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Journal of Virology Feb 1982A uniform nomenclature has been agreed upon for monoclonal antibodies directed against virus-coded proteins of simian virus 40 and polyoma virus. Blocks of numbers from...
A uniform nomenclature has been agreed upon for monoclonal antibodies directed against virus-coded proteins of simian virus 40 and polyoma virus. Blocks of numbers from PAb1 to PAb999 have been allocated to workers involved in the isolation of monoclonal antibodies of this type. The correspondence between PAb numbers and previously used names is given.
Topics: Antibodies, Monoclonal; Antibodies, Viral; Polyomavirus; Simian virus 40; Terminology as Topic; Viral Proteins
PubMed: 6281480
DOI: 10.1128/JVI.41.2.709-709.1982 -
Journal of Virology Sep 1982Three new polyoma mutants were selected for their ability to grow on the embryonal carcinoma cell line F9. These mutants share in common an insertion of two nucleotides,...
Three new polyoma mutants were selected for their ability to grow on the embryonal carcinoma cell line F9. These mutants share in common an insertion of two nucleotides, a thymine and an adenine, in the noncoding region located on the late side of the origin of replication. We have found that these insertions exist in all of the other polyoma virus mutants able to grow on F9 cells (Fujimura et al., Cell 23:809-814, 1981; Katinka et al., Nature (London) 290:720-722, 1981; K. Sekikawa and A. J. Levine, Proc. Natl. Acad. Sci. U.S.A. 78:1100-1104, 1981). The region containing these insertions could be folded into a stable secondary structure which included a guanine plus cytosine (G + C)-rich stem. The adenine and thymine were inserted in such a way that they maintained the palindrome in the G + C-rich stem and were complementary in the putative secondary structure that we present here. Another class of polyoma virus mutants selected on a multipotential carcinoma cell line (PCC4-Aza) were characterized by a more complex rearrangement (deletion and duplication) which occurred in the same region. This arrangement preserved the G + C-rich palindrome and also yielded a sequence which still allowed the folding of another type of stable secondary structure. The significance of these findings is discussed.
Topics: Animals; Base Sequence; Cell Line; Cloning, Molecular; DNA, Viral; Mice; Mutation; Nucleic Acid Conformation; Polyomavirus; Teratoma; Virus Cultivation
PubMed: 6292462
DOI: 10.1128/JVI.43.3.800-808.1982 -
Proceedings of the National Academy of... Jun 1980Rat-1 cells were transfected with the restriction enzyme fragment of polyoma virus DNA that extends clockwise from the Bcl I site ((65.4 map units) to the EcoRI site...
Rat-1 cells were transfected with the restriction enzyme fragment of polyoma virus DNA that extends clockwise from the Bcl I site ((65.4 map units) to the EcoRI site (0/100 map units). Six transformed cell lines were obtained and one of them (BE-1) has been investigated in detail. The viral DNA that is integrated into host DNA in this line appeared to consist of two fragments arranged in a "head-to-tail" tandem with no detectable intervening host sequences. BE-1 cells contained polyoma virus small and middle tumor antigens that were indistinguishable from the corresponding tumor antigens from lytically infected cells. No large tumor antigen was detected but a "new" Mr 34,000 protein, which proved to be a truncated version of large tumor antigen, was immunoprecipitated by anti-tumor-antigen antiserum. After injection of 10(6) BE-1 cells into young syngeneic Fischer rats, tumors appeared within 3--4 weeks. Thus, the coding capacity of the Bcl I/EcoRI fragment of polyoma virus DNA is sufficient to enable the cells to produce all of small and middle tumor antigens and about a third of large tumor antigen, to transform cells stably in culture, and to produce tumors in vivo.
Topics: Animals; Antigens, Neoplasm; Antigens, Viral; Antigens, Viral, Tumor; Base Sequence; Cell Transformation, Viral; Cells, Cultured; Chromosome Mapping; DNA; DNA Restriction Enzymes; DNA, Viral; Genes, Viral; Mice; Neoplasm Transplantation; Polyomavirus; Rats
PubMed: 6251451
DOI: 10.1073/pnas.77.6.3278 -
Journal of Clinical Microbiology Feb 1980An indirect immunofluorescence method was developed and used to detect urinary excretion of abnormal transitional cells infected with JC virus (JCV) or BK virus (BKV)....
An indirect immunofluorescence method was developed and used to detect urinary excretion of abnormal transitional cells infected with JC virus (JCV) or BK virus (BKV). This method was compared with urinary cytology, electron microscopy, viral culture, and viral serology in groups of immunosuppressed renal transplant recipients and normal controls. The indirect immunofluorescence method detected and identified JCV excretion in four persons, BKV excretion in one person, and both JCV and BKV excretion in eight others. Viral antigen was identified only in the nuclei of cytologically abnormal cells. Of these 13 persons, 8 also had polyoma virions detected in the urine by electron microscopy. With repeated study of sequential urine samples, 30% of transplant recipients and 6% of normal controls were positive by one or more microscopy methods. Serological results confirmed a high incidence of both JCV and BKV multiplication in the immunosuppressed patients. However, serology did not correlate directly with urinary virological findings. Urinary cytology and the indirect immunofluorescence method were rapid and sensitive methods for detecting and identifying urinary excretion of JCV and BKV.
Topics: Adult; Animals; Antigens, Viral; BK Virus; Cell Nucleus; Child, Preschool; Female; Fluorescent Antibody Technique; Humans; Male; Middle Aged; Polyomavirus; Tumor Virus Infections; Urinary Tract; Urine
PubMed: 6244330
DOI: 10.1128/jcm.11.2.178-183.1980 -
Proceedings of the National Academy of... Jan 1978The circular genome of the cloned defective polyoma virus D-50 consists of tandemly repeated copies of the DNA sequence between 67 and 84 units on the wild-type polyoma...
The circular genome of the cloned defective polyoma virus D-50 consists of tandemly repeated copies of the DNA sequence between 67 and 84 units on the wild-type polyoma virus DNA map. Each repeated copy thus contains the origin of viral DNA replication, which is located at about 71 map units. Viral RNA was synthesized in vitro using viral transcription complexes extracted late (30 hr) after infection from mouse cells co-infected with D-50 and helper wild-type virus. Both wild-type and D-50 DNA molecules were active as templates for in vitro transcription. Approximately 84% of the RNA transcribed in vitro from wild-type DNA was complementary to the L DNA strand. This is normal for wild-type transcription late after infection. By contrast, at least 90% of the RNA transcribed from D-50 DNA molecules was complementary to the E DNA strand. After normalization of the data to account for the observed molar ratio of D-50 DNA repeated sequences to unit length wild-type DNA, we estimate that transcription of the E DNA strand of each D-50 repeated unit is about 1.4 times as efficient as transcription of the wild-type E DNA strand. Transcription of the D-50 L DNA strand, however, is only 0.03 times as efficient as transcription of the wild-type L DNA strand. The implications of these results concerning the nature and location of promoter sequences in polyoma DNA are discussed.
Topics: Binding Sites; Cell Line; DNA, Circular; DNA, Viral; DNA-Directed RNA Polymerases; Defective Viruses; Helper Viruses; Molecular Weight; Polyomavirus; RNA, Viral; Transcription, Genetic
PubMed: 203943
DOI: 10.1073/pnas.75.1.69 -
Proceedings of the National Academy of... Nov 1976Infection of primary cultures of mouse kidney cells with polyoma virus causes a biphasic increase in the activities of L-ornithine decarboxylase (ODC; L-ornithine...
Infection of primary cultures of mouse kidney cells with polyoma virus causes a biphasic increase in the activities of L-ornithine decarboxylase (ODC; L-ornithine carboxy-lyase; EC 4.1.1.17) and S-adenosyl-L-methionine decarboxylase (SAMD; S-adenoxyl-L-methionine carboxy-lyase; EC 4.1.50), as well as in the level of the polyamines putrescine, spermidine, and spermine. An early peak occurs during the period when early viral mRNA is synthesized and prior to the onset of virus-induced synthesis of host cell DNA. A late peak coincides in time with the maximum rate of virus-induced synthesis of cellular DNA. A similar biphasic stimulation of polyamine synthesis is induced even when DNA synthesis is prevented by 5-fluorodeoxyuridine. Actinomycin D (AMD) in a dose that inhibits rRNA synthesis causes no inhibition of ODC or SAMD. In a dose that inhibits mRNA synthesis as well, short-term AMD treatment causes "superinduction" of ODC but inhibition of SAMD. Prolonged treatment with the high dose of AMD inhibits ODC as well, indicating that late ODC activity may be dependent on mRNA synthesized during early infection. Cycloheximide effectively obliterates the ODC and SAMD activities during the entire infectious cycle. Uncoupling from DNA and rRNA synthesis suggests that polyamine synthesis is regulated independently of these events. The experiments with AMD and cycloheximide suggest that the formation of ODC is subject to post-transcriptional control, whereas that of SAMD is regulated primarily at the transcriptional level.
Topics: Adenosylmethionine Decarboxylase; Cell Line; DNA; Dactinomycin; Enzyme Induction; Floxuridine; Ornithine Decarboxylase; Polyamines; Polyomavirus; Protein Biosynthesis; Putrescine; RNA, Ribosomal; Spermidine; Spermine
PubMed: 186774
DOI: 10.1073/pnas.73.11.4022 -
Journal of Virology May 1974Supercoiled DNA molecules purified from mouse cells infected with high-multiplicity-passaged polyoma virus has a broader size distribution and sediments more slowly than...
Supercoiled DNA molecules purified from mouse cells infected with high-multiplicity-passaged polyoma virus has a broader size distribution and sediments more slowly than DNA derived from low-multiplicity-passaged virus. The shorter DNA molecules are predominately noninfectious. Virus populations containing distinct size classes of defective virus DNA were isolated by growing virus from single cells infected by a defective and nondefective helper virus (infectious center). This technique probably results in the cloning of defective virus particles. By applying the infectious center method to DNA from various fractions of sucrose gradients it has been possible to obtain shorter circular DNA molecules ranging in size from 50 to 95% of the unit-length polyoma DNA molecule. The shorter molecules in any one preparation are homogeneous in size. This class size is retained upon repeated passage of crude viral lysates at high multiplicity. Thus far, all the purified shorter DNA molecules tested appear to be noninfectious and largely resistant to cleavage by the R(1) restriction enzyme. Some of the purified defective molecules have been found to interfere with the production of infectious virus upon co-infection with unit-length infectious polyoma DNA.
Topics: Animals; Cells, Cultured; Centrifugation, Density Gradient; DNA, Viral; Defective Viruses; Mice; Polyomavirus; Viral Plaque Assay; Virus Cultivation
PubMed: 4363255
DOI: 10.1128/JVI.13.5.939-946.1974 -
Journal of Virology Sep 1982To investigate the relation between the polyoma tumor-specific transplantation antigen and the virus-coded proteins, mice were immunized by inoculation of a variety of...
To investigate the relation between the polyoma tumor-specific transplantation antigen and the virus-coded proteins, mice were immunized by inoculation of a variety of viable polyoma virus mutants and then challenged with polyoma virus-induced tumors. Two classes of early region mutants were used. One class produces a normal small T-antigen and truncated middle and large T-antigens. The second class (hr-t mutants) forms a normal large T-antigen together with N-terminal fragments of small and middle T-antigens. All mutants, transforming as well as nontransforming, induced protection against polyoma virus tumors. However, there were quantitive differences between the mutants. The finding that an hr-t mutant could induce tumor rejection suggests that full-length middle and small T-antigens are not necessary for the induction of this response. Since intact middle T-antigen is the only virus-coded protein known to associate with the plasma membrane, the possibility must be considered that the polyoma virus tumor-specific transplantation antigen consists of cellular components.
Topics: Animals; Antigens, Viral; Antigens, Viral, Tumor; Fibrosarcoma; Graft Rejection; Hair; Immunization; Mice; Neoplasm Transplantation; Polyomavirus; Sarcoma, Experimental; Skin Neoplasms
PubMed: 6292460
DOI: 10.1128/JVI.43.3.772-777.1982 -
Journal of Virology Aug 1976The plaque-assay technique was used as a tool to determine the optimal conditions for adsorption of polyoma virions to host cells. Using these optimal conditions of...
The plaque-assay technique was used as a tool to determine the optimal conditions for adsorption of polyoma virions to host cells. Using these optimal conditions of adsorption, an electron microscopy study of the early events of infection was performed. By electron microscopy and autoradiography, it was demonstrated that both the viral coat proteins and DNA arrive simultaneously in the nucleus as early as 15 min postinfection. When horseradish peroxidase-labeled virions, pseudovirions, and capsids were used to infect cells, only the particles with nucleic acid or a factor(s) associated with the nucleic acid, i.e., histones, appeared to enter the nucleus. Moreover, when virions were used to infect either permissive or nonpermissive cells, identical early events of viral infection, i.e., adsorption, penetration, and nuclear transport, were observed, suggesting that these early events of infection are a property of the virion and not the host cell.
Topics: Adsorption; Autoradiography; Cell Line; Cell Membrane; Cell Nucleus; Culture Techniques; DNA, Viral; Horseradish Peroxidase; Microscopy, Electron; Models, Biological; Polyomavirus; Viral Proteins; Virus Replication
PubMed: 183018
DOI: 10.1128/JVI.19.2.620-636.1976 -
Virology Dec 1998Polyoma virus is highly oncogenic when inoculated into immunocompromised adult mice and neonatal mice of specific inbred strains. Although T lymphocytes are known to be...
Polyoma virus is highly oncogenic when inoculated into immunocompromised adult mice and neonatal mice of specific inbred strains. Although T lymphocytes are known to be essential in controlling polyoma virus tumorigenesis, the importance of class I MHC-restricted CD8+ T cells in mediating tumor resistance remains unclear. Here, we investigated the tumorigenicity of polyoma virus in adult mice rendered CD8+ T cell-deficient by homozygous (-/-) disruption of the beta 2-microglobulin (beta 2m) or CD8 alpha (CD8) genes. Nearly all (94%) of the virus-infected adult C57BL/6 beta 2m-/- mice developed tumors, and 20% of the virus-inoculated adult C57BL/6CD8-/- mice developed hindlimb paralysis, which is indicative of vertebral tumors. Only 2 of 20 virus-inoculated adult normal C57BL/6 mice developed tumors. Despite these different tumor susceptibilities, persistent viral DNA was detected in multiple organs of mice of all three strains. Multifocal lymphoplasmacytic interstitial infiltrates were present in the kidneys and lungs of virus-infected C57BL/6 beta 2m-/- and in the lungs of virus-inoculated C57BL/6CD8-/- mice. These infiltrates were composed primarily of B cells and colocalized with staining for the major viral capsid protein, VP1. No infiltrates or VP1 staining was detected in the kidneys of infected C57BL/6 mice. Using a highly sensitive RT-PCR bioluminescence immunoassay, we investigated and detected persistent polyoma T protein and VP1 messages in both C57BL/6 beta 2m-/- and C57BL/6 mice. C57BL/6 beta 2m-/- and C57BL/6 mice had equivalent serum virus-neutralizing antibody titers. These results provide in vivo evidence that class I MHC-restricted CD8+ T cells are involved in mediating protection against polyoma virus tumor development.
Topics: Animals; Antibodies, Viral; Blotting, Southern; Blotting, Western; DNA, Viral; Genetic Predisposition to Disease; Mice; Mice, Inbred C57BL; Mice, Knockout; Papillomavirus Infections; Polymerase Chain Reaction; Polyomavirus; Tumor Virus Infections; beta 2-Microglobulin
PubMed: 9875336
DOI: 10.1006/viro.1998.9455