues by in situ hybridization or in situ polymerase chain reaction (pcr); (ii) detection of hcmv dna in cell or tissue lysates by hybridization with dna or rna probes differently labelled-labels were progressively modified in order to provide an increasing sensitivity (hybridization products we ...19958668932
cytomegalovirus persistence in macrophages and endothelial cells.although human cytomegalovirus (hcmv) rarely causes overt disease in healthy individuals, the virus can be a serious, even life-threatening problem in immunosuppressed or immune-deficient patients and in the setting of maternofetal primary infection. in recent years knowledge about hcmv pathogenesis has increased significantly. identification of the cell types infected by hcmv in vivo has demonstrated that monocytes/macrophages and endothelial cells are key elements both in latent and acute infe ...19958668941
cytomegalovirus latency and latency-specific transcription in hematopoietic progenitors.latency-associated transcripts have been detected in human cytomegalovirus (cmv)-infected granulocyte-macrophage progenitors and have been shown to be encoded from both dna strands in the ie1/ie2 region of the viral genome. one class includes differentially spliced transcripts that extends exon 1 by initiating at two novel start sites 292 and 356 nucleotides in the ie1/ie2 promoter/enhancer region, upstream of the start site used during productive infection. the other class includes an unspliced ...19958668945
the role of cd8+, cd57+ cells in human cytomegalovirus and other viral infections.peripheral blood lymphocytes expressing cd8 and cd57 determinants are a small (1-15%) subset in healthy humans. cd8+, cd57+ peripheral blood lymphocytes may be divided by the level of cd8 expression, into cd8+high (cd57+) t-cells and cd8+low (cd57+) natural killer (nk) cells. cd8+high (cd57+) t-cell numbers are increased in human cytomegalovirus (hcmv)-seropositive subjects, and there is substantial evidence that hcmv is integral in the development of this subset in health and disease. furthermo ...19958668946
deletion mutants in human cytomegalovirus glycoprotein us9 are impaired in cell-cell transmission and in altering tight junctions of polarized human retinal pigment epithelial cells.retinal cytomegalovirus (cmv) disease is one of the major manifestations of viral pathogenesis in immunosuppressed patients with the acquired immunodeficiency syndrome (aids). cmv infection of the retina causes directional destruction which begins at the optic nerve head adjacent to the retinal capillaries and progresses, if untreated, to retinal detachment and blindness. infection does not occur across the basal membrane of the retinal pigment epithelium (rpe), adjacent to the highly vasculariz ...19958668948
successful biolistic transformation of mouse pancreatic islets while preserving cellular function.to utilize gene therapy, we required an efficient method to transfect intact islets before their use in transplantation. the biolistic method transforms cells by bombarding them with microprojectiles coated with dna. once internalized, the dna is solubilized and expressed. we used the firefly luciferase gene driven by the human cytomegalovirus immediate early promoter as a reporter construct in freshly isolated balb/c mouse islets to compare the transfection efficiency using either the biolistic ...19968669098
human cytomegalovirus pp65 lower matrix phosphoprotein harbours two transplantable nuclear localization signals.human cytomegalovirus phosphoprotein pp65 is targeted to the cell nucleus immediately after infection. deletion and point mutation analysis of the pp65 gene expressed in insect cells showed that two hydrophilic regions (hp1 and hp2) within the pp65 c-terminal 40% each harboured an independent nuclear localization signal (nls); strong association to the nuclear stroma also requires the n-terminal domain. either region, when fused to chloramphenicol acetyltransferase, localized the reporter protei ...19968683200
quantitation of human cytomegalovirus (hcmv) dna in cerebrospinal fluid by competitive pcr in aids patients with different hcmv central nervous system diseases.the current study was designed to quantitate human cytomegalovirus (hcmv) dna in cerebrospinal fluid (csf) of persons with aids with specific hcmv-related cns disease. dna present in csf obtained from aids patients was initially detected by a qualitative pcr procedure and then quantitated using a competitive pcr assay. in a group of 21 aids patients with hcmv-related cns disease, 12 patients with hcmv polyradiculopathy had a mean +/- sem of 11,982 +/- 4,480 copies/microliters in their csf compar ...19958685633
structure and functional expression of a complementary dna for porcine parathyroid hormone/parathyroid hormone-related peptide receptor.a complementary dna (cdna) encoding the receptor for porcine parathyroid hormone/parathyroid hormone-related peptide (pth/pthrp) was isolated from a porcine kidney cdna library. the porcine pth/pthrp receptor is a 585 amino acid protein containing seven putative membrane-spanning domains. the porcine pth/pthrp receptor has amino acid identity of 95.6%, 80.4%, and 88.7% with human, opossum, and rat pth/pthrp receptors, respectively and 53.4% identity to the recently cloned human pth2 receptor. th ...19968688470
cytomegalovirus dna level on biopsy specimens during treatment of cytomegalovirus gastrointestinal disease.there is no clear and point for the response to treatment of gastrointestinal human cytomegalovirus (hcmv) disease. hcmv-dna quantitation on gastrointestinal biopsy specimens has proven its value for the diagnosis of gastrointestinal hcmv disease in patients with acquired immunodeficiency syndrome (aids). the aim was to study hcmv-dna levels on gastrointestinal biopsy specimens during the treatment of gastrointestinal hcmv disease.19968690210
mucosal antibodies to human cytomegalovirus glycoprotein b occur following both natural infection and immunization with human cytomegalovirus vaccines.because antibodies against human cytomegalovirus (hcmv) glycoprotein b (gb) neutralize, levels of igg, secretory iga (siga), and mucosal iga1 antibodies to hcmv were measured in saliva and nasal washes. ten seronegative adults lacked these antibodies, but of 10 naturally seropositive adults, 10 had igg to gb, 5 had siga, and 0 had mucosal iga. among 12 recipients of a live hcmv vaccine, 8 had igg to gb, 4 had siga, and 2 had mucosal iga in samples collected 10-20 months after immunization; of 10 ...19968699071
anti-human cytomegalovirus effects of breast milk. 19968699086
determination and analysis of the complete nucleotide sequence of human herpesvirus.human herpesvirus 7 (hhv-7) is a recently isolated betaherpesvirus that is prevalent in the human population, with primary infection usually occurring in early childhood. hhv-7 is related to human herpesvirus 6 (hhv-6) in terms of both biological and, from limited prior dna sequence analysis, genetic criteria. however, extensive analysis of the hhv-7 genome has not been reported, and the precise phylogenetic relationship of hhv-7 to the other human betaherpesviruses hhv-6 and human cytomegalovir ...19968709220
inhibition of herpes simplex virus type 1 immediate-early gene expression by alpha interferon is not vp16 specific.pretreatment of tissue culture cells with alpha interferon (ifn-alpha) inhibits the transcription of herpes simplex virus type 1 (hsv-1) immediate-early (ie) genes, an effect which has been attributed to reduced transactivation of ie promoters by the virion protein vp16. our previous demonstration that ifn-alpha inhibited the replication of the hsv-1 mutant in1814, which has a mutated vp16 unable to activate ie transcription, appeared to be incompatible with ifn-alpha having an effect on vp16 ac ...19968709261
cmv retinitis: ganciclovir/monoclonal antibody. 19968721551
cytomegalovirus as a cause of pancytopenia.human cytomegalovirus, hcmv, infects most of the population by adulthood; the primary infection is often accompanied by transient neutropenia and thrombocytopenia, and is followed by a period asymtomatic viral latency. in the setting of bone marrow transplantation, however, the immunosuppressed state of the recipient enables hcmv to re-activate or to infect the individual and cause serious sequelae. these range from hepatitis and gastrointestinal disease to interstitial pneumonia and hematologic ...19968726402
long-term expression of a fluorescent reporter gene via direct injection of plasmid vector into mouse skeletal muscle: comparison of human creatine kinase and cmv promoter expression levels in vivo.expression of a fluorescent reporter gene has been studied using two alternate promoters to transcribe the green fluorescent protein (gfp) from aequorea victoria. the human cytomegalovirus (cmv) enhancer/ promoter or the human muscle-specific creatine kinase promoter (ckm) were inserted along with the gfp cdna into a plasmid expression vector based on a modified adeno-associated virus genome. naked plasmid dna was injected into the hamstring muscle of mdx mice and gfp gene expression determined ...19968727010
cytomegalovirus antibody avidity in allogeneic bone marrow recipients: evidence for primary or secondary humoral responses depending on donor immune status.the reconstitution of the human cytomegalovirus (cmv) antibody response in cmv seropositive bone marrow transplant patients was investigated by comparing 11 patients whose donors were cmv seropositive with 8 whose donors were cmv seronegative. evidence for primary or secondary responses to cmv was sought by determining igg antibody avidity using an avidity index method, and antibody titre over a period of up to 3 years after transplant. for the patients whose donors were cmv seropositive, the re ...19968732861
enhancement of human nk and lak cytotoxicity against hcmv-infected cells by rhamnogalacturonan: specificity of reaction.human peripheral blood mononuclear cells (pbmc) and their subpopulations obtained from healthy donors were used to study improvement of mhc-unrestricted cytotoxic reactions against cells infected with human cytomegalovirus (hcmv) at different multiplicities of infection. natural killer (nk) and lymphokine-activated killer (lak) cytotoxicity against hcmv-infected cells was greatly enhanced in the presence of rhamnogalacturonan (500 ng/ml). the increase of the multiplicity of infection from moi 0. ...19968733917
gfi-1 encodes a nuclear zinc finger protein that binds dna and functions as a transcriptional repressor.the gfi-1 proto-oncogene encodes a zinc finger protein with six c2h2-type, c-terminal zinc finger motifs and is activated by provirus integration in t-cell lymphoma lines selected for interleukin-2 independence in culture and in primary retrovirus-induced thymomas. gfi-1 expression in adult animals is restricted to the thymus, spleen, and testis and is enhanced in mitogen-stimulated splenocytes. in this report, we show that gfi-1 is a 55-kda nuclear protein that binds dna in a sequence-specific ...19968754800
[human cytomegalovirus infection in the pre- and postnatal period with special regards to routes of infection].the clinical course of hcmv infection in newborns and infants was discussed. different routes and sources of infection were analyzed, especially acquired infection from the mother excreting the virus in breast milk. the usefulness of modern laboratory techniques (pcr to detect hcmv dna and detection of antigen in leucocytes) and the importance of simultaneous tests performed in children and their mothers in stating the diagnosis was also evaluated.19968756770
[polyvalent hyperimmune immunoglobulins prevent the infection of monocytic type cells by human cytomegalovirus].monocyte/macrophage cell types can be infected with human cytomegalovirus (hcmv) could be a reservoir and a vehicle for virus spread in infected patients. we developed a model to study the effects of antiviral molecules on these cells. the monocytic-like cell line thp-1 and the human diploïd cells mrc-5 were used. thp-1 cells were cultivated with a phorbol 12-myristate 13-acetate (pma) for 24h prior to the infection. we studied infection of these cells with reference strain (ad-169). a cell free ...19968758495
the intracellular localization of human cytomegalovirus dna in peripheral blood leukocytes during active infections by high-resolution fluorescence in situ hybridization.although viremia is an integral part of the pathogenesis of human cytomegalovirus (hcmv) disease, the interaction between hcmv and circulating leukocytes of actively infected patients remains an area of uncertainty. it is still a matter of dispute, whether leukocytes support viral replication with subsequent production of infectious virus. in a new approach we developed and applied a sensitive fluorescence in situ hybridization assay for the precise intracellular localization of hcmv genomes in ...19968774688
expression and purification of recombinant, glycosylated human interferon alpha 2b in murine myeloma nso cells.we have expressed recombinant human interferon-alpha 2b in mammalian cells and isolated cell lines constitutively secreting very high levels of biologically active protein. the expression system takes advantage of the strong human cytomegalovirus immediate early promoter in mouse myeloma nso cells and glutamine synthetase as a selectable marker; spontaneous mutants with amplified gene copy numbers were selected by growth of primary transfectants in the presence of methionine sulfoximine. using t ...19968776749
molecular analysis of human cytomegalovirus strains from two lung transplant recipients with the same donor.a total of four cytomegalovirus (cmv) isolates were obtained from two cmv seronegative patients, each of whom received a lung transplant from the same seropositive donor. cmv was isolated from patient 1 from two bronchial alveolar lavage (bal) specimens, one obtained during treatment with ganciclovir (gcv) and a second during later treatment with foscarnet. both of these isolates are sensitive to gcv and foscarnet. cmv was isolated from patient 2 from a blood and a bal specimen obtained during t ...19968781616
a rapid and sensitive radioimmunoassay for the detection of human cytomegalovirus binding and infection of human fibroblasts.a rapid and sensitive radioimmunoassay for the quantitation of hcmv binding and infection of human fibroblasts (hff) was developed. the protocol involves the use of a monoclonal antibody (27-156) reactive with hcmv gb (alpha-gb), followed by an 125i-labeled second antibody to mouse igg. antibody to gb bound specifically to hff inoculated with hcmv when compared to sham inoculated cells or cells inoculated with hsv (strain kos). antibody to gb also bound to hff infected with hcmv 48 h prior to as ...19968783157
synthesis and antiviral evaluation of certain disubstituted benzimidazole ribonucleosides.ribosylation of 2-chloro-5(6)-nitrobenzimidazole (3) gave 2-chloro-5-nitro-1-(2,3,5-tri-o-acetyl-beta-d-ribofuranosyl)benzimidazol e (4a) and 2-chloro-6-nitro-1-(2,3,5-tri-o-acetyl-beta-d-ribofuranosyl)benzimidazol e (4b) as a mixture of positional isomers. subsequent hydrogenation of this mixture over raney nickel afforded the corresponding 5-amino and 6-amino derivatives 5 and 6. at this stage the products were readily resolved via silica column chromatography into pure isomeric forms, and the ...19968784445
gastrin gene expression is required for the proliferation and tumorigenicity of human colon cancer cells.the majority of human colon cancers express the gastrin gene, and a significant percentage bind gastrin-like peptides. however, it is not known if gastrin gene products are physiologically relevant to the growth and proliferation of human colon cancers. to investigate the functional role of gastrin gene expression, we examined the effect of gastrin antisense (as) rna expression on the growth and tumorigenicity of colon cancer cells. the full-length human gastrin cdna was cloned in the as directi ...19968797575
protection of chickens from newcastle disease by vaccination with a linear plasmid dna expressing the f protein of newcastle disease virus.to evaluate the usefulness of a dna vaccine for chickens, we constructed a plasmid vector expressing the newcastle disease virus f protein (ndv-f) under the control of the human cytomegalovirus immediate early enhancer and chicken beta-actin gene promoter. one-week-old chickens injected intramusculary with the circular plasmid dna did not produce significant levels of antibody against ndv-f. however, two of five birds injected with the linearized plasmid dna produced high levels of the antibody. ...19968817820
the effect of ph, hydrogen peroxide and temperature on the stability of human monoclonal antibody.the stability of human monoclonal antibody (c23), which is being developed as a passive immunotherapeutic agent against human cytomegalovirus, was investigated. c23 (about 2 mg ml-1) was incubated under sterile conditions for 14 days in buffers with different ph values (ranging from 4-10), in hydrogen peroxide solutions with different concentrations (0.01% or 0.1%), and in saline at 8 degrees c or 37 degrees c. samples were collected on days 0, 3, 7 and 14, and various physicochemical or biologi ...19968818025
anticytomegaloviral activity of methotrexate associated with preferential accumulation of drug by cytomegalovirus-infected cells.we extend the observation that inhibitors of pyrimidine biosynthesis are active against human cytomegalovirus by demonstrating that methotrexate (mtx) has preferential activity against cytomegalovirus replication. the 50% and 90% inhibitory concentrations of mtx for inhibition of cytomegaloviral dna replication at 3 days postinfection in mrc-5 cells were 0.05 and 0.2 microm, respectively. no cell toxicity was observed in uninfected confluent cells at the highest concentration tested (1 microm). ...19968834893
cidofovir.cidofovir is a nucleotide analogue which inhibits viral dna polymerase and is effective against human cytomegalovirus (cmv) infection. it is phosphorylated to its active form by cellular enzymes. with the long intracellular half-life of its metabolites, cidofovir can be administered weekly during induction and every other week during maintenance therapy. viral resistance has not been documented in patients treated with cidofovir to date, but has developed in vitro. immediate cidofovir therapy de ...19968841740
potent anti-murine cytomegalovirus activity and reduced nephrotoxicity of ganciclovir cyclic phosphonate.ganciclovir cyclic phosphonate (sr3775) is a derivative of the r enantiomer (sr3773) of ganciclovir phosphonate (9-[((+/-)-1-hydroxymethyl-3-phosphono)propyloxymethyl]guanine), both of which are potent inhibitors of human ctyomegalovirus and murine cytomegalovirus (mcmv). against wild-type and four drug-resistant strains of mcmv, sr3773 was 2.3- to 3-fold more potent than sr3775. sr3775 was about half as active as sr3773 against mcmv infections in severe combined immunodeficient mice. however, w ...19968843317
human cytomegalovirus latent gene expression in granulocyte-macrophage progenitors in culture and in seropositive individuals.following infection with cytomegalovirus, human granulocyte-macrophage progenitors carry the viral genome but fail to support productive replication. viral transcripts arise from a region encompassing the major regulatory gene locus; however, their structure differs significantly from productive phase transcripts. one class, sense transcripts, is encoded in the same direction as productive phase transcripts but uses two novel start sites in the ie1/ie2 promoter/enhancer region. these transcripts ...19968855322
the effect of matrix attached regions (mar) and specialized chromatin structure (scs) on the expression of gene constructs in cultured cells and in transgenic mice.the flanking sequences of several genes have been shown to direct a position independent expression of transgenes. attempts to completely identify the insulating sequences have failed so far. some of these sequences contain a matrix attached region (mar) located in the flanking part of the genes. this article will show that the mars in cultured cells located in the 3' oh region of the human apolipoprotein b100 (apo b100) and within the sv40 genome were unable to stimulate and insultate transgene ...19958858571
serological and molecular evidence of enterovirus infection in patients with end-stage dilated cardiomyopathy.to study the relative diagnostic value of enterovirus-specific molecular biological and serological assays in patients with end-stage dilated cardiomyopathy, and to investigate the possible role of other cardiotropic viruses in dilated cardiomyopathy.19968868984
human cytomegalovirus us3 impairs transport and maturation of major histocompatibility complex class i heavy chains.the human cytomegalovirus (hcmv) early glycoprotein products of the us11 and us2 open reading frames cause increased turnover of major histocompatibility complex (mhc) class i heavy chains. since us2 is homologous to another hcmv gene (us3), we hypothesized that the us3 gene product also may affect mhc class i expression. in cells constitutively expressing the hcmv us3 gene, mhc class i heavy chains formed a stable complex with beta 2-microglobulin. however, maturation of the n-linked glycan of ...19968876135
detection of serum immunoglobulin m to human cytomegalovirus by western blotting correlates better with virological data than detection by conventional enzyme immunoassay.western blotting (immunoblotting) with proteins separated from purified human cytomegalovirus (hcmv) particles (viral wb) has repeatedly been shown to be a reliable and sensitive method for detecting hcmv-specific immunoglobulin m (igm). the aim of the present work was to determine whether igm detected by viral wb correlates with virological diagnosis better than conventional enzyme immunoassay (conv-eia). the presence of an active hcmv infection was documented on the basis of isolation of virus ...19968877143
detection of human herpes virus 6 in aids-associated retinitis by means of in situ hybridization, polymerase chain reaction and immunohistochemistry.the ubiquitous nature of hhv-6 and its genomic relationship with cytomegalovirus led us to evaluate an etiological link between hhv-6 and aids-associated retinitis in a prospective study. hhv-6 infection was studied in patients with aids-associated retinitis and in two control populations. eye pairs were obtained at necropsy from nine patients with aids-associated retinitis, four human immunodeficiency virus (hiv)-sero-positive patients with normal fundus examination and three hiv-seronegative p ...19968877761
lymphocyte responses to human cytomegalovirus in different groups of patients in britain and in adults from west africa and the middle east.antibody prevalence and lymphocyte proliferation responses to cytomegalovirus (cmv) and herpes simplex virus (hsv) were compared in several different groups of patients: genitourinary medical (gum) patients, hemophiliacs, men with clinical acquired immunodeficiency syndrome (aids) and cases of primary cmv mononucleosis, and also in adults in the general population (control subjects) comprising separate groups native to britain, west africa, and the middle east. among the british control subjects ...19968877764
a polymerase chain reaction to detect a spliced late transcript of human cytomegalovirus in the blood of bone marrow transplant recipients.a reverse transcription (rt) nested polymerase chain reaction (pcr) procedure is described for detecting rna to a spliced late gene (slg) of human cytomegalovirus (cmv), the product of which (175 bp) is easily differentiated in agarose gels from the product when the target is unspliced viral rna or dna (258 bp). the slg-rt-pcr has been compared against a semi-quantitative pcr for cmv dna in buffy-coat specimens collected weekly after bone marrow transplantation from 3 patients and against the re ...19968882644
evidence for the role of cell protein phosphorylation in human cytomegalovirus/host cell fusion.the mechanism by which human cytomegalovirus (hcmv) enters cells is unknown. we sought evidence that protein phosphorylation plays a role in hcmv infection in two ways: (1) by determining whether the degree of phosphorylation of a constitutively phosphorylated 92.5 kda putative cell membrane receptor for hcmv gh is changed following exposure to hcmv or monoclonal anti-idiotype antibodies (mab2) that antigenically mimic hcmv gh, and (2) by studying the effects of protein kinase inhibition on rece ...19968887496
igg subclass antibodies to human cytomegalovirus (cmv) in normal human plasma samples and immune globulins and their neutralizing activities.an enzyme-linked immunoabsorbent assay (elisa) was developed for quantitation of igg subclass antibodies to human cytomegalovirus (cmv) in human serum or plasma samples and in immune globulin (ig) preparations. the assay was based on the parallel titration of known concentrations of purified igg subclass myeloma proteins and a specific cmv antiserum. the purified igg subclass myeloma proteins were captured on an elisa plate pre-coated with anti-human kappa, anti-human lambda or a mixture of anti ...19968889058
human cytomegalovirus glycoprotein b genotypes in renal transplant recipients.based on sequence variation of the glycoprotein b (gb) gene, human cytomegalovirus (hcmv) strains can be classified into four gb genotypes. in a previous study of bone marrow transplant recipients, infection with the gb type 1 correlated with a more favorable clinical outcome than infection with the gb types 2, 3, or 4. the gb type was determined in 60 renal transplant and in 47 bone marrow transplant recipients using pcr and restriction analysis. all hcmv variants in patient specimens could be ...19968890038
over- and underrepresentation of short dna words in herpesvirus genomes.the relative abundance and rarity of dna words have been recognized in previous biological studies to have implications for the regulation, repair, and evolutionary mechanisms of a genome. in this paper, we review several different measures of abundance and rarity of dna words, including z-scores, representation ratios, and cross-ratios, that have appeared in the recent literature, and examine the concordance among them using the human cytomegalovirus genome sequence. we then rank all words of l ...19968891954
the human cytomegalovirus ie1-72 protein interacts with the cellular p107 protein and relieves p107-mediated transcriptional repression of an e2f-responsive promoter.the rb-related p107 protein has been implicated as an important control element in proper cell cycle progression. the p107 protein is thought to restrict cellular proliferation in part through its interaction with the e2f family of transcription factors and is, therefore, a specific target for regulation by several dna viruses. here, we demonstrate that p107 protein levels are induced in a biphasic manner in human fibroblasts during productive infection by the human cytomegalovirus (hcmv). expre ...19968892909
mutations in the carboxyl-terminal hydrophobic sequence of human cytomegalovirus glycoprotein b alter transport and protein chaperone binding.human cytomegalovirus glycoprotein b (gb) plays a role in the fusion of the virion envelope with the host cell membrane and in syncytium formation in infected cells. hydrophobic sequences at the carboxyl terminus, amino acids (aa) 714 to 771, anchor gb in the lipid bilayer, but the unusual length of this domain suggests that it may serve another role in gb structure. to explore the function(s) of this region, we deleted aa 717 to 747 (gb deltai mutation), aa 751 to 771 (gb deltaii mutation), and ...19968892927
virus-specific interaction between the human cytomegalovirus major capsid protein and the c terminus of the assembly protein precursor.we previously identified a minimal 12-amino-acid domain in the c terminus of the herpes simplex virus type 1 (hsv-1) scaffolding protein which is required for interaction with the hsv-1 major capsid protein. an alpha-helical structure which maximizes the hydropathicity of the minimal domain is required for the interaction. to address whether cytomegalovirus (cmv) utilizes the same strategy for capsid assembly, several glutathione s-transferase fusion proteins to the c terminus of the cmv assembl ...19968892933
cytoplasmic sequestration of p53 in cytomegalovirus-infected human endothelial cells.human umbilical vein endothelial cells were infected with human cytomegalovirus (hcmv) at a multiplicity of infection of 0.1 plaque-forming unit/cell and remained > 95% viable even after 10 days of infection. to induce apoptosis, control human umbilical vein endothelial cells and cells infected with hcmv for 3, 5, and 7 days were serum starved for 48 hours. almost one-half of the uninfected cells lost viability after 48 hours of serum starvation whereas hcmv-infected cells were virtually unaffec ...19968909243
intracellular production of a major cytomegalovirus antigenic protein in the methylotrophic yeast pichia pastoris.we have previously shown that single or multiple epitopes of the major human cytomegalovirus (hcmv) antigens, produced as fusion proteins in prokaryotes can be valuable diagnostic material in the serology of hcmv infection. in this work we moved to a eukaryotic system, to produce one of the most immunogenic hcmv antigens, ppul44 (also called pp52 due to its apparent molecular size on acrylamide gels), as a non-fusion protein, in an attempt to eliminate some non-specific reactivity of human sera ...19968918253
identification of the human cytomegalovirus g protein-coupled receptor homologue encoded by ul33 in infected cells and enveloped virus particles.human cytomegalovirus (hcmv), strain ad169, contains four genes (us27, us28, ul33, and ul78) that encode putative homologues of cellular g protein-coupled receptors (gcrs). gcrs transduce extracellular signals to alter intracellular processes, and there is evidence that hcmv may elicit such changes at early times following infection. the us27, us28, and ul33 genes are transcribed during infection, and the us28 gene product has been found to be a functional receptor for the beta-chemokine class o ...19968918538
partial substitution of the functions of the herpes simplex virus 1 u(l)13 gene by the human cytomegalovirus u(l)97 gene.the predicted amino acid sequence of the human cytomegalovirus u(l)97 protein bears partial homology to the herpes simplex virus 1 u(l)13 protein, especially in regions that are homologous to conserved domains characteristic of protein kinases. earlier studies showed that u(l)13 mediated the posttranslational processing of several herpes simplex virus 1 proteins including icp22. whereas no kinase activity has been specifically attributed to u(l)13, it has been shown that u(l)97 can phosphorylate ...19968918921
gene transfer by dna/glycosylated polylysine complexes into human blood monocyte-derived macrophages.macrophages are putative target cells for expressing an exogenous gene with therapeutical effects. knowing that macrophages express membrane lectins mediating endocytosis of their ligands, dna/glycosylated polylysine complexes were used to transfect human blood monocyte-derived macrophages. monocytes from human peripheral blood were matured in culture for 7 days to differentiate into macrophage-like cells in the presence of granulocyte-macrophage colony-stimulating factor (gm-csf). adherent cell ...19968919594
efficient expression of pro-urokinase by human lymphoblastoid namalwa kjm-1 cells using moloney retroviral promoter.we have compared the level of expression of several enhancer/promoters in human lymphoblastoid namalwa kjm-1 cells when fused to a common reporter gene. a cassette containing the pro-urokinase (pro-uk) coding sequence followed by the rabbit beta-globin and simian virus 40 (sv40) 3' nontranslated region was used for evaluation of the enhancer activity. cells containing moloney murine leukemia virus (mo-mulv) promoter had an average of 10-20 fold higher expression levels of pro-uk than those conta ...19958920107
[detection of human cytomegalovirus-antigen in peripheral blood cells by using flow-cytometer].prophlaxis of cmv-associated disease and/or early initiation of therapy is important in the management of patients with cmv infection. recently, we developed the cmv antigenemia assay: a rapid and quantitative method based on the detection of cmc antigens in peripheral blood leukocytes by flow cytometry. heparinized peripheral blood was obtained from healthy donors, bone marrow transplantation patients, patient with collagen disease and patients with adult t cell leukemia. to determine the pheno ...19968921675
characterization of human cytomegalovirus protease dimerization by analytical centrifugation.human cytomegalovirus, a member of the herpesvirus family, encodes a maturational protease required for processing of the assembly protein and virus replication. the protease is synthesized as a precursor protein that undergoes autoproteolytic cleavage to yield a mature, 28-kda enzyme. it has recently been demonstrated that mature human cytomegalovirus protease is capable of dimerization and that the dimer is the active species [darke, p. l., cole, j. l., waxman, l., hall, d. l., sardana, m. k., ...19968952514
delivery of human interferon-gamma via gene transfer in vitro: prolonged expression and induction of macrophage antimicrobial activity.daily parenteral administration of exogenous interferon-gamma (ifn-gamma) induces or accelerates recovery in experimental and human infections. to develop an alternative delivery system, a replication-defective recombinant adenovirus expressing human ifn-gamma was constructed. the complete coding region of ifn-gamma was amplified by rt-pcr and inserted into an adenovirus cloning vector under the control of a human cytomegalovirus promoter. recombinant adenovirus containing the ifn-gamma minigene ...19968974003
human cytomegalovirus capsid assembly protein precursor (pul80.5) interacts with itself and with the major capsid protein (pul86) through two different domains.we have used the yeast gal4 two-hybrid system to examine interactions between the human cytomegalovirus (hcmv) major capsid protein (mcp, encoded by ul86) and the precursor assembly protein (pap, encoded by ul80.5 and cleaved at its carboxyl end to yield ap) and found that (i) the pap interacts with the mcp through residues located within the carboxy-terminal 21 amino acids of the pap, called the carboxyl conserved domain (ccd); (ii) the pap interacts with itself through a separate region, calle ...19978985337
liposomal delivery of the herpes simplex virus thymidine kinase gene in glioma: improvement of cell sensitization to ganciclovir.in this study, we investigated whether the regulation and the copy number of the herpes simplex virus thymidine kinase (hsvtk) gene increased the sensitization to ganciclovir (gcv) of glioma cell lines (rat c6 and human u118-mg) using liposome-mediated gene transfer. three recombinant plasmids carrying the hsvtk gene driven by the thymidine kinase promoter in single (pago) and double copy (pyed) or by the human cytomegalovirus promoter (pcmvtk) were used for the transfection. the dna delivery wa ...19968988841
effect of a modulator deletion on transcription of the human cytomegalovirus major immediate-early genes in infected undifferentiated and differentiated cells.differentiation-dependent expression of the human cytomegalovirus (hcmv) major immediate-early (mie) genes, encoding ie1 and ie2, may partly govern virus replication in monocytic thp-1 and embryonal carcinoma (tera-2) cells. the modulator of the mie promoter was shown previously in transient transfection assays to repress transcription from promoter segments in undifferentiated thp-1 and tera-2 cells but not in differentiated cells. to determine the biological importance of these findings, we co ...19978995648
identification and characterization of a g protein-coupled receptor homolog encoded by murine cytomegalovirus.this report describes the identification of a murine cytomegalovirus (mcmv) g protein-coupled receptor (gcr) homolog. this open reading frame (m33) is most closely related to, and collinear with, human cytomegalovirus ul33, and homologs are also present in human herpesvirus 6 and 7 (u12 for both viruses). conserved counterparts in the sequenced alpha- or gammaherpesviruses have not been identified to date, suggesting that these genes encode proteins which are important for the biological charact ...19978995678
human immunodeficiency virus type 1 gp120 stimulates cytomegalovirus replication in monocytes: possible role of endogenous interleukin-8.recombinant gp120, but not other human immunodeficiency type 1 (hiv-1) structural proteins, dose-dependently stimulates human cytomegalovirus (hcmv) immediate-early antigen (iea) expression and infectious virus yield in freshly isolated normal monocytes infected with hcmv. monoclonal antibodies (mabs) recognizing the gp120 v3 loop, as well as v3 loop octameric multibranched peptides and antibody to galactocerebroside, but not scd4, abrogate the gp120 stimulation of iea expression, suggesting tha ...19978995686
intracellular localization and expression of the human cytomegalovirus matrix phosphoprotein pp71 (ppul82): evidence for its translocation into the nucleus.a polyclonal antiserum, raised against a pp71 fusion protein, was prepared in order to investigate the biosynthesis and localization of the matrix protein pp71 of human cytomegalovirus (hcmv), the ul82 gene product, during the hcmv infectious cycle in human fibroblasts. transcription of the pp71-specific bicistronic 4.0 kb mrna and pp71 biosynthesis exhibited a biphasic pattern during one round of the hcmv infectious cycle, with a first peak at 12 h and a second at 72 h post-infection (p.i.). cy ...19969000101
a novel western blot test containing both viral and recombinant proteins for anticytomegalovirus immunoglobulin m detection.we devised a novel western blot (wb) test for anti-human cytomegalovirus (hcmv) immunoglobulin m (igm) detection which contains viral structural polypeptides, significant portions of recombinant p150 (ppul32), and a significant portion of the most immunogenic nonstructural protein p52 (ppul44). this new test was evaluated in latently infected blood donors, pregnant women, and transplant recipients with ongoing hcmv infection and shown to be more sensitive and specific than traditional wb and con ...19979003603
initial characterization of autoprocessing and active-center mutants of cmv proteinase.human cytomegalovirus (cmv) encodes a unique serine proteinase that is required in the maturation of the viral capsid. the cmv proteinase can undergo autocatalytic activation and is subject to proteolytic self-inactivation. mutant enzyme forms were prepared to eliminate the initial autoprocessing site and thus form an active single-chain protein for structure-function studies. two mutants of cmv proteinase were cloned and expressed in escherichia coli. the a143v mutant was a conservative substit ...19969008301
polyanion inhibitors of human immunodeficiency virus and other viruses. 5. telomerized anionic surfactants derived from amino acids.omega-acryloyl anionic surfactants, whose polar heads are derived from amino acids, have been telomerized to prepare polyanions of a predetermined molecular weight. the main goal of this study was to verify whether the antiviral activity is influenced by the degree of polymerization of the polyanions. the oligomeric polyanions were evaluated for their activity against human immunodeficiency virus (hiv-1 or hiv-2) and various other rna and dna viruses. with regard to their anti-hiv activity, a mi ...19979022800
detection of human viruses in periodontal pockets using polymerase chain reaction.even though viruses have been implicated in the etiology of several medical and dental disorders, little or no data are available on the possible involvement of human viruses in the pathogenesis of human periodontal disease. this study investigated the presence of human cytomegalovirus, epstein-barr virus, herpes simplex virus, human papillomavirus and human immunodeficiency virus (hiv) in crevicular fluid samples from 30 patients with advanced periodontitis and 26 subjects with gingivitis. vira ...19969028252
the role of atf in regulating the human cytomegalovirus dna polymerase (ul54) promoter during viral infection.previous analysis of the human cytomegalovirus (hcmv) dna polymerase (ul54) early gene promoter demonstrated that transcriptional activation of this gene is dependent upon the interaction of cellular transcription factors with viral transactivators (j. a. kerry, m. a. priddy, t. y. jervey, c. p. kohler, t. l. staley, c. d. vanson, t. r. jones, a. c. iskenderian, d. g. anders, and r. m. stenberg, j. virol. 70:373-382, 1996). a sequence element, ir1, was shown to be the primary regulatory element ...19979032345
comparison of human cytomegalovirus (hcmv) protease sequences among laboratory strains and seven clinical isolates.the nucleotide sequence of the human cytomegalovirus (hcmv) protease gene from two laboratory strains and seven clinical isolates, both ganciclovir-sensitive and -resistant, was examined to determine the genetic variability of the hcmv protease catalytic domain and to identify changes that may alter the efficacy of designed protease inhibitors. the towne strain varied from ad169 at 12 nucleotides and led to one amino acid change at position 12 (ala to thr). the clinical isolates had amino acid s ...19979037377
fibroblasts, epithelial cells, endothelial cells and smooth muscle cells are major targets of human cytomegalovirus infection in lung and gastrointestinal tissues.high titre replication of human cytomegalovirus (hcmv) in cell culture is restricted to primary human fibroblasts. during acute infection in vivo, hcmv nucleic acids and antigens have been found in various organs. using only morphological criteria, inconsistent data have been reported about the cell types that can be infected by hcmv. in particular, the role of fibroblasts in organ infections has remained unclear. to define accurately the target cells of hcmv in vivo, tissue sections from lung a ...19959049319
evaluation of a novel lipid prodrug for intraocular drug delivery: effect of acyclovir diphosphate dimyristoylglycerol in a rabbit model with herpes simplex virus-1 retinitis.acyclovir diphosphate dimyristoylglycerol is a lipid prodrug of acyclovir that forms liposomes and provides substantial activity against herpes simplex virus, acyclovir-resistant strains of herpes simplex virus, and human cytomegalovirus. we therefore tested this promising new drug in a rabbit model of herpes simplex retinitis.19979051844
design, synthesis, and antiviral evaluation of 2-chloro-5,6-dihalo-1-beta-d-ribofuranosylbenzimidazoles as potential agents for human cytomegalovirus infections.2-chloro-5,6-difluorobenzimidazole (8) was prepared from 4,5-difluoro-2-nitroaniline (5) via successive reduction, cyclization, and diazotization reactions. 2-chloro-5,6-dibromobenzimidazole (10) was obtained by a direct bromination of 2-chlorobenzimidazole (9) with bromine-water. 2-chloro-5,6-diiodobenzimidazole (15) was synthesized by a stepwise transformation of the nitro functions of 2-chloro-5,6-dinitrobenzimidazole (11) into iodo groups via diazotization reactions. ribosylation of 8, 10, a ...19979057868
human cytomegalovirus us2 destabilizes major histocompatibility complex class i heavy chains.human cytomegalovirus (hcmv) infection causes down-regulation of major histocompatibility complex class i heavy chains. we determined previously that there are two hcmv loci which encode functions responsible for that phenotype and that us11 is one of these loci (t. r. jones, l. a. hanson, l. sun, j. s. slater, r. m. stenberg, and a. e. campbell, j. virol. 69:4830-4841, 1995). through the construction and analysis of defined viral mutants and stably transfected cell lines, we identify us2 as the ...19979060656
the immediate early gene of human cytomegalovirus stimulates vascular smooth muscle cell proliferation in vitro and in vivo.cytomegalovirus (cmv) infection has been carefully studied regarding the relationship with the vascular smooth muscle cell (vsmc) proliferation in either atherosclerosis or post-angioplasty restenosis, but its role in the vessel wall has yet to be elucidated. to clarify the pathogenic ability of cmv in the vessel wall, we transduced the immediate early (ie) gene of cmv into the vsmcs and endothelial cells (ecs) by hemagglutinating virus of japan-liposome method. the in vitro ie gene transfer dem ...19979070298
ganciclovir-loaded polymer microspheres in rabbit eyes inoculated with human cytomegalovirus.to test the antiviral effect of ganciclovir released from biodegradable polymer microspheres in rabbit eyes inoculated with human cytomegalovirus (hcmv).19979071221
interleukin-10 immunoadhesin production by a replication-defective adenovirus.the present study examined the use of replication-defective adenovirus for in vitro production of an immunoadhesin. a recombinant adenovirus, rendered replication defective by deletion of the e1 gene, was constructed to contain the murine interleukin-10 gene fused in frame with the hinge, ch2, and ch3 domains of the murine immunoglobulin gamma 1 heavy chain constant region gene under the control of the human cytomegalovirus promoter. the resultant recombinant virus, ad5.hcmv.mil-10:hfc, was used ...19979075770
successful quantification of cytomegalovirus dna by competitive pcr and detection with capillary electrophoresis.human cytomegalovirus (hcmv) is responsible for severe infections in immunocompromised patients. viral load has recently been identified as one of the major risk factors for subsequent development of hcmv disease. in this context, we developed a protocol allowing rapid, sensitive and precise quantification of hcmv dna using competitive pcr run to saturation. long primers were used for amplification, and internal dna standard was constructed by pcr, with a primer inducing formation of a loop on t ...19979076710
correlation between human cytomegalovirus quantitative p72 antigenemia and viremia.we present a prospective study of the correlation between the human cytomegalovirus (hcmv) quantitative antigenemia with monoclonal antibody to p72 protein (immediate-early antigen) and the number of infected cell foci detected in the shell-vial culture. a comparative study was made of the value of quantitative antigenemia (pp65 and p72) in 14 patients.19969077431
measurement of human cytomegalovirus-associated dna polymerase activity in patient urine as a potential diagnostic tool.virus-associated dna polymerase activity has recently been proposed for the detection of human cytomegalovirus (hcmv) in urine, a method that should allow rapid and quantitative determination of the viral load. in this report, the virus-associated dna polymerase activity recovered from the urine of a group of patients shedding hcmv was measured using a poly(da) oligo(dt) 12-18 synthetic template after polyethylene glycol precipitation of the virions. detection of virus-associated dna polymerase ...19969078470
the class i mhc homologue of human cytomegalovirus inhibits attack by natural killer cells.recognition and destruction of virus-infected cells by class i major histocompatibility complex (mhc) restricted cytotoxic t lymphocytes (ctl) is a central part of the immune system's attempts to control and eliminate virus infection. it is therefore not surprising that many viruses have evolved strategies to interfere with the processing and presentation of peptide antigen on class i mhc molecules (reviewed in ref. 1). these mechanisms act to prevent or reduce expression of mhc molecules at the ...19979087413
alterations in the expression of elam-1, icam-1 and vcam-1 after in vitro infection of endothelial cells with a clinical isolate of human cytomegalovirus.human cytomegalovirus (hcmv) infection of endothelial cells resulted in increased adhesion of the cells to peripheral blood leukocytes. it was demonstrated by flow cytometry that increased adhesiveness parallels the increased expression of cell surface adhesion molecules (elam-1, icam-1, vcam-1). the increased adhesion of pmn and t-lymphocytes was due to upregulation in the expression of elam-1 and icam-1. the upregulation of vcam-1 resulted in the increased adhesiveness of monocytes and t-lymph ...19979087954
a model system for human cytomegalovirus-mediated modulation of human immunodeficiency virus type 1 long terminal repeat activity in brain cells.previously, our laboratory showed that human cytomegalovirus (hcmv) activates human immunodeficiency virus type 1 (hiv-1) in brain-derived cells with limited hiv-1 gene expression but inhibits hiv-1 in cells fully permissive for replication of both viruses (f. m. jault, s. a. spector, and d. h. spector, j. virol. 68:959-973, 1994). to investigate these effects further, we developed a model system that uncouples hiv-1 gene expression from long terminal repeat (ltr) activity. two monoclonal u373-m ...19979094643
human cytomagalovirus ie1 and ie2 proteins are mutagenic and mediate "hit-and-run" oncogenic transformation in cooperation with the adenovirus e1a proteins.some epidemiological studies have suggested a possible link between human cytomegalovirus (hcmv) infection and various malignancies, and hcmv has been shown to transform cultured cells. however, viral dna is not detected in most transformants, and the mechanism by which hcmv might contribute to oncogenesis has remained obscure. here we show that the hcmv immediate early 1 and 2 genes can cooperate with the adenovirus e1a gene to generate transformed foci of primary baby rat kidney cells. hcmv ge ...19979096395
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