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At least 865 records · Page 48Linked to original sources
PubMed1998

[Expression regulation of major late promoter tripartite leader sequence in adenovirus late infection].

OBJECTIVE: To investigate the influence of adenovirus major late tripartite leader sequence on specific exportation of virus mRNA in the late phase of adenovirus infection. METHODS: We constructed a marked, human beta-actin minigene under the control of the glucocorticoid-inducible enhancer and promoter of mouse mammary tumor virus, and inserted it into the left end of the adenovirus type V genome. The promoter gene, designated MA, as well as of a sibling, which differed only in inclusion of a cDNA copy of the adenovirus major late tripartite leader sequence upstream of beta-actin sequences termed MtplA, in recombinant virus-infected cells was strictly dependent on addition of deoxymethesone to the medium. Nuclear and cytoplasmic reporter RNA species were compared by Northern blotting, primer extension and pulse-chase assay. RESULTS: The high concentrations of newly-synthesized RNA were accumulated in cytoplasm when tripartite leader sequence was present in reporter RNA, despite the equal rates of transcription of the two reporter genes. Nuclear and cytoplasmic reporter RNA species compared by Northern blotting, primer extension, and pulse-chase provided no evidence for altered processing induced by tripartite leader sequence. CONCLUSION: The tripartite leader sequence, known to facilitate translation of mRNA during the late phase of adenovirus infection, can also modulate mRNA export from the nucleus.

5' Untranslated Regions↗
PubMed1997

[Gene expression and distribution in mouse abdominal cavity mediated by adenovirus].

Infection and expression of recombinant human adenovirus in mouse abdominal cavity was reported. After adenovirus vector Ad/RSV-beta-gal harboring the E. coli lacZ marker gene was injected into mice abdominal cavity, the peritoneal surface of jejunum, ileum, colon, uterus, liver, spleen, stomach, bladder, abdominal wall, diaphragm and testis was found large patches of lacZ-positive cells. But the adenovirus vector was not able to penetrate the peritoneum, as demonstrated by histochemical staining. Another adenovirus vector Ad/RSV-tk harboring the HSV-tk gene was injected into mouse abdominal cavity and the mouse was treated with ACV. No acute toxic reaction was observed. Based on these data, the feasibility of gene therapy of malignant tumor within abdominal cavity with adenovirus mediated TK/GCV system was discussed.

Abdomen↗
PubMed1975

Nucleotide sequence homology between a virus-specific 6S RNA and various RNA species from adenovirus-2-infected KB cells.

To investigate the metabolic fate of a 6S RNA (VA-RNA), which appears in human cell lines only after infection with adenovirus, various species of RNA from the cells at different stages of infection were tested for the presence of nucleotide sequence homologous to this RNA. RNA extracted from subvellular fractions of the cells was further fractionated by sucrose gradient centrifugation and analysed by acrylamide gel electrophoresis or by competition against 32P-labeled VA-RNA in hybridization with adenovirus DNA. The results pointed to the following conclusions. (1) VA-RNA is synthesized in the cells throughout the entire period after infections, starting before the onset of viral DNA synthesis. (2) VA-RNA does not participate in protein synthesis as a messenger RNA or a precursor of transfer RNA. (3) The nucleotide sequence homologous to VA-RNA exists as a part of certain species of nuclear high molecular weight RNA in the infected cells.

Adenoviridae↗
PubMedDate not supplied

[B-lymphocytes in chronic herpetic and adenovirus eye infections].

Concentrations of serum immunoglobulins and the number of B-lymphocytes in the peripheral blood were determined in herpes and adenovirus infections of the eyes. In severe forms of ophthalmic herpes the level of IgM was found to be high at the peak of the disease. In ophthalmic adenoviral infection and level of IgA was decreased. At the peak of the disease, the relative and absolute number of B-lymphocytes was decreased in severe forms of herpetic and adenovirus infections of the eyes. In the period between relapses the number of B-lymphocytes is completely restored.

Adenoviridae Infections↗
PubMed1986

Modulation of the level of expression of cellular genes in adenovirus 12-infected and transformed human cells.

The level of expression of thymidine kinase (TK), heat shock protein 70 (HSP70), beta-tubulin and p53 was assessed in human embryo kidney cells (HEKs) infected with adenovirus type 12 (Ad 12) and Ad 12 early region 1 (E1) mutants. HSP70, beta-tubulin and p53 levels were unchanged but TK activity was dramatically increased following wild-type infection. The initial activation of TK required the expression of the product of the E1A 13S mRNA but sustained expression only occurred with those viruses expressing the E1B proteins as well. A number of human cell lines transformed with either Ad 12 or Ad 5 E1 DNA were also assessed for the level of expression of HSP70, beta-tubulin and p53. Both HSP70 and beta-tubulin levels were greatly increased compared with primary human cells although there was considerable variation between lines. p53 was only expressed at high levels in Ad 12-transformed lines expressing E1A and E1B proteins.

Adenoviruses, Human↗
PubMed1996

A new strategy for large-scale preparation of high-titer recombinant adeno-associated virus vectors by using packaging cell lines and sulfonated cellulose column chromatography.

The extensive testing of adeno-associated virus (AAV) as a vector for human gene therapy has been hampered by low efficiency of the current packaging system, which is based on transient transfection with plasmid DNAs and infection with adenovirus in permissive cells. In an effort to resolve this problem, HeLa cell-based packaging cell lines were established. These packaging cells carry multiple copies of the AAV genome lacking the inverted terminal repeat (ITR) sequences. The AAV genes were silent in these cells but inducibly expressed by adenovirus infection. When the AAV vector plasmid containing the neoR gene flanked by the ITRs was also integrated into these cells, efficient production of the recombinant AAV particles occurred after adenovirus infection. AAV vector particles in cell lysates could be concentrated by sulfonated cellulose column chromatography. Using the packaging cells and the column chromatography technique, it is possible to prepare AAV vectors with the titer of higher than 10(8) cfu/ml or 5 x 10(10) particles/ml. This new strategy should be useful for testing AAV vectors in vivo.

Cellulose↗
PubMed1970

Transfer ribonucleic acid in KB cells infected with adenovirus type 2.

Populations of transfer ribonucleic acid (tRNA) extracted from control and type 2 adenovirus (Ad2)-infected KB cells were compared. No consistent differences in acceptor activity for 11 amino acids were observed. Comparison of methylated albumin-kieselguhr (MAK) elution profiles of arginyl-tRNA from control and infected cells revealed a minor modification in that the proportion of arginyl-tRNA eluting at high salt concentration was somewhat greater in infected cells. No similar differences were observed in MAK elution profiles of aspartyl-, isoleucyl-, leucyl-, phenylalanyl-, seryl-, tyrosyl-, and valyl-tRNA. Hybridization of 4S RNA from infected cells labeled by incorporation of (3)H-uridine with Ad2 deoxyribonucleic acid revealed the presence of a complementary species of RNA in this preparation. Hybridization of (3)H-arginyl-tRNA and of (3)H-aminoacyl-tRNA labeled by charging with (3)H-arginine or a (3)H-mixture of amino acids, respectively, failed to detect the presence of virus-specific tRNA in Ad2-infected cells.

Adenoviridae↗
PubMed1984

Shut-off of actin biosynthesis in adenovirus serotype-2-infected cells.

Adenovirus produces a dramatic shut-off of host protein synthesis after infection of HeLa cells. The level of actin messenger RNAs remained relatively unchanged after viral infection, when assayed by in vitro translation and two-dimensional gel electrophoresis analysis of the proteins or hybridization of the total cytoplasmic RNAs to the human actin gene. The distribution of actin mRNA in the polyribosomes is altered after adenovirus infection, with small polyribosomes and monoribosomes of the infected cells occupied by actin messages untranslatable in a rabbit reticulocyte lysate. The large polyribosomes still retain enough functional mRNAs to provide significant levels of actin protein in a rabbit reticulocyte in vitro translation system. In contrast, in homologous infected cell lysates, the translation of exogenous actin mRNA is greatly reduced when compared to uninfected HeLa cell lysates. In nuclease-treated uninfected or infected HeLa cell-free extracts, translation of viral mRNA is equally efficient and higher than that of actin mRNA. Thus, translational regulatory mechanisms which include inactivation of a part of the actin mRNA population accompanied by displacement to small polysomes and/or virus-induced modification of the cellular translational machinery to discriminate against cellular actin mRNA seem to account for the sharp reduction in actin protein synthesis of adenovirus-infected cells.

Actins↗
PubMed1956

Electron microscopy of HeLa cells infected with adenoviruses.

HeLa cells were infected with adenoviruses (types 1-4) and sectioned for electron microscopy after intervals of 20 to 48 hours. Clusters of virus-like particles were found within the nuclei of infected cultures but not in those of uninfected controls. The particles were often arranged in rows as if in crystalline formation. Maximal diameter of particles was approximately 65 mmicro, and internal bodies were demonstrated. Lesions of infected cells included target-like structures of the nuclear membrane, large nuclear vacuoles (type 2), and increased numbers of large irregular electron-dense granules in the cytoplasm 48 hours after infection. Examination of infected cultures by light microscopy, using the Feulgen reaction, showed intranuclear inclusion bodies and a cytopathogenic effect consisting of clumping of cells without pyknosis of nuclei. A lipide stain showed numerous cytoplasmic granules that were not identical with the large, irregular, electron-dense granules of the cytoplasm. Practically all the cells showed the viral cytopathogenic effect, but only a minority of cells were found to contain virus-like particles or intranuclear inclusion bodies.

Adenoviridae↗
PubMed2004

A novel method for detection of virus-infected cells through moving optical gradient fields using adenovirus as a model system.

BACKGROUND: Most methods for cellular analysis require labeling with specific antibodies or dyes and are often destructive. We have developed a technology called Optophoresis trade mark, which measures cell physiology based on the cell's motion in a near-infrared optical gradient. This technique does not require labels, is nondestructive, and involves minimal sample processing. METHODS: We have used Optophoresis to interrogate nonproductive and productive adenovirus-infected cell lines. Using an adenoviral vector containing green fluorescent protein (GFP) as a secondary assay, we show that viral infection can be monitored with Optophoresis. RESULTS: In HeLa cells, adenovirus infection after 24 h caused a 12% to 17% increase in optophoretic motility of the cells. In 293 cells, adenovirus infection resulted in a 40% increase in the optophoretic motility. The P values obtained were 4.5 x 10(-11) between noninfected and infected HeLa cells, and 2.1 x 10(-13) between noninfected and infected 293 cells. Cells infected with adenovirus lacking the GFP reporter gene gave similar shifts. In a time course, we observed an optophoretic shift after 4 h of infection, well before GFP expression. CONCLUSIONS: Optophoresis provides nondestructive, label-free analysis of viral infection. Detection is independent of reporter gene expression and can be observed early in the infection process.

Adenoviridae↗
PubMed1990

Severe chronic active Epstein-Barr virus infection syndrome and adenovirus type-2 infection.

Four patients from 4 to 24 years of age (3 males, 1 female) with generalized lymphadenopathy, hepatosplenomegaly, and intermittent fever associated with chronic active Epstein-Barr virus (EBV) infection were investigated. Laboratory data showed polyclonal gammopathy and a tendency for bone marrow suppression. Noteworthy were the extremely elevated immunoglobulin G (IgG) antibody titers to Epstein-Barr viral capsid antigen (VCA) (range, 10,240-81,920) and early antigen (EA) (range, 1,280-40,960). All patients had IgA antibodies to VCA and EA. Subtle, heterogeneous immune functional defects were observed in all four patients. Another unusual feature was our inability to establish spontaneous or B95-8 EBV-immortalized lymphoblastoid cell lines (LCLs) due to a marked cytopathic effect (CPE). Thus, we investigated for other viruses. Both IgG and IgM antibodies to adenovirus type-2 (Ad-2) were positive by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence (IF) test, suggesting recent or activated Ad-2 infection had occurred. Dual active EBV and Ad-2 infections were likely etiologic in this severe chronic active EBV infection syndrome.

Adenoviridae Infections↗
PubMed1976

Characterization of DNA polymerase associated with nuclear membrane fractions from uninfected and adenovirus 2-infected KB cells.

We have isolated a nuclear membrane fraction from KB cells infected with human adenovirus 2 that synthesizes exclusively small viral DNA chains (approx. 9 S) in vitro (Yamashita, T., Arens, M. and Green, M. (1975) J. Biol. Chem. 250, 3273-3279). The DNA polymerase activity present in the adenovirus 2 DNA-nuclear membrane complex was purified through chromatography on phosphocellulose and DEAE-cellulose, glycerol gradient centrifuation and DNA-cellulose chromatography. A single peak of enzymatic activity sedimented in glycerol gradients at about 6.7 S which corresponds to a molecular weight of 125000. The enzyme preparation in the step of glycerol gradient centrifugation utilized activated calf thymus, KB cell and adenovirus 2 DNA as template-primer in the presence of Mg2+; Km values for these DNAs were 5.5, 4.0, and 0.8 mug/ml, respectively. The partially purified enzyme preparation was characterized by several criteria which were compared to the properties of the three major mammalian DNA polymerases, alpha, beta, and psi. On the basis of template-primer preference, effect of salt, inhibition by N-ethylmaleimide and Km for dTTP, the DNA polymerase activity from the membrane complex can be distinguished from the alpha and beta DNA polymerases. The elution profile from DNA cellulose revealed a minor peak (I) and a major peak (II) of DNA polymerase activity utilizing poly(A) -(dT)10 as template-primer in the presence of Mn2+ - Peak II from DNA cellulose, which contained about 90% of the total DNA polymerase activity eluted from the column, was 2-3 times as active with poly(A) - (dT)10 as template-primer in the presence of Mn2+ than with activated calf thymus DNA in the presence of Mg2+. On the other hand, peak I had a low ratio of poly(A) - (dT)10 to activated calf thymus DNA activity. DNA polymerase was also purified from the nuclear membrane fraction of uninfected KB cells by the same procedures as those used in enzyme purification from the adenovirus 2 DNA-nuclear membrane complex. A minor peak and a major peak of DNA polymerase activity utilizing poly(A) - (dT)10 as template primer in the presence of Mn2+ were again observed that eluted from DNA cellulose at the same KCl concentrations as peak I and II from adenovirus 2-infected cells. The enzymes of the nuclear membrane fraction of uninfected KB cells could not be differentiated from the enzymes of the adenovirus 2 DNA-nuclear membrane complex through any of the purification steps nor by their template specificities. These results indicate that the predominant enzyme in the adenovirus 2 DNA-nuclear membrane complex and in the KB cell nuclear membrane complex belongs to the class of DNA polymerase psi.

Adenoviruses, Human↗
PubMed1994

Adenovirus pulmonary infections identified by PCR and in situ hybridisation in bone marrow transplant recipients.

AIMS: To investigate adenovirus pulmonary infections in bone marrow transplant (BMT) recipients. METHODS: Formalin fixed, paraffin wax embedded lung tissue was examined from 13 necropsy cases after BMT using PCR and in situ hybridisation to detect adenovirus DNA. The E1A region of the adenoviral genome was targeted for PCR. In situ hybridisation was performed only in the PCR positive cases. RESULTS: Of the 13 lung specimens analysed, nine cases were negative for adenoviral nucleic acid. Four (30%) PCR and two (15%) in situ hybridisation positive cases were found. In some of the patients there were clinical and pathological indications that some diseases might be associated with adenovirus infection--haemorrhagic cystitis (three cases); necrotising pneumonia (one case). In necrotising pneumonia in which no pathogenic agents had been shown by conventional histological study, the in situ hybridisation technique showed positive staining for adenovirus. In a patient who died of renal failure caused by adenovirus nephritis, both PCR and in situ hybridisation were positive in the lung as well as in the kidney, although no histological change was found. Two PCR positive cases lacked positive sites for adenovirus by in situ hybridisation. CONCLUSIONS: The combination of PCR and in situ hybridisation could be useful for diagnosing adenovirus infection of the lung in BMT recipients. These results provide a basis for exploring further the clinical use of PCR and in situ hybridisation to diagnose adenovirus infection.

Adenovirus Infections, Human↗
PubMed1982

Immune electron microscopy and a cultural test in the diagnosis of adenovirus ocular infection.

Immune electron microscopy (IEM) and virus isolation in cell culture were compared in the diagnosis of adenovirus ocular infection during an outbreak of the disease in 1979. Eleven of 14 patients with a keratoconjunctivitis clinically indicative of adenovirus infection had IEM evidence of adenovirus infection or had the virus isolated from ocular swabs. The IEM was positive in 8 patients. Virus as isolated from 10 patients. IEM was positive in one culture negative patient. Since IEM provides a rapid and sensitive method for the detection of adenovirus in human tears, it may be a valuable diagnostic tool for the clinician.

Adenoviridae Infections↗