Virus In Tissue Culture Research Articles

The Effect of Tilorone Hydrochloride on the Growth of Several Animal Viruses in Tissue Cultures

Tilorone hydrochloride, at a concentration of 10 mug/ml inhibits the growth of herpes simplex virus type 1 in BS-C-1 cells. The growth of vaccinia virus in BS-C-1 cells is partially inhibited; however, six viruses containing RNA, including four members of the togavirus group grown in chick fibroblasts, are not affected by the drug. The inhibition of the growth of herpes virus by tilorone hydrochloride is greater when the multiplicity of infection is lower than 1 p.f.u/cell and when the drug is added early in the course of infection.

Characteristics of a virus causing a pox disease in sheep and goats in Kenya, with observations on the epidemiology and control

The sheep and goat pox viruses isolated in Kenya are not host specific as in the Middle East and India. The virus strains from one species are of a similar pathogenicity for the other, and the same virus appears to occur in the field outbreaks in mixed flocks. A silent infection with no skin lesions occurs in the field outbreaks. The virus grows well in lamb testis cell cultures, it does not haemadsorb nor haemagglutinate and is ether sensitive. Most strains could not readily be adapted to grow on the chorioallantoic membrane of embryonated eggs; one isolation only was made in this way. Staining of the virus inclusions showed that they were of DNA; the virus resembled that of lumpy skin disease in many characteristics. It was possible to cross-protect both species with virus of sheep or goat origin. A vaccine was made from tissue culture virus adsorped onto aluminium hydroxide gel and treated with formalin.

Comparison of guinea pig cytomegalovirus and guinea pig herpes-like virus: pathogenesis and persistence in experimentally infected animals

The pathogenesis of guinea pig cytomegalovirus (GPCMV) and guinea pig herpes-like virus (GPHLV) in guinea pigs was compared. Animals were inoculated with the two viruses by different routes and sacrificed after varying periods of time. GPCMV was consistently isolated from salivary gland 2 weeks postinoculation and thereafter following intraperitoneal or subcutaneous incoulaton. Virus was less frequently found in other tissues including blood, spleen, and kidney. Intranuclear inclusions were seen in tissue sections of salivary gland after inoculation with GPCMV- infected tissue suspension, but were only rarely found after inoculation with tissue culture virus. In GPHLV-infected guinea pigs, consistent latent infection of leukocytes and other tissues was detected by cocultivation techniques. Intranuclear inclusions were not found in the spleen, salivary gland, or other infected tissues after GPHLV infection with either tissue culture virus or infected tissue suspension. Guinea pigs inoculated with GPCMV produced high titers of specific neutralizing antibody to the homologous virus; those inoculated with GPHLV developed long-term viremia accompanied by minimal neutralizing antibody levels to the virus.

Infection and Replication of Insect Pathogenic Viruses in Tissue Culture

Publisher Summary This chapter discusses the recent findings concerning the use of continuous cell lines for the study of viruses pathogenic to insects. The type of tissue culture medium used for growing cells may have an influence on the development of insect viruses. Insect pathogenic viruses are generally divided into two main groups: occluded and nonoccluded. Virus inoculum of several types can be used to infect insect tissue cultures: (2) infected tissues and tissue extracts; (2) naked viral nucleic acid; (3) virions purified from polyhedra; (4) infectious hemolymph; and (5) cell-free infected tissue culture medium. The latter two types of inoculum are the most commonly used. Several studies on the infection and replication of insect pathogenic viruses are also presented in the chapter. These studies investigate the biochemical events in virus replication and optimum conditions for viral infection and virus development. Alkaline treatment of polyhedral inclusion bodies to release the virus particles is a well-documented and simple procedure. However, investigations have shown that virion suspensions prepared by dissolving polyhedra in dilute sodium carbonate solutions are not reliable inocula for infecting cultures.

VIROLOGIC CONSIDERATIONS ON ANTI-DENGUE VACCINE

Possibilities of developing an anti-dengue vaccine are discussed based on our experimental findings with a dengue virus type 1 (DEN-1) Mochizuki strain. The Mochizuki strain virus was attenuated for human beings in the course of several passages through mouse brains since its isolation from the blood of a dengue patient in the 1943 epidemic in Nagasaki, Japan. This attenuated virus in the living state induced no abnormal symptoms after injection into human volunteers or monkeys, yet it stimulated production of specific anti-DEN-1 NT and HI antibodies and induced a prophylactic effect on the human beings against virulent dengue viruses (in the form of patient's serum) given experimentally. The immunogenic effects of the virus was maintained after passages in primary monkey kidney tissue cultures. Using the tissue culture virus, together with the 17D yellow fever (YF) vaccine, a combined immunization of human beings against DEN and YF was possible; production of the antibodies of significant titers against both viruses was unequivocally demonstrated. The partially purified virus was similarly effective in stimulating production of the anti-DEN antibodies, and the same statements could be made as to the viral components obtained by sucrose density gradient centrifugation. The virus was shown to multiply in the WI-38 cell line cultures, and this suggests application of this cell line to produce a live tissue culture vaccine.Conclusions of this review are : (1) the Mochizuki virus is completely attenuated; (2) yet it stimulates production of specific anti-DEN antibodies and induces protection against DEN-1 infection by virulent human virus; and (3) that possibility of developing a live tissue culture vaccine is suggested; this also makes possible a combined DEN-YF vaccination on the experimental basis. It is stated in the connection that the purified virus or its components can be used for the above purposes.Problems regarding the implication of “hypersensitivity hypothesis” of dengue and dengue hemorrhagic fever-shock syndrome in anti-DEN vaccination are in addition discussed.

Inactivation of African horse-sickness virus by betapropiolactone and by pH

The inactivation of several types of African horse sickness virus (AHSV) by pH and by betapropiolactone (BPL) was studied. At 19 degrees - 22 degrees C the virus was stable between pH 6.0 and 10.4, whether suspended in mouse brain or in serumfree buffer. Below pH 5.6 and above pH 10.9, more than 99 per cent of infectivity was inactivated within 15 minutes. The addition of 50 per cent serum did not influence pH stability. Disinfection in the presence of citric acid and caustic soda is briefly discussed. Inactivation by BPL was complete within 30 minutes at 37 degrees C, yet incomplete after 15 hours at 4 degrees C. Types 3 and 9 virus grown in suckling mouse brain and types 1, 3 and 9 produced in pig kidney cells were equally susceptible to 0.1 per cent BPL, more than 99.9 per cent being inactivated. The effectiveness of BPL was reduced at least 10-fold by the addition of 50 per cent serum. No infective virus was detected following incubation of either tissue culture virus with 0.2 per cent BPL or of mouse brain virus with 0.3 per cent BPL. Virus suspensions exposed to 0.3 per cent BPL required buffering with Tris of at least 0.05 molar strength in order to maintain the pH within an acceptable range. Inactivated antigens prepared with 0.4 per cent or lower concentrations of BPL were immunogenic in guinea pigs.

Temperature-dependent rosette formation by mouse lymphoma cells as a result of viral hemadsorption.

Cells from several mouse lymphomas formed rosettes with nonsensitized foreign erythrocytes through C-type virus particles clustered on the cell surface in serum-free medium held at 4 degrees C. This type of rosetting was found most typically in a lymphoma induced by Rauscher leukemia virus in tissue culture (RD-12), but it also occurred in 23 of 61 spontaneous thymic lymphomas in AKR mice. Chemically or X-ray-induced leukemias and spontaneous reticulum cell sarcomas did not form rosettes. The nature of the rosette formation may be interpreted as viral hemadsorption, with a possible relationship to hemagglutination by murine leukemia viruses. The receptor on virus particles was trypsin sensitive and showed high affinity to serum inhibitors (RIF). Serum rosette-inhibiting activity was assessed by a quantitative rosette inhibition test; rosette inhibition proved widely distributed among species. Physicochemical properties of serum RIF and their function both in vivo and in vitro were described. Rosette formation with similar temperature requirements, previously reported in a mouse lymphoma carrying membrane-bound heterophile cold hemagglutinin, was readily distinguished from viral hemadsorption by its insensitivity to mouse serum RIF.

Efficacy of ribavirin against influenza virus in tissue culture and in mice.

Annals of the New York Academy of SciencesVolume 255, Issue 1 p. 366-371 EFFICACY OF RIBAVIRIN AGAINST INFLUENZA VIRUS IN TISSUE CULTURE AND IN MICE F. E. Durr, F. E. Durr Lederle Laboratories Division American Cyanamid Company Pearl River, New York 10965Search for more papers by this authorH. F. Lindh, H. F. Lindh Lederle Laboratories Division American Cyanamid Company Pearl River, New York 10965Search for more papers by this author F. E. Durr, F. E. Durr Lederle Laboratories Division American Cyanamid Company Pearl River, New York 10965Search for more papers by this authorH. F. Lindh, H. F. Lindh Lederle Laboratories Division American Cyanamid Company Pearl River, New York 10965Search for more papers by this author First published: August 1975 https://doi.org/10.1111/j.1749-6632.1975.tb29244.xCitations: 10AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume255, Issue1Chemistry, Biology, and Clinical Use of Nucleoside AnalogsAugust 1975Pages 366-371 RelatedInformation

Synthesis and antiviral activity of certain 5'-monophosphates of 9-D-arabinofuranosyladenine and 9-D-arabinofuranosylhypoxanthine.

A number of 5'-phosphates of 9-D-arabinofuranosyladenine and 9-D-arabinofuranosylhypoxanthine were prepared and tested against a variety of DNA viruses in tissue culture. The syntheses of the antiviral agent 9-beta-D-arabinofuranosylhypoxanthine 5'-monophosphate (6) and a series of related nucleotides, 9-beta-D-arabinofuranosyladenine 5'-O-methylphosphate (3), 9-beta-D-arabinofuranosylhypoxanthine 5'-O-methylphosphate (7), 9-beta-D-arabinofuranosylhypaxanthine cyclic 3',5'-phosphate (13), and 9-alpha-D-arabinofuranosylhypoxanthine 5'-monophosphate (17), are described. The concepts underlying the development of these antiviral agents are discussed. Comparison of the anti-DNA viral activity is made with 9-beta-D-arabinofuranosyladenine (ara-A). Reproducible antiviral activity against three DNA viruses in vitro at nontoxic dosage levels is demonstrated by 3,6, and other related nucleotides.

Propagation of human wart virus in tissue culture

HUMAN wart (papilloma) virus (HWV) is an authentic human tumour virus. Based on morphology, it has been classified as a member of the papova group1 of animal tumour viruses. Using electron microscopy, HWV has been found to be associated with human skin warts2, laryngeal and venereal papillomas3,4 and epidermodysplasia verruciformis5. Moreover, experiments done with human volunteers at the beginning of this century6–8, showed that filtered extracts of warts, laryngeal and venereal papillomas can cause skin warts in man. Since these early experiments, there has been little increase in our knowledge of HWV as a causative agent of either benign tumours or those which can progress to malignancy5,9, mainly because of the lack of a system in which to propagate HWV in vitro10. This report describes the propagation of HWV in a tissue culture system.

Studies of FBJ Osteosarcoma Virus in Tissue Culture. II. Autoinhibition of Focus Formation

A high-titered non-focus-forming virus, FBJ-MuLV (murine leukemia virus), present in FBJ tumor preparations, inhibited significantly the expression and production of FBJ-MuSV (murine sarcoma virus) in tissue culture. This "autoinhibition" was comparable to that observed when a 3- to 4-log excess of infectious MuLV was added to standard MuSV. The degree of inhibition was influenced by the tropism of the MuLV (or the ease of spread and propagation of MuLV in certain cells), multiplicity of infection by MuLV, amount of excess MuLV, and ability of the MuSV-transformed cells to replicate independently. The FBJ MuLV-MuSV complex may be a model system for the detection of sarcoma viruses in spontaneous tumors in various animals where inhibition by excess nontransforming virus could be an important biologic phenomenon.

Expression and detection of endogenous mouse C-type RNA viruses.

Many naturally occurring C-type RNA viruses are of endogenous origin. The genetic information for synthesizing these RNA viruses is present in the DNA of normal mouse cells, probably as part of their chromosomal DNA. Some C-type viruses infect mouse cells (homotropic virus), while others infect certain tissue culture cells from other species but not mouse fibroblasts (xenotropic virus). All mouse strains studied appear to contain endogenous xenotropic viral genomes. However, based on the regularity with which homotropic virus is detected, inbred mice can be divided into high, low, and nonvirus-yielding strains. Nucleic acid hybridization studies have shown that DNA from high virus strains contains several copies of the homotropic virus genome, while that from low virus strains contains fewer copies, and DNA from nonvirus strains lacks a significant portion of the homotropic virus genome. In vivo and in vitro genetic studies support the nucleic acid hybridization results. In addition, high virus mouse strains are more likely than low virus strains to release virus that will replicate efficiently in their own cells. Methods for the activation and detection of endogenous C-type virus in tissue culture are discussed.

Tissue culture cultivation ofLaunaea nudicaulis Hook f. infected with a mosaic virus disease

Healthy and mosaic infected leaves ofL. nudicaulis have been cultivated onMS and modifiedMS media (MC). Profuse growth and differentiation of tissue have been obtained onMS medium in diseased tissues as compared to the modified medium. It has been found on bioassay that although there is no difference between the concentration of the virus in tissue culture and glasshouse material yet the incubation period of the virus is decreased in tissue cultures.

Effect of folic acid and methotrexate on reproduction of Rauscher leukemia virus.

Folic acid was found to stimulate and its structural analog methotrexate was found to inhibit the reproduction of an oncogenic RNA-containing virus in tissue culture.

Low resistance junctions between normal and between virus transformed fibroblasts in tissue culture

The occurrence of low resistance junctions between normal chick embryo fibroblasts and between fibroblasts transformed with Rous sarcoma virus in tissue culture was studied with intracellular microelectrodes. The results showed that these junctions were present between normal chick fibroblasts in proliferating cultures as well as between cells in ‘density dependent inhibited cultures’. Mechanical injury to a fibroblast within a small group of coupled cells caused the injured fibroblast to uncouple immediately from its neighboring cells without interrupting coupling between the healthy uninjured cells. In the case of fibroblasts transformed with a Rous sarcoma virus, the results further showed that low resistance junctions were present when the transformation appeared in the infected cells and remained present thereafter.

Two viruses from the Lucké tumor isolated in a frog pronephric cell line.

SummaryThe first established frog pronephric cell line was exposed to tumor cell fractions of the oncogenic (nuclear inclusion body) phase of the Lucke renal adenocarcinoma. A mixed infection of the herpes virus and adeno-like virus was obtained. When tested by embryonic bioassay, the original tumor fractions and the first passages of the viruses in tissue culture produced subdermal hemorrhage, turgid edema and total deaths in 3-6 weeks. In subsequent passages the adeno-like virus appeared to abort and the herpes virus predominated. Bioassay of these and later passages produced no trauma within 3 months.

Some observations on the propagation of avian infectious bronchitis virus in tissue culture.

Some observations on the propagation of avian infectious bronchitis virus in tissue culture.

Two-Hour Embedding Procedure for Intracellular Detection of Viruses by Electron Microscopy

An embedding method requiring only 2 h to complete and giving excellent ultrastructural preservation has been used for the rapid detection of viruses in tissue cultures. The method has also been applied successfully to mammalian tissue. It provides a rapid technique for identifying viruses isolated in tissue cultures, for screening cultures for adventitious agents, and for examining tissue biopsies for viruses.

Rapid Identification of Swine Vesicular Disease Virus in Vesicular Epithelium Using the Fluorescent Antibody Technique

The fluorescent antibody technique was used to detect virus in tissue cultures inoculated with suspensions of vesicular epithelium from lesions of known age. Positive results were obtained after 3 h from vesicles less than 5 days old and after 3 to 5 h from lesions 6 to 10 days old. The infectivity of vesicular epithelium declined from approximately 10 10 pfu/g in vesicles 1 day old to 10 6.9 pfu/g in lesions 10 days old.

Complement-fixing antigens produced by varicella-zoster virus in tissue culture.

The complement-fixing antigens present in tissue cultures infected with varicella-zoster virus were separated into four components. There were (i) a cell-free soluble antigen, (ii) a cell-associated soluble antigen, (iii) a cell membrane-associated antigen, and (iv) a virion antigen. All four antigens were reactive with sera from patients with varicella or zoster, and about 90% of the total complement-fixing activity was found to be nonvirion associated.