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At least 1,243 records · Page 69Linked to original sources
PubMed1993

Fate of the fish pathogen Aeromonas salmonicida in the peritoneal cavity of rainbow trout.

A model was developed to study the fate of the fish pathogen Aeromonas salmonicida in vivo, inside a specialized intraperitoneal chamber implanted in rainbow trout, Oncorhynchus mykiss. Although normally recalcitrant to lytic agents in vitro, owing to the presence of its regular surface array (S layer), A. salmonicida was rapidly killed in the peritoneal cavity by a host-derived, soluble lytic activity present in peritoneal fluid. Peritoneal fluid was also found to kill other bacteria and lyse various types of erythrocytes, but was particularly lytic to A. salmonicida. Intraperitoneal survival of injected (free) A. salmonicida cells was several orders of magnitude higher than survival of implanted (restrained) cells. Injected free cells could evade the lytic activity of peritoneal fluid because they readily spread, initiating lethal infections. One evasion strategy was envisioned to be the penetration of peritoneal and (or) tissue macrophages. In spite of the killing mechanisms of these phagocytic cells, A. salmonicida was still able to survive and even replicate inside head kidney macrophages, thereby supporting the notion of A. salmonicida as a facultatively intracellular pathogen. Intraperitoneal chambers in rainbow trout may constitute a valuable experimental tool for studying the in vivo fate of A. salmonicida, and perhaps of other fish pathogens as well.

Aeromonas↗
PubMed1979

Isolation and properties of a phage lytic for non-smooth Brucella organisms.

A phage for non-smooth cells of Brucella abortus was isolated from a mixture of three brucella-phages incubated with rough brucella cells in the presence of N-methyl-N'-nitro-N-nitrosoguanidine. It did not lyse smooth cells of Br. abortus strains nor those of other Brucella species, but during the course of its replication in rough organisms a small proportion of phage mutants were produced which were similar in properties to smooth-specific phages. The rough-specific phage, R, itself resembled Weybridge phage in its morphological and serological properties. Phage R was found to contain DNA and segregated into high and low density fractions, both with plaque-forming activity, on ultra-centrifugation in CsSO4 gradients. Unlike the smooth-specific phages it attached to heat-labile receptors present on non-smooth brucella cells.

Adsorption↗
PubMed1989

Cytomegalovirus enhances lysis of HIV-infected T lymphoblasts.

A T4+ lymphoblastoid cell line (CR-10) persistently infected with the human immunodeficiency virus (HIV) and designated CR-10/NIT was superinfected with cytomegalovirus (CMV) isolated from peripheral blood mononuclear cells of a patient with AIDS. A productive CMV cycle in the CR-10/NIT lymphoblasts was demonstrated by fluorescent antibody staining (IF) using a monoclonal antibody (MAb) to the 150-kDa major capsid protein, by infectivity assays and by electron microscopy (EM). Two-color IF analysis showed that a small percentage of the CR-10/NIT cells were producing both CMV and HIV at any one time. EM studies revealed that all doubly infected cells were lysed whereas most cells infected only with HIV appeared intact. Cell lysis appeared 24 hr after superinfection of the CR-10/NIT cells with CMV and progressed to complete destruction of the cell culture between days 9 and 10. Our results suggest that CMV may convert a mildly cytopathic HIV infection of T lymphoblasts into a highly lytic process.

Antigens, Viral↗
PubMed1982

Propagation of satellite phage P4 as a plasmid.

Satellite phage P4 has two known options for propagation. In its lytic cycle, its regulatory functions can act in trans to alter the actions of a helper virus (P2), which then provides necessary gene products, including capsid proteins. P4 also can be propagated in the absence of a helper as a prophage, with distinct sites for integration within the Escherichia coli chromosome. We determined that a single spontaneous mutation (vir1) of phage P4 allows a third mode of propagation: as a plasmid (along with continued integration into the host chromosome). Hence, the P4 regulatory element is capable of (i) temperate; (ii) lytic, helper-dependent; and (iii) plasmid modes of development. These findings emphasize the close relationship between defective viruses and plasmids.

Cloning, Molecular↗
PubMed2005

Equine herpesvirus-4 kinetics in peripheral blood leukocytes and nasopharyngeal secretions in foals using quantitative real-time TaqMan PCR.

Based on the hypothesis that the viral load of cells infected with EHV-4 will likely change during the course of disease, TaqMan PCR was used to investigate and characterize the kinetics of EHV-4 viral DNA load (glycoprotein B gene) and transcriptional activity (glycoprotein B and latency-associated transcripts) in peripheral blood leukocytes (PBLs) and nasopharyngeal secretions (NSs) collected from 11 foals during a field outbreak of respiratory disease. The EHV-4 DNA load in PBLs was low and of short duration after onset of clinical signs. In contrast, the EHV-4 load in NSs remained high for the majority of the foals over a period of 4 weeks. Viral replication determined by detection of mRNA expression of the structural glycoprotein B was detected only in NSs during the first 7 days after onset of clinical signs for most foals. The majority of foals expressed latency-associated transcripts in NS sonly during the first 7 days after onset of clinical signs. Persistence of the expression of latency-associated transcripts in NS, as a reflection of a latent viral state, was not documented during the 28-day study period. Based on these results, it was concluded that lytic infection with EHV-4 can be diagnosed either by high EHV-4 DNA load of glycoprotein B gene or by detection of transcriptional activity of glycoprotein B.

Animals↗
PubMed2005

Characterization of West Nile viral replication and maturation in peripheral neurons in culture.

The North American West Nile virus (WNV), New York 1999 strain, appears to be highly neurotropic, and its neuroinvasiveness is an important aspect of human disease. The authors have developed an in vitro model to study WNV replication and protein processing in neurons. They compared WNV infection of the dorsal root ganglion (DRG) neurons (sensory neurons) and PC-12 cells (sympathetic neurons) to WNV infection of the mosquito cell line, C6/36, and Vero cells. WNV infection of both neuronal cell types and C6/36 cells was not cytopathic up to 30 days post infection, and continual viral shedding was observed during this period. However, WNV infection of Vero cells was lytic. Interestingly, WNV infection of neurons was not efficient, requiring a high multiplicity of infection of > or = 10. Indirect immunofluorescence assays using normal and confocal microscopy with flavivirus-reactive antibodies and WNV-infected neurons demonstrated viral antigen mostly associated with the plasma membrane and in the neurite processes. Treatment of WNV-infected C6/36, PC-12, or DRG cells with brefeldin A (BFA; a trans-Golgi inhibitor) or nocadazole (a beta-tubulin inhibitor) had little effect on viral maturation and secretion. Treatment of WNV-infected Vero cells with BFA resulted in a 1000-fold decrease in viral titer, but nocodazole had no effect. Our studies suggest that even though PC-12 and DRG neurons are mammalian cells, viral protein processing and maturation in these cells more closely resembles replication in C6/36 insect cells than in mammalian Vero cells.

Animals↗
PubMed2003

Sequence analysis of the genome of the temperate Yersinia enterocolitica phage PY54.

The temperate Yersinia phage PY54 belongs to the unusual group of phages that replicate as linear plasmids with covalently closed ends. Besides Escherichia coli phage N15, PY54 is the only member of this group to be identified. We have determined the complete sequence (46,339 bp) of the PY54 genome. Bioinformatic analyses revealed 67 open reading frames (ORFs) with good coding potential located on both DNA strands. The comparison of the deduced PY54 gene products with known proteins encoded by other phages and bacteria along with functional studies have enabled us to assign the possible functions of 25 ORFs. In the left arm of the PY54 genome, we identified a number of ORFs that obviously code for head and tail proteins. Furthermore, this part of the phage genome contains genes probably involved in plasmid partitioning. Regarding the predicted gene functions and gene order, the PY54 and N15 left arms are similar. However, there are only weak DNA homologies and, in contrast to N15, the Yersinia phage harbours only a few ORFs related to genes found in lambdoid phages. The PY54 right arm comprises mainly regulatory genes as well as genes important for plasmid replication, DNA methylation, and host cell lysis. Out of 36 deduced products of the right arm, 13 revealed strongest database homologies to N15 proteins, of which the protelomerase and the Rep protein are exclusively homologous to their N15 counterparts. A number of PY54 genes essential for the lytic or lysogenic cycle were identified by functional analysis and characterization of phage mutants. In order to study transcription during the lytic and lysogenic stage, we analysed 34 PY54 ORFs by reverse transcriptase (RT)-PCR. The phage transcription patterns in lysogenic bacteria and at the late lytic stage of infection are nearly identical. The reasons for this finding are spontaneous release of phages during lysogeny and a high rate of phages that lysogenize their Yersinia host upon infection.

Amino Acid Sequence↗
PubMed1984

Lentiviruses are naturally resident in a latent form in long-term ovine fibroblast cultures.

Long-term ovine fibroblast cultures contain replicative-competent lentiviruses in a latent form. This in vitro phenomenon, never described previously for lentiviruses, was clearly demonstrated by activating the expression of latent viruses with various inducing cell treatments, some of which were efficient in inducing endogenous retroviruses or latent herpesviruses. Activated lentiviruses were highly lytic in ovine fibroblasts (type I), or they established persistent infections (type II) as described previously for field isolates from sheep with progressive pneumonia (Quérat et al., J. Virol. 52:671-678, 1984).

Animals↗
PubMed1989

Relative rates of RNA synthesis across the genome of Epstein-Barr virus are highest near oriP and oriLyt.

The rates of Epstein-Barr virus transcription were measured in isolated nuclei from marmoset and human lymphoblasts transformed in vitro. In B95-8, a marmoset B-lymphoid cell line, the most frequently transcribed viral genes are the EBERs (small nuclear RNAs) and BHLF-1 (encoding a lytic-phase gene product). The EBERs and BHLF-1 genes are separated by nearly 50 kilobase pairs on the Epstein-Barr virus genome and lie adjacent to (less than 300 base pairs from) oriP and oriLyt, respectively. oriP and oriLyt are putative origins of viral DNA replication, and each is associated with a transcriptional enhancer element. Among the human B-lymphoblastoid cell lines tested, only the transcription of EBERs predominates.

Cell Line↗
PubMedDate not supplied

[Epstein-Barr virus].

Epstein-Barr Virus (EBV) is the prototype member of the human herpesvirus subfamily Gammaherpesviridae. EBV establishes a latent infection in lymphoïd cell and replicates in epithelial cells. Infection of individuals with EBV is widespread in all human populations, as shown by the high proportion of individual with specific antibodies in their serum. EBV is the etiological agent of the infectious mononucleosis and has been implicated in the pathogenesis of an increasing number of human malignancies, the best characterized being endemic Burkitt's lymphomas, nasopharyngeal carcinomas (NPC), and polyclonal lymphomas in immunocompromised patients. EBV infection in vivo is a complex mixture of latent, reactivated, transforming, or replicative type of infection. EBV infection is characterized by sporadic viral excretion in the oropharynx and persistent latent infection in the bone marrow and peripheral blood lymphocytes. EBV infection includes an intense immune response. During primary infection, the humoral response is primarily directed toward antigens of the lytic cycle, membrane antigen complex, early antigen (EA), viral capsid antigen (VCA) while the antibodies response to EBNAs is delayed. The cell mediated response controls the proliferation of EBV-infected lymphocytes through two classes of cytotoxic cells, namely, natural killer and T8 cytotoxic cells. Laboratory diagnosis of EBV infection is recently based upon molecular biology techniques which provides a useful tool for direct identification of EBV and may allow to better understand the role of the virus in the pathogenesis of EBV associated disorders.

Antibody Formation↗
PubMed2005

Induction of Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen by the lytic transactivator RTA: a novel mechanism for establishment of latency.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent contributing to development of Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman desease. Following primary infection, latency is typically established. However, the mechanism by which KSHV establishes latency is not understood. We have reported that the latency-associated nuclear antigen (LANA) can repress RTA (for replication and transcription activator) expression by down-regulating its promoter. In this study, we show that RTA is associated with the virion particle. We also show that RTA can activate the LANA promoter and induce LANA expression in transient reporter assays. Additionally, the transcription of RTA correlates with LANA expression in the early stages of de novo infection of KSHV, and induction of LANA transcription is responsive to induction of RTA with an inducible system. This induction in LANA transcription was dependent on recombination signal sequence binding protein Jkappa (RBP-Jkappa), as a RBP-Jkappa-deficient cell line was significantly delayed and inefficient in LANA transcription with expression of RTA. These studies suggest that RTA contributes to establishment of KSHV latency by activating LANA expression in the early stages of infection by utilizing the major effector of the Notch signaling pathway RBP-Jkappa. This describes a feedback mechanism by which LANA and RTA can regulate each other and is likely to be a key event in the establishment of KSHV latency.

Antigens, Viral↗
PubMed1999

Cloning and expression of a gene encoding the lytic functions of Bacillus amyloliquefaciens phage: Evidence of an auxiliary lysis system.

A bacteriophage specific to Bacillus amyloliquefaciens, a gram-positive bacterium, was isolated from a local sewage treatment center. Using a lysis assay, a gene, lys1521, was isolated and its nucleotide sequence revealed one open reading frame of 375 bp. Homology studies showed amino acid alignment similarity with gene 5A of Bacillus subtilis phages PZA and phi29. Overexpression of the cloned gene yielded a 13 kDa protein corresponding to the predicted gene product. Despite the fact that no significant homology with known cell wall lytic enzymes was apparent, the lytic profile obtained in an in vivo expression assay showed that lys1521 had cell wall hydrolysis activity. This is a significant revelation since the function of the homologous gene 5A product of phage phi29 has been suggested to be required for the in vivo elongation of phage DNA replication. The lys1521 gene could be evidence of the presence in gram-positive bacteriophages of a third lysis gene in addition to the well characterized two-step lysis system.

Journal Article↗
PubMed1981

Pleiotropic expression of Epstein--Barr virus DNA in human epithelial cells.

We have attempted to establish a system that can be used to study the association of Epstein--Barr virus (EBV) with epithelial cells. Attempts were made to transfect human carcinoma cells with EBV DNA. Successful transfection was confirmed by the expression of EBV-specific early antigen (EA), virus capsid antigen, and the presence of virus DNA. The transfecting preparation contained a mixture of EBV and cellular DNA extracted from two producer cell lines, P3HR-1 and AG-876. Our data suggest that virus DNA obtained from the P3HR-1 nontransforming, EA-inducing strain of EBV was lytically expressed in the epithelial tumor cells. The DNA derived from AG-876 cells, which produce a transforming, non-EA-inducing strain of EBV, also produced a lytic infection.

Carcinoma↗
PubMed1981

The plaque-forming factor for mink lung cells present in cytomegalovirus and herpes-zoster virus stocks identified as Mycoplasma hyorhinis.

Previous investigation of the ability of cytomegalovirus and varicella-zoster virus to replicate in a variety of cell lines suggested that both virus types plaqued with high efficiency in mink lung cells. However, many of the virus isolates used appeared to be contaminated with mycoplasma. We now report that the observed cytopathic effect is due to a mycoplasma which grows lytically to high titre in mink lung cells, but is difficult to cultivate in cell-free media. The mycoplasma was plaque-purified and shown to contain DNA with a buoyant density of 1.684 g/ml, with restriction endonuclease patterns identical to the porcine mycoplasma M. hyorhinis. This was confirmed by serological identification.

Animals↗
PubMed1995

Identification of an epithelial cell differentiation responsive region within the BZLF1 promoter of the Epstein-Barr virus.

The Epstein-Barr virus (EBV) BZLF1 gene product, Zta is able to trigger the viral lytic cycle in latently infected B lymphocytes. Investigations into regulation of the promoter, Zp for the BZLF1 gene in B cells have identified 12-O-tetradecanoylphorbol 13-acetate, anti-immunoglobulin and Zta responsive elements as well as a negative regulatory element within Zp. EBV infects and replicates within squamous epithelial cells and there is evidence to indicate that EBV gene expression is linked to the differentiation status of epithelial cells. We have investigated regulation of Zp in undifferentiated and differentiated cells of the human squamous epithelial cell line SCC12F. Zp was found to be active in SCC12F cells and activity was increased approximately 7-fold upon induction of epithelial terminal differentiation. Sequences responsive to epithelial cell differentiation were contained within the region of Zp from -86 to +12 bp, a region previously shown to contain an AP-1-like binding site, designated the ZII domain. In addition, enhancing sequences were detected in the region -221 to -86 bp. These studies will lead to a greater understanding of differentiation-linked EBV gene expression in human squamous epithelial cells.

Animals↗
PubMed1976

Genetic analysis of heterogeneous DNA circles formed after prophage Mu induction.

Induction of a Mu prophage in Escherichia coli Hfr strains lyosgenic for Mu cts62 leads to the generation of F' episomes. Each episome thus formed carries at least one copy of the Mu genome. These results suggest that integration of Mu is mandatory for the formation of the heterogeneous circles during the lytic cycle. The circles may be precursors for phage maturation.

Coliphages↗