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Journal of Virology May 1976Purified defective interfering (DI) particles of vesicular stomatitis virus (VSV) inhibit the replication of a heterologous virus, pseudorabies virus (PSR), in hamster... (Comparative Study)
Comparative Study
Purified defective interfering (DI) particles of vesicular stomatitis virus (VSV) inhibit the replication of a heterologous virus, pseudorabies virus (PSR), in hamster (BHK-21) and rabbit (RC-60) cell lines. In contrast to infectious B particles of VSV, UV irradiation of DI particles does not reduce their ability to inhibit PSR replication. However, UV irradiation progressively reduces the ability of DI particles to cause homologous interference with B particle replication. Pretreatment with interferon does not affect the ability of DI particles to inhibit PSR replication in a rabbit cell line (RC-60) in which RNA, but not DNA, viruses are sensitive to the action of interferon. Under similar conditions of interferon pretreatment, the inhibition of PSR by B particles is blocked. These data suggest that de novo VSV RNA or protein synthesis is not required for the inhibition of PSR replication by DI particles. DI particles that inhibit PSR replication also inhibit host RNA and protein synthesis in BHK-21 and RC-60 cells. Based on the results described and data in the literature, it is proposed that the same component of VSV B and DI particles is responsible for most, if not all, of the inhibitory activities of VSV, except homologous interference.
Topics: Cell Line; Defective Viruses; Herpesviridae; Herpesvirus 1, Suid; Interferons; Protein Biosynthesis; RNA; Radiation Effects; Ultraviolet Rays; Vesicular stomatitis Indiana virus; Viral Interference; Virus Replication
PubMed: 178895
DOI: 10.1128/JVI.18.2.534-541.1976 -
Methods in Cell Biology 1994
Review
Topics: Animals; Chlorocebus aethiops; Cloning, Molecular; DNA, Recombinant; Defective Viruses; Escherichia coli; Gene Expression; Gene Transfer Techniques; Genetic Vectors; Helper Viruses; Humans; Liposomes; Neurons; Plasmids; Recombinant Fusion Proteins; Simplexvirus; Vero Cells; Virus Cultivation
PubMed: 7823862
DOI: 10.1016/s0091-679x(08)60604-4 -
Biochemical and Biophysical Research... Jul 1992We are interested in using retroviral vectors to trace cell lineage and to introduce exogenous genes in chicken skin explant cultures. Here the LZ10 virus carrying the...
We are interested in using retroviral vectors to trace cell lineage and to introduce exogenous genes in chicken skin explant cultures. Here the LZ10 virus carrying the gene encoding beta-galactosidase was introduced to the skin explants by two different means: a) the virus was added to the media or b) the virus was microinjected into regions of the developing feather buds. Infection by microinjection led to localized expression of beta-galactosidase in the developing feather bud, while, surprisingly, infection by adding the virus to the culture media led to localized band of beta-galactosidase expression in the middle of the feather filament. The significance of this finding in skin morphogenesis and as a tool for experimental embryology is discussed.
Topics: Animals; Avian Sarcoma Viruses; Cell Transformation, Viral; Chick Embryo; Defective Viruses; Organ Culture Techniques; Skin; Virus Replication; beta-Galactosidase
PubMed: 1323281
DOI: 10.1016/0006-291x(92)90848-f -
Journal of Virology Feb 2013Virus-like particles (VLPs) from hepatitis B and human papillomaviruses have been successfully used as preventative vaccines against these infectious agents. These VLPs...
Virus-like particles (VLPs) from hepatitis B and human papillomaviruses have been successfully used as preventative vaccines against these infectious agents. These VLPs consist of a self-associating capsid polymer formed from a single structure protein and are devoid of viral DNA. Since virions from herpesviruses consist of a large number of molecules of viral and cellular origin, generating VLPs from a subset of these would be a particularly arduous task. Therefore, we have adopted an alternative strategy that consists of producing DNA-free defective virus particles in a cell line infected by a herpesvirus mutant incapable of packaging DNA. We previously reported that an Epstein-Barr virus (EBV) mutant devoid of the terminal repeats (ΔTR) that act as packaging signals in herpesviruses produces substantial amounts of VLPs and of light particles (LPs). However, ΔTR virions retained some infectious genomes, and although these mutants had lost their transforming abilities, this poses potential concerns for clinical applications. Therefore, we have constructed a series of mutants that lack proteins involved in maturation and assessed their ability to produce viral DNA-free VLP/LPs. Some of the introduced mutations were deleterious for capsid maturation and virus production. However, deletion of BFLF1/BFRF1A or of BBRF1 resulted in the production of DNA-free VLPs/LPs. The ΔBFLF1/BFRF1A viruses elicited a potent CD4(+) T-cell response that was indistinguishable from the one obtained with wild-type controls. In summary, the defective particles produced by the ΔBFLF1/BFRF1A mutant fulfill the criteria of efficacy and safety expected from a preventative vaccine.
Topics: CD4-Positive T-Lymphocytes; Cells, Cultured; DNA, Viral; Defective Viruses; Gene Deletion; Herpesvirus 4, Human; Humans; Membrane Proteins; Vaccines, Virus-Like Particle; Viral Proteins; Virus Assembly
PubMed: 23236073
DOI: 10.1128/JVI.02533-12 -
Archives of Virology Mar 2012Short defective RNAs (D-RNAs) associated with tomato black ring virus (TBRV) were isolated, cloned and sequenced. As a result, two types of D-RNAs associated with...
Short defective RNAs (D-RNAs) associated with tomato black ring virus (TBRV) were isolated, cloned and sequenced. As a result, two types of D-RNAs associated with different TBRV isolates were identified. Both types were derived from RNA1. The first one contained sequences from the 5' and 3' untranslated regions (UTR) and from the 5' region of a single large open reading frame. The second one included a portion of the coding region for the RNA-dependent RNA polymerase flanked by a short fragment of the 5' UTR and the entire 3' UTR. The possible nature and origin of these RNA species is discussed.
Topics: 3' Untranslated Regions; 5' Untranslated Regions; Base Sequence; Cloning, Molecular; Defective Viruses; Solanum lycopersicum; Molecular Sequence Data; Nepovirus; RNA, Viral; Sequence Analysis, DNA; Viral Proteins
PubMed: 22203315
DOI: 10.1007/s00705-011-1200-z -
Journal of Virology Oct 2011The reverse genetics technology for bluetongue virus (BTV) has been used in combination with complementing cell lines to recover defective BTV-1 mutants. To generate a...
The reverse genetics technology for bluetongue virus (BTV) has been used in combination with complementing cell lines to recover defective BTV-1 mutants. To generate a potential disabled infectious single cycle (DISC) vaccine strain, we used a reverse genetics system to rescue defective virus strains with large deletions in an essential BTV gene that encodes the VP6 protein (segment S9) of the internal core. Four VP6-deficient BTV-1 mutants were generated by using a complementing cell line that provided the VP6 protein in trans. Characterization of the growth properties of mutant viruses showed that each mutant has the necessary characteristics for a potential vaccine strain: (i) viral protein expression in noncomplementing mammalian cells, (ii) no infectious virus generated in noncomplementing cells, and (iii) efficient replication in the complementing VP6 cell line. Further, a defective BTV-8 strain was made by reassorting the two RNA segments that encode the two outer capsid proteins (VP2 and VP5) of a highly pathogenic BTV-8 with the remaining eight RNA segments of one of the BTV-1 DISC viruses. The protective capabilities of BTV-1 and BTV-8 DISC viruses were assessed in sheep by challenge with specific virulent strains using several assay systems. The data obtained from these studies demonstrated that the DISC viruses are highly protective and could offer a promising alternative to the currently available attenuated and killed virus vaccines and are also compliant as DIVA (differentiating infected from vaccinated animals) vaccines.
Topics: Animals; Antibodies, Neutralizing; Antibodies, Viral; Bluetongue; Bluetongue virus; Cell Culture Techniques; Defective Viruses; Female; Male; Reassortant Viruses; Sheep; Viral Vaccines; Viremia
PubMed: 21795358
DOI: 10.1128/JVI.05412-11 -
The Journal of General Virology Jul 1997A naturally occurring defective RNA of 2379 nt (D2.3) from the VT strain of citrus tristeza closterovirus (CTV) was cloned and sequenced. The D2.3 RNA is a fusion of two...
A naturally occurring defective RNA of 2379 nt (D2.3) from the VT strain of citrus tristeza closterovirus (CTV) was cloned and sequenced. The D2.3 RNA is a fusion of two regions of 1521 and 858 nt from the 5' and 3' ends of the CTV genome, respectively. A cDNA clone of D2.3 RNA was tagged by the insertion of a 0.47 kb chimeric DNA fragment and the recombinant cDNA was inserted downstream of the cauliflower mosaic virus 35S promoter. The resulting construct was bombarded into CTV-infected tissue, which was then grafted onto virus-free plants. The presence of recombinant RNA in systemically infected leaves was demonstrated by RT-PCR. Sequencing the RT-PCR products synthesized from double-stranded RNA confirmed the presence of the chimeric segment used for tagging. This is the first report of an infectious cDNA molecule derived from CTV D-RNA.
Topics: Base Sequence; Citrus; Cloning, Molecular; Closterovirus; DNA, Complementary; DNA, Viral; Defective Viruses; Helper Viruses; Molecular Sequence Data; RNA, Viral
PubMed: 9225053
DOI: 10.1099/0022-1317-78-7-1765 -
Journal of Virology Aug 1982Defective genomes present in serially passaged virus stocks derived from the tsLB2 mutant of herpes simplex virus type 1 were found to consist of repeat units in which...
Defective genomes present in serially passaged virus stocks derived from the tsLB2 mutant of herpes simplex virus type 1 were found to consist of repeat units in which sequences from the U(L) region, within map coordinates 0.356 and 0.429 of standard herpes simplex virus DNA, were covalently linked to sequences from the end of the S component. The major defective genome species consisted of repeat units which were 4.9 x 10(6) in molecular weight and contained a specific deletion within the U(L) segment. These tsLB2 defective genomes were stable through more than 35 sequential virus passages. The ratios of defective virus genomes to helper virus genomes present in different passages fluctuated in synchrony with the capacity of the passages to interfere with standard virus replication. Cells infected with passages enriched for defective genomes overproduced the infected cell polypeptide number 8, which had previously been mapped within the U(L) sequences present in the tsLB2 defective genomes. In contrast, the synthesis of most other infected cell polypeptides was delayed and reduced. The abundant synthesis of infected cell polypeptide number 8 followed the beta regulatory pattern, as evident from kinetic studies and from experiments in which cycloheximide, canavanine, and phosphonoacetate were used. However, in contrast to many beta (early) and gamma (late) viral polypeptides, the synthesis of infected cell polypeptide number 8 was only minimally reduced when cells infected with serially passaged tsLB2 were incubated at 39 degrees C. The tsLB2 mutation had previously been mapped within the domains of the gene encoding infected cell polypeptide number 4, the function of which was shown to be required for beta and gamma viral gene expression. It is thus possible that the tsLB2 mutation affects the synthesis of only a subset of the beta and gamma viral polypeptides. An additional polypeptide, 74.5 x 10(3) in molecular weight, was abundantly produced in cells infected with a number of tsLB2 passages. This polypeptide was most likely expressed from truncated gene templates within the most abundant, deleted repeats of tsLB2 defective virus DNA.
Topics: Cell Line; DNA, Viral; Defective Viruses; Gene Expression Regulation; Genes, Viral; Humans; Mutation; Repetitive Sequences, Nucleic Acid; Simplexvirus; Temperature; Viral Proteins
PubMed: 6287032
DOI: 10.1128/JVI.43.2.574-593.1982 -
Virology Jun 1987A cyclic pattern of virus production was observed when human parainfluenza virus 3 (HPIV3) was serially passaged nine times in LLC-MK2 cells. Viruses produced from...
A cyclic pattern of virus production was observed when human parainfluenza virus 3 (HPIV3) was serially passaged nine times in LLC-MK2 cells. Viruses produced from serial passages 8 and 9 interfered with the replication of standard HPIV3. Three subgenomic RNA species (DI-1, DI-2, and DI-3) and virus genomic RNA were detected in the progeny virions produced from cells mixedly infected with standard virus and virus from either serial passages 5 or 8. Northern blot analysis with probes representing all six HPIV3 structural protein genes revealed that DI-1 and DI-2 RNAs contain sequences from the 5' end of the standard virus genome. DI-1 RNA contains L, HN, and F specific sequences, while DI-2 RNA contains only L and HN sequences. DI-3 RNA did not hybridize with any of the probes used. The possibility that DI-3 RNA contains sequences from the 5' end of the standard virus genome is discussed. These results demonstrate that 5' defective interfering particles are generated during serial passage of HPIV3.
Topics: Defective Viruses; Genes, Viral; Parainfluenza Virus 3, Human; RNA, Messenger; RNA, Viral; Respirovirus; Viral Proteins; Virus Replication
PubMed: 3035791
DOI: 10.1016/0042-6822(87)90217-0 -
Virology Mar 2014Defective RNAs (D RNAs) are small RNA replicons derived from viral RNA genomes. No D RNAs have been found associated with members of the plus-strand RNA virus genus...
Defective RNAs (D RNAs) are small RNA replicons derived from viral RNA genomes. No D RNAs have been found associated with members of the plus-strand RNA virus genus Aureusvirus (family Tombusviridae). Accordingly, we sought to construct a D RNA for the aureusvirus Cucumber leaf spot virus (CLSV) using the known structure of tombusvirus defective interfering RNAs as a guide. An efficiently accumulating CLSV D RNA was generated that contained four non-contiguous regions of the viral genome and this replicon was used as a tool to studying viral cis-acting RNA elements. The results of structural and functional analyses indicated that CLSV contains counterparts for several of the major RNA elements found in tombusviruses. However, although similar, the CLSV D RNA and its components are distinct and provide insights into RNA-based specificity and mechanisms of function.
Topics: Base Sequence; Defective Viruses; Genome, Viral; Molecular Sequence Data; Nucleic Acid Conformation; RNA, Viral; Tombusviridae; Tombusvirus; Viral Proteins; Virus Replication
PubMed: 24606684
DOI: 10.1016/j.virol.2013.12.033