Survival of UV-irradiated herpes simplex type 1 virus in herpes simplex type 2-transformed hamster cells.
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Cell Line↗
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Induction of deoxycytidine deaminase activity in mammalian cell lines by infection with herpes simplex virus type 1.
Herpes simplex virus type 1 induces deoxycytidine deaminase (cytidine/deoxycytidine aminohydrolase, EC 3.5.4.5) activity when it lytically infects a number of mammalian cell lines. The deaminase activity is induced in a mouse cell line that is deficient in this enzyme. The induction of the enzyme in this mutant cell line does not occur in the presence of actinomycin D and the induced enzyme is more thermolabile than the enzyme of the wild-type mouse cell line. Furthermore, a new deoxycytidine deaminase species with a characteristic electrophoretic mobility that is different from that of the host cell enzyme is found in cell extracts prepared from a human cell line infected with herpesvirus. These results strongly suggest that the virus-induced deoxycytidine deaminase is coded by the viral genome. Because a deficiency in this enzyme is conditionally lethal for cells growing in a medium containing 5-methyldeoxycytidine as the sole source of thymidylate, this enzyme can be utilized as a selective marker for selecting mutant cells that have regained deoxycytidine deaminase activity as the result of infection by ultraviolet-inactivated herpes simplex virus.
Cell Line↗
Herpes simplex encephalitis: an autopsy case with isolation of type 1 herpes simplex virus.
An adult case of herpes simplex encephalitis was studied after autopsy. Postmortem examination revealed necrotizing encephalitis associated with Cowdry type A intranuclear inclusion bodies in glial cells. Herper simplex virus type 1 was isolated from the removed brain. Herpes simplex virus antigens were detected diffusely in wide areas of the brain by immunofluorescent test and viral particles characteristic to herpes simplex virus were demonstrated by electron microscopy. There was an apparent discrepancy between severity of histological changes and distribution of virus antigen.
Adult↗
Syntheses of virus-induced thymidine kinase and viral DNA in herpes simplex type 1 virus-infected chick embryo fibroblasts.
Herpes simplex virus type 1, strain Kupka, did not replicate in chick embryo fibroblasts (CEF), but the infection was followed by the development of cytopathic changes. This effect could be abolished by UV irradiation of the virus. Virus-induced thymidine kinase was synthesized in the infected cells reaching a maximum level at 24 hours post infection (p.i.). In the presence of cytosine arabinoside, thymidine kinase synthesis was enhanced. This suggested that the late (post-replicative) viral function, which turns off thymidine kinase synthesis, was expressed in the infected CEF untreated with the drug. HSV type 1 laboratory strains Kupka and KOS were capable of inducing the synthesis of virus-specific DNA in CEF. But in CEF infected with fresh type 1 virus isolates replication of viral DNA was not observed.
Animals↗
Strain characteristics and features of ocular infection of herpes simplex virus type 1 isolates.
Herpes simplex virus type 1 isolates from 63 patients with herpetic keratoconjunctivitis were investigated. For the purpose of detecting strain differences neurovirulence was determined in mice, and viral DNA's were analyzed by digestion with EcoRI and Hind III restriction endonucleases. Neurovirulence differed by a factor up to 10(5) between individual strains and proved to be independent of the ratio between infectious and noninfectious particles in the stocks used. The endonuclease cleavage patterns revealed differences of the viral DNA structure which permitted us to distinguish seven clusters of strains. Correlations between neurovirulence and DNA markers could not be established nor could correlations be found between these markers and features of disease. The main reason for not finding a relationship in the latter case may be attributed to the significant role of host factors in the course of herpetic diseases.
Adolescent↗
Cloning of herpes simplex type 1 DNA fragments in a bacteriophage lambda vector.
DNA isolated from defective and nondefective virions of herpes simplex type 1 (HSV-1) (strain Patton) was digested with restriction endonucleases, and the resulting DNA fragments were inserted in the EK2 coliphage vector lambdagtWES . lambdaB. The recombinant DNA was encapsidated in vitro under P4 maximum containment conditions. These lambda-HSV1 hybrids were purified and amplified, and the DNA was isolated in the P4 facility. DNA, free of viable phage and bacteria, was removed from P4 conditions and analyzed. Represented among the hybrids studied to date are DNA fragments from about 50 percent of the normal HSV-1 genome. The hybrids derived from defective HSV-1 DNA fragments demonstrate the existence of many similar but not identical classes of defective genomes.
Chromosome Mapping↗
Serum and CSF antibody levels to herpes simplex type 1, measles and rubella viruses in patients with schizophrenia.
Serum and CSF specimens from 12 schizophrenic patients and 10 non-psychiatric controls were tested for herpes simplex type 1 virus neutralizing antibody and for measles and rubella haemagglutination inhibiting antibodies. There were no significant differences in the distribution of virus antibody titres in serum or CSF specimens between the patients and the controls. The possible aetiological role of viruses or virus-like agents in schizophrenia and some methodological aspects are discussed.
Antibodies, Viral↗
Viremia with herpes simplex type 1 in adults. Four nonfatal cases, one with features of chicken pox.
Recovery of herpes simplex virus (HSV) Type 1 from the blood buffy coat of four adults is reported for the first time. All of the patients had vesicular stomatitis and facial vesicles; two also had either keratoconjunctivitis or disseminated skin lesions. The infection was not the primary one with HSV in any of them. Two of three patients who had renal failure were receiving immunosuppressive drugs; one patient was normal except for alcoholism and diabetes. None developed signs of visceral organ infection and all recovered within 2 to 4 weeks. The findings demonstrate the occurrence of heretofore unrecognized nonfatal HSV Type 1 viremia in both healthy and immunosuppressed leukocytes, can occur regardless of the presence of serum antibody, and may or may not be associated with the disseminated lesions.
Adult↗
Detection by complementation of defective or uninducible (herpes simplex type 1) virus genomes latent in human ganglia.
Reconstruction experiments have shown that temperature-sensitive (ts) mutants of herpes simplex virus type 1 (HSV-1)(Glasgow strain 17) grow, complement, and recombine with similar efficiency in human nerve ganglion cells, human brain cells, normal human fibroblasts (WI38), and baby hamster kidney (BHK) 21/C13 hamster cells. Cultures of human trigeminal, superior cervical, and vagus ganglia that had failed to release herpes simplex virus spontaneously were superinfected with a range of ts mutants of HSV-1 and incubated at both permissive (31 degrees C) and nonpermissive (38.5 degrees C) temperatures. Progeny virus was assayed at both temperatures to determine if complementation of or recombination with the input genomes had occurred. The results showed that the ganglia from 8 of 14 individuals, which had been consistently negative for spontaneous release of virus, contained information that could be detected or rescued following superinfection with ts mutants of herpes simplex virus. In two additional cases, positive results were obtained after the superinfection of negative ganglia explants, but in each of these herpes simplex virus had previously been spontaneously released from one of six ganglia explanted.
DNA, Viral↗
Immunoglobulin G antibodies to herpes simplex type 1 virus detected by radioimmunoassay in serum and cerebrospinal fluid of patients with schizophrenia.
Serum and CSF specimens from 16 schizophrenic patients and 18 nonpsychiatric controls were tested by radioimmunoassay for immunoglobulin G antibody of capsid, envelope and excreted antigens of herpes simplex type 1 virus. There were no significant differences in the antibody levels between the schizophrenic patients and the controls. The etiological role of viruses and virus-like agents in schizophrenia and some methodological aspects are discussed.
Adult↗
Effects of cytosine arabinoside on the incorporation of pyrimidine nucleosides into hamster cells transformed by herpes simplex type 1 and type 2 viruses and by human cytomegalovirus.
The incorporation of 3H-thymidine and 3H-deoxycytidine into acidoprecipitable fraction of hamster cells transformed by herpes simplex viruses type 1 and type 2 and of 3H-thymidine into hamster cells transformed by human cytomegalovirus was found to be resistant to the action of cytosine arabinoside. More 3H-thymidine was incorporated into these cells in the presence than in the absence of the drug. Similar stimulaton of 3H-thymidine uptake could be achieved by using unlabelled deoxycytidine instead of cytosine arabinoside. Incorporation of both nucleosides into spontaneously and SV40 transformed cells was efficiently inhibited by the drug.
Animals↗
Construction and characterization of a recombinant plasmid encoding the gene for the thymidine kinase of Herpes simplex type 1 virus.
We have constructed a hybrid plasmid by insertion of the thymidine kinase (TK) gene of Herpes simplex virus (HSV) type I at the BamHI site on Escherichia coli plasmid pBR322. The restriction endonuclease cleavage site map for the viral DNA fragment was determined for ten nucleases, and the insert in the recombinant plasmid has the same restriction nuclease digestion pattern as bona fide viral DNA. This result indicates that the plasmid contains an accurate copy of the viral DNA. The viral TK gene carried on the plasmid can be introduced into mammalian cells where it is expressed. This source of DNA with a selectable marker should be of considerable practical use in gene-transfer experiments in mammalian cells.
Chromosome Mapping↗
Differentiation of herpes simplex virus type 1 and type 2 by immunofluorescence: discriminative staining by labelled IgG preparations.
While evaluating herpes simplex virus (HSV) typing by indirect immunofluorescence staining, an undesired specific staining pattern turned out to be a reliable marker for herpes simplex type 1. Cells infected with herpes simplex type 1 displayed clear staining with a FITC-conjugated antiglobulin preparation, also in the absence of herpes simplex-specific antibodies. Using the same conjugate, herpes simplex type-2-infected cells exhibited no fluorescence. The particular type of staining observed was influenced by neither the anatomical site of origin of the virus isolate nor the cell type used for virus preparation. Herpes simplex type 1-specific fluorescence was only obtained with the use of FITC-conjugates possessing anti-IgG activity. Both reliability and specificity of this discriminating procedure as a diagnostic tool has been established by typing 282 virus isolates over a period of 4 years.
Antibodies, Anti-Idiotypic↗
Isolation of herpes simplex type 1 from ventricular fluid of an infant with encephalitis.
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Cerebral Ventricles↗
Isolation and localization of herpes simplex virus type 1 mRNA.
Herpes simplex virus (HSV) DNA bound to cellulose has been used as a reagent to isolate viral mRNA for size analysis on denaturing agarose gels. Total viral polysomal polyadenylated RNA was isolated from cells late after infection when such RNA has sequences encoded by approximately 45% of the HSV DNA. This RNA has a size range of from 1.5 to greater than or equal to 8 kilobases, with certain sizes, such as 1.7 to 1.9 kilobases, being favored. We have used the restriction endonucleases HindIII and XbaI singly and together to generate various sized fragments covering the entire HSV-1 genome. These fragments have been bound to cellulose to allow isolation of HSV-1 mRNA annealing to different regions of the viral genome. Discrete sizes of viral mRNA are associated with certain regions of the genome, but the mRNA population hybridizing to even the smallest restriction fragments is complex. We used hybridization of size-fractionated RNA to Southern blots of restriction fragments of HSV-1 DNA generated by the BglII as well as HindIII and XbaI endonucleases to confirm the preparative hybridization data and to provide some overlap data for positioning transcripts. The data of blot and preparative hybridization agreed very well and were combined to construct a preliminary transcription map of HSV-1. Such a map revealed at least two areas of the long unique region of the HSV-1 genome which annealed to a large number of HSV-1 transcripts. Furthermore, discrete-sized mRNA species larger than 5 kilobases in length were found only in the middle of the long unique region. The implications of these data are discussed.
DNA Restriction Enzymes↗
Deoxyribonucleoside triphosphate pools in herpes simplex type 1 infected cells.
Deoxyribonucleoside triphosphate pools were analysed in both exponentially growing and serum starved wild type BHK C13 cells and in a derivative of this cell line which lacks both thymidine kinase and deoxycytidine kinase activities, before and after infection with herpes simplex virus. Serum starved BHK cells had low levels of all four deoxyribonucleoside triphosphates. In exponentially growing cells all pools were expanded, the pool of dCTP being largest and dGTP the smallest. The dATP and dTTP pools were of intermediate sizes. In exponentially growing deoxypyrimidine kinase free cells the pools, with respect to level and distribution, were the same as those observed in wild type cells. After infection with herpes simplex virus there were marked changes in the levels of all deoxyribonucleoside triphosphate pools; the most predominant being a 25- to 50-fold expansion of dTTP pool. The pools of dCTP and dGTP also increased while the pool of dATP was very much reduced. These effects could be observed in both wild type and mutant cells.
Animals↗
Heterozygosis and genetic recombination in herpes simplex type 1 virus.
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DNA, Viral↗
Comparison of the development of resistant strains of Type 1 herpes simplex virus to in vitro antiviral activity of 5-iodo-2'-deoxyuridine or ribavirin.
Exposure of HSV/1 to low concentrations of ribavirin during 4--5 passages does not produce ribavirin-resistant virus. IDU resistance was developed by HSV/1 while it was being passed simultaneously. This resistance was seen to develop in both KB and Vero cells. The IDU-resistant virus is also resistant to ribavirin in KB cells.
Drug Resistance, Microbial↗