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Biomedical subjects

J Daillie

Publications and source records attributed to J Daillie.

At least 19 recordsLinked to original sources
PubMed1989

Relationship between nasopharyngeal carcinoma and high antibody titers to Epstein-Barr virus-specific thymidine kinase.

Epstein-Barr virus (EBV) has long been implicated in nasopharyngeal carcinoma (NPC). Recent studies in our and other laboratories have shown a correlation between the disease and high antibody titers to EBV-specific DNase. These data led us to also examine serial sera from healthy adults and patients with infectious mononucleosis or NPC, for their capacity to neutralize the EBV-specific thymidine kinase (TK) activity from chemically induced EBV-carrying human lymphoblastoid cells. Our results were the following: (i) sera were found that efficiently blocked the EBV-specific TK activity of induced-Raji TK- cell extracts, but not the host-cell TK activity from EBV-negative BJAB cells; (ii) a relationship appeared between high levels of EBV-specific TK-neutralizing activity in sera and NPC pathology, even though in this preliminary study the degrees of EBV-induced TK-blocking activity detected in sera were not significantly correlated with EBV-specific antibody titers; (iii) the EBV-induced TK-neutralizing activity was found in the main IgG fraction derived from NPC sera. These data must be compared with other known antibody responses to EBV for their clinical interest in NPC control.

Antibodies↗
PubMed1988

Altered expression of two Epstein-Barr virus early genes localized in BamHI-A in nonproducer Raji cells.

The Epstein-Barr virus-carrying lymphoblastoid cell line Raji has two major genomic deletions and is incapable of virus production. Two cDNA clones, c70 and c55, were constructed from early mRNA of P3HR-1 cells and localized, respectively, in BALF-2 and BARF-1 open reading frames where one of the major genomic deletion in Raji cells is situated. These were used to search the different early viral transcripts in producer P3HR-1 and nonproducer Raji lines. c70 and c55 hybridized with their corresponding mRNAs only in producer lines. Analysis with in vitro-synthesized RNA probes showed quite a different transcriptional profile in Raji cells than in P3HR-1 cells. In the P3HR-1 line, BALF-2 encodes a 3.4-kilobase (kb) mRNA during the early phase and a 3.3-kb mRNA during the late phase, and in the Raji line, the probe corresponding to BALF-2 hybridized with three mRNAs of 5.0, 3.1, and 2.4 kb; in P3HR-1 cells, BARF-1 encodes a group of 3'-conterminal transcripts (0.8, 1.2, 1.7, 2.7, 3.2, and 5.0 kb) during both the early and late stages; in Raji cells, however, 0.8-, 1.2-, and 1.7-kb mRNAs are absent, the only mRNAs transcribed being upstream of the deletion and of 5.0, 2.6, and 2.0 kb in size. In vivo and in vitro experiments demonstrated that the BALF-2 open reading frame encodes an early 135-kilodalton (kDa) protein which possesses DNA-binding ability and can be recognized by a herpes simplex virus ICP-8 antiserum. The BARF-1 open reading frame encodes in vitro a 26- to 33-kDa early protein recognized by anti-EA serum. The proteins of both two genes expressed in psi AM 22b cells were localized in nuclei. According to their properties, both proteins, particularly the BALF-2-encoded 135-kDa DNA-binding protein, could play a role in virus replication.

Cell Line↗
PubMed1987

Identification of an Epstein-Barr virus-specific desoxyribonuclease gene using complementary DNA.

We have recently obtained 18 distinct cDNA clones representing different genes expressed in the early phase of EBV infection. One of them, c37, which is situated at the position 12907-122451 in the B95-8 viral genome, is shown here to code for a viral desoxyribonuclease [DNase]. Cell free translation of c37-selected messenger RNA yielded a protein of about 52 KDa which was immunoprecipitated by a high EA titer serum from nasopharyngeal carcinoma patient. This protein showed a DNase activity which was resistant to high salt concentrations (150 to 300 mM KCl) and was specifically neutralized by EA positive serum. These properties are typical of the EBV-specific DNase activity that we recently described in chemically induced EBV-transformed lymphoid cells. The same results were obtained on cell-free translation of the native RNA synthesized in vitro from pGEM-37 plasmid containing the entire c37 cDNA sequence (1.53 Kb). These data indicate that the BGLF5 open reading frame contained in c37 encodes for the EBV-specific DNase.

Base Sequence↗
PubMed1987

Localization polymorphism of EBV DNA genomes in the chromosomes of Burkitt lymphoma cell lines.

The localization of Epstein-Barr virus (EBV) genomes in nuclei of the human lymphoblastoid cell lines Raji, Jijoye, P3HR-1, Daudi and Ramos was investigated by in situ hybridization with biotinylated EBV DNA probes. We found that all sites of hybridization were associated with the chromosomes. Only some of these sites were present on both chromatids and these had a non-random distribution; these sites could represent EBV sequences integrated at specific points on the chromosomes. The total mean site number corresponded with the number of viral DNA copies estimated in the different cell lines by other techniques, but the copy number was highly variable from cell to cell in a given line.

Burkitt Lymphoma↗
PubMed1986

Both Epstein-Barr virus (EBV)-encoded trans-acting factors, EB1 and EB2, are required to activate transcription from an EBV early promoter.

We have identified two Epstein--Barr virus (EBV) transacting factors which are involved in the transcriptional activation of EBV early promoters in latently infected Raji cells. In Raji cells, expression of the factor EB1 encoded by the open reading frame (ORF) BZLF1 is necessary and sufficient to disrupt latency. However, factor EB2 encoded by the ORF BMLF1- BSLF2 does not disrupt latency when expressed alone in Raji cells. Expression of an EBV activatable early promoter depends on the presence of both EB1 and EB2.

Cell Line↗
PubMed1986

Characterization of an Epstein-Barr virus-induced thymidine kinase.

Previous work from our laboratory suggested that the selective inhibition of Epstein-Barr virus (EBV) replication by 1-beta-D-arabinofuranosylthymine in human lymphoid cell lines involved the induction of a new thymidine kinase (TK) able to phosphorylate the thymidine analog. We further characterized this enzyme induced in various EBV-positive cell lines after viral genome activation with a combination of sodium butyrate and 12-O-tetradecanoylphorbol-13-acetate. The following results confirmed the existence of an EBV-specific deoxypyrimidine kinase: induction of EBV-related TK was connected with the appearance of viral early antigens in EBV-carrying cells; unexpected behaviors of the enzyme activity upon different fractionating treatments led to the conclusion that EBV-induced TK was extracted as a complex molecular form, larger than other known cellular or viral isozymes; enzymatic properties distinguished EBV-induced TK from host lymphoid cell isozymes but made it resemble other herpesvirus-specific deoxypyrimidine kinases, i.e., by partial inhibition by dTTP or ammonium sulfate, insensitiveness to dCTP, and nonstringent specificity for normal TK substrates. Genetic evidence is required to definitively ensure that EBV-specific TK actually is virus coded in EBV-transformed human lymphoid cells.

Antigens, Viral↗
PubMed1986

Epstein-Barr virus-specific DNA polymerase in virus-nonproducer Raji cells.

Virus-nonproducer Raji cells, when induced to early antigen synthesis by 12-O-tetradecanoyl-phorbol-13-acetate and sodium butyrate, showed an increase in DNA polymerase activity. This enzyme has the characteristics of a typical Epstein-Barr virus DNA polymerase with regard to chromatographical pattern and biological properties: it is eluted from DEAE-cellulose at 0.08 M NaCl, has a high salt resistance, is sensitive to phosphonoacetic acid and phosphonoformate, and shows a substrate preference for poly(dC)-oligo(dG12-18). The resistance of Epstein-Barr virus polymerase activity to aphidicolin is a property distinct from that of HSV DNA polymerase. Viral DNA polymerase activity increases in the absence of Epstein-Barr virus DNA replication, indicating that this enzyme is an early viral protein.

Antigens, Viral↗
PubMed1985

Construction and use of cDNA clones for the mapping and identification of Epstein-Barr virus early P3HR-1 mRNAs.

cDNA clones, specific for early Epstein-Barr virus (EBV) RNAs, were constructed from total cytoplasmic RNA of P3HR-1 TK- cells. From 10,000 cDNA clones screened, 22 virus-specific cDNA clones were selected by hybridization with a total EBV DNA. These clones were then precisely mapped on the EBV genome and the corresponding mRNAs were identified by Northern blot hybridizations. Most of them are clearly related to some of the open reading frames described by Baer et al. (Nature [London] 310:207-211, 1984). They represent at least 18 different genes active during the early viral cycle. The transcriptional activity of the virus during the early stage was also studied by dot blot hybridization of total early cDNA probe to EBV genomic fragments. Three main regions showed very strong hybridization with the cDNA probe: BamHI a, M, and L fragments, BamHI K, B, and G fragments, and BamHI B1 fragment (deleted in strain B95-8) and the adjacent right end of the DNA molecule. Seventeen of the cDNA clones were localized in these highly transcribed regions. The five others were dispersed all along the EBV genome.

Animals↗
PubMed1984

Epstein-Barr virus-specific DNase activity in nonproducer Raji cells after treatment with 12-o-tetradecanoylphorbol-13-acetate and sodium butyrate.

An Epstein-Barr virus (EBV)-specific DNase was induced in EBV nonproducer Raji cells after treatment with 12-O-tetradecanoylphorbol-13-acetate and sodium butyrate. The increase in EBV DNase activity was related to the appearance of early antigen-positive cells. The enzyme had a sedimentation coefficient of 4S and was resistant to 300 mM KCl, and its induction did not depend on viral DNA synthesis. The EBV-specific DNase activity was specifically inhibited by sera from patients who had nasopharyngeal carcinoma with high early antigen activities but not by sera from normal, healthy individuals. There was a correlation between the degree of anti-EBV DNase activity and the titers of early antigen antibody.

Antigen-Antibody Complex↗
PubMed1983

Effect of arabinofuranosylthymine on the replication of Epstein-Barr virus and relationship with a new induced thymidine kinase activity.

1-beta-D-Arabinofuranosylthymine (araT) is a selective inhibitor of Epstein-Barr virus replication induced in both thymidine kinase (TK)-negative (TK-) and TK+ variants of the lymphoid cell line P3HR-I. This analog has no effect on the growth of noninduced cells (T. Ooka and A. Calender, Virology 104:219-223, 1980). The synthesis of early antigens is not affected by the analog, whereas that of late viral capsid antigens is completely inhibited, as demonstrated by the indirect immunofluorescence technique; kinetic reassociation experiments have also shown that araT strongly inhibits replication of viral DNA. Phosphorylation of the tritiated form of the analog ([3H]araT) was analyzed by thin-layer chromatography in cultures of control and induced cells, and the results demonstrated that only induced cells can convert the analog to the triphosphate form. These results indicate that the selective effect of araT in induced cells is probably related to a new virally induced TK activity. Preliminary characterization of this new activity has shown that it is able to phosphorylate the analog specifically, whereas cellular TKs cannot. araTTP, a final phosphorylation product of araT, is a potent inhibitor of Epstein-Barr virus-specific DNA polymerase, suggesting a possible inhibitory action of this product on Epstein-Barr virus replication.

Antigens, Viral↗
PubMed1979

Juvenile hormone modifies larvae and silk gland development in Bombyx mori.

Topical application of a Juvenile Hormone Analogue (ZR 515) during the first half of the first half of the last (fifth) larval instar of Bombyx mori induces a prolongation of this instar and increases larvae weight and silk secretion. Later treatment can lead to an extra larval molt. Moreover, the hormone analogue stops either the development or the secretory activity of the silk gland, depending on the period of treatments. When Juvenile Hormone is administered at the beginning of the fifth instar, silk gland growth as well as RNA and protein syntheses are first inhibited and later resumed and amplified. Finaly silk production increases up to 150 per cent of controls. One of the primarly effects of treatments seems to be RNA synthesis. Thus Juvenile Hormone analogue application could be a new tool for studying RNA transcription and/or processing.

Animals↗
PubMed1979

Characterization of an Epstein-Barr virus-induced DNA polymerase.

The addition of iododeoxyuridine to P3HR-I cell cultures led to a large increase in both Epstein-Barr virus (EBV)-induced DNA polymerase activity and early antigen-positive cells. This EBV-induced DNA polymerase was separated from the cellular alpha- and beta-polymerases by sequential column chromatography on Sepharose 6B, DEAE-cellulose, and phosphocellulose, resulting in partial purification of about 320-fold. The partially purified-EBV DNA polymerase could be distinguished from the cellular DNA polymerases by its activation by salts, its catalytic properties, and its degree of sensitivity to N-ethylmaleimide, phosphonoacetic acid, araATP, and araCTP. The viral polymerase showed properteis similar to those reported for other herpesvirus DNA polymerases. The enzyme exhibited optimal activity for copying activated calf DNA in the presence of 50 mH (NH4)2SO4 and was resistant to 150 mM (NH4)2SO4. It utilized with high efficiency template-primer poly(dC)-oligo(dG)12-18 or poly(dA)-oligo(dT)12-18, but failed to copy poly(rA)-oligo(dT)10 and oligo(dT)10, indicating that this enzyme has characters distinct from DNA polymerase gamma, reverse transcriptase, and terminal deoxynucleotidyl transferase. Phosphonacetic acid inhibited not only EBV DNA polymerase, but also, to a lesser degree, the cellular polymerase alpha. AraATP did not severely inhibit viral activity, whereas the polymerase alpha was inhibited most effectively. Both EBV polymerase and polymerase alpha were inhibited at a comparable level by araCTP.

Ammonium Sulfate↗
PubMed1977

RNA polysomes and RNA synthesis in the silk glands of the silkworm Bombyx mori;.

Total RNA polymerase activity, as well as the proportion of alpha-amantin-sensitive and resistant during activity, have been measured in the posterior silk glands of the silkworm as a function of growth the fifth larval instar. During the first 5 days, termed the growth phase, the total enzyme activity and particularly the portion that is alpha-amantin-resistant increases to reach a peak value and thereafter declines during the secretory phase, Much of the enzyme remains firmly bound and insoluble. Heparin only only does not inhibit this insoluble and probably chromatin-bound activity which would indicate lack of initiation, but it enhances the activity. A large proportion of newly transcribed RNA is released from the transcription complex. The synthesis of RNA has been studied both qualitatively and quantitatively during the same period. RNA synthesis becomes important on the second day of the fifth instar, as does the RNA polymerase, and stays at a high level for several more days. The results from these studied as well as those with incorporation of 32P indicate interference of varying precursor pools in quantitatively measured RNA synthesis. However, RNA content as well as RNA synthesis in vitro show a close correlation with RNA polymerase activity. The labeled RNAs extracted at different days of the fifth instar have been fractioned on sucrose gradients; this demonstrated that the predominant product of RNA synthesis, as followed by [3H]uridine incorporation at short time intervals, is 45-S preribosomal RNA and 4-5 S RNA. The 45-S RNA is transformed to 19-S and 30-S ribosomal RNA as time progresses or after a chase with unlabeled and/or actinomycin D. There also exists a component heavier than 45S which is fairly rapidly labeled to a small extent.

Amanitins↗