Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.2.1.31 (beta-glucuronidase)
7,680 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A vaccinia virus-specific cytotoxic protein (3--10 X 10(4) daltons) had previously been detected in extracts of infected HeLa cells. In this study the protein has been shown to be a late gene product and a virion component--almost certainly the monomer of the surface tubules of the vaccinia virion. The relationship of this cytotoxin to a number of early vaccinia-induced cytopathic effects was examined: it was not the mediator of vaccinia-induced early cell rounding, did not inhibit protein or RNA synthesis in cell-free systems or intact cells (after uptake-induction by hypertonic MgSO4), or cause the release of beta-glucuronidase from a crude preparation of HeLa cell lysosomes. Its possible role in vaccinia-induced cell degeneration, late in infection, is discussed.
...
PMID:Further studies on a vaccinia virus cytotoxin present in infected cell extracts: identification as surface tubule monomer and possible mode of action. 42 31

It was reported that neurotropin (NSP), an extract isolated from the inflamed skins of rabbits inoculated with vaccinia virus, activates murine T cell functions participating in cell-mediated immunity. The present study was undertaken to examine the effect of NSP on plastic dish-adherent macrophages (M phi) from ddY mice in vitro. Total activities of beta-glucuronidase and N-acetyl-beta-D-glucosaminidase in resident peritoneal M phi was slightly enhanced when the M phi were cultured with NSP (10-1000 micrograms/ml) for 48 and 96 hr, but no enhancement was noted in 24 hr culture. Intracellular activity of lactate dehydrogenase (LDH) was also strongly enhanced in a dose-dependent manner by culturing with NSP for 48 and 96 hr. The enhanced LDH activity in the M phi cultured with NSP for 96 hr was completely inhibited by cycloheximide, an inhibitor of protein synthesis. In addition, consumption of glucose in the culture media by the M phi was also enhanced by culturing with NSP for 96 hr. Intracellular activity of LDH and glucose consumption of plastic dish-nonadherent cells from normal mouse peritoneal cells, however, was not enhanced by NSP in 96 hr culture. In regard to allogeneic M phi-mediated cytostatic activity to P815-X2 mastocytoma, NSP had no effect on cytostatic activities of the resident and thioglycollate-induced M phi, although NSP by itself dose-dependently inhibited the growth of P815-X2 mastocytoma without affecting cell viability. These results suggest that NSP biochemically activates mouse peritoneal M phi in vitro, but the M phi activated by NSP can not inhibit the growth of P815-X2 mastocytoma.
...
PMID:[Immunopharmacological actions of neurotropin (4). Effect of neurotropin on mouse peritoneal macrophages]. 644 Aug 35

The role of the cap-binding complex, eIF4F, in the translation of vaccinia virus mRNAs has been analyzed within infected cells. Plasmid DNAs, which express dicistronic mRNAs containing a picornavirus internal ribosome entry site, produced within vaccinia virus-infected cells both beta-glucuronidase and a cell surface-targeted single-chain antibody (sFv). Cells expressing sFv were selected from nonexpressing cells, enabling analysis of protein synthesis specifically within the transfected cells. Coexpression of poliovirus 2A or foot-and-mouth disease virus Lb proteases, which cleaved translation initiation factor eIF4G, greatly inhibited cap-dependent protein (beta-glucuronidase) synthesis. Under these conditions, internal ribosome entry site-directed expression of sFv continued and cell selection was maintained. Furthermore, vaccinia virus protein synthesis persisted in the selected cells containing cleaved eIF4G. Thus, late vaccinia virus protein synthesis has a low requirement for the intact cap-binding complex eIF4F. This may be attributed to the short unstructured 5' noncoding regions of the vaccinia virus mRNAs, possibly aided by the presence of poly(A) at both 5' and 3' termini.
...
PMID:Vaccinia virus protein synthesis has a low requirement for the intact translation initiation factor eIF4F, the cap-binding complex, within infected cells. 976 26

Vaccinia virus (VV) is a useful expression vector for many laboratory applications. To date, approximately 60% ( approximately 120) of the VV genes remain uncharacterized. The thought of smallpox being used as a biological weapon has gained attention. In light of this, it is imperative that we continue to study the basic replication of VV, a poxvirus that is closely related to smallpox. A set of plasmid vectors were constructed to generate gene deletions (pZIPPY-NEO/GUS) in VV or for foreign gene expression (pBR-EXPRESS). The vectors contain the Escherichia coli neomycin resistance (neo) and beta-glucuronidase (gusA) genes as selectable markers to facilitate isolation of recombinant viruses. These are the first transfer vectors to use a neo/gusA selection system. We used these vectors to successfully generate a recombinant D9R deletion mutant of VV and to express E. coli lacZ gene. Results indicate that both vectors are highly suited for their designed purpose.
...
PMID:Construction of a novel set of transfer vectors to study vaccinia virus replication and foreign gene expression. 1258 96

Activities of the enzymes beta-glucuronidase, acid phosphatase, acid DNAase, acid RNAase, and acid protease have been measured in the lysosomal and supernatant fractions of mouse liver cells and monkey kidney cells before and after infection with mouse hepatitis virus and vaccinia virus, respectively. In the infected cells there was easily measurable release of lysosomal enzymes into the supernatant fraction. Evidence was presented that this is not an artefact of homogenization and precedes cell degeneration demonstrable histologically. It is suggested that release of lysosomal enzymes may explain some of the biochemical changes found in infected cells and may contribute to the cytopathic effects of some viruses.
...
PMID:Activation of lysosomal enzymes in virus-infected cells and its possible relationship to cytopathic effects. 1401 75

Downregulation of MHC class I molecules is believed to be often the cause of tumor immune escape and at the same time it is the major obstacle to T-cell based immunotherapy of tumors. In our experimental model, the C57BL/6 mice bearing tumors induced by TC-1/A9 cells characterized by expression of HPV16 oncogenes and downregulation of H-2b molecules were immunized with highly immunogenic E7GGG.GUS DNA vaccine expressing the fused gene of modified HPV16 E7 (E7GGG) with E.coli beta-glucuronidase (GUS). The DNA vaccine was administered by gene gun on days 7 and 14 after s.c. injection of tumor cells. The tumors in situ were injected with recombinant vaccinia virus MVA expressing the gene for murine granulocyte-macrophage colony-stimulating factor (MVA-GM-CSF). Two doses of the DNA vaccine combined with at least two consecutive local treatments with MVA-GM-CSF were able to inhibit significantly the growth of tumors. We have shown by ELISPOT-IFNgamma that in situ expression of the GM-CSF gene did not enhance the E7 specific systemic Tcell response. We found that local injections of MVA-GM-CSF induced an increase of intratumoral CD3+ T cell counts and that the DNA vaccination resulted in up-regulation of MHC type I molecules on tumor cells in vivo. We suppose that i.t. delivery of MVA-GM-CSF changed the local tumor microenvironment and rendered tumors more attractive and better accessible to effector T cells.
...
PMID:Combination of intratumoral injections of vaccinia virus MVA expressing GM-CSF and immunization with DNA vaccine prolongs the survival of mice bearing HPV16 induced tumors with downregulated expression of MHC class I molecules. 1782 23

Previously, we reported that a recombinant vaccinia virus (VACV) carrying a light-emitting fusion gene enters, replicates in, and reveals the locations of tumors in mice. A new recombinant VACV, GLV-1h68, as a simultaneous diagnostic and therapeutic agent, was constructed by inserting three expression cassettes (encoding Renilla luciferase-Aequorea green fluorescent protein fusion, beta-galactosidase, and beta-glucuronidase) into the F14.5L, J2R (encoding thymidine kinase) and A56R (encoding hemagglutinin) loci of the viral genome, respectively. I.v. injections of GLV-1h68 (1x10(7) plaque-forming unit per mouse) into nude mice with established (approximately 300-500 mm3) s.c. GI-101A human breast tumors were used to evaluate its toxicity, tumor targeting specificity, and oncolytic efficacy. GLV-1h68 showed an enhanced tumor targeting specificity and much reduced toxicity compared with its parental LIVP strains. The tumors colonized by GLV-1h68 exhibited growth, inhibition, and regression phases followed by tumor eradication within 130 days in 95% of the mice tested. Tumor regression in live animals was monitored in real time based on decreasing light emission, hence demonstrating the concept of a combined oncolytic virus-mediated tumor diagnosis and therapy system. Transcriptional profiling of regressing tumors based on a mouse-specific platform revealed gene expression signatures consistent with immune defense activation, inclusive of IFN-stimulated genes (STAT-1 and IRF-7), cytokines, chemokines, and innate immune effector function. These findings suggest that immune activation may combine with viral oncolysis to induce tumor eradication in this model, providing a novel perspective for the design of oncolytic viral therapies for human cancers.
...
PMID:Eradication of solid human breast tumors in nude mice with an intravenously injected light-emitting oncolytic vaccinia virus. 1794 38

Modified Vaccinia virus Ankara (MVA) constitutes a good candidate for the development of non-replicative expression viral vectors because it does not replicate in most of mammalian cells. It is essential, for the production of recombinant MVA, the availability of transfer vectors which allow the introduction of desired genes into non-essential regions for in vitro viral replication, by homologous recombination with the viral genome. In the present work, the transfer vectors named VT-MHA and VT-MTK were designed and obtained. They carried genomic regions corresponding to 1-303 and 608-948 positions of the MVA165R gene and 1-244 and 325-534 of the MVA086R gene, respectively, which flank a multiple cloning site for the insertion of foreign genes. In these vectors, the cassettes for the expression of lac Z or uidA genes were cloned, and the activity of the marker enzymes beta-galactosidase and beta-glucuronidase was confirmed in situ. Furthermore, the vector named VT-MTK-GUS was used to obtain and isolate pure recombinant MVA, which carried and expressed the uid A gene. The results herein constitute the basic tools for establishing the methodology to obtain recombinant MVA with the purpose of locally developing non-replicative viral vectors as candidate vaccines.
...
PMID:[Design and construction of transfer vectors in order to obtain recombinant modified vaccinia virus Ankara (MVA)]. 1798 47

Malignant pleural mesothelioma (MPM) is a fatal disease with a median survival of less than 14 months. For the first time, a genetically engineered vaccinia virus is shown to produce efficient infection, replication, and oncolytic effect against MPM. GLV-1h68 is a replication-competent engineered vaccinia virus carrying transgenes encoding Renilla luciferase, green fluorescent protein (both inserted at the F14.5L locus), beta-galactosidase (inserted at the J2R locus, which encodes thymidine kinase), and beta-glucuronidase (at the A56R locus, which encodes hemagglutinin). This virus was tested in six human MPM cell lines (MSTO-211H, VAMT, JMN, H-2373, H-2452, and H-2052). GLV-1h68 successfully infected all cell lines. For the most sensitive line, MSTO-211H, expression of green fluorescent protein (GFP) started within 4 hr with increasing intensity over time until nearly 100% of cells expressed GFP at 24 hr. All cell lines were sensitive to killing by GLV-1h68, with the degree of sensitivity predictable by infectivity assay. Even the most resistant cell line exhibited 44 +/- 3.8% cell survival by day 7 when infected at a multiplicity of infection of 1.0. Viral proliferation assays demonstrated 2-to 4-fold logarithmic replication of GLV-1h68 in the cell lines tested. In an orthotopic model, GLV-1h68 effectively prevented development of cachexia and tumor-related morbidity, reduced tumor burden, and cured MPM in both early and late treatment groups. GLV-1h68 was successfully used to treat MPM in vitro and in an orthotopic model (in vivo). These promising results warrant clinical investigation of GLV-1h68 as a novel agent in the treatment of MPM.
...
PMID:Novel oncolytic agent GLV-1h68 is effective against malignant pleural mesothelioma. 1875 10

Modified vaccinia virus Ankara (MVA) is a safe vector for high-level expression of proteins in mammalian cells. To simplify the molecular cloning procedures for shuttling genes into the MVA genome, we constructed generic destination plasmids that allow in vitro recombinational cloning (Gateway) and quick isolation of expression plasmids for any gene to be incorporated into the virus. Downstream purification steps were simplified by including N-terminal peptide tags (His, Strep, and Flag) in the generic plasmids. We demonstrate the ability to produce 10mg of beta-glucuronidase from 10(8) hamster cells and to purify tagged proteins with affinity gels.
...
PMID:Vectors for recombinational cloning and gene expression in mammalian cells using modified vaccinia virus Ankara. 2045 Aug 73


1

---