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PubMed1983

Temperature-sensitive mutants of newcastle disease virus affecting interferon induction.

Temperature-sensitive (ts) mutants of Newcastle disease virus (NDV) were isolated and studied for interferon (IFN) induction in primary chick embryo (CE) cells. At the non-permissive temperature (41 degrees C), there was no viral RNA synthesis or IFN induction by u.v.-treated virions except for ts-3 (RNA+), which did synthesize RNA at 41 degrees C, and whose u.v.-treated virions did induce IFN at this temperature. Another mutant (ts-4) induced IFN without irradiation, at the permissive temperature (37 degrees C). The minimum u.v. target size for IFN inducibility was unaffected by the mutation and corresponded to about 5% of the genome required for the expression of infectivity. These results support the hypothesis that the appearance of NDV RNA immediately after infection (primary transcription) plays a key role in IFN induction.

Animals↗
PubMed1975

Relationships among the polypeptides of Newcastle disease virus.

We have studied the relationships among the polypeptides of Newcastle disease virus by using both kinetic and tryptic peptide analyses. The results of our tryptic peptide analyses suggest that there are at least six unique viral polypeptides--L, HN, FO(F), NP, M, and a 47,000-dalton polypeptide. The small virion glycopolypeptide F is related to FO, a glycopolypeptide found only in infected cells. In addition, several smaller polypeptides, including a 53,000-dalton polypeptide found both in purified virions and in infected cells, are related to the nucleocaspid protein. Kinetic analysis of each viral polypeptide reveals that all of the major viral polypeptides, with the possible exception of L, are stable after an amino acid chase. A precursor-product relationship between FO and F was not demonstrable by pulse-chase experiments. Also, almost the same relative amount of F, the putative product, was present in infected cultures after either 5 or 30 min of radioisotopic labeling. These results suggest that FO is processed rapidly.

Glycopeptides↗
PubMed1970

Growth in chick chorioallantoic membranes of strains of Newcastle disease virus of differing virulence.

The growth of eight strains of Newcastle disease virus in chick embryo chorioallantoic membranes was studied by comparing, at different times after infection, the amounts of haemagglutinin released into the allantoic fluid (extracellular haemagglutinin) with that associated with the membrane (cell-associated haemagglutinin). The virulence of the strains examined differed in that some killed chick embryos more rapidly than others. All strains released similar amounts of extracellular haemagglutinin and maximum titres were achieved about 12 hr. after infection. With virulent strains cell-associated haemagglutinin titres increased exponentially until the death of the host and maximum titres were much higher than those of extracellular haemagglutinin. With avirulent strains cell-associated haemagglutinin titres increased exponentially for only a limited time and titres were always lower than the titres of extracellular haemagglutinin.Similar results were obtained when the titres of neuraminidase and viral ribo-nucleoprotein were measured during the growth of two virulent and two avirulent strains. Virulence appears to be associated with the continued intracellular accumulation of viral antigens.

Allantoin↗
PubMed1980

Synthesis, stability, and cleavage of Newcastle disease virus glycoproteins in the absence of glycosylation.

Polypeptides synthesized in Newcastle disease virus (NDV)-infected CHO cells in the absence of glycosylation were characterized. Incorporation of either [3H]mannose of [3H]glucosamine into NDV polypeptides was inhibited to greater than 99% by the antibiotic tunicamycin. Under these conditions, infected cells synthesized proteins which comigrated on polyacrylamide gels with the viral L protein, nucleocapsid protein, membrane protein, and a polypeptide with a molecular weight of 55,000 (P55). These cells did not synthesize polypeptides with the size of the hemagglutinin-neuraminidase (HN) protein or the fusion (F0) protein. They did, however, synthesize new polypeptides with molecular weights of 75,000 (P75), 67,000 (P67), and 52,000 (P52). Peptide analysis revealed that P75 was a host cell protein whose synthesis is enhanced by tunicamycin. P67 corresponded to the unglycosylated forms of the glycoproteins were found to be relatively stable in infected cells. P55, previously thought to correspond to the cleaved form of F0, was found to be a unique viral protein which is associated with intracellular nucleocapsid structures.

Animals↗
PubMedDate not supplied

Characterization of field isolates of Newcastle disease virus using antipeptide antibodies.

A recent Australian field isolate of Newcastle disease virus (NDV) was analyzed with antipeptide antibodies capable of differentiating between the sequences at the cleavage activation sites of fusion proteins of different NDV pathotypes. The isolate was found to have the same fusion protein cleavage activation signal as the V4 isolate of the Queensland strain of NDV. However, the isolate failed to react with an antibody specific for the carboxyl-terminal extension on the hemagglutinin/neuraminidase (HN)-protein precursor (HNo-protein) of the V4 isolate and other similar strains (e.g., Ulster and D26). Identity of the fusion protein cleavage activation motif of the isolate was confirmed by chemical analysis of purified fusion protein subunits of the isolate. The combination of a V4-like fusion protein cleavage activation motif but lack of an HNo-protein has not been described before, and the findings indicate that the isolate is a distinct strain of NDV. Analysis of a range of isolates from the state of Victoria, Australia, indicated that similar strains have been present in Australia since at least 1976. Other isolates examined appeared to have fusion protein cleavage activation motifs that could not be defined with the fusion-protein-targeted antibodies. These isolates also appeared to lack the HNo-protein characteristic of the Queensland strain.

Amino Acid Sequence↗
PubMed1989

Differential expression of interferon alpha and beta induced with Newcastle disease virus in mouse macrophage cultures.

Stimulation of mouse macrophages with Newcastle disease virus (NDV) leads to a rapid and high interferon (IFN) response. The magnitude of this response is influenced by the mouse genotype. We have analysed NDV-induced IFN production at the protein and mRNA levels in two different populations of macrophages derived from 'high producer' C57BL/6 and 'low producer' BALB/c mice in vitro. The data indicate that bone marrow and peritoneal macrophages from both strains grown in the presence of L cell conditioned medium (CM) as a source of macrophage colony-stimulating factor 1 (M-CSF) or purified murine M-CSF produce 10- to 50-fold more IFN on a per cell basis than cultures of resident peritoneal macrophages. These differences were also found when steady state levels of IFN mRNA were analysed. Differential analysis for the ratios of IFN-alpha and IFN-beta showed that CM- or M-CSF-cultured macrophages produced equal amounts of both IFN species as determined by specific monoclonal antibodies and hybridization experiments using IFN-alpha and IFN-beta DNA probes, whereas resident peritoneal macrophages induced under identical conditions produced almost exclusively IFN-beta. This suggests a stimulating effect of M-CSF on IFN synthesis in NDV-induced cultures of mouse macrophages, which is in part due to additional activation of IFN-alpha gene expression.

2',5'-Oligoadenylate Synthetase↗
PubMed2005

The use of in situ hybridization and immunohistochemistry to study the pathogenesis of various Newcastle disease virus strains and recombinants in embryonated chicken eggs.

Avian paramyxovirus type 1, commonly referred to as Newcastle disease virus (NDV), is a serious pathogen of significant economic importance to the industry. To investigate the role of the fusion (F), hemagglutinin-neuraminidase (HN), and (P) phosphoprotein gene sequences in virulence, six strains of Newcastle disease virus (NDV) representing all pathotypes and seven recombinant strains created by reverse genetics were inoculated into 9-day-old chicken embryos. Tissues and chorioallantoic membranes (CAM) were harvested at 24-hour intervals post-inoculation. Riboprobe in situ hybridization and immunohistochemistry highlighted distinct tissue tropisms among the viruses. Presence of F and/or HN from virulent viruses inserted into lentogenic backbones caused dissemination of virus in a manner similar to wild type virulent viruses. Disruption of P gene decreased dissemination of velogeinic infectious clones. It is concluded that each of these genes contributes to pathogenicity.

Animals↗
PubMed1983

Monoclonal antibodies to newcastle disease virus: delineation of four epitopes on the HN glycoprotein.

Eighteen independent hybridomas producing monoclonal antibodies to Newcastle disease virus have been prepared by fusion of SP2 cells with spleen lymphocytes from a BALB/c mouse immunized with intact UV-inactivated Newcastle disease virus strain Australia-Victoria. They have been divided into three groups on the basis of radioimmunoprecipitation, infected cell surface and cytoplasmic fluorescence, and isotype. The anti-HN group is made up of nine antibodies which give surface fluorescence on infected cells and immunoprecipitate the HN glycoprotein. These antibodies bind to HN in nitrocellulose transfers of sodium dodecyl sulfate gels, but only if it has been neither reduced nor boiled. To varying degrees, all of these HN antibodies neutralize infectivity. These results suggest that they recognize exposed determinants of a conformational nature on the native HN molecule. They have been used in competition antibody-binding radioimmunoassays and additive neutralization assays, and on the basis of these studies the epitopes they recognize have been subdivided into four domains, two of which are overlapping, on the HN glycoprotein. The relatively weaker neutralizing activity observed with some of these antibodies cannot be explained by lower avidities for their epitopes because there is not an inverse correlation between estimated binding constant and neutralizing activity. The four antibodies in the second group all give a predominantly cytoplasmic fluorescence pattern, immunoprecipitate the nucleocapsid protein, and bind to nucleocapsid protein in nitrocellulose transfers of reduced and nonreduced sodium dodecyl sulfate-polyacrylamide gels. All five of the antibodies in the third group are of the immunoglobulin M class, unlike the others which are all immunoglobulin G antibodies. Members of this group show variable fluorescence patterns, but none is able to immunoprecipitate or bind to a specific viral antigen transferred to nitrocellulose paper from sodium dodecyl sulfate-polyacrylamide gels.

Animals↗
PubMed1982

Newcastle disease virus stimulates the cellular accumulation of stress (heat shock) mRNAs and proteins.

A biological agent, Newcastle disease virus, stimulated the synthesis of stress proteins in cultured chicken embryo cells. Previously, only physical and chemical agents were known to induce these proteins. The levels of translatable stress mRNAs were elevated in cells infected with avirulent or virulent strains; however, stress protein synthesis was stimulated strongly only in cells infected by avirulent strains. As did several other paramyxoviruses, avirulent strains of Newcastle disease virus stimulated the synthesis of glucose-regulated proteins as well as stress proteins. Possible stimuli of the synthesis of these two sets of proteins in paramyxovirus-infected cells are considered.

Animals↗
PubMed1964

STUDIES ON PERSISTENT INFECTIONS OF TISSUE CULTURES. V. THE INITIAL STAGES OF INFECTION OF L(MCN) CELLS BY NEWCASTLE DISEASE VIRUS.

The initial stages of infection of L(MCN) cell populations with standard Newcastle disease virus (NDV(ST)) were analyzed in an effort to elucidate the steps leading to survival of the cultures and to the indefinite persistence of the infectious process at a low level. Cells were exposed in suspension to NDV at varying multiplicities and the monolayer cultures derived from such cells assayed at intervals for cellular growth rates, percentage of infected cells as determined by immunofluorescence, yields of viral progeny and of interferon, and, on occasion, resistance to superinfection with vesicular stomatitis virus. The percentage of cells calculated to be initially infected on the basis of adsorption data was found to match closely the percentage of immunofluorescent cells resulting from the first infectious cycle (up to 24 hours). Cells initially infected with NDV(ST) produced a mixed progeny of infectious virus (from 15 to 40 pfu/cell) and about 10 times as many non-infectious particles in 24 hours [NDV(L(MCN))], but little or no interferon. If all cells were infected the cultures ultimately died. At multiplicities of infection (m) of 2 or less the cultures survived with increasing ease as the percentage of infected cells was reduced. The number of pfu per infected cell was of the above order during the first 3 days; it declined thereafter. Limited secondary spread of the infection was noted by 48 hours and no further cycling was noted thereafter. As m decreased from 2.0 to 0.1 there was an increase in the yields of interferon and the time at which peak titers were reached. Addition of anti-NDV serum 2 hours after infection prevented measurable production of interferon. In contrast, following exposure of cells to NDV(L(MCN)) at multiplicities ranging from 20.0 to 0.2 (based on infectious virus) all cultures survived, no secondary spread was noted, the number of pfu per infected cells was reduced at the higher multiplicities, and the yields of interferon were similar and maximal by 24 hours and not affected by anti-NDV serum added after an adsorption period of 2 hours. It is concluded that the non-infectious virus particles in the progeny released from NDV(ST)-infected cells induce resistance in remaining cells or, if adsorbed simultaneously with infectious virus, abort the intracellular infectious process. In both instances interferon is produced which may then render additional cells resistant. The non-infectious component is considered an incomplete or defective product of viral replication and not merely thermally inactivated virus. NDV(ST) partially or completely inactivated at 37 degrees C induced neither cellular resistance nor synthesis of interferon. The incomplete viral component behaved in all respects like ultraviolet-inactivated NDV(ST) except that it was significantly more efficient in inducing interferon synthesis. On the basis of the presented data a scheme has been devised and discussed which appears to explain satisfactorily the events which take place on initial infection of L(MCN) cells with NDV and which lead to the persistence of the infectious process.

Animals↗
PubMed2001

[Study of Newcastle disease virus in the treatment of human laryngeal squamous carcinoma].

OBJECTIVE: To explore the biological value of newcastle disease virus in the therapy of human laryngocarcinoma. METHODS: Nude mice model bearing laryngocarcinoma were established using human laryngeal squamous carcinoma cell line (Hep-2). Large amounts of Newcastle disease virus(NDV) were injected into the tumor. Changes in carcinoma administrated with the NDV were observed under light and electron microscopes. Isolation of NDV was attempted and sera were examined using the method of enzyme-linked immunosorbent assay (ELISA). RESULTS: The difference between experimental and control groups was statistically significant, including the average weight of the mice: t' = 2.397 (P < 0.05), the average weight of the tumor tissue: t' = 2.852 (P < 0.05) and the average volume of the tumor tissue: t' = 6.058 (P < 0.01). In the experimental group, the NDV was isolated in the tumor tissue and ELISA was positive. The necrosis of the tumor cell and inflammatory cellular infiltration were found under light and electron microscopes. CONCLUSION: These results indicated that NDV was effective in the treatment of laryngocarcinoma and had no damage to normal tissue.

Animals↗
PubMed1980

Collection and processing of blood samples dried on paper for microassay of Newcastle disease virus and avian influenza virus antibodies.

A practical method for collection and processing of dried whole blood samples on filter paper was developed to facilitate large-scale testing programs for Newcastle disease virus and avian influenza virus antibodies. A modified paper punch was used to cut and place dried blood samples simultaneously in a standard 96-well microlate for elution of antibody. Twelve eluted samples were simultaneously transferred to another microplate for the hemagglutination-inhibition (HI) microtest. Similar HI titers were obtained with simultaneously collected serum and dried blood samples. Minor differences were not considered of practical importance in diagnostic serologic studies. Dried blood titers were not markedly affected by method of drying (37 C for 2 hours or 26 C for 4 hours), by storage for 24 hours before drying, or by storage of dried samples at 4 C for 28 days or 30 C for 14 days. Blood dried on paper was a satisfactory sample for assay of HI antibodies to Newcastle disease virus and avian influenza virus.

Animals↗
PubMed1961

Enumeration of cell-infecting particles of Newcastle disease virus by the fluorescent antibody technique.

A procedure has been developed for the determination of the concentration of infective Newcastle disease virus (NDV) based on the enumeration of singly infected and distributed HeLa cells which are visualized by staining with fluorescent antibody. Infective virus assayed by the fluorescent cell-counting procedure is expressed in terms of cell-infecting units (CIU). Adsorption of NDV to HeLa cell monolayers reached a plateau 1 to 1.5 hours after inoculation of coverslip cultures, and 12 per cent of the infective particles inoculated failed to adsorb. The half-life of NDV in protein-free Eagle's medium at 37 degrees C. was 2.1 hours. There was a linear relationship between virus concentration and the number of infected cells. The coefficient of variation of the mean of replicate determinations of infective NDV was 8.2 per cent. The distribution of single infected HeLa cells in the monolayer corresponded to the Poisson distribution. With NDV the cell-infecting unit (CIU) determined in HeLa cells is equivalent to the plaque-forming unit in chick embryo cells and the egg infective dose. In experiments on the mechanism of dissemination of NDV in monolayer cultures of HeLa cells, NDV was found to spread from cell to cell through the extracellular milieu.

Animals↗
PubMed1985

[Effect of proteolytic enzymes on the enhanced sensitivity of a cell culture of goat embryonic kidney to Newcastle disease virus].

The influence of proteolytic enzymes on the increase of the sensitivity of cell cultures to Newcastle disease virus was tested in the continuous culture of goat embryo kidney cells. In this investigation the dose of trypsin and the time of incubation for the treatment of the cell culture was selected. The preliminary treatment of the monolayer culture of goat embryo kidney cells with trypsin by incubation at a temperature of 18-20 degrees C for 30 minutes made it possible to increase the sensitivity of the culture to Newcastle disease virus 10-fold.

Animals↗
PubMed1977

Butylated hydroxytoluene protects chickens exposed to Newcastle disease virus.

Dietary butylated hydroxytoluene, an antioxidant widely used in food and feed processing, prevents mortality of chickens exposed to virulent Newcastle disease virus and prevents the serological response of chickens exposed to avirulent Newcastle disease virus. This chemoprophylactic effect is evident when chickens are fed diets containing concentrations of butylated hydroxytoluene normally used for antioxidant purposes (100 to 200 parts per million of total diet).

Animals↗