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PubMedDate not supplied

Genetic stability of Newcastle disease virus CG179: a strain containing only clear plaque subpopulations.

Two cultures of the CG179 strain of Newcastle disease virus were maintained apart in two laboratories for over 30 years. One culture had been passaged 45 times through embryonated chicken eggs, yet this virus had the same virulence, thermostability of hemagglutinin, plaque types, and RNA fingerprint as a culture lyophilized in 1948. The genetic stability observed here has not been reported for strains containing both red and clear plaques and may occur only in strains like CG179 that do not have red plaques.

Animals↗
PubMed1993

Unexpected isolation of virulent Newcastle disease virus from commercial embryonated fowls' eggs.

The authors report a case of causal isolation of virulent Newcastle disease virus (NDV) from embryonated eggs. The virus was isolated from uninfected chicken embryo liver and fibroblast cultures prepared from commercial embryonated fowls' eggs. A serological and virological investigation carried out on the breeders which had laid those eggs showed high titres against NDV, and virus isolation from cloacal swabs. The virus was also isolated from the progeny of the same breeders which showed no clinical signs of ND, following episodes of mortality. Vertical transmission of the virus is discussed.

Animals↗
PubMed1983

The characterization of monoclonal antibodies to Newcastle disease virus.

Monoclonal antibodies to the haemagglutinin-neuraminidase (HN), fusion (F), polymerase and nucleocapsid polypeptides of Newcastle disease virus were prepared. Two epitopes were recognized on the HN polypeptide: one was associated with inhibition of haemagglutination and poor neutralization and the other with good neutralization and no inhibition of haemagglutination. The most effective neutralizing antibody was that produced against the F polypeptide. The poorer neutralization associated with the antibody against the HN epitope was augmented by antiglobulin or complement. The monoclonal antibodies that inhibited haemagglutination also inhibited neuraminidase activity when fetuin but not neuraminyl lactose was the substrate.

Antibodies, Monoclonal↗
PubMedDate not supplied

Use of egg yolk in serological tests (ELISA and HI) to detect antibody to Newcastle disease, infectious bronchitis, and Mycoplasma gallisepticum.

Serum and yolks from commercial flocks and from hens exposed to Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and Mycoplasma gallisepticum (MG) were tested for immunoglobulin G antibody by the enzyme-linked immunosorbent assay (ELISA) and the hemagglutination-inhibition (HI) test. Yolks prepared by chloroform extraction and low-speed centrifugation performed well in the serological tests used and were a suitable alternative to serum for antibody determination by the ELISA for NDV, IBV, and MG and by HI test for NDV.

Animals↗
PubMedDate not supplied

Differentiation of exotic strains of Newcastle disease virus by oligonucleotide fingerprinting.

The oligonucleotide fingerprints of five of six strains of velogenic-viscerotropic Newcastle disease (VVND) virus were readily distinguishable, even though the strains could not be differentiated by a series of in vivo and in vitro procedures. Two of the VVND virus strains tested (10333-0 and 8231412) showed marked similarity to the prototype strain (MIL-0). Two of the remaining three strains (80-27489 and 80-28634) had identical oligonucleotide fingerprints, even though they had a different isolation history and exhibited small differences in their virulence for chickens. Fingerprints of the sixth VVND strain, 80-12927, demonstrated some similarities when it was compared with the prototype virus fingerprint, but it was quite different from strains 80-27489, 10333-0 and 8231412.

Newcastle disease virus↗
PubMed1972

Hemolytic interaction of Newcastle disease virus and chicken erythrocytes. I. Quantitative comparison procedure.

The extent to which erythrocytes are hemolyzed by Newcastle disease virus is a function of the relative concentrations of both virus and erythrocytes. Under proper conditions, the interaction of a single virus particle with an erythrocyte is sufficient to cause lysis. The extent of hemolysis is directly proportional to virus concentration only when the virus-erythrocyte ratio is very low. At the higher virus-erythrocyte ratios usually employed in hemolysis experiments, the extent of hemolysis is proportional to the logarithm of the virus concentration. Thus, quantitative comparisons of hemolytic activities of different virus preparations cannot be made by directly comparing the extent of hemolysis. Relative hemolytic activities must be determined by comparing virus concentrations which yield equivalent amounts of hemolysis (the quantitative comparison procedure).

Animals↗
PubMed1988

Location of a neutralizing epitope for the haemagglutinin-neuraminidase glycoprotein of Newcastle disease virus.

The binding site of a monoclonal antibody to the haemagglutinin-neuraminidase (HN) polypeptide of Newcastle disease virus (NDV) has been located. Complementary DNA or synthetic oligonucleotides corresponding to portions of the HN gene were cloned into the Escherichia coli vector pUC19 and fragments of the HN protein were thereby fused to the alpha-peptide of beta-galactosidase. Western blot analysis of E. coli lysates containing expressed fragments of the HN cDNA or synthetic oligonucleotides identified an antibody-binding peptide (Asp-Glu-Gln-Asp-Tyr-Gln-Ile-Arg; amino acid residues 346 to 353). Nucleotide sequence analysis of an antibody-resistant mutant of NDV revealed a Glu (wild-type) to Lys (mutant) substitution within the above sequence. The methods described could be useful for the location of continuous epitopes of other polypeptides.

Animals↗
PubMed1984

[Newcastle disease virus in tissue cultures studied by an immunofluorescence method].

Studied was the opportunity to demonstrate the Newcastle Disease Virus in tissue cultures with the employment of an immunofluorescence method. The development was followed up of four strains--the vaccinal ones Hitchner B1, La Sota, and Komarov and the velogenic one, Texas GB, using chick fibroblasts. It was found that the velogenic strain cumulated earlier and more intensely than the vaccinal ones, where the virus was seen to explicitly locate perivascularly. The most dependable demonstration of the virus proved possible with the velogenic strain at the 4th hour, and with the vaccinal strains--from the 6 th to the 10 th hour following the infection of the cultures. The virus antigen cumulated progressively, and with the velogenic strain it was accompanied with the production of a cytopathic effect which destroyed the cells by the 20th hour; with the vaccinal strains the cells remained intact up to the 48th hour following infection.

Animals↗
PubMed1972

Temperature-sensitive mutants isolated from L cells persistently infected with Newcastle disease virus.

Virus mutants (NDV(pi)) isolated from L cells persistently infected with the Herts strain of Newcastle disease virus have been previously reported by this laboratory to differ from the wild-type virus (NDV(o)) in several physical and biological properties. It has now been determined that, in addition to these differences, the NDV(pi) mutants are also spontaneously selected temperature-sensitive mutants. The temperature sensitivity of 10 NDV(pi) clones was confirmed by temperature inhibition, plaquing efficiency, and single-cycle yield experiments. The cut-off temperature, at which more than 90% of virus replication is inhibited was between 41 and 42 C. All 10 NDV(pi) clones were also found to be defective in virus-specific ribonucleic acid (RNA) synthesis in infected chick embryo cells at 42 C and are tentatively classified as RNA(-). The possible relationships of the temperature sensitivity, the other NDV(pi) properties, and the maintenance of the persistently infected state are discussed.

Animals↗
PubMed2003

On the origins and relationships of Newcastle disease virus vaccine strains Hertfordshire and Mukteswar, and virulent strain Herts'33.

The origins and relationships of Newcastle disease virus (NDV) vaccine strains Hertfordshire (H) and Mukteswar, and the virulent Herts'33 were studied using partial sequence analysis of the fusion protein gene. The mesogenic strain H was obtained by egg passages of a field virus isolated in England in 1933 (later known as Herts'33). Different lines of the strain Herts'33, however, divided into two distinct groups: genotype IV, and a hitherto undescribed lineage, which comprised the Weybridge line (Herts'33/56). Vaccine strain H and the two clusters comprising viruses designated Herts'33 displayed 6.5 to 6.8% and 15.6 to 16.3% mutational distances, respectively, which precluded parent-offspring relationships with either of them. In contrast, the different lines of the vaccine strain Mukteswar, which was reportedly derived from an Indian field isolate in the mid-1940s, showed 98.9 to 100% sequence similarity to strain H. It is therefore probable that the two vaccines were derived from the same virus stock.

Evolution, Molecular↗
PubMed1983

Effects of a heat-labile factor(s) in normal serum on the interferon-inducing activity of Newcastle disease virus (NDV). Brief report.

The effect of fresh serum on the interferon-inducing activity of Newcastle disease virus (NDV) was studied. Infectivity in fertile eggs, hemolytic activity in human erythrocytes and interferon-inducing activity in mouse L cells and mouse spleen cells were all reduced by treatment of NDV with fresh serum from humans or guinea pigs, while hemagglutinating (HA) activity remained unchanged. The interferon-inducing ability of UV-inactivated NDV was also similarly reduced after treatment with serum. The serum effects were decreased by heating at 56 degrees C for 30 minutes or the addition of 10 mM EDTA. These results suggest that a heat-labile serum factor(s) such as complement or certain enzyme(s) reduced the hemolytic activity and consequently the penetrating activity of NDV, and that interferon-inducing activity was related to the hemolytic but not to the HA activity of NDV particles. Thus, it was concluded that the hemolytic activity of NDV was necessary for interferon induction in both mouse L cells and mouse spleen cells.

Animals↗
PubMed1997

Dynamic properties of Newcastle Disease Virus envelope and their relations with viral hemagglutinin-neuraminidase membrane glycoprotein.

The lipid composition of Newcastle Disease Virus (NDV) Clone-30 strain shows a low lipid/protein ratio, a high cholesterol/phospholipid molar ratio, and major phospholipids being qualitatively different to other NDV strains. The major fatty acyl constituents are palmitic, stearic, oleic, and linoleic acids; cerebrosides, sulfatides and two kinds of gangliosides are also found in the NDV membrane. It is reported for the first time in NDV that phospholipid classes are asymmetrically distributed over the two leaflets of the membrane: 60 +/- 4.5% of the phosphatidylcholine and 70 +/- 5.0% of the sphingomyelin are in the outer monolayer. Intact viral membranes and reconstituted NDV envelopes showed similar dynamic properties. Hemagglutinin-neuraminidase (HN) and fusion (F) proteins of NDV membrane affect the lipid thermotropic behaviour in reconstituted proteoliposomes made up of a single class of phospholipids. It is shown that the lipid composition is more important than the bulk membrane fluidity/order for both sialidase (neuraminidase) and hemagglutinating HN activities. Sialidase and hemagglutinating activities requires the presence of definite phospholipids (phosphatidylethanolamine) in its environment.

Fatty Acids↗
PubMed1986

Antigenic mapping and functional analysis of the F protein of Newcastle disease virus using monoclonal antibodies.

Twelve monoclonal antibodies to the F protein of a velogenic strain of Newcastle disease virus (NDV) were established. Each of these antibodies inhibited virus-induced plaque formation in BHK-21 cells. Seven antibodies neutralized viral infectivity in eggs; thus, antigenic variants could be selected with these antibodies and used for antigenic mapping. Based on the reactivity of the antigenic variants with the antibodies used in selection, 4 distinct antigenic sites (I-IV) were defined on the F protein molecule. In competitive binding assay, sites II and III were found to be spatially close to each other. Each antibody to sites I, II and III inhibited both virus-induced hemolysis of chicken erythrocytes and syncytium formation of BHK-21 cells. On the other hand, some of the antibodies to site IV selectively inhibited either hemolysis or cell fusion. This finding may indicate that the fusion of the viral envelope with erythrocytes and host cell membrane is modulated through different ways. Comparative analysis of different NDV strains using monoclonal antibodies to each of the different antigenic sites showed that the antigenicity of the F protein is highly conserved.

Animals↗
PubMedDate not supplied

Pathological changes of tracheal mucosa in chickens infected with lentogenic Newcastle disease virus.

Forty-day-old specific-pathogen-free chickens were exposed by aerosol to lentogenic Newcastle disease virus (NDV) and observed for 24 days for pathological changes in the tracheal mucociliary system. Specific fluorescence of NDV antigen was observed through day 5 postexposure (PE) in the cytoplasm of the tracheal epithelium and desquamated epithelium in the lumen. On day 1 PE, scanning electron microscopy revealed hypertrophy of goblet cells and small patches of the deciliated epithelium scattered mainly around the openings of mucous glands. The deciliated area of tracheal surface increased through day 4 PE. Light microscopy showed small vacuoles containing lymphocytes and heterophils in the epithelial layer. Immature epithelium proliferated in some areas. On days 5 and 6 PE, ciliated areas of the trachea tended to increase as a result of regeneration of the epithelium, still leaving many nonciliated patches of various sizes. On and after day 8 PE, there remained plaques with nonciliated flat epithelium, but most areas were covered with well-ciliated epithelium. Non-ciliated plaques were observed until day 24 PE, but they gradually decreased in size. These plaques were covered by a single layer of flat epithelium and were formed upon lymph follicles in subepithelial tissue.

Animals↗
PubMed2005

Structural analysis of peptides that interact with Newcastle disease virus.

A peptide with the sequence CTLTTKLYC has previously been identified to inhibit the propagation of Newcastle disease virus (NDV) in embryonated chicken eggs and tissue culture. NDV has been classified into two main groups: the velogenic group, and mesogenic with lentogenic strains as the other group based on its dissociation constants. In this study the peptide, CTLTTKLYC, displayed on the pIII protein of a filamentous M13 phage was synthesized and mutated in order to identify the amino acid residues involved in the interactions with NDV. Mutations of C1 and K6 to A1 and A6 did not affect the binding significantly, but substitution of Y8 with A8 dramatically reduced the interaction. This suggests that Y8 plays an important role in the peptide-virus interaction. The three-dimensional structure of the peptide was determined using circular dichroism (CD), nuclear magnetic resonance (NMR), and molecular modeling. The peptide exhibited two possible conformers. One that consists of consecutive beta-turns around T2-L3-T4-T5 and K6-L7-Y8-C9. The other conformer exhibited a beta-hairpin bend type of structure with a bend around L3-T4-T5-K6.

Antiviral Agents↗
PubMed1975

Modification of normal cell surface by smooth membrane preparations from BHK-21 cells infected with Newcastle disease virus.

Smooth membrane fractions were prepared from the cytoplasmic extract of BHK-21 cells infected with Newcastle disease virus (NDV). These membranes exhibited high hemagglutinating, neuraminidase, and hemolytic activity but little infectivity, suggesting that they might be precursors for viral envelope. When such membranes were adsorbed to the monolayers of uninfected BHK-21 cells at 4 degrees C and then incubated at elevated temperature for a couple of hours, the cells became highly hemadsorptive even in the presence of cycloheximide. This phenomenon occurred between 15 degrees C and 25 degrees C, and was maximal at 31 degrees C, where approximately 4 times more erythrocytes were adsorbed than to the cells incubated at 4 degrees C. Immunofluorescent staining suggested that diffusion of viral antigens might occurred rapidly over the entire surface of the cells. Cell fractions containing virions induced hemadsorption in uninfected cells, too. However, induction occurred now at 31 degrees C and was maximal at 37 degrees C, and erythrocytes appeared to be adsorbed not to the entire surface of the monolayer but restricted areas of the cells. The diffusion of viral antigens on the cell surface was not so significant under these conditions. On the basis of these findings the possible role of the membranes of the smooth endoplasmic reticulum in virus replication is discussed.

Antigens, Viral↗